universiti putra malaysia growth and …psasir.upm.edu.my/32028/1/ita 2012 4r.pdf · august 2012...

UNIVERSITI PUTRA MALAYSIA

GROWTH AND DEVELOPMENT OF TOMATO (Lycopersicon esculentum Mill.) CULTIVARS IN VITRO UNDER SALINITY CONDITIONN

ABDALMAJID NASHER AHMED MOHAMED

ITA 2012 4

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GROWTH AND DEVELOPMENT OF TOMATO

(Lycopersicon esculentum Mill.) CULTIVARS IN

VITRO UNDER SALINITY CONDITION


DOCTOR OF PHILOSOPHY

UNIVERSITI PUTRA MALAYSIA

2012

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GROWTH AND DEVELOPMENT OF TOMATO (Lycopersicon esculentum

Mill.) CULTIVARS IN VITRO UNDER SALINITY CONDITION

By


Thesis Submitted to the School of Graduate Studies, University Putra

Malaysia, in Fulfillment of the Requirements for the Degree of Doctor

Philosophy

August 2012

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بسم اللـه الرحـن الرحيم

﴿ رب أوزعن أن أشكر نعمتك الت أن عمت علي وعلى والدي وأن أعمل صالا الي ﴾ ت رضاه وأدخلن برحتك ف عبادك الص

"My Lord, enable me to be grateful for Your favor which You have bestowed

upon me and upon my parents and to do righteousness of which You

approve. And admit me by Your mercy into [the ranks of] Your righteous

servants."

صدق هللا العظيم

سورة النمل-االية)91(

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DEDICATION

Dedicated to my parents,

To my wife,

To my kids,

And to all my brothers and sisters.

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Abstract of thesis presented to the Senate of University Putra Malaysia in

fulfillment of the requirement for the degree of Doctor of Philosophy

GOWTH AND DEVELOPMENT OF TOMATO (Lycopersicon esculentum.

Mill) CULTIVARS IN VITRO UNDER SALINITY CONDITION

By


August 2012

Chairman: Professor Mohd Razi Bin Ismail, PhD

Faculty: Agriculture

The salinity of soil and irrigation water present one of the greatest

environmental strains affecting plant growth and development. Salinity

influences agricultural production in most parts on the world. The tissue

culture technique provides a unique chance for studying many aspects of

plant growth and development, and studying the physiological and

biochemical processes under controlled environment, which contribute to

salinity tolerance. In addition, it offers greater control than in vivo growth

conditions and has the benefit of small scale with clear visibility for

observation of shoot and root responses in the presence of enforced stress.

This study was carried out in the Agrobiotechnology laboratory- Faculty of

Agriculture and plant physiology in ITA-MTDC, UPM, and achieved the

following objectives: Establishment of a protocol for shoot regeneration from

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hypocotyl and cotyledon explants by different concentrations of BAP

hormone, response hypocotyl and cotyledon explants to shoot formation

under NaCl concentration, study of the changes in organic and inorganic

solutes in shoots grown from hypocotyl, and cotyledon explants under NaCl

stress and increase for NaCl tolerance by shoot apices sub-culture under

different levels of NaCl stress.

In vitro, response of hypocotyl and cotyledon explants to shoots regeneration

in MS medium supplemented different levels of 6-benzylaminopurine (0. 1,

2, 4) mg/l was studied. The cultures harvested after six weeks from culture

explants. Different responses were recorded in tomato explants under

different levels of BAP. The formation of roots from hypocotyl explants

found in the control treatment, whereas the cotyledons did not show any

response. The media supplemented with 4 mg/l BAP resulted in callus in

both explants. The medium supplemented 2 mg/l BAP was most effective for

the induction of adventitious shoots for both the hypocotyl, and cotyledon

explants in tomato cultivars; where the number of shoots/explant was 4 and

4.5 in hypocotyls and cotyledons Pearl cultivar and 2.25 and 3

shoots/explants in hypocotyls and cotyledons Beril cultivars respectively.

The best shoots length achieved by hypocotyl explants in Pearl cultivar;

where it was 1.62 cm in medium was supplemented 2 mg/l BAP.

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In vitro, evaluations of sodium chloride (NaCl) effects on two tomato

cultivars (Pearl and Beril) were investigated with four NaCl levels (0, 25, 50

and 75 mM) using hypocotyl and cotyledon explants. The explants were

cultured in MS media having 2.0 mg/l BAP (the suitable medium from the

previous experiment) along with different concentrations of NaCl. The

cultures harvested after eight weeks from culture explants. The growth

characteristics (shoots number, shoots length, fresh weight and dry weight)

decreased with increasing NaCl in the growth medium. The reduction

percentage in shoots length was 85% in high level NaCl (75 mM) in

comparison with control; whereas the reduction in shoots number, fresh

weight and dry weight was 65, 64 and 54% in high level NaCl. In addition,

there was greater degradation of chlorophyll b than chlorophyll a and

different responses of tomato cultivars and explants for salt tolerance in vitro;

where the reduction percentage in chlorophyll a and b was 69%, 76% in high

level NaCl. The growth characteristics and chlorophyll content were superior

in the Beril compared to the Pearl cultivar, and the shoots induced from

cotyledon explants showed best in growth characteristics and chlorophyll

content than the shoots induced from hypocotyl explants under NaCl stress.

The findings may contribute to best multiplication from cotyledon explants

in Beril cultivar to raise a salt-tolerant line for future use.

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Changes of carbohydrate, soluble protein, proline and ions content (Na+, K+,

Ca2+, and Mg2+) were studied in shoots induced from hypocotyl, and

cotyledon explants of Pearl and Beril cultivars in vitro under salt stress. The

explants were cultured in MS+2 mg BAP supplemented different levels NaCl

(0, 25, 50 and 75 mM). The cultures harvested after eight weeks from culture

explants. The accumulation of organic and inorganic solutes is significantly

affected by NaCl stress. An increase of NaCl in the growth medium led to

increased accumulation of carbohydrates, proline and Na+ in the shoots

induced from hypocotyl and cotyledon explants, whereas the accumulation

of protein, K+, Ca+2 and Mg+2 decreased by the increase of NaCl in the growth

medium. The high amount of carbohydrate and proline (20.2 mg/g and 13.69

µmoles/g) achieved by shoots induced from cotyledon explant in Beril

cultivar in high level NaCl. The content of potassium in high level NaCl

reached to 2.46 and 2.43 mg/g by shoots induced from cotyledon explants in

both Beril and Pearl cultivars respectively. Different responses were seen

between the tomato cultivars for the accumulation of organic and inorganic

solutes in shoots induced from hypocotyl, and cotyledon explants in vitro

under salt stress. The Beril cultivars and cotyledon explant were better than

Pearl and hypocotyl explant in almost all organic and inorganic solutes.

In vitro, response of shoot apices subculture of Pearl and Beril tomato

cultivars under different concentration NaCl (0, 25, 50 and 75 mM) to

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enhance of salt tolerance in tomato plantlets was studied. The shoot apices

were cultured in MS+0.1 mg/l IBA medium for four weeks, and the shoot

apices isolated and subculture in the same medium for four weeks. The

growth characteristics were evaluated in both first and subculture shoot

apices included (Shoots length, roots length, plantlets fresh and dry weight,

shoots and roots carbohydrates contents, shoots and roots protein contents,

shoots and roots proline contents and shoots and roots (Na+, K+) ions contents

were recorded. In respect of the growth of tomato plantlets grown from

shoot apices explants affected by NaCl stress, an increase in NaCl in the

growth medium led to a decrease of the growth characteristics of tomato

plantlets. The shoots length increased after shoot apices subculture to 5.38

and 5.78 cm, while it was 4.30 and 4.50 cm in first culture in both Pearl and

Beril cultivar in high level NaCl, and the same trend shown in roots length,

fresh and dry weight characteristics. The accumulation of carbohydrates,

proline and Na+ in the roots and shoots of tomato plantlets increased with

increased salinity in both the first and subculture, whereas the protein and K+

content decreased. Shoot apices subculture enhanced the accumulation of

organic solutes and K+ in the roots and shoots, as well as a decrease of Na+

after the shoot apices subculture. The accumulation of carbohydrate folded

two times in roots and shoots after subculture of shoot apices. The high

carbohydrate and proline content achieved in roots and shoots of Beril

cultivar, where it was the carbohydrate content 18.5 and 14.7 (mg/g. FW)

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and the proline content 16.02 and 13.85 (µmoles/g. FW) after shoot apices

subculture. The results suggest that the tissue culture technique is an

effective tool for developing a new strategy to improve the salt tolerance of

tomato plants for regions that are affected by the salinity of soil and water

irrigation.

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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia

sebagai memenuhi keperluan untuk ijazah Doktor Falsafah

PERTUMBUHAN DAN PERKEMBANGAN KULTIVAR TOMATO

(Lycopersicon esculentum Mill.) SECARA IN-VITRO DI BAWAH

KONDISI SALINITI

Oleh


Ogos 2012

Pengerusi: Profesor Mohd Razi Bin Ismail, PhD

Fakulti: Pertanian

Saliniti pada tanah dan pengairan merupakan antara faktor kekangan alam

sekitar yang sangat mempengaruhi pertumbuhan dan perkembangan

tumbuhan. Saliniti mempengaruhi pengeluaran pertanian di kebanyakan

tempat di dunia. Teknik kultur tisu merupakan medium terbaik untuk

mengkaji pelbagai aspek berkaitan pertumbuhan dan perkembangan, dan

proses fisiologi dan biokimia tumbuhan di bawah persekitaran terkawal

yang menyumbang kepada sifat toleran saliniti. Di samping itu, skala yang

lebih kecil memberikan kawalan dan pemerhatian yang menyeluruh ke atas

tindakbalas pucuk dan akar terhadap tekanan persekitaran berbanding

kajian dalam persekitaran in-vivo. Kajian ini dijalankan di Makmal

Agrobiotechnology, Fakulti Pertanian dan Makmal Fisiologi Tumbuhan di

ITA-MTDC, UPM. Kajian ini telah mencapai objektif seperti berikut:

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penentuan protokol untuk pembentukan semula pucuk daripada eksplan

hipokotil dan kotiledon pada kepekatan hormon BAP yang berlainan,

tindakbalas eksplan hipokotil dan kotiledon dalam pembentukan pucuk di

bawah persekitaran NaCl, kajian perubahan kimia dan biokimia yang

berlaku semasa proses pertumbuhan pucuk dari eksplan hipokotil dan

kotiledon di bawah persekitaran NaCl dan peningkatan toleransi subkultur

apical pucuk terhadap NaCl pada kepekatan NaCl berlainan.

Sebagai tanggapan vitro dari hipokotil dan kotiledon eksplan untuk tunas

regenerasi dalam media MS ditambah berbagai tingkat 6-benzylaminopurine

(0 1, 2, 4) mg/l dipelajari. Kultur dipanen setelah enam minggu dari eksplan

budaya. Tindakbalas yang berbeza telah direkodkan pada eksplan tomato di

bawah kepekatan BAP yang berbeza. Pada rawatan kawalan, pembentukan

akar dari eksplan hipokotil telah dicatatkan, manakala tiada sebarang

tindakbalas pada eksplan kotiledon. Media yang ditambah dengan 4mg/l

BAP menghasilkan kalus pada kedua-dua eksplan. Media yang diberikan

rawatan 2 mg/l BAP sangat efektif dalam merangsang pembentukan pucuk

adventitus pada kedua-dua sumber eksplan dalam kultivar tomato; mana

bilangan pucuk/eksplan adalah 4 dan 4.5 dalam hypocotyls dan kotiledon

kultivar Pearl dan 2.25 dan 3 pucuk/eksplant dalam hypocotyls dan kultivar

Beril kotiledon masing-masing. Panjang pucuk terbaik dicapai oleh eksplant

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hypocotyl dalam kultivar Pearl, di mana ia telah 1.62 cm dalam medium

ditambah 2 mg / l BAP.

Dalam evaluasi vitro efek natrium klorida (NaCl) pada dua kultivar tomat

(Pearl dan Beril) diselidiki dengan empat tingkat NaCl (0, 25, 50 dan 75 mM)

dengan menggunakan eksplan hypocotyl dan kotiledon. Eksplan dikultur

dalam media MS memiliki 2 mg/l BAP (media yang sesuai dari percobaan

sebelumnya) bersama dengan konsentrasi yang berbeda NaCl. Kultur

dipanen setelah delapan minggu dari eksplan budaya. Sifat-sifat

pertumbuhan (pucuk nombor, panjang pucuk, berat badan yang segar dan

berat kering) menurun dengan peningkatan kepekatan NaCl di dalam

media. Peratusan pengurangan panjang pucuk adalah 85% di peringkat

tinggi NaCl (75 mM) dalam perbandingan dengan kawalan; manakala

pengurangan dalam bilangan pucuk, berat segar dan berat kering adalah 65,

64 dan 54% di peringkat tinggi NaCl. Di samping itu, terdapat kemerosotan

yang lebih besar b klorofil daripada klorofil dan jawapan yang berbeza

kultivar tomato dan eksplant untuk toleransi garam dalam vitro. di mana

peratusan pengurangan dalam klorofil a dan b ialah 69%, 76% di peringkat

tinggi NaCl. Dalam keadaan bergaram, sifat-sifat pertumbuhan dan

kandungan klorofil sangat tinggi pada kultivar Beril berbanding kultivar

Pearl manakala eksplan kotiledon juga menunjukkan pembentukan pucuk

dan kandungan klorofil yang tinggi berbanding eksplan hypocotyl. Temuan

dapat menyebabkan multiplikasi yang lebih baik dari eksplan kotiledon

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dalam kultivar Beril untuk meningkatkan garis garam toleran untuk

penggunaan masa depan.

Perubahan dari karbohidrat, protein terlarut, prolin dan konten ion (Na+, K+,

Ca2+, dan Mg2+) yang belajar di pucuk diinduksi dari eksplan hypocotyl dan

kotiledon dari Pearl dan kultivar Beril in vitro di bawah tekanan garam.

Eksplan dikultur dalam MS+2 mg BAP dilengkapi tingkat yang berbeda

NaCl (0, 25, 50 dan 75 mM). Kultur dipanen setelah delapan minggu dari

eksplan budaya. Akumulasi bahan larut organik dan bukan organik

dipengaruhi secara signifikan oleh kepekatan NaCl. Peningkatan kepekatan

NaCl dalam media pertumbuhan yang membawa kepada peningkatan

akumulasi karbohidrat, proline dan Na+ di dalam pucuk yang terbentuk dari

eksplan hypocotyl dan kotiledon, manakala akumulasi protein, K+, Ca2+ dan

Mg2+ menurun dengan peningkatan NaCl dalam media. Jumlah yang tinggi

karbohidrat dan prolin (20.2 mg/g dan 13.69 μmoles/g) yang dicapai oleh

pucuk disebabkan daripada eksplan kotiledon dalam kultivar Beril di

peringkat tinggi NaCl. Kandungan kalium di peringkat tinggi NaCl

mencapai 2.46 dan 2.43 mg/g oleh pucuk disebabkan dari eksplant kotiledon

Beril dan kultivar Mutiara masing-masing. Di bawah persekitaran bersaliniti,

terdapat perbezaan kadar akumulasi bahan larut organik dan tidak organik

pada pucuk yang terbentuk dari eksplan hypocotyl dan kotiledon yang

diperolehi daripada kultivar tomato yang berlainan.

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Di Dalam tindak balas vitro menembak apices cabang kebudayaan Pearl dan

kultivar tomato Beril bawah berbeza kepekatan NaCl (0, 25, 50 dan 75 mM)

untuk meningkatkan toleransi garam dalam anak pokok tomato telah dikaji.

The apices menembak telah berbudaya dalam MS+0.1 mg/l sederhana IBA

selama empat minggu, dan menembak apices diasingkan dan cabang

kebudayaan dalam medium yang sama selama empat minggu. The ciri-ciri

pertumbuhan telah dinilai dalam kedua-dua apices dan cabang kebudayaan

menembak dimasukkan (panjang Pucuk, panjang akar, berat anak pokok

segar dan kering, pucuk dan akar karbohidrat kandungan, pucuk dan

kandungan akar protein, pucuk dan akar kandungan prolin dan pucuk dan

akar (Na+, K+) ion kandungan telah direkodkan. Berhubung pertumbuhan

anak pokok tomato ditanam dari eksplant apices menembak terjejas oleh

tekanan NaCl, satu peningkatan dalam NaCl dalam medium pertumbuhan

yang membawa kepada penurunan itu pertumbuhan sifat-sifat tomato anak

pokok. The panjang pucuk meningkat selepas apices menembak cabang

kebudayaan kepada 5.38 dan 5.78 cm, manakala ia adalah 4.30 dan 4.50 cm

dalam budaya pertama dalam kedua-dua Pearl dan kultivar Beril di

peringkat tinggi NaCl, dan trend yang sama yang ditunjukkan dalam

panjang akar, segar dan ciri-ciri berat kering. pengumpulan karbohidrat,

prolin dan Na+ yang akar dan pucuk anak pokok tomato meningkat dengan

kemasinan meningkat dalam apices kedua-dua pertama dan cabang

kebudayaan, sedangkan yang protein dan K+ kandungan menurun. Tembak

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cabang kebudayaan dipertingkatkan pengumpulan larutan organik dan K+

akar dan pucuk, serta penurunan Na+ selepas yang cabang kebudayaan

apices menembak Pengumpulan karbohidrat dilipat dua kali dalam akar dan

pucuk selepas cabang kebudayaan apices menembak. The karbohidrat yang

tinggi dan kandungan proline dicapai dalam akar dan pucuk kultivar Beril,

di mana ia adalah yang kandungan karbohidrat 18.5 dan 14.7 mg/g. FW dan

kandungan proline 16.02 dan 13.85 μmoles/g.FW selepas apices cabang

kebudayaan menembak. Keputusan mencadangkan bahawa teknik kultur

tisu adalah satu alat yang berkesan untuk membangunkan satu strategi baru

untuk meningkatkan toleransi garam tanaman tomato untuk kawasan yang

dipengaruhi oleh kemasinan pengairan tanah dan air.

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ACKNOWLEDGEMENT

First and foremost, Alhamdulillah for giving me the strength, patience, and

determination in completing this work. All grace and thanks belongs to

Almighty ALLAH.

My deep gratitude goes to the chairman of my supervisory committee,

Professor Dr. Mohd Razi Bin Ismail for his invaluable advice, constant

guidance and encouragement throughout execution of the experiments and

write-up of the manuscript. This work would not have been possible without

the expert guidance of my esteemed supervisor, Prof. Dr. Mohd Razi Bin

Ismail. Not only was he always available for me, as he so generously is for all

of these students, but also his comments were always extremely, helpful, and

appropriate.

In addition, my deepest appreciation to my committee members Assoc. Prof.

Dr. Halimi Mohd Saud and Assoc. Prof. Dr. Mihdzar Abdul Kadir for their

cooperation, effort, and valuable comments.

Sincere and heartfelt thanks to the Institute Tropical of Agriculture and the

staff of the postgraduate office, Library and University Putra Malaysia, for

providing a study and research environment.

I am grateful to the staff and technicians of Plant Physiology in MTDC

Laboratory Section of Department of Food Crops, Institute Tropical of

Agriculture, UPM, and staff and technicians of Agrobiotechnology

Laboratory, Faculty of Agriculture, UPM.

I would like also to extend my sincere thanks to all colleagues who helped

me and give me advice during me study.

Last but of course not least, this work not have been achieved without the

support and understanding of my wife and my kids. They have always

supported me in everyway I have chosen in life.

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I certify that an Examination Committee has met on 14 August 2012 of viva

voce to conduct the final examination of Abdalmajid Nasher Ahmed

Mohamed on his degree thesis entitled “Growth and development of tomato

(Lycopersicon esculentum. Mill) cultivars in vitro under salinity condition" in

accordance with University Pertanian Malaysia (Higher Degree) Act 1980

and Universiti Pertanian Malaysia (Higher Degree) Regulations 1981.The

Committee recommends that the student be awarded the degree of Doctor of

Philosophy.

Members of the Examination Committee were as follows:

Siti Nor Akmar binti Abdullah, PhD

Professor

Faculty of Agriculture

Universiti Putra Malaysia

(Chairman)

Puteri Edaroyati bt Megat Wahab, PhD

Associate Professor



(Internal Examiner)

Maziah binti Mahmood, PhD

Professor

Faculty of Biotechnology and Bimolecular Sciences


(Internal Examiner)

Hamide Gubbuk, PhD

Professor

Department of Horticulture, Faculty of Agriculture

University Akdeniz

Turkey (External Examiner)

Seow Heng Fong, PhD

Professor and Deputy Dean

School of Graduate Studies


Date:

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This thesis was submitted to the Senate of Universiti Putra Malaysia and has

been accepted as fulfillment of the requirement for the degree of type of

degree. The members of the Supervisory Committee were as follows:

Mohd Razi Bin Ismail

Professor



(Chairman)

Halimi Mohd Saud

Associate Professor



(Member)

Mihdzar Abdul Kadir

Associate Professor



(Member)

BUJANG BIN KIM HUAT, PhD

Professor and Dean

School of Graduate Studies

University Putra Malaysia

Date:

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DECLARATION

I declare that the thesis is my original work except for quotations and

citations which have been duly acknowledged. I also declare that it has not

been previously, and is not concurrently, submitted for any other degree at

University Putra Malaysia or at any other institution.


Date: 14 August 2012

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TABLE OF CONTENTS

Page

DEDICTION

ABSTRACT

ABSTRAKT

ACKNOWLEDGEMENTS

APPROVAL

DECLARATION

LIST OF TABLES

LIST OF FIGUERS

LIST OF APPENDIX

LIST OF ABBREVIATIONS

iv

v

Xi

Xvii

Xviii

Xx

Xxvi

Xxviii

Xxxi

Xxxii

CHAPTER

1 INTRODUCTION 1

2 LITERATURE REVIEW 6

2.1. Tomato plant importance 6

2.2. Salinity problems 7

2.3. Salinity and plant growth 8

2.3.1. Effects of salinity on plants growth 8

2.3.2. Effect of salinity on tomato growth 10

2.4. Micropropagation of tomato plants 14

2.5. Effect of salt stress on tomato growth in vitro 16

2.6. Organic solutes accumulation under salt stress 19

2.6.1. Carbohydrate accumulation 19

2.6.2. Protein accumulation 22

2.6.3. Proline accumulation 23

2.7. Ion Homeostasis under salinity 25

3 IN VITRO RESPONSE FROM COTYLEDONS AND

HYPOCOTYLS EXPLANTS IN TOMATO BY

INDUCING 6-BENZYLAMINOPURINE

29

3.1. Introduction 29

3.2. Materials and methods 31

3.2.1. Tomato seeds obtaining 31

3.2.2. Seed sterilization culture 32

3.2.3. Isolation and culture of explants 32

3.2.4. Experimental design and statistic analysis 33

3.3. Result 33

3.4. Discussion 41

3.5. Conclusion 45

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4 IN VITRO PERFORMANCE OF HYPOCOTYLE AND

COTYLEDON EXPLANTS OF TOMATO CULTIVARS

UNDER SODIUM CHLORIDE STRESS

46



4.2.1. Sterilization and germination of seeds 48

4.2.2. Growth parameters 49

4.2.3. Estimation of chlorophyll content 49

4.2.4. Statistic analysis 50

4.3. Results 51

4.3.1. Shoots number and Shoot length 51

4.3.2. Fresh and dry weight 58

4.3.3. Chlorophyll content 62

4.3.4. Correlation coefficient between growth

characteristics and chlorophyll content

69

4.4. Discussion 70

4.5. Conclusion 74

5 CHANGES IN ORGANIC AND INORGANIC

SOLUTES OF IN VITRO TOMATO CULTIVARS

UNDER NaCl STRESS

76



5.2.1. Sterilization and germination of seeds 79

5.2.2. Explants isolation and culture 80

5.2.3. Determination of organic content 80

5.2.4. Determination of inorganic content 82

5.2.5. Statistical analysis 83

5.3. Results 83

5.3.1. Total soluble carbohydrates 83

5.3.2. Total soluble protein 84

5.3.3. Proline content 85

5.3.4. Inorganic contents 90

5.3.5. Correlation coefficient between organic and

inorganic solutes

96

5.4. Discussion 97

5.5. Conclusion 103

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6 IN VITRO SHOOT APICES SUB-CULTURE FOR THE

DEVELOPMENT OF SALT TOLERANT TOMATO

PLANTLETS

104



6.2.1. Seed sterilization and germination 107

6.2.2. Shoot apices isolate and culture 107

6.2.3. Growth parameters 108

6.2.3.1. Growth characteristics 108

6.2.3.2. Carbohydrates contents in roots and

shoots

108

6.2.3.3. Protein content in roots and shoots 109

6.2.3.4. Proline contents in roots and shoots 109

6.2.3.5. Inorganic contents in roots and shoots 109

6.2.4. Statistical analysis 109

6.3. Results 110

6.3.1. Root and shoots length 110

6.3.2. Plantlets fresh and dry weight 114

6.3.3. Roots and shoots total soluble

carbohydrates

117

6.3.4. Roots and shoots total soluble protein 120

6.3.5. Roots and shoots Proline contents 123

6.3.6. Roots and shoots Na+ contents 126

6.3.7. Roots and shoots K+ contents 129

6.3.8. Correlation between organic and inorganic

solutes of roots and shoots tomato plantlets

under NaCl stress

129

6.4. Discussion 133

6.4.1. Growth characteristics 133

6.4.2. Carbohydrates 136

6.4.3. Soluble proteins 137

6.4.4. Proline 139

6.4.5. Na+ and K+ ions 141

6.5. Conclusion 143

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7 SUMMARY, GENERAL CONCLUSION AND

RECOMMENDATIONS FOR FUTURE RESEARCH

144

7.1. General summary 144

7.2. General conclusion 148

7.3. Recommendations for future research 151

REFERENCES 152

APPENDIX 180

BIODATA OF STUDENT 185

LIST OF PUBLICATIONS 186

universiti putra malaysia growth and …psasir.upm.edu.my/32028/1/ita 2012 4r.pdf · august 2012...

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