international journal on coconut r & d - vol. 31 no. 1, 2015 · 2019-07-02 · asian and...

Asian and Pacific Coconut Community

International Journal on Coconut R & D - Vol. 31 No. 1, 2015

ISSN – 0215-1162

Antimicrobial Properties of Cocos nucifera: A Review -Lalitha Ramaswamy, R. Rajendran, U. Saraswathi, R. Sughanyaand C. Geethadevi

Formulation of Zinc Rich Coconut Nutribar Designed forAthletes - Anusha Priyadarsini. K and Dr. Lalitha Ramaswamy

Ecofriendly Organosolv Process for Pulping of TenderCoconut Fibre - Jincy P.J., Anita Das Ravindranath and U.S.Sarma

Low Temperature Grafting of MMA on to Coir Fibre -Lakshmi N.S., Sarika Babu, Sumy Sebastian and P.K. Ravi

Characterisation of Silver Deposited Coir Fibers byMagnetron Sputtering - Melvi Chandy, U.S. Sarma, M.S. Lathaand K. Shreekrishna Kumar

An Investigation of the Tender Nut Potential of DiverseCoconut (Cocos nucifera L.) Varieties/Forms in Sri Lanka -S.A.C.N. Perera, G.K. Ekanayake and H.M.N.B. Herath

Pollen Dispersal and Pollination Patterns Studies in PatiKopyor Coconut using Molecular Markers - Siti HalimahLarekeng, Ismail Maskromo, Agus Purwito, Nurhayati AnshoriMatjik and S. Sudarsono

Cord

Cord is a semi-annual Journal of the Asian and Pacific Coconut Community (APCC)

devoted to coconut research and development (R & D).

The APCC is the first commodity based organization established under the auspices of

United Nations-Economic and Social Commission for Asia and the Pacific (UN-ESCAP) in

1969. The APCC is an independent intergovernmental organization, currently consisting of

eighteen member countries, namely: Federated States of Micronesia, Fiji, India, Indonesia,

Kiribati, Malaysia, Marshall Islands, Papua New Guinea, Philippines, Samoa, Solomon

Islands, Sri Lanka, Thailand, Tonga, Vanuatu, and Vietnam. Jamaica and Kenya are

associate members of the APCC. The objectives of the APCC are to promote, coordinate and

harmonize all activities of the coconut industry to achieve the maximum socio-economic

development of the industry.

In addition to Cord, the APCC publishes The Cocommunity (monthly newsletter),

Coconut Statistical Yearbook (yearly) and Cocoinfo International (semi-annual popular Journal

on the coconut industry) and other ad-hoc publications.

Cord welcomes original research articles on any aspect of the coconut industry. The

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responsibility for the opinions expressed and for the accuracy of statements rests with the

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CC oo rr dd IInntteerrnnaattiioonnaall JJoouurrnnaall oonn CCooccoonnuutt RR && DD

EDITORIAL BOARD

URON N. SALUM

Editor-in-Chief and Executive Director

Asian and Pacific Coconut Community (APCC)

D.L. JAYARATNE

Department of Microbiology

Faculty of Science

University of Kelaniya

Kelaniya 11600, Sri Lanka

Email: [email protected]

DINA B. MASA

Manager

Product Development Department

Philippine Coconut Authority

152 Chico St., Project 2

Quezon City, Philippines


DIVINA D. BAWALAN

Free Lance Consultant on Coconut Processing and

Utilization

B-44 Far East Asia Village

Marcos Highway, Bo. Mambugan

1870 Antipolo City, Philippines


DOMINIQUE BERRY

Director

Tree Crops Department

CIRAD-Cultures Perennes

TA 80/PS3, Boulevard de la Lironde

34398 Montpellier

Cedex 5, France


ERLINDA P. RILLO

Chief, Tissue Culture Division


Albay Research Center

Banao, Guinobatan

Albay 4503, Philippines


GEORGE V. THOMAS

Formerly Director

Central Plantation Crops Research Institute

(CPCRI)

Vettimoottil House, Vakayar P.O.

Konni 689698, Kerala, India


H.A. TILLEKERATNE

Consultant in Development of Coconut Processing &

Marketing

(Former Director General, Coconut Development

Authority, Sri Lanka)

No. 18D, Temple Road

Kalubowila, Sri Lanka


HENGKY NOVARIANTO

Coconut Breeders

Indonesian Palm Research Institute (IPRI)

P.O. Box 1004, Manado 95001

North Sulawesi Province, Indonesia


H.P. SINGH

Deputy Director General (Horticulture)

Indian Council of Agriculture Research (ICAR)

Krishi Anusandhan Bhavan-II

Pusa, New Delhi 110012, India


HUGH HARRIES

Moderator

Coconut Information Exchange and Time Line

2 Beech Road, Broadway

Weymouth, Dorset DT3 5NP

United Kingdom


JAMES V. KAIULO Managing Director KOKONAS Indastri Koporesen (KIK) P.O. Box. 81 Port Moresby Papua New Guinea Email: [email protected]

JOCELYN E. EUSEBIO

Director

Crops Research Division (CRD)

Philippine Council for Agriculture, Forestry and Natural

Resources Research and Development (PCARRD)

Department of Science and Technology (DOST)

Paseo de Valmayor, Los Banos

Laguna 4030, Philippines


mailto:[email protected]


K.V.A. BAVAPPA

Karooth Villa

P. O. KAPPUR

Kumaranellur, Palakkad - 679 552

Kerala, India


L.C. PRIYANTHIE FERNANDO

Additional Director and Head

Crop Protection Division

Coconut Research Institute

Lunuwila 61150

Sri Lanka


LEX A.J.THOMSON

EU-FACT Team Leader

Agri-Forestry Export Production Specialist

Secretariat of the Pacific Community (SPC)

Private Mail Bag, Suva

Fiji Islands


LUISITO J. PENAMORA

Division Chief lll

Timber Utilization Division

Zamboanga Research Center

San Ramon, Zamboanga City

Philippines


M. SYAKIR

Director

Central Research Institute for Estate Crops

Jl. Tentara Pelajar No. 1

Bogor 1611

Indonesia


MICHEL DOLLET

CIRAD

Campus International de Baillarguest

TA-A-98/F

34398 Montpellier Cedex 5

France


MIKE FOALE

Honorary Research Consultant

The School of Land Crops and Food Sciences

The University of Queensland

4 Carinya Close, Maleny

Queensland 4552

Australia


MILLICENT WALLACE

Botanist/Plant Breeder

Coconut Industry Board

Ministry of Agriculture

18 Waterloo Road, Kingston

Jamaica


NANTARAT SUPAKAMNERD

Senior Scientist (Soil Science)

Horticulture Research Institute

Department of Agriculture

50 Phaholyothin Road

Bangkok, Thailand


NARONG CHOMCHALOW

Chairman of the Conservation and Development of

Coconut Oil Forum of Thailand and

Former FAO-RAP

Production Officer (Industrial Crops)

770 Phahonyothin 32, Chatuchak

Bangkok 10900

Thailand


P.K. THAMPAN

President

Peekay Tree Crops Development Foundation

M.I.G. 141, Gandhi Nagar

Cochin 682 020, India Email: [email protected]

PEYANOOT NAKA

Senior Scientist (Post Harvest and Processing Specialist)



Ministry of Agriculture and Cooperatives

Phaholyothin Road, Chatuchak

Bangkok 10900, Thailand


PISAWAT BUARA

Former Director




Bangkok, Thailand


PONCIANO A. BATUGAL

Chairman,

APCC Technical Working Committee (TWC) on Climate

Change and Poverty Reduction Project Proposal

Block 5 Sacay Grand Villas

Los Banos, Laguna 4031

Philippines


PONNIAH RETHINAM

Plantation Crops Management Specialist

18, Lakshmi Nagar S. N. Palayam

Coimbatore 641007

Tamil Nadu, India


PRASERT ANUPHAN

Former Deputy Director General



Bangkok, Thailand








RAMON L. RIVERA

Deputy Administrator for Research and

Development Branch


Don Mariano Marcos Ave., cor Elliptical Road

Diliman, Quazon City

Philippines


RICHARD MARKHAM

Research Program Manager - Pacific Crops

Australian Centre for International Agricultural

Research (ACIAR)

C/o Secretariat of the Pacific Community (SPC)


Fiji Islands


ROLAND BOURDEIX COGENT Coordinator

Bioversity HRF, Cirad Umr CEFE

Montpellier. 1990, Bd de la Lironde

Parc Scientifiques Agropolis II

34397 Montpellier

France Email: [email protected]

ROSA S. ROLLE

Senior Agro-Industry and Post Harvest Officer

FAO Regional Office for Asia and the Pacific

39 Phra Atit Road



SOMCHAI WATANAYOTHIN

Senior Agriculture Scientist


Phaholyothin Road, Chatuchak



EDITORIAL STAFF

TEVITA KETE

Export Processing and Marketing Officer

(Value-Added Coconut Products)

Land Resources Division

Secretariat of the Pacific Community (SPC)


Fiji Islands


U.S. SARMA

Director

Central Coir Research Institute

P.O.-Kalavoor, Dist.-Alappuzha

Kerala-688 522

India

Email: [email protected] VIVENCIO C. GALLEGO Scientist I Crop Protection Division Philippine Coconut Authority Davao Research Center Bago Oshiro, Davao City 8000 Philippines Email: [email protected] WALTER L. BRADLEY Distinguished Professor of Mechanical Engineering School of Engineering and Computer Science Baylor University Waco, TX 76798-7356 Texas USA Email: [email protected]

YUPIN KASINKASAEMPONG

Agricultural Senior Scientist

Chumphon Horticultural Research Center

70, Moo 2, Wisaitai Subdistrict

Sawee District, Chumphon 86130

Thailand


Associate Editors ………………………………………....

Editorial Assistant……………………………………….

DEEPTHI NAIR, S.

Assistant Director, APCC

MUHARTOYO

Librarian / Documentalist, APCC

TATI M. KABIB

Office Assistant, APCC



Cord 2015, 31 (1)

Table of Contents

Page:

Antimicrobial Properties of Cocos nucifera: A Review - Lalitha Ramaswamy, R.

Rajendran, U. Saraswathi, R. Sughanya and C. Geethadevi

Formulation of Zinc Rich Coconut Nutribar Designed for Athletes - Anusha

Priyadarsini. K and Dr. Lalitha Ramaswamy

Ecofriendly Organosolv Process for Pulping of Tender Coconut Fibre - Jincy P.J.,.,

Anita Das Ravindranath and U.S. Sarma

Low Temperature Grafting of MMA on to Coir Fibre - Lakshmi N.S., Sarika Babu,

Sumy Sebastian and P.K. Ravi

Characterisation of Silver Deposited Coir Fibers by Magnetron Sputtering - Melvi

Chandy, U.S. Sarma, M.S. Latha and K. Shreekrishna Kumar

An Investigation of the Tender Nut Potential of Diverse Coconut (Cocos nucifera

L.) Varieties/Forms in Sri Lanka - S.A.C.N. Perera, G.K. Ekanayake and H.M.N.B.

Herath

Pollen Dispersal and Pollination Patterns Studies in Pati Kopyor Coconut using

Molecular Markers - Siti Halimah Larekeng, Ismail Maskromo, Agus Purwito,

Nurhayati Anshori Matjik and S. Sudarsono

1-6

7-12

13-23

24-31

32-38

39-45

46-60

Cord 2015, 31 (1)

1

Antimicrobial Properties of Cocos nucifera: A Review

Lalitha Ramaswamy1, R. Rajendran

2, U. Saraswathi

3, R. Sughanya

4 and C. Geethadevi

4

Abstract

Coconut is known as the ―wonder food‖ and is regarded as perfect diet because it contains almost

all essential nutrients needed by the human body. The various products of coconut include tender

coconut water, copra, coconut oil, raw kernel, coconut cake, coconut toddy, coconut shell and wood

based products, coconut leaves, coir pith etc. Coconut is a unique source of various natural products for

the development of medicines against various diseases and also for the development of industrial

products. Almost all the parts of the palm have medicinal properties such as antibacterial, antifungal,

antiviral, antiparasitic, antidermatophytic, antioxidant, hypoglycemic, hepatoprotective and

immunostimulant property. The various medicinal properties of coconut and its products are

summarized in this paper.

Keywords: coconut, antimicrobial properties

____________________________________

1Head of the Department and Associate professor, Department of Nutrition and Dietetics, PSG College of Arts

and Science, Coimbatore, India.


2Principal, PSG College of Arts and Science, Coimbatore, India.

3Associate professor, Department of Biochemistry PSG College of Arts and Science, Coimbatore, India.

4Junior Research Fellow, Department of Nutrition and Dietetics, PSG College of Arts and Science,

Coimbatore, India.

Cord 2015, 31 (1)

2

Introduction

Plants have always played a major role in

the treatment of human and animal diseases.

Medicinal plants can be used in different forms:

(i) as raw materials for extraction of active

compounds or (ii) for extraction of abundant but

inactive constituents which can be transformed

by partial synthesis into active compounds as

such as extracts or as traditional preparations

(Mukherjee, 2008).

Coconut is a drupe borne by the coconut

palm (Cocos nucifera), a member of the

monocotyledonous family Palmae. It is known as

the ―wonder food‖ and is regarded as perfect diet

because it contains almost all essential nutrients

needed by the human body. Coconut palms are

grown in more than 80 countries of the world,

with a total production of 61 million tonnes per

year (FAO, 2010). India is the third largest

coconut producing country, having an area of

about 1.78 million hectares under the crop. In

India, the four south Indian states namely Kerala,

Tamilnadu, Karnataka and Andhra Pradesh

account for around 90% of the coconut

production in the country (NMCE, 2007).

Coconut is a very versatile and indispensable

fruit for most people under the tropical belt. It is

a complete food is rich in calories, vitamins, and

minerals. It is nourishing, strengthening and

fattening food. It has high oil content. The

protein is of high quality and contains all amino

acids essential for the growth and maintenance

of the body. It is rich in K, Na, Mg and S. The

energy value of the dried coconut is 662 calories

per 100 g (Bakhru, 2000). The nutrient content

of nuts varies by species, but in general they

provide rich sources of vegetable protein,

monosaturated and polyunsaturated fatty acids,

dietary fiber, vitamins E & K, folate,

magnesium, copper, selenium and potassium.

Nuts are also naturally low in saturated fatty

acids and sodium (O’Neil et al, 2012). Nuts also

provide phenols, phytosterols, flavanoids,

proanthocyanidins, resveratrol and arginine;

these bioactive compounds, coupled with

micronutrients such as vitamin E and selenium,

serve as antioxidants and are anti-inflammatory

(Bolling et al., 2010).

All parts of coconut tree besides being

used as food and commercially, are also rich in

medicinal properties. Most of the parts of

coconut tree such as endosperm, coconut oil,

tender coconut water, inflorescence and root are

being used in the Ayurvedic medicine for the

treatment of several clinical conditions. In view

of its utilitarian value of the entire tree it is often

referred as Kalpavriksha (Pushpan et al., 2013).

It is the unique source of various natural

products for the development of medicines

against various diseases and also for the

development of industrial products. The parts of

its fruit like coconut kernel and tender coconut

water have numerous medicinal properties such

as antibacterial, antifungal, antiviral,

antiparasitic, antidermatophytic, antioxidant,

hypoglycemic, hepatoprotective and

immunostimulant property. Coconut water and

coconut kernel contain microminerals and

nutrients, which are essential to human health,

and hence coconut is used as food by the peoples

in the globe, mainly in the tropical countries

(Deb Mandal and Mandal 2011).

Anti-bacterial activity of coconut

The antibacterial activities of coconut

endocarp extracts using methanolic and aqueous

extracts showed a strong activity against Bacillus

subtilis, Pseudomonas aeruginosa,

Staphylococcus aureus, and Micrococcus luteus

(Singla et al., 2011). The antimicrobial property

of coconut shell showed strong antibacterial

activity on Ecsherichia coli and Salmonella typhi

(Verma et al., 2012). The antibacterial potentials

of crude aqueous and n-Hexane extracts of the

husk of Cocos nucifera against forty-five strains

of Vibrio pathogens and twenty-five other

bacterial isolates those normally implicated in

food and wound infections were studied. The

aqueous extract was active against 17 of the

tested bacterial and 37 of the Vibrio isolates;

while the n-Hexane extract showed antibacterial

activity against 21 of the test bacteria and 38 of

the test Vibrio species (Akinyele et al., 2011).

Tender coconut water is given to cholera

patients because of its saline and albumen

content (Effiong et al., 2010). It is generally used

to treat urinary infection and diarrhea. Three

http://www.ncbi.nlm.nih.gov/pubmed?term=Mandal%20S%5BAuthor%5D&cauthor=true&cauthor_uid=21771462

Cord 2015, 31 (1)

3

peptides lower than 3kDa were purified and

identified from green coconut water by using

reversed phase-high performance liquid

chromatography (RP-HPLC), showing molecular

masses of 858Da, 1249Da and 950Da. These

peptides have remarkable potential to contribute

in the development of novel antibiotics from

natural sources (Mandal et al., 2009).Bacterial

isolates obtained from fermented toddy were

checked for the activity against B. cereus,

Listeria monocytogenes and E. coli which are

common food borne pathogens that infect the

gastro intestinal tract. The results showed that

two of the ten isolates could inhibit the indicator

organisms, however, at different inhibition levels

(Krishnamoorthy and Arjun 2012).

Antimicrobial property of coconut leaf

extracts against Acinetobacter spp., B. cereus,

E. coli, S. typhi, Shigella dysenteriae, S. aureus,

Aspergillus flavus and A. niger were

investigated. The results showed that the leaf

extract were active against all the organisms

except S. aureus and A. flavus (Ifesan et al.,

2013). The phytochemical screening of coconut

flowers demonstrated the presence of alkaloids,

flavonoids, phenols, phytosterols, tannins,

aminoacids and carbohydrates (Dyana and

Kanchana, 2012). These phytochemicals present

in coconut flowers have well known curative

activity against several human pathogens.

Antibacterial properties of aqueous and

methanolic extracts of 26 medicinal plants used

in Mexico to treat gastrointestinal disorders were

studied. The results showed that coconut being

one of the plants which possessed a strong

bactericidal activity against tested species

(Alanis et al., 2005).

The effects of oil-pulling against oral

microorganisms in biofilm models using

different edible oils were investigated. The study

proved that oil-pulling using coconut oil

exhibited antimicrobial activity against S.

mutans and C. albicans which are considered to

be the predominant microorganisms found in

dental caries (Thaweboon et al., 2011). Owing to

the high Lauric acid content in coconut flour, it

has used as a medication for oral sores (Taheri et

al., 2010). Husk fibers extract of coconut was

proved to have potential cure for oral diseases

(Alviano et al., 2008).

Lauric acid is a natural compound that is

the main acid in coconut oil and also resides in

human breast milk. Studies have been done to

prove the potentiality of lauric acid against many

harmful pathogens. Batovska et al., in 2009

worked on antibacterial activity of medium chain

fatty acids and their 1- monoglycerids. In the

study they have reported that among all

monoglycerides, monolaurin displayed the

greatest anti bacterial activity towards various

Gram positive strains. The antimicrobial

property of lauric acid against

Propionibacterium acnes both in vitro and in

vivo was studied. The in vitro studies proved that

lauric acid inhibited the growth of P. acnes, and

did not have much effect on resident flora of

skin. The cytotoxicity effect of lauric acid

reported that higher concentrations of lauric acid

also did not affect viability of the cells, instead

killed the acne causing bacteria. (Nakatsuji et al.,

2009).

Anti-fungal activity

Heating the coconut shells gives oil that is

used against ringworm infections in the popular

medicine of India. The alcoholic extract of ripe

dried coconut shell has antifungal activity

against Microsporum canis, M. gypseum, M.

audouinii, Trichophyton mentagrophytes, T.

rubrum, T. tonsurans and T. violaceum. The

extract showed antifungal activity against all

dermatophytes tested with twice the

concentration needed against Epidermophyton

flocossum (200 ug/ml. The activity was mainly

attributed to the high content of phenolic

compounds (Venkataraman et al., 1980). An

extensively study carried out by Ogbolu et al.,

(2007) revealed that the antifungal activity of

coconut oil against 6 Candida sp. obtained from

clinical settings. They further reported that

coconut oil was active against species of

Candida at 100% concentration compared to

fluconazole.

Antiviral activity

Coconut oil is very effective against a

variety of viruses that are lipid-coated. The

Cord 2015, 31 (1)

4

medium chain fatty acids in coconut oil destroy

the viruses by disrupting their membranes,

interfering virus assembly and maturation (Arora

et al., 2011). Coconut oil as an anti- HIV

medication was administered to 15 HIV positive

patients at different concentrations for 6 months.

By the end of three months 50% of the patients

showed reduced viral load and by the end of 6th

month, eight patients showed reduced viral load

and favorable CD4/ CD8 count (Conrado 2000).

Extraction of polyphenols from husk fiber was

done and the extract was checked for

antimicrobial and antiviral activities. The

selective antibacterial activity of C. nucifera

against S. aureus and Herpes Simplex Virus -

1(HSV-1) suggests that this plant may be useful

for topical application in wound healing

(Esquenazi et al., 2002).

Antiprotozoan activity

The antihelmintic assay was performed on

chironomus larvae by Mariselvam et al., 2013.

The crude extract of the coconut inflorescence

not only confirmed inactivation of helminthes,

but also caused death in the shorter time as

compared to standard drug albendazole. The in

vitro leishmanicidal effects of coconut husk

extracts on Leishmania amazonensis were

evaluated. These results showed that extract of

coconut husk at 10 µg/ml was a strikingly potent

leishmanicidal substance which inhibited the

growth of both promastigote and amastigote

developmental stages of L. amazonensis after 60

min, presenting no in vivo allergenic reactions or

in vitro cytotoxic effects in mammalian systems.

A combination of specially prepared

extracts of onion (Allium cepa) and coconut was

tested against the organisms causing

gastrointestinal infection in animals. The sheep

with gastrointestinal helminthic infection were

fed with extract for 8 days containing each 60 g

coconut and onion extract. The results showed

that the worm stages disappeared from the feces

and were also not found 9 and 20 days after the

end of the feeding with the extract (Mehlhorn et

al., 2011).

Conclusion

Coconut has a potent antimicrobial activity

against various microflora. Besides being

antimicrobial in nature, coconut and its products

have other health benefits such as improving

heart health, digestion, management of diabetes

mellitus and some other diseases. The utilization

of coconuts on a regular basis can be

encouraged. Food formulations with coconut and

its products can be made commercially viable

and popularized as a functional food. Drugs and

supplements can be formulated with coconut

products for prophylactic and therapeutic

purposes.

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rich extract from husk fiber of Cocos

nucifera Linn. (Palmae) Research in

Microbiology 155: 136–143.

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(2008). Plant Made Pharmaceuticals

(PMPs)- Development of Natural Health

Products from Bio-Diversity. Indian J.

Pharm Educ. Res 42(2), 113-121.

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C.C., Zhang, L, Gallo R.L. and Huang

C.M. 2009. Antimicrobial Property of

Lauric Acid against Propionibacterium

acnes: Its Therapeutic Potential for

Inflammatory Acne Vulgaris, Journal of

Investigative Dermatology, Volume 129.

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Commodity Exchange of India Limited;

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Fulgoni V.L. 2012. Out of-hand nut

consumption is associated with improve

nutrient intake and health risk markers in

Cord 2015, 31 (1)

6

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and Nutrition Examination Survey 1999-

2004. Nutr Res 32: 185–194.

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Oloko 2007. In Vitro Antimicrobial

Properties of Coconut Oil on Candida

Species in Ibadan, Nigeria, J Med Food 10

(2), 384–387.

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Vishwanathan. 2013. Preliminary

Phytochemcial Screening of

Narikelapushpa (Flower of Cocos nucifera

L.) Global Journal of Traditional

Medicinal Systems, 2(2): 1-3.

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Jagani 2011. Antioxidant & Antimicrobial

Activities of Cocos Nucifera Linn.

(Arecaceae) Endocarp Extracts Indo

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Sciences; 1(4): 354-361.

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Bakshi M, Nakhostin M.R 2010.

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oral microflora: An in vitro study. Res J

Biol Scs. 5(6): 456-459.

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Microorganisms in Biofilm Models, Asia

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activity of the alcoholic extract of coconut

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2012. A Potential Antimicrobial Agent

from Cocos nucifera mesocarp extract;

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Sciences. Vol. 1(2), 48-54.

Cord 2015, 31 (1)

7

Formulation of Zinc Rich Coconut Nutribar Designed for Athletes

Anusha Priyadarsini. K1 and Dr. Lalitha Ramaswamy

2

Abstract

The main dietary goal for athletes is to obtain competent nutrition to improve health, fitness and

sports performance. Athletes require nutritious convenient foods like nutribars to provide instant

energy and other nutrients. Zinc supplementation has been shown to increase anabolic hormone status

to meet catabolic activity in athletes which is profound during training. In the current study nutribar

was formulated using zinc rich food sources namely dehydrated coconut meat, cashewnuts, pumpkin

seeds and cocoa. Two variations of the nutribar were prepared variation I using dark chocolate and

variation II with milk chocolate. The samples were subjected to organoleptic evaluation using a 9 point

Hedonic scale by semi trained panel members. The nutribars were also analyzed for selected nutrients

using standardized procedures. The mean score obtained for over all acceptability was 8.0±0.632,

8.7±0.458 and 8.5±0.5 by the control, variations I and II respectively. Variations I and II had obtained

a mean score of more than 8.0 for flavour and taste, while the control sample had obtained lesser

scores. The zinc content of variations I and II was 7.38 mg & 5.2 mg being greater than the control.

Keywords: Dehydrated coconut meat, Cashew nuts, Cocoa, Pumpkin seeds, Athletes, Zinc, Nutribar

____________________________________

1AnushaPriyadarsini. K, Research Scholar, Department of Nutrition and Dietetics, PSG College of Arts &

Science, Avinashi road, Civil Aerodrome, Coimbatore-641014, Tamilnadu, India.

Email: [email protected].

2Dr. LalithaRamaswamy, Associate professor and Head, Department of Nutrition and Dietetics, PSG College

of Arts & Science, Avinashi road, Civil Aerodrome, Coimbatore-641014, Tamilnadu, India.

Email: [email protected].

Cord 2015, 31 (1)

8

Introduction

Sports nutrition is a part of nutrition

science that relates to the interaction of nutrition

and physical activity (Maughan, 2001). Nutrition

plays a vital role in sports performance. Dawn

Jackson, RD (2003) spokeswoman for the

American Dietetic Association, concurs;

nutribars are convenient, especially for

physically active individuals. The coconut has

been a traditional food in almost all the countries

where it is grown. It enters the diet of the people

many ways, the water from the tender coconut is

drunk and the mature nuts are used in cooking in

the preparation of sweetmeats and for house hold

oil production (Grimwood, 1975). Increasingly,

there is an emphasis on the synergistic relation

between diet and exercise for well being and a

growing awareness of the beneficial role that

mineral element nutrition may play in achieving

good health and enhanced physiologic function.

In contrast with the role of macro nutrients in the

body, micronutrients despite their relative

paucity in the diet and the body, perform

important roles in regulating whole body

metabolism, including energy utilization and

work performance. The importance of these

micro nutrients is revealed by the diversity of

metabolic process they help to regulate. Zinc is a

ubiquitous element, that plays a fundamental role

in many cellular reactions. It is also an

intracellular cation, and is required to serve

either a catalytic or structural role by 300

enzymes in mammals. (Ralf, 2003). Zinc

containing enzymes participate in many

components of macro nutrient metabolism and

cell replication. In addition, some zinc

containing enzymes, such as carbonic anhydrase

and lactate dehydrogenase, are involved in

intermediary metabolism during exercise (Henry,

2000). Zinc is essential in keeping the immune

system strong. For athletes, this is especially

important, since being side lined with a viral

infection, or other illness, can force one to miss

valuable workout time. Zinc deficiency is

problematic as plasma testosterone is regulated

in part by zinc. Therefore, zinc deficiency may

adversely affect this hormone, causing muscular

mass and strength to suffer. Exercise increases

losses from the human body, and severe zinc

deficiency can compromise muscle function

(Lukaski HC, 2000). Dietary zinc intake and

plasma zinc each have a positive association

with BMD in men leading to osteoporosis

(Tiasun H Hyun, 2004). Therefore in order to

augment zinc deficiency in athletes and to

enhance their endurance activity level, this

nutribar was designed. The objective of the study

is to formulate zinc rich coconut nutribar for

athletes and finding out its organoleptic

acceptability and nutrient content.

Materials and methods

Selection of ingredients

There are several zinc rich food sources

like pumpkin seeds, sesame seeds, squash seeds,

watermelon seeds, dark chocolate, wheat germ

(Edward, 2013). The researcher selected

dehydrated coconut meat, cashew nut and

pumpkin seeds as zinc sources as they have 5mg,

6mg and 6.6mg/100gm respectively. These three

ingredients were also found to blend well in the

final product after a pilot study. The chocolate

coating given to the product also enhanced the

zinc content dehydrated coconut meat is dried

coconut flesh which is widely used for extracting

coconut oil and the remaining coconut meal is

used for cattle feeding. In India, dehydrated

coconut meat is used for cooking food, apart

from using coconut oil. Dehydrated coconut

meat meets 42% of daily requirement of Zinc for

an adult. The recommended dietary allowance of

Zinc for an adult per day is 12 mg (Sesikeran,

2010). The nutrient content of dehydrated

coconut meat is presented in Table 1.

Table 1. Nutrient Content of Dehydrated

coconut meat per 100gm

S. No Nutrients Values

1 Energy 662 kcal

2 Carbohydrate 18.4 gm

3 Protein 6.8 gm

4 Fat 62.3gm

5 Crude fibre 6.6 gm

6 Calcium 400 mg

7 Phosphorus 210 mg

8 Iron 7.8 mg

9 Zinc 5.0 mg

Source: Nutritional value of Indian Foods, National

Institute of Nutrition

Cord 2015, 31 (1)

9

Preparation of nutribar

Dehydrated coconut meat, cashew nut and

pumpkin seeds were purchased in a pure form

from a reputed shop known for selling these

products. Other ingredients namely cocoa

powder, sugar and honey were purchased from a

reputed departmental store. 100gm of the

dehydrated coconut meat is taken and the brown

outer skin was peeled and discarded as it was

found to interfere with the organoleptic

characteristics. The pulp was flaked and toasted

in a frying pan till a fine aroma emanated from

it. 20 gm of pumpkin seeds and 10 gm of cashew

nuts were also dry roasted separately. All the

ingredients were coarsely powdered and kept

aside. Sugar syrup was prepared up to the thread

stage in 1:1 ratio. The other ingredients were

added to the syrup and cooked in a low flame

with constant stirring till the thick mass did not

stick to the sides of the pan. The mass was then

allowed to set on a greased plate and cut into

uniform sized pieces each in the shape of a bar

weighing approximately 50gm each and allowed

to cool. One half (variation I) was coated with

dark chocolate and other half (variation II) with

milk chocolate as the preference for the dark and

milk chocolate varies between individual.

(Figure 1 shows a display of the formulated

products). A control sample was also prepared

replacing dehydrated coconut meat, cashew nut,

pumpkin seeds and chocolate with oats and the

same method of preparation was followed.

Figure 1. Formulated Coconut nutribar

Nutrient analysis

The nutribars were analyzed for their fiber,

protein, zinc calcium and iron. These nutrients

were selected for analysis as they affect the

bioavailability of zinc. Protein was estimated by

Kjeldahl method, fiber by AOAC method, zinc

by atomic absorption spectrophotometer, iron by

Wongs method and calcium was estimated by

precipitating it as calcium oxalate. The

estimations were carried out in triplicates.

Sensory evaluation

Sensory evaluation is defined as a

scientific discipline used to evoke, measure,

analyze, and interpret those responses to

products that are perceived by the senses of

sight, smell, touch, taste, and hearing (Stone and

Sidel,1993). The nutribars thus prepared were

evaluated for the sensory attributes. Each of the

qualities namely appearance, texture, flavour,

taste and over all acceptability was assessed

using a 9 point Hedonic scale by a team of 10

semi trained panel members. The scores given

by the panelists were statistically analyzed by

ANOVA.

Results and discussion

Nutritional composition of Nutribar

The nutrient content of the prepared

nutribar is presented in Table 2. The nutribar had

a protein content of 13.86, 15.60 and 15.82g%

for the control, variations I and variation II

respectively. The protein is essential for muscle

building, and wear and tear of the athletes and it

also enhances the absorption of zinc. The Zinc

content of the nutribar was higher in variation I

(7.38 mg %) due to the addition of dark

chocolate, as it has more cocoa in it. Cocoa is

one of the zinc sources. In dark chocolate, the

amount of cocoa added being greater has

enhanced the zinc content. The amount of zinc in

variation II was 5.32mg of which was nearly

2mg lesser than the variation I while that of

control was even lesser (3.08 mg %), as there

was no zinc food source added to it. The fiber

content of the both the variations was almost

same (7.10g and7.14g% for variations I and II

respectively) and was higher than the control

Cord 2015, 31 (1)

10

Table 2. Nutrient composition of Nutribar per 100gm

S. No Nutrients Control Variation I

(Dark chocolate)

Variation II

(Milk Chocolate)

1 Protein 13.86gm 15.60gm 15.82mg

2 Zinc 3.08mg 7.38mg 5.32mg

3 Fibre 6.70gm 7.10gm 7.14gm

4 Calcium 310.0mg 330.0mg 342.0mg

5 Iron 4.80mg 6.0mg 6.20mg

Table 3. Mean Scores of the organoleptic evaluation of nutribars

Sensory

attributes

Control Variation I

(Dark Chocolate)

Variation II

(Milk Chocolate)

P value

Colour 8.4±0.663 8.0± 0.4 8.5± 0.670 0.342774

Texture 8.7 ± 0.458 8.5 ±0.670 8.3± 0.640 0.377798

Taste 7.8 ± 0.6 8.4± 0.663 8.4± 0.663 0.091761

Flavour 7.9 ± 0.7 8.7± 0.458 8.5± 0.670 0.028525

Over all

acceptability 8.0 ± 0.632 8.7 ±0.458 8.5± 0.670 0.0282674

Figure 2. Mean Scores of the organoleptic evaluation of nutribars

7.2

7.4

7.6

7.8

8

8.2

8.4

8.6

8.8

9

Control(Varies ±0.458 to 0.7)

Variation I ( Dark Chocolate) varies from ± 0.4 to 0.67

Variation II( Milk Chocolate)varies ± 0.5 to 0.67

Cord 2015, 31 (1)

11

(6.7g %). Addition of dehydrated coconut meat

added to the variation may be the reason for this.

The fiber concentration is high in dehydrated

coconut meat and palm kernel products

(Stephaine, 2013). Calcium in the control group

was found to be 310mg whereas in variations I

and II it was 330.0mg and 342.0mg respectively.

The iron content was found to be 4.80mg in the

control and comparatively high in variations I

and II (6.0mg and 6.20mg respectively).

Sensory evaluation of nutribar

The sensory attributes and the mean scores

of the nutribar are indicated in Table 3. The

mean scores obtained for over all acceptability

was 8.0±0.632, 8.7±0.458 and 8.5±0.5

respectively by the control, variation I and II

respectively. Variations I and II had obtained a

mean score of more than 8.0 for flavour and

taste, while the control sample had obtained

lesser scores. Results of ANOVA indicate no

significant difference (p≥0.05) in colour, texture

and taste between the three samples, while it was

significant (p≤0.05) for flavour and over all

acceptability. This is due to the addition of

chocolate and dehydrated coconut meat as it has

good flavour (Gohl, 1982) in the variations

which have mutually enhanced the acceptability

of the nutribar. Graphical representation of

organoleptic scores are shown in figure 2.

Conclusion

The control sample had obtained highest

mean score in the texture criteria compared to

the variations I and II. This is because the texture

of the oats being crispy and crunchy, whereas the

variations I and II had obtained higher mean

scores in over all acceptability compared to the

control sample. Statistically it was found out that

there was no significant difference (p≥0.05) in

the colour, texture and taste between the three

samples, while it was significant (p≤0.05) for

flavor and over all acceptability. The addition of

dehydrated coconut meat, pumpkin seeds,

cashew nuts and chocolate in the variations are

not only nutritious, but also highly acceptable.

The nutribar provide almost one half of the daily

zinc requirement of athletes.

Consumption of the formulated nutribar

(100 gm) will help in contributing to the zinc

requirement, thereby enhancing the muscle

power and endurance capacity of athletes. The

designed nutribar is organoleptically accepted

and nutritious in terms of protein, zinc, iron,

calcium and fiber. The formulated nutribar can

be recommended as healthy nurturing snack for

athletes to increase their endurance performance.

Acknowledgments

My in-depth thanks to my guide Dr.

Lalitha Ramaswamy M.Sc., Ph.D Associate

Professor and Head, Department of Nutrition and

Dietetics for her guide ship and support

throughout this research and Dr. R. Rajendran

M.Sc., PGDEM., MBA., Ph.D Principal PSG

Arts & Science for permitting me to perform this

research. I acknowledge PSG Institute of

Medical Science & Research Institutional

Human Ethics Committee recognized by

SIDCER for approving my proposal.

References

Brain. E Grimwood., (1975) “Coconut palm

products, Their processing in developing

countries. Food and agricultural

organization of united nations”.

Dawn Jackson, RD., (2003). Speaker for the

American Dietetic Association.

Edward., (2013) “8 Foods high in zinc”, Global

healing centre. www.globalhealingcenter.

com/natural-health/foods-high-in-zinc

Göhl., (1982) FAO, Division de Production et

Santé Animale, Roma, Italy.

Gopalan. C, Rama Sastri B.V and

Balasubramaniam S.C, Narasinga Rao,

Deosthale Y.G, Pant K.C., (2012)

“Nutritive value of Indian Foods”.

National Institute of Nutrition, Indian

council of Medical Research. Hyderabad-

500007, India.

Henry C Lukaski., (2000) “Magnesium, zinc,

and chromium nutriture and physical

activity”. American Society for Clinical

Nutrition, Vol. No.72 (6), 585-593.

http://www.globalhealingcenter.com/natural-health/foods-high-in-zinc

http://www.globalhealingcenter.com/natural-health/foods-high-in-zinc

http://ajcn.nutrition.org/search?author1=Henry+C+Lukaski&sortspec=date&submit=Submit

Cord 2015, 31 (1)

12

Lukaski HC., (2000) “Magnesium, zinc, and

chromium nutriture and physical activity”.

American Society for Clinical Nutrition

Vol. No.72 (6), 585-593.

Maughan R.J., (2001) “Clinical sports nutrition”

Roseville, NSW: McGraw-Hill Book

company Australia Pty Lit. (pp.369-390).

Ralf R. Henkel1

, Kerstin Defosse1

, Hans-Wilhelm

Koyro2

, Norbert Weissmann3

, Wolf-

Bernhard Schill1

., (2003) “Estimate of

oxygen consumption and intracellular zinc

concentration of human spermatozoa in

relation to motility” Asian. J. Androl Mar;

5: 3-8

Sesikeran B., (2010). Revised RDA for Indians,

National Institute of Nutrition ICMR,

Hyderabad.

Stephanie Henry., (2013) “Establishing

nutritional value in dehydrated coconut

meat and palm products fed to pigs”.

http://www.journalofanimalscience.org/co

ntent/91/3/1391.

Stone Herbert, Sidel Joel., (1993) “Sensory

Evaluation Practices”, Academic Press

Incorporated, Jan 1, Science pp. 338.

Taisun H Hyun, Elizabeth Barrett-Connor, and

David B Milne., (2004) “Zinc intakes and

plasma concentrations in men with

osteoporosis: the Rancho Bernardo Study”.

American Journal of Clinical Nutrition,

vol. (80) no. 3, pp. 715-721.


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http://ajcn.nutrition.org/search?author1=David+B+Milne&sortspec=date&submit=Submit

Cord 2015, 31 (1)

13

Ecofriendly Organosolv Process for Pulping of Tender Coconut Fibre

Jincy P.J.1, Anita Das Ravindranath

2 and U.S. Sarma

3

Abstract

The huge biomass generated by vendors of tender coconut is the broken husks refuse dumped

along roads and highways in Kerala. These dumps become breeding grounds for diseased causing

germs and carrier mosquitoes causing threat to human life. In order to avoid pollution and find use of

the rejected biomass of tender husks, a study was carried out on pulping of tender coconut husk fibre

which could be used for papermaking. The optimum pulping condition, the quality of the pulp and its

yield was evaluated using different variables like time and temperature. It was observed that the

organosolv process could efficiently remove lignin from the tender coconut fibre yielding maximum

cellulose. During the traditional pulping processes such as Kraft pulping to isolate the cellulose fibers

for the production of paper, the hemicellulose and lignin fractions are degraded, limiting their

valorization possibilities. Organosolv pulping has been advocated as the environmentally benign

version of the kraft process. Unlike other pretreatment methods, organic solvents can easily be recycled

and reused. The lignin dissolved by organosolv pulping is easily recovered by dilution and is

unsulphonated and relatively unmodified. Products like handmade paper, egg cartons, handicraft items,

garden articles like paper pots could be made from the organasolv pulp of tender coconut husk fibre.

Keywords: tender coconut, organosolv pulping, morphology, handmade paper, organosolv lignin

____________________________________

1Department of Environmental Engineering, National Chung Hsing University, Taichung 402, Taiwan, ROC.


2Microbiology Department, Central Coir Research Institute, Kalavoor P.O, Alleppey Dist, PIN-688522 Kerala,

India. Email: [email protected]

3Ex-Director, Research (RDTE), Central Coir Research Institute, Kalavoor P.O, Alleppey Dist, PIN-688522

Kerala, India. Email: [email protected]

https://mail.google.com/mail/u/0/h/qgurbp9mcusb/?&v=b&cs=wh&[email protected]

Cord 2015, 31 (1)

14

Introduction

The coconut palm (Cocos nucifera), an

important member of the family Arecaceae

(palm family) is a tropical tree that is cultivated

in nearly 90 different countries (Pires et al.

2004). It is estimated that out of total production

of coconuts in the country 10% are plucked as

tender coconuts. The water of the tender

coconut, technically called the liquid endosperm,

is enjoyed by people in tropical regions of the

world, especially in Tropical Asia as well as

Central and South America. It was discovered

that this water has medicinal characteristics as an

antioxidant, replenish fluid hydration, provide

parenteral nutrition, etc. Despite its benefits, the

market for coconut water causes solid waste

environmental problems and is one of the major

agro-industrial waste generators in some

developing countries. It is a lignocellulosic

material which is resistant to easy decomposition

due to its high lignin content and causes

environmental pollution. The husk containing

about 80-85% moisture acts as breeding sites for

insectsmosquitoes, flies etc. With the objective

of utilization of the tender coconut

huskagrowaste and reducing the environmental

impact of its disposal, an ecofriendly organosolv

process for pulping of tender coconut fibre could

be developed.

Tender coconut fibre is a lignocellulosic

material and is composed of carbohydrate

polymers (cellulose and hemicellulose), lignin

and a remaining smaller part (extractives, acids

and salts and minerals). Cellulose is a polymer

composed of chains of six carbon sugars

(primarily glucose). These chains are bundled

into strong fibers that on close inspection are

seen to have an organized crystalline structure.

Lignin is a complex, three dimensional polymer

composed of linked six-carbon phenolic rings

with various carbon chains and other chemical

functionalities. Lignin is non-crystalline, and its

structure has been described as analogous to a

gel or foam. The lignin serves to bind the

cellulose fibers. It is degradable by only few

organisms, into several higher value products

such as organic acids, phenols and vanillin. Via

chemical processes valuable fuel additives may

be produced. Hemicellulose is a very complex

polymer composed of mainly xylose (five-

carbon) and further arabinose (five- carbon),

galactose, glucose and mannose (all six carbon);

it also contains smaller amounts of non-sugars

such as acetyl groups. Hemicellulose, because of

its amorphous nature, is relatively easy

tohydrolyzes. Hemicellulose attaches weakly to

both cellulose and lignin and fills the intervening

spaces. Extractives are non-structural

components of biomass samples that is soluble

either in water or ethanol. It includes, but is not

limited to sucrose, nitrates, nitrites, protein,

chlorophyll, and waxes (fatty acids). The ash

content is a measure of the mineral content and

other inorganic matter in biomass.

Regardless of the application of

lignocellulosic materials, it is required a

preliminary processing to separate the three

macromolecular fractions, particularly the lignin,

which can be considered the main physical

barrier for making the fibers (mainly cellulose)

cemented together. These processes modify the

lignocellulosic material by disruption of cell wall

structure of plant biomass, removing,

solubilizing or depolymerizing lignin. The kinds

of processes depend on the material used and the

proposed purpose of lignocellulosic fractions

utilization and may be mechanical, physical,

biological or chemical. The development of

pretreatment processes strong enough as to

separate the cell wall arrangement and mild

enough as to avoid a significant chemical

degradation of biomass components is a

challenge for today’s chemical industry

(Canetieri et al, 2007). For the novel

pretreatment methods it is advisable to use cheap

and easily recoverable chemicals and low-cost

equipment. The use of environmentally friendly

and low energy-intensive approaches is highly

desired. The chemical cooking process is the

most efficient and most used to perform the

separation of lignocellulosic components of

vegetal biomass (Fernandez, 1996). It results in

enlargement of the inner surface area of substrate

particles, accomplished by solubilization and/or

degradation of hemicellulose and lignin.

The shortage of raw materials and the

large energy and water consumptions are

presently the main concerns of pulp and paper

Cord 2015, 31 (1)

15

industry. Considerable research effort has been

made to introduce alternative species as raw

materials, like biomass from agricultural and

forestry residues. Organosolv processes were

experimented based on the use of organic

solvents as delignification agents, to degrade the

lignocellulosic biomass to obtain cellulose fibers

for paper making, high quality hemicelluloses

and lignin degradation products avoiding

emissions and effluents. Organosolv

pretreatment has been evaluated as an effective

pretreatment method for high lignin

lignocellulosic biomass. The treatment can break

down internal lignin and hemicellulose bonds

and thus remove all of the lignin from biomass.

A strong inorganic acid is usually applied in the

organosolv pretreatment as a catalyst to

hydrolyze the lignin-lignin and lignin-

carbohydrate bonds in biomass. Inorganic acids

including hydrochloric acid, sulfuric acid and

phosphoric acid have been frequently chosen.

Generating high quality lignin is one of the

unique advantages of the organosolv

pretreatment over alternative methods, such as

steam explosion, dilute acid treatment and hot

water treatment, where the only proposed use for

the lignin is as a boiler fuel. The organosolv

spent liquor mixed with water could precipitate

the dissolved lignin. In contrast to lignin

produced by other technical processes, such as

Kraft pulping, organosolv lignin is a sulfur free,

low molecular weight product of high purity.

High quality lignin can be used as a substitute

for polymeric materials, such as phenolic powder

resins, polyurethane foams and epoxy resins (Li

et al, 2012). The other main advantages of the

organosolv processes over the conventional ones

are the following: low environmental impact,

higher pulp yield, ease of bleaching, easy solvent

recovery, low capital for a new plant and

recovery of lignin and sugars for profitable

utilization (Saberikhah et al, 2011).

In this investigation, the feasibility of an

organosolv pulping process for the production of

pulp from depithed tender coconut husk fiber

was evaluated. Acetic acid was selected as an

organic solvent for tender coconut husk fibre

pulping, considering the results obtained in

preliminary experiments and due to its easy

availability. In the present study, a 70% aqueous

solution of acetic acid was selected for tender

coconut fibre pulping. Dilute sulfuric acid was

applied as the catalyst for organosolv pulping.

The effect of cooking time and temperature on

pulp properties like yield, Kappa number and %

delignification were investigated. The bleaching

ability of organosolv pulp by totally chlorine free

(TCF) bleaching using alkaline hydrogen

peroxide was also examined.

Conventional methods of bleaching

involve treatment with molecular chlorine or

chlorine based chemicals. Effluent from these

processes produces large amounts of chlorinated

organic compounds and releases into the

environment. The interest in totally chlorine free

bleaching processes has led to the development

of peroxide based bleaching. One of the biggest

advantage of hydrogen peroxide is that it is

environmentally friendly throughout its whole

life cycle. The perhydroxyl anion (HOO-) is the

principal active agent in peroxide bleaching.

This anion is a strong nucleophile which during

bleaching converts electron rich chromophores

to their non chromophoric counterparts. The

reaction of lignin with peroxide is not reversible

and lead to the permanent removal of most of the

chromophoric groups in the lignin molecule

(Presley and Hill, 1996).


Substrate preparation

Tender coconut husk used in this work was

collected from the local vendors in Alleppey

district. Husk was depithed using shredder and

the fibers were separated. After the separation of

fiber from pith, the fibers were chopped into

uniform sized pieces and air dried. The fibre had

an average moisture content of 11.7%.

Chemical analysis of tender coconut fibre

The specimens were sampled and

characterized according to the standard methods.

Chemical composition, given on an oven dry

weight basis, was the following: 27.02%

Cellulose (Uppdegraf, 1969), 56.82%

Holocellulose (ASTM D1104-56(1978), 36.9%

Acid insoluble lignin ( TAPPI T 222 om-98),

3.62% extractives in alcohol-benzene (TAPPI

Cord 2015, 31 (1)

16

T204 cm-97) and 3.1% Ashes at 5250C (TAPPI

T211 om-02).

Organosolv pulping of tender coconut fiber

Organosolv pulping involves contacting a

lignocellulosic feedstock with an aqueous

organic solvent at temperatures ranging from

140°C to 220°C. This causes lignin to break

down by hydrolytic cleavage of alpha aryl-ether

links into fragments that are soluble in the

solvent system. So in order to achieve high

temperature, acetic acid cooking was performed

under different conditions in a laboratory

pulping unit provided with six autoclaves, each

of 2.5 liter rotate in a heated poly glycol bath.

The ratio of liquor/ fibre, acetic acid and H2SO4

catalyst concentration was maintained constant

at 10:1, 70% and 0.5% (v/v) respectively.

Pulping was done at four temperatures (1500C,

1600C, 170

0C and 180

0C) and five cooking times

5,10,15,20 and 25 min respectively at each

temperature. After the digester was loaded with

tender coconut fibre and cooking liquor, the

temperature was allowed to rise. The rising

period to the operating temperature is maintained

constant as 70 minutes. After pulping for

different time intervals the digester was kept for

10 min before opening. The spent liquor was

immediately separated from the pulp by filtering

in order to avoid lignin precipitation. The

resultant pulp was then washed with fresh

corresponding acetic acid and finally with water

and air- dried overnight. The performance of the

pulping process was evaluated by Kappa number

and yield. The Kappa number (TAPPI T236 cm-

85) is the volume (mL) of 0.1 N potassium

permanganate solution consumed by 1 g of oven

dry fibrous material under the conditions

specified in this standard method.

Bleaching of organosolv pulp

Bleaching of organosolv pulp was carried

out using 15% hydrogen peroxide by weight of

the dry pulp at a pH of 11.5 at 1: 15 solid to

liquid ratio. 3% Sodium silicate was added to

stabilize hydrogen peroxide. The material was

treated for 4 hours at 80oC. After treatment, the

pulp was taken out, washed with cold water and

air dried in the shade.

Fiber morphology using Scanning electron

microscopy (SEM)

The changes in the morphology of the

fibers were examined microscopically. For

scanning electron microscopy (SEM) analysis,

fiber was cut into small samples, mounted on

stubs with adhesive, and then they were placed

under vacuum, evacuated and sputter-coated

with platinum using auto fine coater (JEOL JFC-

1600). After preparation of samples, the samples

were observed under the Scanning Electron

Microscope (JEOL JSM-6380LV).

Thermal stability using thermogravimetric

analysis (TGA)

The thermal stability and decomposition

analyses of the fibre and pulp were evaluated by

thermogravimetric analysis (TGA) using a

Mettler Toledo TGA/SDTA 851e

Thermogravimetric Analyser. A sample mass of

2-3 mg were heated from 50 º C up to 700 º C at

a rate of 5º C/min under nitrogen atmosphere in

a constant flow of 80 cm3 min-1.


Optimization of organosolv pulping of tender

coconut fibre

The pulp yield, Kappa number, and %

delignification of organosolv pulp obtained from

tender coconut husk fibre by different cooking

conditions have been furnished in Figures 1,2

and 3 respectively.

The first assessment after any stage of

lignocellulosic material treatment is the yield

measurement (which is a parameter for the

process classification). The total yield is

measured as the ratio between the mass of

material obtained after the treatment stage and

the initial mass used to perform the same.

(Candido et al, 2012).

The results indicate that the temperature

and reaction time had a significant influence on

delignification. It was found that with increase in

temperature and time there was a decrease in

pulp yield, Kappa number and residual lignin.

However, it was seen that at all temperatures the

Cord 2015, 31 (1)

17

delignification reactions were slow down after

the initial 15 minutes reaction time.

Figure 1. Effect of Temperature and Reaction

Time on Pulp Yield


Time on Kappa Number


Time on Delignification

It was seen that the pulping at 1800C

resulted in an increased delignification. However

the pulp yield was lowered to an unacceptable

level. From the studies carried out it could be

concluded that pulping at 1700C for 15 minutes

was the most suitable condition for digestion of

the fibre. Pulping at these conditions gave higher

pulp yield and % delignification with low Kappa

number. When the reaction time increased from

15 to 25 minutes the delignification didn’t

increase significantly and the pulp yield was

reduced from 39.94% to 34.49%.The pulp yield

at 180oCis very low compared to the pulp yield

at170oC which is undesirable for the pulp and

paper production. Increase in reaction time from

5 to 25 minutes resulted in the decrease of pulp

yield from 30.97 to 21.08. This indicates that at

higher temperature the loss of weight of tender

coconut fiber was more due to the loss of more

amounts of cellulose and hemicellulose content

along with lignin.

Therefore, the optimum cooking

conditions for a bleachable pulp of tender

coconut husk fibre are as follows: acetic acid 70

%, temperature 1700C, cooking time 15 minutes

and catalyst concentration 0.5%. The pulp yield,

% delignification and Kappa number values at

this condition were 39.94%, 5.2 and 85.91

respectively.

Tender coconut fiber before and after

organosolv pulping

Figure 4. Raw fiber

Cord 2015, 31 (1)

18

Figure 5. Fiber after organosolv treatment

at 1700C and 15 min

The material tends to get a dark colour

after organosolv treatment (Figure 5) in contrast

to raw fiber which is brown in colour (Figure 4).

Therefore, the pulp has to be subjected to

bleaching for acceptable brightness. In addition

the structure of tender coconut fiber after

organosolv treatment changed dramatically

compared to the raw material (from fibers to a

cardboard like appearance). The cellulose and

hemicelluloses present in lignocellulosic fibers

do not contribute significantly to coloration, due

to their naturally white color characteristics. On

the other hand, lignin and other extractives

contribute to its darkness and therefore can be

removed by bleaching.

After pulping, acetic acid can be

regenerated from the black liquor by simple

distillation and can be re-used in the cooking.

The dissolved lignin and hemicelluloses in the

acetic acid are collected as concentrated slurry

after the distillation. The concentrated slurry is

mixed with water, lignin is separated as

precipitate and the hemicelluloses remain in

water solution. The hemicelluloses are

precipitated in ethanol. Therefore, we can utilize

all the fractions of tender coconut in the acetic

acid pulping.

Bleaching of the organosolv pulp was

done by hydrogen peroxide. Physical and optical

properties of the organosolv pulp were improved

during hydrogen peroxide bleaching. After

bleaching the pulp yield, % delignification and

Kappa number of the organosolv pulp at 1700C

for 15 minutes was 92.6, 94.31 and 16.2

respectively.

Surface morphology of tender coconut fibre

and pulp

Microscopic examination reveals that a

single fiber comprises a bundle of multiple cells

called ultimates. These ultimates in aggregate

bundle are bound together by cementing

materials like lignin and oriented roughly

parallel to one another (Figure 6). The fibre

shows globular structures in regular intervals

(porous) and other constituents, probably organic

residues (Mohanty and Nayak 2004; Ameida et

al 2006) of the extraction of the fibres. The

fibrils are covered by a cuticle layer, i.e. wax of

aliphatic origin (Ratta 1999). Each cell is

roughly polygonal in shape, with a central hole

or lumen, comprising about 10% of the cell area

of cross section.

Figure 6. Cross–section of Raw coir fiber shown

in SEM

After Organosolv treatment, the impurities

and wax cuticle layers of the fibres surface were

removed. The fibrils separate from each other

due to removal of the lignin, the cementing

component, by the action of the organic solvent,

leading to an increase of the surface area (Figure

7a, b&d).

Cord 2015, 31 (1)

19

(a)

(b)

(c)

(d)

Figure 7. (a) & (b) micrograph of fiber after

organosolv pulping, (c) surface of the

fiber after organosolv pulping, (d)

micrograph of pulp after blending

The treatment also leads to changes in the

hydrogen bonding interactions of hydroxyl

groups of cellulose, resulting in the deformation

of individual micro fibrils (Ganan and

Mondragoni 2005). The treatment with acetic

acid showed a more effective result in the

lixiviation of the cuticle layer in removal of

bonding material. Under this condition, the most

external layer of the fibre was eliminated,

revealing the fibrillar structures (Figure 7a&b).

Grinding separates the individual fibers (figure 7

d). The surface morphology of the unbleached

pulp (Figure 7 c) showed globular protrusions,

identified as silicate stegmata. The cavities

became more defined, and the surface fibres

became rougher than in the untreated fibre.

(a)

Cord 2015, 31 (1)

20

(b)

Figure 8. (a) & (b) micrograph of the organosolv

pulp after hydrogen peroxide

bleaching

After hydrogen peroxide treatment the

surface of the organosolv pulp was free of silica

bodies, exposing the visible pits (Figure 8a&b).

The surface became smoother than the

unbleached pulp. The pits measured about 2.44

µm in diameter.

Thermo gravimetric Analysis / Simultaneous

Differential Thermal Analysis (TGA/SDTA)

of raw fibre and pulp

The thermo gravimetric (TG) and

derivative of the thermogravimetry curve(DTG)

curves for the raw tender coconut fibres and

organosolv treated fibres are shown in Figure 9

and 10. The results indicate that the range of

thermal degradation decreased after organosolv

treatment ( Karnani 2004, Mohanty 2000). The

DTG curves show the first peak below 1000C as

a result of evaporation of residual moisture.

(Pothan et al. 1997; Pothan and Thomas 2003;

Eichorn et al. 2001).

Thermal degradation in N2 atmosphere was

characterized by an overlapping of processes that

may occur in the same range of temperature,

which can be separated only by observation of

one of the peaks in the DTG output.

The treated fibre showed a single

degradation peak in contrast to the double peaks

observed in untreated fibres results. This is a

further indication of a higher homogeneity

resulting from organosolv treatment which also

indicates that the macrocomponents were

removed by the organosolv treatment.

(Esmeraldo et al 2010).

The degradation in the treated fibres is

attributed to the pyrolysis of cellulose and to

residual hemicelluloses and this is shifted to

lower temperature when the amount of lignin is

reduced. Reported by Mitra et al (1998), Varma

et al (1986) and Van Dam et al (2004) state that

weight losses above 2000C can be attributed to

the oxidation and condensation of carbohydrates,

while the phenolic components of lignin are

considered to be more stable. Also Vazquez-

Torres et al (1992) after extraction and

characterization of lignin samples from coconut

fibre, verified that their decomposition starts

from a temperature of approximately 3800C.

Figure 9. TG/DTG curve of raw tender coconut

fibre

Figure 10. TG/DTG curve of organosolv pulp

treated at 1700 C, 15 min

Cord 2015, 31 (1)

21

Handmade papers and garden articles made

from tender coconut husk organosolv pulp

The organosolv pulp was diluted with

water and then poured uniformly over a screen.

The wet pulp was then transferred onto a cloth to

remove the excess water. As the water drains

out, the fibers come closer and closer together

forming a tightly bonded mesh. The wet sheet

was pressed to remove the remaining moisture.

The pressed sheets were then laid on a smooth

surface and dried under the sun. After the drying,

calendaring of the handmade paper was done

between stainless steel sheets. It was observed

that blending bleached organosolv pulp with

waste paper pulp in suitable proportions

improves strength and surface smoothness of the

handmade paper.

(a)

(b)

Figure 11. (a) Seedling pots made from

unbleached organosolv plp, (b)

Handmade paper made from

bleached organosolv pulp

The pulp can also be moulded into various

disposable articles like handmade paper, egg

cartons, paper plates, cups, glass covers, garden

articles like paper pots for seedling, and

packaging materials.

Conclusions

The potential of tender coconut fibre for

making diversified products by an ecofriendly

organosolv process could be confirmed.

Optimum digestion conditions to obtain a

bleachable pulp of tender coconut husk fibre

were as follows: using acetic acid 70 %, at a

temperature of 1700C and cooking time 15

minutes in presence of a catalyst (0.5%). The

pulp yield, % delignification and Kappa number

values at this condition were 39.94, 85.91 and 40

respectively. After bleaching the %

delignification and Kappa number of the

organosolv pulp has become 94.31 and 16.2

respectively. The drawbacks of kraft pulping

relate to bad-odor problems, sulfur usage and

bleaching problems could be eliminated

considerably by applying the ecofriendly

organosolv pulping processes. The recovery of

the solvent from black liquor is easy and

effective. Lignin is precipitated and can be

separated from the black liquor after distillation.

Therefore, complete utilization of the tender

coconut husk fiber is possible by this

environment friendly pulping method.

Acknowledgment

The authors gratefully acknowledge the

financial support of the Coconut Development

Board, Ministry of Agriculture, Government of

India. Thanks are due extend to Prof. G.

Balachandran, Ex-Chairman, Coir Board for his

keen interest and guidance in this work.

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http://onlinelibrary.wiley.com/doi/10.1002/(SICI)1439-2054(20000301)276:1%3C1::AID-MAME1%3E3.0.CO;2-W/abstract




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Cord 2015, 31 (1)

24

Low Temperature Grafting of MMA on to Coir Fibre

Lakshmi N. S.1, Sarika Babu

2, Sumy Sebastian

3 and P.K. Ravi

4

Abstract

Low temperature grafting of methyl methacrylate (MMA) on to coir fibre was carried out in

aqueous medium using Potassium per sulphate (PPS) as an initiator under the catalytic influence of

Ferrous ammonium sulphate (FAS). Optimization of various parameters of grafting viz. monomer,

initiator and catalyst concentration, time and temperature was carried out to obtain the maximum

tensile properties. Evidence of grafting was characterized from Fourier Transform Infrared

spectroscopy (FTIR), Scanning Electron Microscopy (SEM) and Thermal analysis (TGA). The

maximum breaking stress (BS) of control and grafted coir fibre were 213.08 and 365.00 N/mm2

respectively. Hence the percentage of improvement of grafted coir fibre was found to be 71.30%.

Increase in tensile properties with maximum BS observed under monomer (25%), initiator (0.75%) and

catalyst (0.75%) concentration, time (150min) and temperature (500C) respectively. The t-test and

Analysis of variance (ANOVA) were studied for statistical significance and the P values obtained were

less than 0.05 which revealed that the value was highly significant for the improvement of mechanical

strength on coir fibre by graft Co- polymerization.

Keywords: Methyl Methacrylate, Potassium persulphate, Ferrous ammonium sulphate, grafting, Co-

polymerization.

____________________________________

1Incubation Assistant, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey,

Kerala. Email: [email protected]

2Project Assistant, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey, Kerala.


3Scientific Assistant, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey, Kerala.


4Senior Scientific Officer, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey,

Kerala. Email: [email protected]

Cord 2015, 31 (1)

25

Introduction

Natural fibers are abundant, versatile,

renewable, cheap and biodegradable raw

material which can be used for making wide

variety of products. Natural fibers have a number

of advantages such as low cost, less toxicity, non

abrasive nature and easy process ability. Among

the natural fibres, Coir occupies a prominent

position owing to its qualities such as natural

resilience, biodegradability, salt resistance,

mechanical strength and toughness which

enhances its commercial and industrial

usefulness. (Singha and Raj, 2010)

Coir is a multicellular biopolymer in

which crystalline cellulose is arranged helically

in a matrix consisting of non crystalline

lignocellulosic complex. (Ott et. al., 1954). Due

to hydrophilic nature and low thermal stability

coir fibers have got certain limitations. There are

several methods such as acetylation,

benzoylation, grafting, silane treatment, plasma

treatment etc. which have been employed to

improve the properties of the natural fibers

through surface modification. Graft copolymers

are assuming increasing importance because of

their tremendous industrial potential. Graft

copolymerization is one of the best techniques

for modifying the properties of natural polymers.

It is a convenient and clean means for altering

the properties of numerous polymer backbones.

As the natural fibres bear hydroxyl groups from

cellulose and lignin, they are amendable to

chemical modifications. Chemical modification

may activate these groups or can introduce new

moieties that can effectively interlock with

matrix. Modifying the properties of natural

polymers through graft copolymerization has

been reported by various workers. (Renu et. al.,

2010). Desirable and targeted properties can be

imparted to the natural polymers through graft

copolymerization in order to meet out the

requirement of specialized applications. Grafted

fibers have been found to increase the strength

and thermal stability than raw fibers.

In the present study coir fibre was grafted

with Methyl Methacrylate (MMA) for improving

its properties such as tensile strength, thermal

stability, light fastness and resistance to

chemicals. In order to make modification process

more economic, effort has been made to find out

the optimum modification condition depending

on the concentration of monomer, initiator and

catalyst, reaction time and temperature. Graft

yield was determined on the basis of the weight

increase of the coir fibre treated. Some

instrumental analysis such as Infrared

spectroscopy (IR), Scanning Electron

Microscopy (SEM) and Thermo Gravimetric

Analysis (TGA) were done to identify graft on

coir fibre.


Materials

Mechanically extracted coir fibres were

collected from Alleppey, Kerala. Monomer:

Methyl Methacrylate (MMA), Initiator:

Potassium per sulphate (PPS) and Catalyst:

Ferrous ammonium sulphate (FAS) used was of

analytical grade.

Methods

Purification of materials

The fibres were cleaned first in Willowing

machine and sorted using combing board. The

fibres were soaked in water over night. Then it

was washed with water under stirring for 8 hrs

and dried in hot air oven at 700C.

MMA was washed with 5% NaOH

solution followed by water and dried over

anhydrous sodium sulphate. Catalyst ferrous

ammonium sulphate was recrystalised from hot

water. Initiator potassium per sulphate was used

as received.

Graft Co-polymerization

Fibres were immersed in distilled water for

24 hrs prior to the reaction. The material to

liquor ratio was maintained at 1:50. A known

amount of the initiator, catalyst and monomer

were then added to the reaction vessel

maintained at required temperature. Grafting of

MMA on to coir fibre was carried out under

constant stirring. At the end of the desired

reaction period, the coir fibre was thoroughly

washed with acetone to remove any

homopolymer generated during the reaction.

Cord 2015, 31 (1)

26

Fibres were then washed with distilled water and

dried in a hot air oven at 700C till a constant

weight was obtained. Polymerization reaction

were done at varying conditions of 5-75%

monomer, 0.0-1.5% initiator, 0.0-1.25% catalyst

based on the weight of the fibre, 300C-80

0C of

temperature and 30-210 minute reaction time on

to the coir fibre. The reaction conditions were

optimized so as to get maximum Grafting Yield

(GY) and BS. Percentage GY was calculated by

the equation

GY = (W2-W1/W1) 100 (1)

Where W1 and W2 are the weights of coir

fibre and grafted fibre respectively.

Scanning Electron Microscopy (SEM)

Scanning Electron Microscopy studies of

ungrafted coir fibre and grafted coir fibre was

carried out on Scanning Electron Microscope

(SEM) (JEOL JSM-6390 LV, Tokyo, Japan)

system with an accelerating voltage of 10 kV.

The SEM images are studied under 10µm

magnification.

Infrared Spectroscopy (IR)

Infrared spectra of the ungrafted and

grafted coir fibre were taken on Perkin Elemer

Spectrophotometer.

Thermo Gravimetric Analysis (TGA)

The Thermo Gravimetric Analysis (TGA)

of the coir fibre was done using Mettler Toledo

TGA/SDTA 851e, Japan. The test was carried

out in an inert atmosphere under nitrogen flow

rate at 10ml/min and heating rate throughout the

test was 100C/min. The weight change was

recorded as a function of the heating

temperature.

Tensile properties

Tensile properties of coir fibres were

determined using UTM (Universal Testing

Machine) Shimadzu AG-X/R. At test

parameters; strain rate = 10mm/min, gripping

length=5cm at atmospheric temperature.


Mechanism

It has been observed that C2, C3, C6

hydroxyls and C-H sites of cellulose back bone

are the active sites of coir fibre for grafting of

MMA. PPS takes part in the redox reaction with

Fe2+

to produce SO4-*.

Fe2+

+ -O3S-O-O-SO3

- Fe

3+ + SO4

2- + SO4

-* (2)

Interaction of SO4-* with H2O generates

OH free radicals (OH*) and these free radicals

are responsible for free radical generation on coir

and the monomer as well as further chain

propagation there by resulting in the formation

of graft copolymer.

Optimization of different reaction parameters

for grafting Methyl Methacrylate on to Coir

Fibre

Optimization of various reaction

parameters such as concentration of the

monomer, initiator, catalyst, reaction time and

temperature were carried out for graft co-

polymerization of methyl methacrylate on to coir

fibre.

i. Effect of monomer concentration

Grafting increases with increase in

monomer concentration up to 25% and further

increase in monomer concentration results in

decrease of grafting yield (fig 1). This may be

due to the formation of more homo polymer as

compared to graft co-polymer at higher

monomer concentration. Due to homo

polymerization, viscosity of the reaction medium

increases which creates hindrance in the

movement of the free radical towards the active

sides, there by resulting in decrease of grafting

yield (Singha et.al., 2007 ).

ii. Effect of initiator

The initiator plays an important role in

obtaining higher graft yield. It is observed (fig 2)

that the percentage of grafting increases with

increase in initiator concentration PPS up to

0.75% and there after the percentage of grafting

starts decreasing.

Cord 2015, 31 (1)

27

-2

0

2

4

6

8

10

12

14

100

150

200

250

300

0 5 15 25 35 45 55 65 75G

Y (

%)

BS

(N

/mm

2)

Monomer concentration (%)

BSGY

Figure 1.

-8

-7

-6

-5

-4

-3

-2

-1

0

1

2

100

150

200

250

300

0 0 0.25 0.5 0.75 1 1.25 1.5

GY

(%)

BS

(N

/mm

2)

Initiator concentration (%)

BS

GY

Figure 2.

Initiator PPS generates activated

monomers, which show a sufficient affinity to

cellulose macro radicals that are produced by the

action of catalyst FAS on the surface of coir

fibre and enhances the grafting up to a certain

initiator concentration (Arifuzzaman Khan et.

al., 2009). But at a high initiator concentration

large number of activated monomers

accumulates and forms the homo polymer,

thereby reducing the active monomers available

in the vicinity of cellulose macro radicals and

hence the percentage of grafting starts

decreasing. Decrease in grafting may also be

explained due to the termination of growing

grafted chain by primary free radical resulting

from the decomposition of excess initiator.

iii. Effect of catalyst

It is observed that (fig 3) grafting yield

increased with increasing catalyst concentration.

As the catalyst concentration increases, more

numbers of Fe (III) ions as well as more reactive

site on the coir fibre surface are produced,

thereby increasing the grafting up to a certain

concentration of catalyst. But beyond this

concentration the grafting yield declined. This is

probably due to the formation of large number of

Fe (III) ions which promote the premature

termination of the growing grafted chain.

-8

-6

-4

-2

0

2

4

100

150

200

250

300

350

0 0 0.25 0.5 0.75 1 1.25

GY

(%

)

BS

(N

mm

2)

Catalyst concentration (%)

BSGY

Figure 3.

iv. Effect of reaction time

The percentage of grafting increases with

increase in reaction time up to 150min for MMA

and there after the percentage of grafting starts

decreasing (fig 4). The initial sharp increase in

grafting is due to the high co-polymerization

reaction between the reactive sites of coir fibre

and the activated monomer molecules. After

150min the homo polymerization reaction

prevails over the co polymerization possibly due

to the decrease in reactive sites on the coir fibre

(Salam, M.A. 2005). So the percentage of

grafting becomes steady. On prolonged exposure

Cord 2015, 31 (1)

28

to heat, the percentage of grafting decreases due

to the partial dissolution of grafted fibre.

Figure 4.

v. Effect of temperature

It is observed that at ordinary temperature,

the grafting is slow. But at 500C the percentage

of grafting is high (fig 5). The increasing

percentage of grafting up to optimum

temperature may be due to increase in the rate of

production of active free radical which increase

the number of grafting sites at a higher rate, the

rate of initiation by coir fibre radicals is thereby

increased. The increase in temperature increases

the rate of diffusion of monomers into the coir

fibre matrix. The decrease in percentage of

grafting beyond optimum temperature may be

attributed to the increase in the rate of homo

polymerization.

Figure 5.

The t-test was studied for statistical

significance and all the P values obtained were

less than 0.05 (table). Since it shows that the

values were statistically significant. Analysis of

variance (ANOVA) of BS of grafted Coir fibre

(P<0.0021) was also calculated and the p value is

(p<0.05) revealed that this was highly significant

for the improvement of mechanical strength on

Coir fibre by graft Co-polymerization.

Characterisation Study

Scanning Electron Microscopy (SEM)

It is quite evident from figures that there

has been a sufficient deposition of MMA on to

coir fibre. Comparison of the Scanning electron

micrographs of ungrafted coir fibre Fig. 6(a) and

MMA grafted coir fibre; Fig. 6(b) reveals a clear

cut distinction between the ungrafted and grafted

fibers.

Figure 6. SEM images of (a) Ungrafted

Coir fibre (b) MMA grafted Coir fibre

Figure 6(a)

Figure 6(b)

-7

-6

-5

-4

-3

-2

-1

0

1

2

100

150

200

250

300

350

400

0 30 60 90 120 150 180 210

GY

(%

)

BS

(N

/mm

2)

Reaction time (min)

BSGY

-1

-0.5

0

0.5

1

1.5

2

2.5

3

100

150

200

250

300

350

400

0 30 40 50 60 70 80

GY

(%

)

BS

(N

mm

2)

Reaction temperature (0C)

BS

GY

Cord 2015, 31 (1)

29

The pits observed in ungrafted coir fibre

are no longer observed in grafted fibres. The

surface becomes more or less uniform owing to

deposition of PMMA grafts along the direction

of fibre axis. This orderly arrangement of

PMMA changes the porous surface of coir fibre

resulted in an appreciable increase in the strength

of grafted fibres compared with ungrafted fibres

(Khullar et. al., 2008).

FTIR Spectra

Figure 7. FTIR spectrums of the (a) ungrafted

coir and (b) MMA grafted Coir fibre

Figure 7(a)

Figure 7(b)

The peak assignments of the absorption

band corresponding to various groups are

summarized in table 1 (Zuraida et. al., 2011)

Table 1. FTIR spectral data of ungrafted Coir

fibre

Position / cm Possible Assignment

~3600-3200 ν (OH) broad, strong band from

the cellulose, hemicellulose and

lignin of coir

~3000-2900 ν (C-H) in aromatic rings and

alkanes

~1732.7 ν (C=O) most probably from the

lignin and hemicelluloses

~1608.5 ν (C=C) aromatic in-plane

~1512.6 ν (C=C) aromatic skeletal ring

vibration due to lignin

~1464.2 δ (C-H); δ (C-OH) 10 & 20

alcohol

~1426.6 δ (C-H)

~1375.4 δ (C-H)

~1267.4 δ (C-OH) out-of-plane

~1046.0 ν (C-OH) 20 alcohol

~898.5 ν (C-O-C) in plane symmetric

FTIR spectrum of ungrafted coir and

MMA grafted coir fibre are shown in fig.7 (a-b).

The ungrafted coir fiber shows the characteristic

absorption of -OH group ~3600-3200cm-1

(Zuraida et. al., 2011). But in MMA grafted coir

fibre, broadening of absorption band takes place

at ~3600-3200cm-1

. The intensity of –OH peaks

in MMA grafted fibres reduced as a result of

grafting, since it is the probable site of grafting.

Grafted fibre shows an additional peak at

~1732cm-1 may be due to the presence of (C=O)

of MMA.( Balamurugan et. al., 2004; Jogeswari

et. al., 1999a).

Thermo gravimetric analysis (TGA)

Thermal behavior of ungrafted coir fibre

and its MMA grafted coir fibre were examined

by a study of their TGA thermograms. From the

results, listed in Table 2, it is evident that initial

and final decomposition temperature of

ungrafted coir fibre are 80 and 4500C where as in

the case of MMA grafted coir fibre the initial

and final decomposition temperature are 900C

and 6700C respectively (Fig. 8 (a & b ). From the

97.9

90

80

70

60

52

Wavenumbers (cm-1

)

% T

ransm

itta

nce

4000 3000 2000 1500 1000 400

97.9

85

75

65

55

Wavenumbers (cm-1

)

% T

ransm

itta

nce

4000 3000 2000 1500 1000 400

Cord 2015, 31 (1)

30

results it is observed that the thermal stability of

MMA- grafted fibre is higher than that of

ungrafted fibre. (Jayaraj et. al., 2014)

Figure 8(a)

Figure 8(b)

Table 2. TGA of ungrafted and MMA grafted

Coir fibre

Tensile Properties

The tensile properties of ungrafted and

grafted coir fibre were determined by taking a

minimum of 50 fibres from each optimization

fig. 1 to 5.

It clearly shows that there is an appreciable

change in the tensile properties under maximum

stress at break. Grafting of coir fibres with MMA

from control to complete optimization brings

about a substantial increase in maximum stress at

break. The increase in maximum stress in the

case of completely optimized fibre from 213.08

N/mm2

to 365 N/mm2

may be due to the increase

in the degree of crystallinity due to the orderly

arrangement of the short grafts of the PMMA

units on the cellulosic backbone. In all the cases

of grafting it has been found that the maximum

stress at break is higher than ungrafted fibre Fig.

9 (a). (Jogeswari et. al., 1999b)

Figure 9. Tensile properties of Optimized

Condition

Figure 9(a)

Conclusion

Low temperature Graft Copolymerization

of MMA on to coir fibre was done at an

optimum temperature of 50OC. Percent of

grafting increases with the increase in each of

the reaction parameters, such as monomer

concentrations, initiator concentrations, catalyst

concentrations, polymerization time and

temperature up to a limited extent and then

decreases. During graft copolymerization

Sample T10

(°C)

T50

(°C)

T90

(°C)

Untreated coir fibre

80

310

450

MMA grafted coir

fibre

270

350

630

213.08

284.96 289325

360

365

0

50

100

150

200

250

300

350

400

C MGf Igf CGf TGf °CGf

Bre

ak

lo

ad

(N

/m

m²)

Optimized conditions

Maximum stress at break

Cord 2015, 31 (1)

31

keeping all other variables constant, better graft

yield were obtained at 50OC, 2.30 hr, 25%

monomer, 0.75% initiator, 0.75% catalyst. The

polymer grafted on to coir fibre improves

magnificently the tensile strength. FTIR, SEM

and TGA measurements characterized the extent

of grafting.

Acknowledgments

The authors are very much grateful to Coir

Board, Cochin and Central Coir Research

Institute (CCRI), Kalavoor for permitting to

conduct the studies and publication of this

article.

References

Arifuzzaman Khan, G.M., Sheruzzaman., S.M.

Abdur Razzaque., Sakinul Islam and

Shamsul Alam. 2009. Grafting of

acrylonitrile monomer on to bleached okra

bast fibre and its textile properties. Indian

J. Fibre and Textile Research 34: 321-327.

Balamurugan, A., Kannan, S., Selvaraj, V. and

Rajeswari, S. 2004. Development and

Spectral Characterization of Poly (Methyl

Methacrylate) /Hydroxyapatite Composite

for Biomedical Applications. Trends

Biomater. Artif. Organs, Vol. 18 (1), 41-

45.

Jayaraj A.P., Anitha Das Ravindranath and U.S.

Sarma. 2014. Nanocellulose from diseased

coconut wood. Cord 30 (1), 1-10.

Jogeswari R., Manjusri M. and Amar K.M. 1999.

Surface Modification of Coir Fibers I:

Studies on Graft Copolymerization of

Methyl Methacrylate on to Chemically

Modified Coir Fibers. Polym. Adv.

Technol. 10: 336-344.

Kaith, V.S and S. Kalia. 2008. Graft

Copolymerization of MMA on to flax

under different reaction conditions: a

comparative study. eXPRESS Polymer

Letters 2(2): 93-100.

Khullar, R., V.K. Varshney., S. Naithani and

P.L. Soni. 2008. Grafting of acrylonitrile

on to cellulosic material derived from

bamboo (Dendrocalamus strictus).

eXPRESS Polymer Letters 2 (1): 12-18.

Ott E, Spurline H M & Graffin M W, 1954.

Cellulose and cellulose derivatives, Part II

(Interscience, Newyork), 863

Renu S., S Vashistha and S. Kalia. 2010.

Biologically and chemically modification

of ramie fibers: A comparative study.

International Conference on Advance in

polymer Technology.239

Salam, M.A. 2005. Graft co polymerization of

methyl acrylonitrile monomer on to

sulfonated jute- cotton blended fabric. J.

Textile and Apparel technology and

Management 4(4).

Singha, A.S., S. Anjali and V.K. Thakur. 2007.

Pressure induced graft copolymerization of

acrylonitrile on to Saccharum cilliare fibre

and evaluation of some properties of

grafted fibre. Bull. Master. Sci. 31(1):7-

13.

Singha A.S and Raj K.R. 2010. Graft

Copolymerization of Methyl Methacrylate

(MMA) onto Agave americana Fibers and

Evaluation of their Physicochemical

Properties. International Journal of

Polymer Analysis and Characterization

11(1):27-42.

Zuraida, A., S. Norshahida., Sopyan and H.

Zahurin. 2011. Effect of fibre length

variation on mechanical and physical

properties of coir fibre reinforced cement-

albumin composite (CFRCC). J. IIUM

Engineering 12(1).

Cord 2015, 31 (1)

32

Characterisation of Silver Deposited Coir Fibers by

Magnetron Sputtering

Melvi Chandy1, U.S. Sarma

2, M.S. Latha

3 and K. Shreekrishna Kumar

1*

Abstract

Silver thin films are extensively used due to their superior optical, electrical and antimicrobial

properties. Recent development in the incorporation silver thin films on natural fibers makes it possible

to utilize its excellent physical and chemical properties in the field of textiles. Present study focuses on

the surface functionalization of natural coir fiber with silver thin film by magnetron sputtering. This

will help to widen the use of natural coir fibers. The surface morphology of the coated coir fibers are

investigated by employing a scanning electron microscope. The results show that the surface

functionalization of silver-coated coir fibers are highly versatile, and it possess excellent protection

against ultraviolet radiation, exhibit excellent hydrophobicity (contact angle=105.2°) and good

antibacterial effects. This study demonstrates that treatment, which uses silver thin films by magnetron

sputtering, is a promising method for achieving multifunctional coir fabrics.

Keywords: Silver thin film/magnetron sputtering/hydrophobicity

____________________________________

1School of Technology and Applied Sciences, Mahatma Gandhi University, Malloossery P.O, Kottayam

District, Kerala-686 041, India. E-mail: [email protected]

2Central Coir Research Institute, Kalavoor Post Office, Alapuzha District, Kerala-688522, India.

3S.N. College, Neduvaramcode P. O., Alappuzha District, Kerala-689 508, India.

*To whom correspondence should be addressed. E-mail: [email protected]


Cord 2015, 31 (1)

33

Introduction

Natural fibres are environmentally friendly

materials, with many advantages like eco-

friendliness, biodegradability, user friendly etc.

Even though they have some markable

advantages, inherent drawbacks of the natural

fibres are high hydrophilic nature Bledski &

Gassan (1999), low resistance to microbial

attack, the tendency to form aggregates during

processing and poor surface adhesion for

association with a polymer matrix Cho et.al

(2002). Various physical and chemical methods

have been employed to improve the properties of

natural fibers. The chemical method involves

dewaxing, delignification, bleaching, acetylation,

mercerization, salinization and chemical grafting

Mohanty et.al (2001) and physical methods

involves plasma treatment, corona treatment,

thin film coating and irradiation Seong and Hae

(2010).

The coir (Cocos nucifera) is an important

lignocellulosic hard and stiff fibre obtained from

coconut husk, by the process of retting or

mechanical combing. Because of its hardwearing

quality, durability and other advantages, it is

used for making a wide variety of floor

furnishing materials, yarn, rope etc Silver et. al

(2001). However, these traditional coir products

consume only a small percentage of the total

world production of coconut husk and these

products are facing stiff completion with the

synthetic fiber alternatives. Hence, research and

development efforts have been underway to find

some diversified application areas for coir fibres

including utilization of coir fibres as

reinforcement in polymer composites Moteiro et.

al (2008), desiccant materials Ratanakaran et. al

(2010) geotextiles, and sound absorbents.

Wassilief (1996).

In recent years, natural fiber industries

have been focussing on improving the functional

properties of fibers such as UV protection, fire

retardency, and antibacterial properties.

Tremendous effort has been made to apply silver

thin films on fibers, including electroless plating,

electroplating, and vacuum deposition. However,

such conventional techniques generally use wet-

chemical processing and produce much waste

water. On the other hand, as eco friendly and dry

technology, sputtering is an attractive alternative

by adding new functionalities such as

hydrophobicity, UV protection and antibacterial

properties, making it a promising technique for

future applications. The ability to deposit a well-

controlled silver coating on textiles would extent

the applications of coir fibers, based on changes

to both the physical and chemical properties of

textiles. Not much study has been reported on

the surface functionalisation of coir fibers.

This investigation is directed towards

coating of coir fibers with silver thin films by

magnetron sputtering technique. Their

morphological characterisation is studied by

using SEM. Moreover the coated coir fibers are

studied to characterise the functional properties

such as hydrophobicity and antibacterial activity.

The thermal stability of the coated fibers are also

studied.

Experimental

Silver target with high purity (purity:

99.99%) is used to deposit silver thin films onto

coir fiber by using a magnetron sputtering

system. The dimensions of the silver target are

50 mm in diameter and 5 mm in thickness. In the

present study acetone is used for removing the

fats and oils on the coir fiber substrates. Prior to

deposition, the coir fiber samples are first

immersed into an acetone solution for 2 min, and

then washed thrice with deionised water.

Following that, the coir samples are dried at a

temperature of 80°C for 24 hours.

The pressure before evaporation is kept at

2* 10-6

Torr and then it is increased to about

7*10-6

Torr when argon is introduced into the

vacuum chamber as the bombardment gas. The

deposition rate is 1Ǻ/sec for all coating. The

thickness of the coating was monitored by using

a quartz crystal monitor. The sputtering current

and voltage are maintained at 0.15A and 115V

respectively. Since the size and shape of the coir

fibers may vary from fiber to fiber, we had taken

a set of 20 samples to produce reliable results.

The deposition time is 10min and 20 min on the

samples.

Cord 2015, 31 (1)

34

The surface morphology and chemical

composition analysis of the silver coated coir

fibers are performed by a scanning electron

microscope (SEM). The thermal stability of the

uncoated (UC) and silver coated (SC) are

evaluated by means of a thermo gravimetric

analyser. Water contact angle (WCA) of the

samples is measured at ambient temperature

using a SEO Phoenix optical contact angle

system. A volume of 5µl water is applied in this

test. The hydrophobicity of the silver coated coir

fabrics are further confirmed by water drops test,

floatation and swelling tests.

Antibacterial tests on UC and SC coir

fibers are carried out following the Standard

‘SNV 195920-1992’. The antibacterial activity

of the samples is assessed through a diffusion

test in Agar Aldrich). In this method, an E. Coli

colony is positioned in a petri dish filled with

agar gel. UC and SC fibres are placed over the

colony and the whole dish is incubated in oven at

37oC for 24 h. After this period of time, the dish

is removed from the oven and the area covered

by the bacterial colony with respect to the

samples is evaluated. If a growth inhibition zone

is observed close to the sample (characteristic

zone size ≥1 mm), antibacterial property is

indicated as ‘good’. If the sample is totally

rehabitated by the bacteria, the antibacterial

property is indicated as ‘not sufficient’.


The surface morphology of the coated and

uncoated coir fibers is shown in the Fig.1 The

SEM study confirms that silver particles are

grafted on to the fiber suraface. The surface of

uncoated coir fiber shows the presence of

micro pores. Metal coated fiber, on the other

hand is smooth and without any pores

indicating the formation of metal layer on the

surface. After sputtering for 10min, a thin layer

of silver is obtained on the fiber surface. With

an increase in sputtering time homogenous silver

depositions are observed on the coir fibers.

(a) untreated

(b) silver coated (10min)

(c) silver coated (20min)

Figure 1.

The hydrophobicity of the silver coated and

uncoated coir fibers is examined by WCA

measurement. Fig. 2 shows the microscopic view

of the contact angle. Generally coir fibers are

highly hydrophilic due to the presence of

abundant hydroxyl groups on the surface. When

metal particles are coated on the surface of the

fabric, the fabric becomes hydrophobic. The easy

rolling of the water droplet on the fabric surface is

the primary evidence for that. The

hydrophobisation occurs due to the layer of metal

coated on the surface of the coir fiber. The WCA

on the surface of the coir fabric coated with silver

for ten selected areas is measured as 105.2.2o

+/-

2.5oC for a 5µl water droplet on the silver coated

Cord 2015, 31 (1)

35

fabric. The WCA measurement of silver coated

fabric (20 minutes) is 106.3o

C. All the samples

showed almost same WCA measurements. After

5 washes the metal coated coir fabrics shows 25%

decrease in the WCA as compared to the uncoated

one. The reason for this decrease in WCA of

coated fabric is due to the slow removal of metal

particles from the surface of the coir fabric during

washing.

Figure 2. Microscopic view of water contact

angle

The water repellent property is further

confirmed by water drops test and floatation tests.

In water drops tests, the drops of controlled size

are placed at a constant rate upon the fabric

surface and duration of time required for them to

penetrate the fabrics are measured. The results are

shown in Table 1, in which the absorption time is

recorded for different metal coated coir fabrics.

From the results it is clear that the water

repellency of the coir fabric is increased

considerably with the metal coating. In the case of

uncoated coir fabric the water droplet disappears

immediately within seconds, with water spreading

over large areas. In the case of coated fabric the

droplet remained spherical even after 30min with

no spreading, indicating that all the modified

fabrics are water repellent. From the floatation

tests (Fig 3) it is found that the coated fabric

floated on the surface of water for more than 100

minutes, whereas the uncoated coir fabrics

gradually sink to the bottom, when they are

placed on the water surface. Fig 4 shows the

variation of percentage water absorption coated

and uncoated coir fabric with time. More

absorption of water is obtained in untreated coir

fabric and lesser absorption in metal coated coir

fabrics. The samples with 20minutes sputtering

time showed lesser absorption of water at ambient

temperature than the respective untreated sample.

Figure 3. Floatation tests for a) uncoated and b)

silver coated coir fabric

Cord 2015, 31 (1)

36

Figure 4. Water absorption versus time

untreated coir fiber Silver

coated (10min sputter coated

Silver coated (20 min sputter

coated

Table 1. Absorption time of untreated and silver

coated coir fibers

Samples Absorption time

Untreated

Ag coated sample for 10min

Ag coated sample for 20min

3s

>100min

>100min

The thermal stability of the uncoated and

coated fibers is found out by using TGA. The

TGA of all the coated and uncoated samples are

shown in Fig 5a & 5b. From the figure it is

evident that the thermal stability increased by

metal coating. In the case of untreated coir fabrics

there are three stages of degradation. The first one

is attributed to the evaporation of water and

occurs between room temperature and 1500

C.

The second step, which corresponds to the

hemicelluloses degradation, starts at about 1900 C;

the third step occurs between 290 and 3600

C,

corresponding to thermal degradation of cellulose.

Lignin presents a broad peak throughout the

range, degrading between 280 and 5000

C. The

untreated coir fibers are completely oxidised to

form char residue at 7000C Alvaraz & Wasquez

(2004), Tomzac et. al (2007). Metal coating is

found to have a considerable effect on the thermal

degradation behaviour of the coir fibers,

promoting an increase in the temperature at which

the thermal degradation take place These changes

may be attributed to the quick distribution of heat

on the surface of the coated coir fabric when

compared to uncoated one.

(a)

(b)

Figure 5. TGA analysis of a) untreated fiber b)

silver coated (10min)

Table 2. Thermogravimetric analysis of coir

fibers and silver coated coir fibers

No. Sample Transition

temperature

range (0C)

Temperature

of maximum

Weight

loss

(%)

1

2

Untreated

Silver

coated

30-150

150-297

297-500

30-170

170-320

320-500

53

286

334

55

301

360

5

26

45

5

26

49

Cord 2015, 31 (1)

37

Figure 7. Antibacterial Activity Test

Table 3. Characteristic antibacterial zone for coated and uncoated coir fiber

No. Sample Sputtering

Time (min)

Bacteria Inhibition

Zone

Level of

antibacterial

capacity. Pollini

et. al (2009)

1

2

3

Untreated

(3845)

Silver Coated

(3847 & 3849)

Silver Coated

(3846 & 3848)

10

20

E. Coli

E. Coli

E. Coli

------

<1mm

>1mm

Not sufficient

Fairly good

Good

In literature it is reported that, either

polyester or cotton fibers coated by silver exhibit

antibacterial capability against E. coli. In this

study antibacterial tests are performed on an

uncoated and silver coated coir fiber. The effect

of silver compared with the control samples is

determined by measuring the size of the area of

inhibition growth close to each sample. As clearly

shown in Fig. 3, the E. coli colony seeded in the

petri dish filled with agar grows on the untreated

coir sample after incubation in oven at 370C for

24 h: untreated coir fiber does not display any

antibacterial activity. The characteristic

antibacterial zone of uncoated and coated coir

fibers are given in Table 3. Silver in its metallic

state is inert, but it reacts with moisture on the

skin and fluid on a wound, and become ionized.

The ionized silver is highly reactive as it binds to

tissue proteins and brings structural changes to the

bacterial cell wall and nuclear membrane leading

to cell distortion and death. Jiang et. al (2010). It

can be observed a clearly defined bacterial free

zone around each coated sample which confirms

the growth inhibition effect induced by silver

ions.

Cord 2015, 31 (1)

38

Conclusion

Silver thin films on coir fibers are prepared

by magnetron sputtering technique. SEM anlysis

shows a uniform layer of silver over the coir

fiber surface. The results obtained from water

contact angle confirms that the hydrophobicity

of the silver coated coir fabric is higher than that

of the uncoated fabric. Silver coated coir fabrics

show excellent antibactrial activity against E.

Coli. as confirmed by the antimicrobial tests.

Such silver coated coir fabric could therefore be

used in water repellent coating and antibacterial

applications. The investigation described

suggests that the silver coated coir fabrics will be

promising multifunctional material for military

and medical application.

References

Alvaraz V.A, Vazquez. 2004. Thermal

degradation of cellulose derivatives/starch

blends ans sisal fiber biocomposites,

Polymer degradation and stability, 84; 13-

21.

Bledski AK, Gassan J. Composites reinforced

with cellulose based fibers. 1999. Progress

in Polym. Sci.; 24: 221-274.

Cho A, Lee SG, Park WH, Han SO. 2002. Eco-

friendly biocomposite materials using

biofibrils. Polym. Sci. Technol. I; 13; 460-

476.

Mohanty AK, Misra M, Drsal LT. 2001. Surface

modification of natural fibers and

performance of the resulting composites;

an overview, Comp Interfaces, 8: 313-43.

Moteiro SN, Terrones LAH, Almeida JRM.

2008. Mechanical performance of coir

fiber/polyester composites, Polymer

Testing, 591-595.

M. Pollini, M. Russo A. Licciulli, A. Sannino, A.

Maffezzoli. 2009. Characterization of

antibacterial silver coated yarns, Journal

of Material Science, 20, 2361–2366.

Ratanakorn Rawangkula, Joseph Khedaria,

Jongjit Hirunlabhb, Belkacem Zeghmatic.

2010. Characteristics and performance

analysis of a natural desiccant prepared

from coconut coir, Science Asia, 36; 216-

222.

Seong OH, Hae YC. 2010. Morphology and

surface properties of natural fibers treated

with electron beam. Microscopy: Science

Tachnology, Applications and education,

1880-1888

Silva GC, Souza DA, Machado JC, Hourston DJ.

2001. Mechanical and thermal

charecerization of Brazilia coir fibers,

Journal of Applied Polymer science, 76,

1197-1206

S.X. Jiang, W.F. Qin, R.H. Guo, L. Zhang. 2010.

Surface functionalisation of

nanostructured silver coated polyester

fabric by magnetron sputtering, Surface

Coating and Technology, 204, 3662-3667.

Tomzac, F., Satyanarayana, K.G., Sydenstricker,

T.H.D. 2007. Studies on lignocellulosic

fibers of Brazil: Part II- morphology and

properties of Brazilian coconut fibers,

Composites: Part A, Applied Science and

Manufacturing, 38, 1710-1721.

Wassilief C.1996. Sound absorption of wood

based materilas, Applied acoustics, 48;

339-356.

Cord 2015, 31 (1)

39

An Investigation of the Tender Nut Potential of Diverse Coconut

(Cocos nucifera L.) Varieties/Forms in Sri Lanka

S.A.C.N. Perera1, G.K. Ekanayake

2 and H.M.N.B. Herath

3

Abstract

There is a rising demand in the world for coconut water as a healthy natural beverage. Different

coconut varieties are used in different countries to be processed as a natural drink. The coconut form

“King coconut” has long been used in Sri Lanka as the ideal coconut variety for this purpose. However,

with the expansion in the local and export beverage coconut market, the supply does not meet with the

demand at present. Therefore, this study was conducted to identify potential coconut varieties/forms

mainly relating to the yield of beverage. The study was conducted in the main coconut triangle and

Southern Sri Lanka. Eleven coconut forms, namely, King coconut and Bothal thembili in the variety

Aurantiaca, Red, Yellow, Green and Brown dwarf and Murusi in the variety Nana, Bodiri, Dothalu,

and Sri Lanka tall. These are included in the variety Typica along with Nipol, which is a natural hybrid

being identified to be suitable as beverage coconuts. Out of them the yielding ability was high in King

coconut, Red dwarf and Yellow dwarf while Bodiri, Nipol and Dothalu fared well. Many coconut

forms suitable for beverage purpose were observed to be having seasonality in nut production.

Ensuring adequate soil fertility and prevention of water deficit over prolonged periods were observed

to help sustain yield and reduce seasonality in bunch emission. There were no large scale plantations

for beverage coconuts in Sri Lanka, and the entire collection was from home gardens and very small

scale holdings. It is recommended to establish medium scale holdings with coconut forms having high

yield potential as identified in the current study, with proper management guidance to ensure a steady

supply of beverage coconuts to the local as well as export market.

Keywords: Beverage, Coconut, Cocos nucifera, King coconut, Varieties

____________________________________

Coconut Research Institute, Lunuwila, Sri Lanka.

[email protected]

[email protected]

[email protected]



Cord 2015, 31 (1)

40

Introduction

Currently, there is a worldwide demand for

coconut as a natural beverage. Coconut water

became a popular natural beverage firstly in the

coconut producing countries in Asia and in

Brazil and later in western countries, where a

huge demand has been created at present. There

is a very high international demand for coconut

water, especially in the United States of

America, and spreading in the Europe and

Middle Eastern Countries as well.

Coconut water has gained recognition as a

healthy non-alcoholic natural beverage, suitable

as a restorative energy and sports drink. Coconut

water is rich in ions and minerals containing,

Potassium (at high concentration) and

Magnesium, Calcium, Phosphorous, Iron,

Copper, Sulphur, Chlorides, Sodium as well as

amino acids and vitamins B and C. Coconut

water is equally beneficial for everyone

including children and the aged. It is of pleasant

taste and has the added bonus of a low

concentration of sugar in comparison to many

other natural beverages.

Almost all of the main coconut growing

countries are in Asia except Brazil. Indonesia,

the Philippines, India, Brazil and Sri Lanka are

the major coconut growing countries. Out of

them the main economic product in Brazil is

tender coconut water, while kernel and copra

products have been economically more

important in Asia. Despite this, there are

expanded tender coconut industries in Asian

countries such as India and Thailand. In these

countries different varieties and hybrids are used

for beverage purposes. In India, Chowghat

Orange Dwarf has been found to be the best

tender nut variety followed by Malayan Orange

Dwarf and Philippine Ordinary (Dhamodaran et

al., 1993). An aromatic and other dwarf varieties

are popular in Thailand and other South East

Asian countries. Although there may be several

varieties suitable as tender coconuts there is

competition for them to be used in other

industries especially kernel based products such

as oil and copra.

Sri Lanka is the 5th largest coconut

producer and has the highest per capita

consumption of coconut in the world. The

estimated per capita consumption of coconut in

Sri Lanka is around 110 nuts/year (Anon, 2009)

and 80% of the production is consumed

domestically. Much of the production is used in

kernel-derived products: oil; copra; and

desiccated coconut; in addition to the use of

mature green nuts in daily cuisine. However,

consumption of both tender and mature coconut

water as a beverage has been a tradition in Sri

Lanka over a long period of time and there are

certain varieties preferred for this purpose.

The semi tall coconut variety King

Coconut (locally known as Thembili or Rath

thembili) has long been known in Sri Lanka as

the ideal variety for tender coconut beverage

purpose (Liyanage, 1958; Bandaranayake and

Fernando, 1999). King coconut is intermediate in

form between tall and dwarf. It is semi tall, has

a medium height growth rate, produces a root

bole and is predominantly self pollinating. It

bears profusely fruit having an orange epicarp.

King coconut production in Sri Lanka is almost

exclusively for tender coconut water. However,

there are several other varieties which are less

commonly used for beverage purposes and yet

with a higher potential productivity as beverage

coconuts. Therefore this study was conducted

with the objective of identifying varieties having

a potential for beverage production and studying

some of their specific characteristics which make

them suitable for tender nut purposes.


A field survey was conducted in Puttalum,

Chillaw and Gampaha in the main coconut

triangle in the North Western province and

Galle, Unawatuna area in the Southern Sri

Lanka. The coconut varieties which were used

for beverage purposes were identified.

Approximately 30 palms (except for

Dothalu and Nipol in which the sample sizes

were smaller) of each of the identified varieties

were used for recording the variables: number of

bunches; number of nuts in each bunch; nut

water volume at 7-8 month maturity stage;

seasonality in bunch emission; and nut setting.

Observations were made over a period of one

year and data were analysed by deriving means

Cord 2015, 31 (1)

41

Table 01. Coconut varieties used as beverage coconuts at the tender nut stage

Common name Variety/form (in Sri

Lankan classification)

Distribution and occurrence of palms

Thembili

Bothal Thembili

Red Dwarf

Green Dwarf

Brown Dwarf

Yellow Dwarf

Murusi

Bodiri

Dothalu

Nipol*

Sri Lanka Tall

Aurantiaca/King coconut

Aurantiaca/Bothal thembili

Nana/Red Dwarf

Nana/Green Dwarf

Nana/Brown Dwarf

Nana/Yellow Dwarf

Nana/Green Dwarf „Murusi‟

Typica/Bodiri

Typica/Dothalu

A natural hybrid

Typica/Typica

Throughout the coconut growing areas.

Only in the Southern Province of Sri Lanka

Throughout the country. More common in the Southern Province.







Throughout the country

Throughout the country

*Nipol is a natural hybrid between King coconut and Sri Lanka tall

Figure 01. Coconut forms used as tender nut beverage in Sri Lanka

Cord 2015, 31 (1)

42

and performing an analysis of variance and mean

separation in statistical software SAS v8.


Varieties used for beverage purpose in Sri

Lanka

The results revealed the presence of

several varieties/forms which are used for

beverage at the tender nut stage (Figure 01). Out

of them, King Coconut was identified to be the

dominant variety for beverage purpose in Sri

Lanka. The other varieties were used in more

limited quantities at the tender nut stage (Table

01).

Thembili (King coconut) exceeded 75% of

the tender nut market in Sri Lanka. Also the

main purpose of growing Thembili was to be

used as beverage at the tender nut stage; being 7-

8 months of developmental from flowering. The

coconut form Red Dwarf was also highly valued

as beverage coconuts. The general public could

not distinguish the fruit of Thembili and Red

dwarf due to the close morphological similarity

of the fruit forms. The pericarp colour of both

was orange while the only visual difference was

the unique button-like protrusion at the tip of

Thembili fruit. The remaining dwarf forms;

green, yellow and brown, were used in limited

scale as beverage being less common, especially

in the main coconut triangle of the country. The

main commercially grown cultivar, Sri Lanka

tall was occasionally used for beverage purposes

all over the country. The other listed two tall

forms, Dothalu and Bodiri were also used

because their smaller nut size made them less

desirable for processing into kernel products.

Both were rare and found only in the Southern

Province of Sri Lanka. Murusi has the phenotype

of the Green Dwarf very common in the home

gardens of Southern province. It was a very

popular beverage form in the south of Sri Lanka.

Bothal thembili was also found only in the south

being a form related to the variety Aurantiaca,

but with a different nut shape (Figure 01). Bothal

thembili was also a source of beverage. Among

the various forms used for beverage in Sri

Lanka, are several different phenotypes which

are not found in the main coconut triangle, but

only in the south. The wide occurrence together

of the Tall along with King variety in home

gardens has simultaneously given rise to Nipol, a

natural hybrid. While the King coconut is

naturally self-pollinating there is some out-

crossing. Therefore, when the farmers collect

seednuts of the King coconut there would be a

low percentage of Nipol hybrid seedlings due to

pollen shed by the surrounding Tall palms.

Evaluation of the potential of different

varieties/forms for beverage purpose

The yield variables related to quantity of

nut water measured at the 7-8 month

developmental stage of the nuts are presented in

Table 02.

Number of bunches emitted during a given

period, number of nuts produced in each bunch

and the volume of nut water at the appropriate

stage are the main yield components. These

variables were significantly different among the

studied coconut forms (Table 02). Bodiri

recorded the highest values for both bunch

emission and the nut production. However, the

nuts of Bodiri were smaller resulting in a lesser

amount of water per nut. It was observed that

when the number of nuts per bunch was higher

the size of the nuts became smaller and vice-

versa in prolific bearers: Green Dwarf; Brown

Dwarf; Murusi; Bodiri; and Dothalu, resulting in

lesser volume of water per nut. This factor was

not prominent in the other phenotypes. Clearly

the above values need be considered in

combination rather than in isolation to determine

the yield potential of different varieties and

forms for beverage purpose. The total volume of

nut water calculated in this manner in each

coconut form/variety is presented in Figure 02.

As presented in Figure 02, Red Dwarf, King

Coconut and Yellow Dwarf were the highest

yielders in that order, each recording volumes

exceeding 50 litres per palm in the annum.

Bodiri, Nipol, Sri Lanka Tall and Dothalu were

also higher yielders while the rest of the dwarf

varieties along with Bothal thembili recording

volumes in the range of 30-40 litres of nut water

per palm in the annum.

Cord 2015, 31 (1)

43

Table 02. Beverage related yield variables of different coconut varieties/forms

Common name Mean number of

bunches in the

year

Total nut

production in the

year

No. of

nuts/bunch

Volume of

water/nut (mL)

Thembili

Bothal thembili

Red Dwarf

Green Dwarf

Brown Dwarf

Yellow Dwarf

Murusi

Bodiri

Dothalu

Nipol

Sri Lanka Tall

15.6b

13.6cd

15.5b

14.6bc

13.2d

14.1c

14.2c

16.5a

14.2c

13.5d

13.5d

156c

108.8e

164.3c

150.2cd

137.3d

138.2d

149.1cd

221.1a

184.6b

140.4d

91.8e

10.0

8.0

10.6

10.3

10.4

9.8

10.5

13.4

13.0

10.4

6.8

352bc

316d

346c

231ef

236ef

378b

252e

224f

218f

336c

448a

Figure 02. Annual volume of nut water in diverse coconut varieties/forms

Cord 2015, 31 (1)

44

Seasonality in nut production

Seasonality in bunch emission was

observed to varying degrees in all the coconut

forms studied. In general bunch emission was

low in the months of December to February in

all the forms including the widely grown

commercial Sri Lanka Tall, which is known to

be hardy and more tolerant to variation in

environmental conditions. During this period the

number of bunches, number of nuts per bunch,

as well as the nut size, were observed to be less.

In addition, the dwarfs (except Yellow Dwarf),

Bodiri and Dothalu, showed clear lean periods in

both bunch emission and nut production. This

seasonality was less common in King Coconut,

Red Dwarf and Nipol when they were under

good management and when the palms were not

subjected to prolonged periods of water deficit.

Potential varieties/forms as tender nut

beverage coconuts

In Sri Lanka the predominantly grown

tender nut beverage coconut variety is King

Coconut or Thembili, in local dialect. The

present study provided evidence for the equal

potential of Red Dwarf and Yellow dwarf with

respect to yielding ability and hence need to be

promoted in the development of tender nut

beverage industry within Sri Lanka as well as to

provide for the export market.

In the current study Nipol is the only

hybrid that was tested. Nipol is not a

recommended coconut hybrid in Sri Lanka.

However, this natural hybrid proved to have a

high potential for beverage production.

Similarly, the recommended hybrids between tall

and dwarf coconuts have been known to have a

high potential or beverage (Dhamodaran et al.,

1993) However, in Sri Lanka, there is a very

high competition for the nuts of these

recommended hybrids for different uses mainly

for kernel products, limiting their availability for

beverage. This situation is more so for the most

expansively grown commercial coconut cultivar,

Sri Lanka Tall, which is almost entirely

harvested at the mature stage and used in kernel

based products. Therefore, in Sri Lanka it is

advisable to concentrate on the coconut forms

which have been identified as having a high

potential as beverage coconuts. This will avoid

competition with processors seeking supplies of

mature. Collection of coconut water from the

mature nut at the processing plant of kernel

products is also done. Alternative techniques are

needed in processing this mature nut water for

beverage purpose instead of discarding it. In this

method a smaller amount of water will be

obtained per nut but there will be the twin

advantage of deriving kernel products and saving

the mature nut water for a separate beverage

market.

The composition of nut water is an

important quality issue in beverage coconuts that

has not been considered in the current study.

However, earlier studies have reported that the

composition of nut water depended on the stage

of maturity of the nut (Nathanael, 1970). The

desirable palatability of King coconut and other

beverage forms mainly depends on the sugars

and mineral components present. The maximum

concentration of sugar and optimum levels of

minerals and vitamins are found at the 7-8

months stage from the pollination

(Gunawardena, 1973; Ediriweera, 1996). The

sugar component comprises a larger portion of

reducing sugars compared to non-reducing

sugars (Mohandas, 1982). With the ripening of

the nuts major changes in composition of sugars

takes place. There may be genetic differences in

these factors in the potential beverage varieties

identified in this study and future research efforts

on the biochemical aspects could help in

developing the quality of beverage coconuts.

Beverage coconuts are not grown in

plantation scale in Sri Lanka but only in home

gardens or as small blocks adjacent to larger

plantations. However, with the huge increase in

local and international beverage there is an

urgent need for establishing larger scale coconut

holdings for beverage coconuts. With the

information derived in the current study suitable

varieties for this purpose can be selected to

provide a generically diverse population of nut

water producing palms.

The coconut forms King coconut, Red

Dwarf and Yellow dwarf are the best suited

forms throughout the country. In addition, Bodiri

Cord 2015, 31 (1)

45

and Nipol will also be very high yielders. Using

Sri Lanka tall and recommended coconut hybrids

will not be practical due to the demand for kernel

products. There is a need to rationalise

management of the tender nut beverage coconut

holdings. Regional diversity could avoid

localised periods of water deficit and overcome

seasonality and to stabilise the supply of nuts.

With adequate soil fertility and soil moisture

supply, all the dwarf forms can be promoted as

suitable for tender nut beverage purposes in Sri

Lanka.

Acknowledgments

This research was conducted with the

financial assistance from the Coconut CESS

Fund in Sri Lanka. Authors are grateful to Dr. L.

Perera, Head of the Genetics and Plant Breeding

Division and Dr. J.M.D.T. Everard, former

Deputy Director of the Coconut Research

Institute of Sri Lanka, for their assistance and

comments.

References

Anonymous 2009. Annual Report of the Central

Bank of Sri Lanka.

Bandaranayake, C.K. and Fernando, W.M.U.

1999. Genetic improvement of King

coconut, Cocos nucifera var. aurantiaca in

Sri Lanka. Plant Genetic Resources

Newsletter No. 118:30-33.

Dhamodaran, S., Ratnambal, M.J., Chempakam,

B., Pillai, R.V. and Viraktamath, B.C.

1993. Evaluation of tender nut water in

coconut cultivars. In: Proceedings of the

International Symposium on Advances in

Coconut Research and Development

ISOCRAD II (Eds: M.K. Nair, H.H. Kan,

P.

Ediriweera, N.D. 1996. King coconut. CORD,

XII No.2:43-47.

Gopalasundaram and E.V. Bhaskara Rao).

Oxford & IBH Publishing Co. Pvt. Ltd.,

New Delhi. pp. 123-128.

Gunawardena, M. 1973. A study of the free

amino acids in the liquid endosperm of

coconut. Ceylon Coconut Quarterly

24:102-106.

Liyanage, D.V. 1958. Varieties and forms of the

coconut varieties grown in Ceylon. Ceylon

Coconut Quarterly 11:43-51.

Mohandas, S. 1982. Report of the coconut

processing division. Ceylon Coconut

Quarterly 33:48-56

Nathanael, W.R.N. 1970. Non-conventional uses

and processing techniques for coconut

products. Ceylon Coconut Quarterly

21:99-106.

Cord 2015, 31 (1)

46

Pollen Dispersal and Pollination Patterns Studies in Pati Kopyor

Coconut using Molecular Markers

Siti Halimah Larekeng1,2

, Ismail Maskromo1,3

, Agus Purwito1, Nurhayati Anshori Matjik

1,

and S. Sudarsono1

Abstract

Parentage analysis has been used to evaluate pollen dispersal in Kopyor coconut (Cocos nucifera

L.). Investigations were undertaken to elucidate (i) the dispersal of pollen, (ii) the rate of self and out-

crossing pollination, and (iii) the distance of pollen travel in Pati kopyor coconut population. The

finding of this activities should be beneficial to kopyor coconut farmers to increase their kopyor fruit

harvest and to support breeding of this unique coconut mutant. As many as 84 progenies were

harvested from 15 female parents. As many as 95 adults coconut provenances surrounding the female

parents were analyses as the potential male parents for the progenies. The adult coconut palms were

mapped according to their GPS position. All samples were genotyped using six SSR and four SNAP

marker loci. Parentage analysis was done using CERVUS version 2.0 software. Results of the analysis

indicated that evaluated markers were effective for assigning candidate male parents to all evaluated

seedlings. There is no specific direction of donated pollen movement from assigned donor parents to

the female ones. The donated pollens could come from assigned male parents in any directions relative

to the female parent positions. Cross pollination occured in as many as 82.1% of the progenies

analyzed. Outcrossing among tall by tall (TxT), dwarf by dwarf (DxD), hybrid by hybrid (HxH), TxD,

DxT, TxH, DxH, and HxD were observed. Self-pollination (TxT and DxD) occurred in as many as

17.9% of the progenies. The dwarf coconut was not always self pollinated. The presence of DxD, TxD,

and HxD outcrossing was also observed. The donated pollens could come from pollen donor in a range

of at least 0-58 m apart from the evaluated female recipients. Therefore, in addition to the wind, insect

pollinators may have played an important role in Kopyor coconut pollination.

Keywords: Coconut mutant, abnormal endosperm, parentage analysis, pollen movement, outcrossing

rate, self pollination, SSR marker, SNAP marker

____________________________________

1PMB Lab., Department of Agronomy, Fac. of Agriculture, IPB, Bogor, Indonesia.

2Fac. Of Forestry, Hasanuddin University, Makasar, Indonesia.

3Indonesian Palm Research Institute, Agency for Agricultural Research and Development (AARD), Manado,

Indonesia.

Corresponding author:

S. Sudarsono, PMB Lab., Department of Agronomy, Fac. of Agriculture, IPB, Jl. Meranti – Darmaga Kampus,

Bogor 16680, Indonesia. Ph./Fax. 62-251-8629353; Email: [email protected]

Cord 2015, 31 (1)

47

Introduction

Kopyor coconuts are natural coconut

mutants having abnormal endosperm and only

exist in Indonesia. The endosperm is soft,

crumbly and detached from the shell, forming

flakes filling up the shell (Maskromo et al. 2007;

Novarianto et al. 2014). The Makapuno coconut

grown in the Philipines and other Asian countries

is another example of coconut mutant exhibiting

endosperm abnormality (Samonthe et al. 1989;

Wattanayothin, 2010). This mutant has been used

as parent for hybridizations in coconut breeding

(Wattanayothin, 2005). The Macapuno coconut

exhibits a soft and jelly-like endosperm (Santos,

1999) that is phenotypically different to

Indonesian Kopyor coconut.

The kopyor coconut mutant phenotype is

genetically inherited from parents to their

progenies (Sukendah 2009) and most probably is

controlled by a single locus (K locus) regulating

the endosperm development of coconut

(Sudarsono et al. 2014). However, the identity of

the regulatory locus has not yet been resolved.

The abnormal endosperm phenotype in

kopyor coconut is controlled by the recessive k

allele; therefore, the genotype of kopyor fruit of

coconut would be homozygous kk for the zygotic

embryos and homozygous kkk for the endosperm.

On the other hand, the genotype of the normal

fruit of coconut would either be a homozygous

KK or a heterozygous Kk for the zygotic embryo

and either a homozygous KKK, heterozygous

KKk, or heterozygous Kkk, respectively.

The origin of Kopyor coconut mutant is

not well documented; however, currently the

kopyor palms are found in a number of areas in

Java and southern part of Sumatera (Novarianto

and Miftahorrachman 2000). The district of Pati,

Central Java Province is recognized as one of the

Kopyor coconut production centers. Kopyor

coconuts have existed in this region for

generations, especially the dwarf type of Kopyor

coconuts. Although only in a limited numbers,

Kopyor Tall and Kopyor Hybrid coconut types

also exist along side of the dwarf one.

The tall and dwarf coconut have different

morphological characters and pollination

strategy. Tall coconuts are generally outcrossing

since male flower mature earlier than the female

counterpart in the same inflorescence. Dwarf

coconut tends to self-pollinate because of an

overlapping maturation period between male and

female flowers (Deb Mandal and Shyamapada

2011).

Pollination in coconut most probably is

assisted by insect pollinators or by the wind

(Ramirez et. al. 2004). The family of Diptera,

Coleoptera and Hymenoptera are reported as

effective pollinators of coconut (Ramirez et al.

2004). Distances of pollen transfer between male

and female parents may be used to predict the

type of pollinator assisting pollination in coconut.

Such question may be answerred by studying

pollen dispersal.

Evaluating pollen dispersal in various

plant species usually use an approach based on

the parent – progeny genotype genotype

(Austerlitz et al. 2004). Evaluations have been

done in pines (Schuster and Mitoon 2000),

Dinizia excels - Fabaceae (Dick et al. 2003),

Quercus garryana - Fagaceae (Marsico et al.

2009) and teak (Prabha et al. 2011). Availability

of molecular markers capable of identifying

genotype of parents and their progenies should

assist the pollen dispersal studies. Using such

markers, it should also be possible to estimate

the self-pollination and outcrossing rates in a

certain population (Milleron et al. 2012).

To our understanding, pollen dispersal

analysis has not been evaluated in coconuts.

With the development of kopyor coconut in

Indonesia, availability of information associated

with pollen dispersal should be beneficial

considering the recessive nature of the kopyor

character. Such coconut pollen dispersal

evaluation requires availability of some coconut

progeny arrays and polymorphic loci for

molecular markers of coconut genome.

Co-dominant markers, such as SSR and

SNAP markers for coconut have been developed

and routinely evaluated at PMB Lab, Department

of Agronomy and Horticulture, Faculty of

Agriculture, Bogor Agricultural University

(IPB), Bogor, Indonesia for a number of plant

species. These include coconut (Sudarsono et al.

Cord 2015, 31 (1)

48

2014), cacao (Kurniasih 2012), and nut meg –

Myristica sp. (Soenarsih 2012). Moreover, the

gene specific SNAP markers have also been

developed and used successfully in coconut

(Sudarsono el al. 2014).

The SSR markers have successfully been

used in gene flow analysis of pines (Lian et al.

2001; Burczyk and Koralewski. 2005). SNAP

marker have also been reported as an effective

co-dominant marker for plant analysis (Morin et

al. 2004, Sutanto et al. 2013) and proven to

generate better data quality for the majority of

samples on plant genetic studies (Brumfield et

al. 2003) and population structure analysis

(Herrera et al. 2007).

The objectives of this research were is to

evaluate (i) the dispersal of pollen, (ii) the rate of

self and out-crossing pollination, and (iii) the

distance of pollen travel in Pati kopyor coconut

population. The finding of these activities should

be beneficial to kopyor coconut farmers to

increase their kopyor fruit yield and to support

breeding and cultivar development of this unique

mutant.


Time and Location of Research

This research was conducted during the

period of July 2012 up to January 2014. The

field activities were at the Kopyor coconut

plantation belonging to local farmer’s at

Sambiroto, Pati District, Central Java, Indonesia.

The research site was at the following GPS

location: S 6 32.182 E 11 03.354. The soil in the

evaluated Kopyor coconut plantation is sandy

soil. The laboratory activities were done at Plant

Molecular Biology Laboratory (PMB Lab),

Department of Agronomy and Horticulture,

Faculty of Agriculture, Bogor Agricultural

University, Bogor, Indonesia.

Selection of Parents and Progeny Arrays

There were 164 adult coconut trees in the

field research site, consisted of a mixture of both

kopyor heterozygous Kk and normal

homozygous KK coconut trees. Only 95 out of

164 adult coconut trees in one block of 100x100

m2 were sampled in this evaluation. Based on the

coconut type, the sampled population consisted

of 68 dwarf, 14 tall and 13 hybrid coconuts.

Moreover, based on their phenotype, they were

recognized as 22 normal homozygous KK and

73 kopyor heterozygous Kk coconuts. Map of

the existing coconuts in the research site was

generated using the GPS position of all

individuals.

Six dwarf, seven tall, and two hybrid

coconuts among the kopyor heterozygous Kk

trees were selected as female parents. They were

selected using purposive random sampling to

represent different sites in the sampled

population. A single fruit bunch from each

female parent containing 2-10 fruits/bunch was

harvested 10-11 months after pollination. The

total harvested fruits were collected and

identified as either kopyor or normal fruits. The

identified normal fruits were germinated and

DNA was isolated from leaf tissue of the

germinated seedlings (63 seedlings of normal

fruits). The kopyor fruits are not able to naturally

germinate since this character is lethal. Zygotic

embryos were isolated from the identified

kopyor fruits and DNA was isolated directly

from the whole zygotic embryo tissues (21

zygotic embryos). Among the 84 DNA samples,

26 samples were from tall, 45 from dwarf, and

13 from hybrid female parents.

Genotyping of Parents and Progenies

DNA isolation was conducted using the

CTAB method (Rohde et al. 1995). Either young

coconut leaf or zygotic embryo (0.3-0.4 g) was

homogenized in 2 ml of lysis buffer, containing

0.007 g PVP and 10 μl2-mercaptoetanol. The

homogenized tissues were then incubated in

65°C waterbath for 60 minutes and the mixtures

were centrifuged at 11000 rpm for 10 minutes

using using the Eppendorf 5416 centrifuge.

Supernatant was then transferred to an

Eppendorf tube and an equal volume of

chloroform:isoamyl-alcohol (24:1) was added.

The mixtures were mixed well; centrifuged at

11000 rpm for 10 minutes and the supernatant

was transferred into new microtube.

Cold isopropanol (0.8 volume of

supernatant) and sodium acetate (0.1 volume of

supernatant) were added into the supernatant.

Cord 2015, 31 (1)

49

After overnight incubation, the mixture was

centrifuged at 11000 rpm for 10 minutes and

DNA pellet was retained. The DNA pellet was

washed using 500 μl of cold 70% ethanol,

centrifuged and air dried before it was diluted

into100 μl aquabidest. RNA contaminants were

remove using RNase treatment following

standard procedures (Sambrook and Russel

2001).

SSR marker at 37 loci (Lebrun et al. 2001)

were evaluated for their polymorphism 6

polimorphic loci were selected. In addition, four

SNAP marker loci developed based on

nucleotide sequence variabilities of both SUS

and WRKY genes were also used to genotype all

of the parents and progeny arrays. To generate

markers, PCR amplifications were conducted

using the following reaction mixtures: 2µl of

DNA; 0.625µl of primers, 6.25µl PCR mix

(KAPA Biosystem), and 3µl ddH20.

Amplifications were conducted using the

following steps: one cycle of pre-amplification at

95°C for 3 minutes, 35 cycles of amplification

steps at 95°C for 15 seconds (template

denaturation), annealing temperature for 15

seconds (primer annealing), and 72°C for 5

seconds (primer extension), and one cycle of

final extention at 72°C for 10 minutes as

suggested by KAPA Biosystem kit.

The generated SSR markers were

separated using vertical 6% polyacrilamide gel

electrophoresis (PAGE) using SB 1x buffer

(Brody and Kern 2004) and stained using silver

staining. The silver staining was done following

methods developed by Creste et al. (2001).

Electrophoregrams were visualized over the light

table and used to determine the genotype of the

evaluated samples.

The generated SNAP markers were

separated using 1% agarose gel electrophoresis

using TBE 1x buffer and stained using standard

DNA staining procedures (Sambrook and Russel

2001). The electrophoregrams were visualized

over the UV transluminescence table and

recorded using digital camera. The recorded

pictures were used to determine the genotype of

the evaluated samples.

Identification of the Candidate Male Parents

Each sample of the progeny arrays has a

known female parent but unknown pollen donor

(the male parent). The candidate male parents

could be any one of the sampled adult population

including the female parents. Studies were

conducted to determine the assigned male parent

donating pollen to generate any fruit in the

progeny arrays.

Identification of the assigned male parent

was done by analyzing genotype of progeny and

the respective female parent versus the genotype

of all adult trees in the selected samples. The ID

of the potential male parent for any progeny was

determine based on the results of parentage

analysis. Simulation was conducted to determine

the threshold level for confidence interval of

80% (relax) and 95% (strict) levels before the

final parentage analysis steps. Parentage analysis

using the genotype of progenies, female parents,

and potential male parents was done using

CERVUS version 2.0 software (Marshall et al.

1998). Most likely approach (potential male

parent with the highest LOD score) based on the

matching genotype of progeny, female parent

and potential male parent were used as the basis

for assigning certain adult individual as the

potential male parent or pollen donor of a

progeny. The progeny and female parent

genotype were compared with those of other

adult trees and the assigned male parent was

selected based on the output of CERVUS version

2.0 analysis results (Marshall et al. 1998).

Pattern of Pollen Dispersal

The location of the female and the

assigned male parents were plotted in the map of

adult individuals generated by Garmin

MapSource GPS mapping software version

76C5x. The distance between the known female

parent and the assigned male parent was

calculated using the same software. The

distances and positions of both female and male

parents in the generated map was then used to

ilustrate pattern of pollen dispersal in the

location. Self pollination was defined if the

assigned male parent was the same as the female

parent. Otherwise, they were assigned as

outcrossing. The outcrossing were further

Cord 2015, 31 (1)

50

grouped as outcrossing between either dwarf

(dwarf parent pollinated other dwarf), tall (tall

parent pollinated other tall), or hybrid (hybrid

parent pollinated other hybrid); outcrossing

between dwarf and either tall or hybrid (either

tall or hybrid parent pollinated by dwarf);

outcrossing between tall by hybrid coconuts (tall

parent pollinated hybrid) or vice versa. The

numbers of both self pollination and the

respective cross pollination were calculated.


The Parents and Progeny Arrays

Map of the existing coconut palms in the

research site are presented in Fig. 1. As

indicated, the sample coconut population

consists of a mixture of normal homozygous KK

and kopyor heterozygous Kk individuals and a

mixture of dwarf, tall and hybrid coconuts. All

of these adult trees were used as potential male

parents capable of donating pollens to and

pollinating the selected female parents and

generating the evaluated progeny arrays. The

position of the selected female parents (6 dwarf,

7 tall, and 2 hybrid kopyor heterozygous Kk

coconuts) are indicated in Fig. 1. The harvested

progenies from selected female parents ranged

from 2-10 progenies per female parent. Out of 84

selected progenies, 21 were kopyor nuts and 63

were normal ones. They were harvested from tall

(26 progenies), dwarf (45 progenies), and hybrid

(13 progenies) female parents, respectively.

Genotyping of Parents and Progeny Arrays

The selected SSR and SNAP marker loci

generated polymorphic markers in the evaluated

coconut population. An example of the

polymorphic marker generated by either the

selected SSR (CnCir_56 locus) and SNAP (SUS

1_3 locus) primer pairs producing polimorphic

markers is presented in Fig. 2. and 3. In Fig. 2,

the evaluated individuals are either homozygous

cc (sample #1), bb (sample #7-10), heterozygous

bc (sample # 2-6, and 11), or heterozygous ab

(sample # 12) for the CnCir_56 SSR locus. On

the other hand, the evaluated individuals (sample

# 1, 3, 4, 6) are heterozygous for reference and

alternate SNAP alleles and the other two (sample

# 2 and 5) are homozygous for the reference

allele (Fig. 3). All individuals were genotyped

using the same approaches. The summary of

genotping results for a total of 179 individuals

using six SSR and four SNAP marker loci are

presented in Table 1. The marker loci generated

a range of 2-4 alleles per locus (Table 1).

Mean number of alleles per locus is 3.4

and mean PIC for all marker loci was 0.47. The

polymorphic information content (PIC) for SSR

marker loci ranges from 0.31-0.68 while that of

SNAP markers ranges from 0.28-0.37 (Table 1).

The PIC values represents measures of

polimorphism between genotypes in a locus

using information of the allele numbers (Sajib et

al. 2012). Total exclusionary power using the ten

marker loci is either 0.85 (first parent) or 0.97

(second parent), indicating the SSR and SNAP

markers should be informative enough for

analyzing the evaluated coconut population.

Figure1. Map of study site with the existed

coconut palms at Pati, Central Java,

Indonesia. The marks in this map

indicated the positions of the coconut

provenances. The sampled

provenances in an approximately one

hectare area are in the square box. The

flags indicate the positions of the

selected female parents.

Note: The marks indicate the position

of ( ) normal tall, ( ) kopyor tall, ( )

normal dwarf, ( ) kopyor dwarf, ( )

normal hybrid, and ( ) kopyor hybrid

coconuts, respectively.

Cord 2015, 31 (1)

51

1 2 3 4 5 6 M R A R A R A R A R A R A

1 2 3 4 5 6 7 8 9 10 11 12 M

Figure 2. Polymorphism of SSR markers generated by PCR of the genomic DNA

sample #1-12 with a pair of CnCir_56 SSR primers. M: 100 bp DNA ladder

markers. The a, b, and c are the three specific alleles of the CnCir_56 locus

Figure 3. Polymorphism of SNAP markers generated by PCR of the genomic DNA

samples 1-6 with two pairs (the R: reference and the A: alternate primer

pairs) of the SUS 1_3 SNAP locus. M: 100 bp DNA ladder markers. The R–

PCR product of the R primer pairs and the A – PCR product of the A primer

pairs. The occurrences of both R and A PCR products indicating the

evaluated individuals are heterozygous, while if either onlyA or B indicating

they were homozygous

Cord 2015, 31 (1)

52

Table 1. Number of alleles and individuals, heterozygous and homozygous individuals,

observed (O) and expected (E) heterozygosity, and polymorphic information

content (PIC) at 10 molecular marker loci of Kopyor coconut

Locus Name No. of

alleles

No. of

individual

No. Of Heterozygosity PIC

Hetero-

zygous

Homozy-

gous O E

CnCir_87 2 179 27 152 0.15 0.39 0.31

CnCir_86 4 179 100 79 0.56 0.72 0.67

CnZ-18 4 179 74 105 0.41 0.61 0.57

CnZ_51 5 179 72 107 0.40 0.58 0.54

CnCir_B12 6 176 64 112 0.37 0.72 0.68

CnCir_56 5 179 78 101 0.44 0.64 0.57

CnSus1#14 2 179 143 36 0.80 0.49 0.37

CnSus1#3 2 174 147 27 0.85 0.50 0.37

WRKY19#1 2 176 111 65 0.63 0.47 0.36

WRKY 6#3 2 179 76 103 0.43 0.34 0.28

Table 2. Crossing schemes and pollination types identified based on results of

pollen dispersal analysis of the progeny arrays

Crossing Scheme Pollination types Event numbers Percentage

TxT Self 2 2.4%

DxD Self 13 15.7%

HxH Self 0 0

Sub-total Self 15 17.9

TxT Outcross 4 4.8%

TxD Outcross 16 19.3%

TxH Outcross 4 4.8%

DxD Outcross 15 18.1%

DxT Outcross 6 7.2%

DxH Outcross 11 13.3%

HxH Outcross 2 2.4%

HxT Outcross 0 0

HxD Outcross 11 13.1%

Sub-total Outcross 69 82.1%

Total progenies 84 100.0%

Note: T - tall coconut, D - dwarf coconut, and H - hybrid coconut

Cord 2015, 31 (1)

53

Identification of the Candidate Male Parents

Results of simulation analysis using

10.000 iterations, 95 candidate male parents, and

the known female parent for each progeny,

predicted the rate of success in identifying male

parents at 95% (strict) was 32% and at 80%

(relax) confidence interval was 62%. Parentage

analysis was able to resolve the identity of the

male parent for every individual in the 84

progeny arrays using the most likely parent

approach. Moreover, the results of analysis also

indicated that assignment of the predicted male

parents for the 20% (17 individuals) progenies

are at least in the minimum of 95% confidence

and 43% (36 individuals) were at least in the

minimum of 80% confidence. The assignment

for the male parents of other 57% (48

individuals) progenies were at the level of less

than 80 % confidence. Although the confidence

level was below 80 %, the male parent

assignment for these progenies shows LOD

(likelihood of odds) value higher than 0. A

positive LOD value indicates the suspected male

parent might be the true parents. According to

Marshall et al. (1998), the higher the LOD value

the higher the possibility for the assigned male

parent to be the actual parent (Marshal et al.

1998).

Cross pollination is pollination of female

flower by male pollen from different parents.

Cross pollination produces half-sib progenies.

The tall, dwarf and hybrid coconuts could

reciprocally donate their pollens. Based on the

assigned male parent of the 84 progeny arrays,

cross pollination occured in as many as 69

events (82.1 %). Among those identified as

outcrossing, 4 events are cross pollination

between tall x tall (TxT), 16 tall by dwarf (TxD),

and 4 tall by hybrid (TxH) parents. Moreover,

outcrossing among DxD (15 events), DxT (6

events), DxH (11 events), HxH (2 event) and

HxD coconuts (11 events) were also observed.

Complete scheme and pollination types

identified based on results of pollen dispersal

analysis are presented in Table 2.

The general understanding stated that

because of the open flower morphology and the

differences in flower maturation, tall coconut is

probably always cross pollinated (Ramirez et al.

2004; Maskromo et al. 2011). However, our data

indicated there are at least 2.38% of self

pollination among the tall coconut (Table 2).

Self pollination is characterized by the

pollination of female flower by male pollen of

the same parent. Self pollination produces full-

sib progenies. Total numbers of self pollination

are observed in as many as 15 events (17.9 %) in

the evaluated progeny arrays (Table 2). They

consist of two self pollination events in the tall

kopyor coconut (2.38 %) and 13 self pollination

events in the dwarf kopyor one (15.48%). Based

on 13 progeny arrays harvested from the hybrid

parents, no self pollination in the hybrid coconut

is recorded (Table 2).

The general understanding stated that

because of the overlapping period between male

and female flower maturation, dwarf coconut is

always self pollinated (Maskromo et al. 2011).

However, our data indicated the dwarf coconut is

not always self pollinated. Contrary to the basic

understanding, our data indicated the presence of

more dwarf to dwarf (15 events, 17.86%), dwarf

to tall (6 events, 7.14%) and dwarf to hybrid (11

events, 13.1%) outcrossing (Table 2).

Finding by Rajesh et al. (2008) has

previously indicated that cross pollination did

occur in dwarf coconuts. Availability of new

tools, such as molecular markers, for analyzing

outcrossing rate may change the previous

understanding. Such changes have been shown

in Hymenaea coubaril which was previously

reported as more cross pollinated because of self

incompatibility (Dunphy et al. 2004). However,

more recent pollen dispersal studies indicated

that H. coubaril is more self pollinated (Carneiro

et al. 2011).

Other alternative explanation for this

findings is that it is just a special case in the

evaluated site. In the study site, coconut palms

were planted in high density. Moreover,

population of honey bees exist in the coconut

plantation. Honey bees are known to roam

around the male and female flowers and function

as effective pollinators for coconuts. The high

density planting and the availability of

pollinators may have caused the unexpected

Cord 2015, 31 (1)

54

Figure 4. Numbers of assigned male parent donating different numbers of pollen to evaluated

female parents

Figure 5. Numbers of female parent receiving donated pollens from different number of assigned

male parents

Figure 6. Numbers of pollination events for each distance class from the assigned male to the female

coconut parents

Nu

mb

er

of

ev

en

ts

Class distance between male to female (m)

Number of assigned male parents

Nu

mb

er

of

fem

ale

pa

ren

t N

um

ber

of

male

pa

ren

t

Number of donated pollen

Cord 2015, 31 (1)

55

outcrossing rate. However, those are subjects of

further investigations.

One assigned male parent may donate one

or more pollens to the evaluated female coconut

parent, with a range of 1-5 pollens per assigned

male one. Number of assigned male parents

donating certain numbers of pollen to the

evaluated female parents is presented in Fig. 4.

The data indicate that most of the assigned male

parents contribute only one pollen to the

evaluated female parents. Only three assigned

male parents (two dwarf, and one hybrid

coconuts) donated 4 or 5 pollens to the

surrounding female parents.

The same female parents may receive

donated pollens from different numbers of

assigned male parents, with a ranged of 1-7

assigned male parents donated pollen to the same

female one. The numbers female parents

receiving donated pollens from different number

of assigned male parent iss presented in Fig. 5.

The data indicated a single female parent most

frequently received pollens from 2, 4 or 5

different assigned male parents. Only three

female parents evaluated in this experiment (two

dwarf and one hybrid coconuts) are found

receiving pollens from at least 6 assigned male

parents (Fig. 5).

Pattern of Pollen Dispersal

The distances between female to the

assigned male parents have been determined

based on their GPS positions. The distance of

pollen travel between assigned male to female

parents as measured in this evaluation ranged

from 0 - 58 m. Numbers of pollination events of

each distance class from the assigned male to the

female coconut parents are presented in Fig. 6.

The assigned male parents are distributed almost

evenly in the different class distances from the

female parents. The 0 m distance between

parents indicates self pollination events.

To evaluate pattern of pollen dispersal

among the assigned male parent to the female,

the positions of assigned male parents as pollen

donors to one female parent are plotted to a map

using their GPS positions. Representative

samples of the assigned male parent positions to

a single female recipient parent are presented in

Figures 7-11.

As the female parent, Hybrid kopyor # 059

(Fig. 7) received 6 donated pollens from six

different assigned pollen donors. The pollen

contributors to the progeny array harvested from

Hybrid kopyor # 059 female parent were all

kopyor heterozygous Kk coconuts. However, the

seven progenies harvested from this female

parent were all phenotypically normal, i.e.

genetically either a normal heterozygous Kk or

homozygous KK. The positions of the assigned

male parents relative to the female parent # 059

in the study site are presented in Fig. 7.

Figure 7. Pattern of pollen movement to female

parent #059 inferred from parentage

analysis. The mark indicates position

of ( ) Dwarf kopyor, ( ) Hybrid

kopyor as the assigned male (pollen

donor) parents, and ( ) hybrid

kopyor #59 as the female recipient,

respectively

The Dwarf kopyor # 067 (Fig. 8) received

10 donated pollens from eight different assigned

male parents. The assigned pollen contributors to

the Dwarf kopyor # 067 female parent were all

kopyor heterozygous Kk coconuts. Only one out

of the 10 progenies harvested from this female

parent was phenotypically kopyor. The assigned

male parent for the harvested kopyor fruit was

the tall kopyor # 089. The positions of the

assigned male parents relative to the female

parent # 067 are presented in Fig. 8.

Cord 2015, 31 (1)

56



analysis. The marks indicate position

of ( ) Dwaf kopyor, ( ) Hybrid

kopyor, ( ) Tall kopyor as the

assigned male (pollen donor), and

( ) Dwarf kopyor #067 as the donor

pollens and female recipient,

respectively

Dwarf kopyor # 068 (Fig. 9) received 9

donated pollens from four assigned male parents.

The four progenies were the result of outcross

with either hybrid (# 59) or dwarf (#87 or # 90)

and from self pollination. The assigned pollen

contributors to the Dwarf kopyor # 068 were all

kopyor heterozygous Kk coconuts. Three out of

the 9 progenies harvested from Dwarf kopyor #

069 were phenotypically kopyor. These kopyor

fruits received one donated pollen from either

the tall kopyor # 059, dwarf kopyor # 68 or # 87.

The positions of the assigned male parents

relative to the female parent # 68 are presented

in Fig. 9.



analysis. The marks indicate position

of ( ) Dwarf kopyor and ( ) Hybrid

kopyor as the assigned male parents

(pollen donors), and ( ) Dwarf

kopyor #068 as the donor pollens and

female recipient, respectively


donated pollens from surrounding pollen donors.

The pollen contributors to the Dwarf kopyor #

084 female parent were all kopyor coconuts.

Only two out of the 8 progenies harvested from

Dwarf kopyor # 084 were phenotypically

kopyor. These two kopyor fruits received

donated pollens from either The two assigned

male parents, either dwarf kopyor # 056 (one

pollen) and hybrid kopyor # 057 (one pollen),

each contributed a one pollen to the evaluated

progenies. Moreover, assigned male parent # 32

is the most distance pollen contributor among the

evaluated trees. The positions of the assigned

male parents (pollen contributors) relative to the

female parent # 084 are presented in Fig. 10.

Figure 10. Patterns of pollen movement to

female parent #084 inferred from

parentage analysis. The marks

indicate positions of ( ) Dwarf

kopyor, ( ) Hybrid kopyor, ( )

the Tall kopyor as the assigned

male - pollen donors, and ( )

Dwarf kopyor #84 as the female

recipient, respectively


donated pollens from surrounding pollen donors.

The pollen contributors to the Dwarf kopyor #

089 female parent were all kopyor coconuts.

None of the 7 progenies harvested from Dwarf

kopyor # 089 was phenotypically kopyor. The

Cord 2015, 31 (1)

57

positions of the assigned male parents relative to

the female parent # 089 are presented in Fig. 11.

Figure 11. Pattern of pollen dispersal plotted

based on the assigned male parent

as pollen donor to female parent

#089. The marks indicate position

of ( ) Dwarf kopyor, ( ) the Tall

kopyor as the assigned pollen

donor, and ( ) Tall kopyor #89 as

the donor pollens and female

recipient, respectively

Fig. 7-11 indicated that there is no specific

direction of donated pollen movement from

assigned male parents to the female parents. The

donated pollen could come from assigned male

parents in any directions relative to the female

parent positions.

In the reseach location, wind blows from

left to right during the night and from right to

left during the day. If the wind is the major

pollinators, there should be a specific pattern of

pollen movement. Moreover, the distance of

pollen dispersals should be close to the pollen

donors. Our data did not support the wind as the

only major pollinator in Kopyor coconut since

pollens disperse in random directions and the

assigned male parents are as far as 58 m apart

from the evaluated female recipients. Our data

also indicated that insect pollinators may play an

important role in Kopyor coconut pollination.

Numbers of insects are associated with

inflorescence of kopyor coconuts. Such insects

may aid pollination and promote cross

pollination in kopyor coconuts, as it happens to

other plant species (Bown 1988). These findings,

however, do not rule out the role of wind in the

Kopyor coconut pollination, especially from

closely spacing male pollen donors.

This might have been the first report of

using molecular marker to study pollen dispersal

in coconut. Results of this study point to new

finding about pollen dispersal and pollination,

selfing and out-crossing rates among dwarf,

hybrid, and tall coconuts, respectively. However,

further research and evaluation are necessary to

generalize the finding since the present study is

specific for the current study site.

Conclusion

The evaluated markers were effective for

assigning candidate male parents to all evaluated

seedlings. There is no specific direction of

donated pollen movement from assigned donor

parents to the female ones. The donated pollens

could come from assigned male parents in any

directions relative to the female parent positions.

Based on the assigned male parent of the 84

progeny arrays, cross pollination occured in as

many as 69 events (82.1%) including one among

tall by tall (TxT), dwarf by dwarf (DxD) and

hybrid by hybrid (HxH) cross pollination events.

Moreover, outcrossing among TxD, TxH, DxH

and vice-versa were also observed. This finding

also indicated the dwarf coconut is not always

self pollinated. The presence of 17.86 DxD,

19.05% TxD and 13.10% HxD were also

observed. In Kopyor coconut, the pollens could

travel from pollen donors as far as 58 m apart

from the evaluated female recipients. Therefore,

insect pollinators may have played an important

role in long distance pollen dispersal in Kopyor

coconut.

Acknowledgment

This research was supported by DGHE,

MoEC, Republic of Indonesia through “HI-

LINK Kopyor Coconut Project – 2012-2014”

entitled “Increasing Kopyor Fruit Yield with the

Help of Honey Bee Pollinators and Production of

Kopyor true-to-type Seedlings through Control

Pollination” coordinated by SDR. This research

is used by SHL for her PhD dissertation. SHL

was also supported by Graduate Scholarship

Program (BPPS) and Hibah Doktor Program,

DGHE, MoEC, Republic of Indonesia.

Cord 2015, 31 (1)

58

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Guidelines to the Contributors of CORD

The Asian and Pacific Coconut Community (APCC) publishes semiannually (in April and October) an

international journal on coconut research and development known as Cord. High standard articles written in English

are published in Cord. The manuscript, complete in all respects, should be sent through email (high resolution scanned

tables, photographs, graphs, etc may be sent in text boxes), addressed to the Chief Editor, Cord, Asian and Pacific

Coconut Community, 3rd

Floor, Lina Building, J1. H.R. Rasuna Said, Kav. B-7, Kuningan, Jakarta 12920, Indonesia.

Email address: [email protected].

Hereunder are the guidelines/editorial style in preparing the article for CORD:

Type the manuscript in A4 size paper with double line spacing using MS word. The title of the article should be

short and specific. It should be typed in bold letters, for scientific name use italics.

Below the title, type author's (s) name (s), each name should be superscripted as 1, 2 .… Each superscripted

number should be explained by providing the complete mailing address of the author (s) with all details including pin

code, e-mail etc at the bottom of first page.

Below the author (s') name (s), provide an abstract emphasizing the key aspects of results but without reference

to the body of the text of the full articles. After the abstract, provide a list of the Keywords: (4-6).

The introduction should have a brief statement of the problem and explain the aim or the objectives of the

investigation. The materials and methods should be very clear with all details of experimental design, treatments,

location, period of study, methods adopted etc. Methods should be clearly written so that the reader of your article

should be able to use it in pursuing her/his studies elsewhere.

The results and discussions should provide data organized into tables, figures and photographs, suitably

compared and discussed with earlier published findings. All data must be presented in metric units.

For abbreviations, consult the B.S. (British Standards) on "Letter symbols, signs, and abbreviations".

Abbreviations are alike in singular and plural. The placing of a dot (stop) after a standard abbreviations should be

avoided unless the abbreviation form is likely to be taken for another word, e.g., in for inch, where a full stop would be

necessary.

In the text, references should be quoted according to the Harvard system, in which the author's name and date are

given in parenthesis, e.g. (Magat, 2003) except when the author's name is part of a sentence, e.g. Magat (2003) reported

…. When there are two authors, both names should be given, e.g. (Magat and Margate, 1990). If there are more than

two authors, give the first author's name only followed by the words et al. e.g. Magat et al. (1989).

The table should carry appropriate titles, which should be typed in bold letters. Each table should be

self-explanatory, numbered consecutively. Try to avoid presenting table that is too large to print across the page. Use -

(dash) when no observation was taken and 0 for zero reading. Express values less than unity as 0.25 and not .25.

The figures should be restricted to the display of results when large number of values can be comprehended

more easily; tables and figures should not reproduce the same data. Numbering and lettering should be kept to an

absolute minimum. The photos and plates should be of high quality and be able to make a definite contribution to the

value of the paper. The acknowledgment should follow the section on discussion and immediately precede references.

Only references quoted in the text should be listed. References in the text should be arranged chronologically.

Reference to publications in periodicals should be cited in the following order surname of author, followed by the

initials, year of publication, title of paper, name of journal (italics), volume (in bold Arabic letters), and number in

bracket and after a colon the first and last pages of the reference (without the prefix "pp.") e.g. Morin, J.P, Sudharto,

P.S., Purba, R, Desmier de Chenon, Kakul, T., Laup, S., Beaudoin-Ollivier, L. and Rochat, D. 2001. A new type of trap

for capturing Oryctes rhinoceros (Scarabaeidae: Dynastinae), the main pest in young oil palm and coconut plantings.

Cord 17(2): 13-21.

Reference to books and monographs should include author and/or editor's name full title, number of pages,

edition, the publisher's name and place of publication. The title should be in italics. A list of references should be given

at the end of the text and be arranged alphabetically on authors' name. If an author's name in the list is also mentioned

with co-authors the following order should be used: publications of the single author, arranged according to

publications dates, publications of the same author with one co-author and publications of the author with more than

one co-author. If there are several references by the same author in one year, they are distinguished by the addition of

small letters a, b, c, etc., e.g. Magat, 2003a, Magat, 2003b.

Abbreviations of journal titles should be according to the "World list of Scientific Periodicals.”

Note: The author(s) of selected paper will receive a token honorarium, as well as one-year free subscription of Cord for

senior author if there is more than one author. Articles not accepted will not be sent back to the author.

For each published article, only the senior author will receive one complimentary copy of the journal. For more

copies, the cost of the journal has to be borne by the authors.

Papers published in the Cord may be reprinted or published with prior permission from the APCC Secretariat.


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