international journal of innovative pharmaceutical ...recorded as kuruvichi, or kuruvichi poondu in...

RESEARCH ARTICLE Mageswari et.al / IJIPSR / 3 (5), 2015, 551-559

Department of Botany ISSN (online) 2347-2154

Available online: www.ijipsr.com May Issue 551

MACRO AND MICROSCOPICAL EVALUATION OF ROOT OF

Carmona retusa (Vahl.) MASAM

1S Mageswari*,

2S Karpagam

1Research Scholar, Department of Botany, Queen Mary’s College, Chennai 600004, INDIA

2Associate Professor, Queen Mary’s College, Chennai 600004, INDIA

Corresponding Author

S Mageswari

Department of Botany

Research Scholar,

Queen Mary’s College, Chennai, INDIA

E-mail: [email protected]

Phone: +91 9940401263

International Journal of Innovative

Pharmaceutical Sciences and Research www.ijipsr.com

Abstract

The study deals with the macro and microscopical evaluation of Carmona retusa (Vahl.)

Masam. belong to the family Boaraginaceae. It is an important medicinal plant used in Indian

traditional system of medicine. The root was collected freshly and subjected to macro and

microscopical evaluation and photomicrographs were taken to fix quality standards for this

plant. The microscopical studies have shown styloid crystals and prismatic crystals of calcium

oxalate crystals. The styloids crystals are long scale like parallel with oblique ends upto 200µ in

the bark. This study was helpful in the identity and establishing the authentication of the plant.

Keywords: Pharmacognostic studies, Carmona retusa, Styloid crystals.




INTRODUCTION

Carmona retusa (Vahl.) Masam., Family – Boraginaceae, previously known as Ehretia

microphylla Lam. Carmona is a monotypic genus [1]. Synonyms: Ehretia microphylla Lam.,

Ehretia buxifolia Roxb., Carmona microphylla (Lam.) G. Don., Cordia retusa Vahl., [2].

Carmona retusa is a beautiful shrub (small tree) with many erect branches covered with glossy

dark green coarse leaves. C. retusa is a sub-shrub to 1.5m; leaves alternate, clustered with

axillary leaves, obovate-spathulate, white glandular above, each with a bristle; flowers axillary,

solitary bracteates [3]; calyx lobes unequal elliptic-oblong, herbaceous (5mm); corolla white or

cream, sub-rotate, lobes 5, oblong elliptic, imbricate, herbaceous, obtuse; stamens (4)5, anthers

oblong; ovary globose; stigma capitellate; fruit drupes, globose of 4 pyrenes, when it ripes

become brownish orange. The leaves are medicinally used in the Philippines to treat cough, colic

diarrhea and dysentery [4]. Carmona retusa (Vahl.) Masam (Ehretia microphylla Lam.) is

reported to be medicinally useful in Indigenous System of Medicine [5]. This plant is also

recorded as Kuruvichi, or Kuruvichi poondu in Siddha Materia Medica [6, 7]. It is used for

leprosy, eczema, venereal diseases, chronic dysentery, infertility and toxic diarrhea in children.

A novel natural product microphyllone has been isolated from Ehretia microphylla together with

baurenol and ursolic acid [8]. The plant also contains flavonoids, phytosterols and alkaloids like

astragalin, nicotoflorin, bauerenol, α-amyrin, β-amyrin were also isolated from this plant [9]. It

has anti-inflammatory, antibacterial, analgesic, anti-allergic, anti-mutagen, anti-diarrheal,

antimicrobial and anti-tumor activity. The leaves are used as a stomachic, in the ailments of

cough, fever and constitutional syphilis. The roots of the plants are used in southern India for

Cachexia and syphilis and as an antidote for certain plant poisons [10, 11]. E. microphylla

promote the pituitary-ovary axis activities and cause an elevation in the serum concentrations of

LH, FSH and estradiol hormones as well as increase the mean numbers of follicles and eventually

ovarian weight [12]. The pharmacognostical studies on aerial part of Ehretia microphylla Lam.

was also reported [13]. As there is no detailed study on the root Carmona retusa, hence the

present study attempts to develop the macro-microscopical studies on root of C. retusa.

MATERIAL AND METHODS

The root of the plant C. retusa (Fig. A) was collected from Chengalpattu District, Tamil Nadu,

India. The plant was identified by Dr. P. Jayaraman, Plant Anatomy Research Centre, West

Tambaram, Chennai-45. The root sample of the plant were cut and fixed in FAA for

http://en.wikipedia.org/wiki/Shrub

http://en.wikipedia.org/wiki/Drupe

http://en.wikipedia.org/wiki/Philippines

http://en.wikipedia.org/wiki/Cough

http://en.wikipedia.org/wiki/Colic

http://en.wikipedia.org/wiki/Diarrhea

http://en.wikipedia.org/wiki/Dysentery




pharmacognostical studies. The samples in FAA were dehydrated with graded series of tertiary –

butyl alcohol. Paraffin infiltrated specimens were cast into paraffin blocks. The paraffin

embedded specimens were sectioned using Rotary Microtome. Dewaxing of the sections and

staining of sections with Toluidine blue were carried out as per the standard methods [14-16].

Photomicrographs

Microscopic descriptions of tissues are supplemented with micrographs wherever necessary.

Photographs of different magnifications were taken with Nikon labphoto 2 microscopic Unit. For

normal observations bright field was used. For the study of crystals, starch grains and lignified

cells, polarized light was employed.

The powder microscopical studies were also carried out as per the standard method described by

Wallis [17].

RESULTS AND DISCUSSION

Identification and authentication of the plant

The plant specimen was identified by Botanical Survey of India, Coimbatore vide Letter No.

BSI/SRC/5/23/2012-13/Tech/1840 dated 12th

February 2013. The Basionym of the species is

Cordia retusa Vahl. and Synonym of the species is Ehretia microphylla Lam.

Macroscopy

The root was cylindrical (Fig. B) about 10-40mm diameter, bark is approximately 3mm in

thickness. The root was externally earthy brown in colour, its surface was fissured and the

fracture was fibrous. The bark is externally rough and light brown in colour and exfoliating into

very thin papery phellem. It has no specific odour and taste.

Microscopy

Root

The root consists of very wide periderm and thick solid secondary xylem surrounded by

secondary phloem (Fig. C). The periderm has replaced the epidermis and forms a thick

homogeneous tangentially elongated tabular suberized cell. The periderm is 200µ thick. The

periderm includes entirely phellem cells and phelloderm cells are not evident. Inner to the

periderm is a narrow cylinder of cortex which is 2 or 3 layered and parenchymatous. Secondary

phloem occurs in thick cylinder around the secondary xylem. The secondary phloem elements are

in short radial lines (Fig. D), in the central part while in the peripheral part the xylem elements are

distributed in random circular zone (Fig. C). The xylem elements are vessels which are circular or




elliptical, narrow and thick walled. They measure 10-50µ in diameter. The xylem fibres are very

thick walled with reduced lumen and lignified walls.

Root Bark

The surface of the bark is smooth and even. The outermost part of the periderm undergoes

exfoliation in the form of thick membrane (Fig. E). The total thickness of the bark is 1.6mm. The

bark consists of 2 major zones namely outer periderm and inner secondary phloem. In between

these 2 zones is a narrow cortex (Fig. E & F). The periderm is 580µ thick. The outer part of the

periderm consists of thin walled, tabular, homogeneous, supersized phellem cells. Towards the

inner part of the periderm the cells are wider and gradually become square shaped. The periderm

becomes sharply delimited by a line of cortical parenchyma tissue. The cortical cells are

polyhedral, fairly thick walled and darkly stained (Fig. F). The cortical zone gradually transits

into wide secondary phloem zone. This part is the major portion of the bark. The secondary

phloem consists of outer, wider zone of collapsed secondary phloem and inner narrow non

collapsed secondary phloem. There is no distinct border separating the collapsed and non-

collapsed phloem. The non collapsed phloem consists of radial rows of wide, angular, thick

walled sieve elements and parenchyma cells (Fig. G). Calcium oxalate prismatic crystal are

frequently seen both in the collapsed and non collapsed phloem. The collapsed phloem exhibits

thin, dark, tangential lines which represents the crushed sieve elements. The phloem parenchyma

cells in this region are dilated. Calcium oxalate crystals are abundant in the phloem tissue. They

are prominent prismatic type (Fig. H). They are mostly cuboidal in shape. The crystals are located

in regular, radial lines and are located in the phloem parenchyma cells. The crystals are solitary in

each cell and occupy the entire lumen on the parenchyma cells (Fig. K). The crystals are 12x12µ

in size,

Tangential longitudinal section of the phloem

In TLS view the phloem rays appear non-storied. They are biseriate or less frequently uniseriate.

The rays are heterocellular possessing middle procumbent cells and marginal upright cells. The

rays are spindle shaped and thick walled. The uniseriate rays mostly possess upright cells. The

rays are 110 to 200µ in height and 30 to 40µ in thickness. The axile parenchyma cells are

vertically elongated thick walled cells arranged one below the other in vertical strands (Fig. I).

When TLS sections of the phloem viewed under polarized light large number of calcium oxalate

crystal called styloids are seen located in the parenchyma cells. The styloids are long, scale like

parallel crystals with oblique ends (Fig. J). The styloids are vertically oriented and occupy the




entire length of the parenchyma cells. The styloids are upto 200µ long and about 10µ wide (Fig.

L).

Radial longitudinal section of the phloem

In RLS view the phloem rays appear in horizontal ribbon like bands (Fig. M). The rays extend

from inner phloem upto the inner border of the periderm. The phloem parenchyma cells are in

vertical rows. The periderm cells appear in compact radial files. The phloem rays consists of two

types of cells those in the middle of the ray are square shaped or horizontally rectangular and

these are called procumbent cells. The cells in the marginal part are vertically oblong. These cells

are called upright cells. So, these rays are called heterocellular rays. The procumbent cells are 20

x 25µ in size. The upright cells are 20 x 30µ in size (Fig. M & N).

Powder Microscopy

The powder preparation exhibits the following elements:

(i) Vessel elements: The xylem vessel elements are abundant in the powder. They are long,

narrow, cylindrical cells, 350–450µ long and 30µ wide (Fig. O & P) with circular,

wide, end wall perforations. Some vessels elements have long, narrow tails at one end

or at both ends.

(ii) Fibres: Fibres are very long, narrow, thick walled cells with tapering ends and upto 680µ

long and 10µ thick (Fig. P & Q).

(iii) Parenchyma cells: Vertically elongated, parallel, rectangular, thin walled parenchyma

cells are seen mixed with other elements (Fig. O, P & Q). The parenchyma cells are

200–400µ long and 25µ wide with wide lumen.

A - Habit profile of the plant B - Root collected and dried




Pe – Periderm; SPh – Secondary phloem; Sx – Secondary xylem; Co – Cortex; NcPh-Non collapsed phloem

C - T. S. of Root - Entire view D - T. S. of Root – Periderm and

secondary phloem portion enlarged

E - T. S. of Bark - Entire view

F - T.S. of bark periderm and cortical zones

enlarged

G - T.S. of bark non collapsed phloem elements

enlarged

H - T.S. of secondary phloem with radial

rows

of crystal bearing parenchyma cells

I -Tangential longitudinal view of the phloem showing

non-storied biseriate, occasionally uniseriate phloem

rays.

350µ 1000µ

1000µ

100µ

250µ

250µ 250µ




Pe – Periderm; VE – Vessel; CPh – Collapsed phloem; PhP – Phloem parenchyma;

Pa – Parenchyma: PhR – Phloem ray; Cr – Crystals; St – Styloid crystals;

T – Tail; Fi – Fibre; PC - Procumbent cell; UC - Upright cell.

CONCLUSION

The study evidenced the presence of stratified phellem, styloid crystals, prismatic crystals,

phloem ray cells, collapsed and non collapsed secondary phloem cells which are some of the

salient microscopic features of C. retusa plant roots. The macroscopy, microscopy and powder

microscopical analysis helps in the identification and authentication of the plant Carmona retusa.

M - RLS view of the phloem

showing

horizontal bands of ray cells.

N - Horizontal procumbent and upright cells

of the phloem rays enlarged.

Powder

350µ 100µ

350µ 250µ 250µ O Q P

K & L - Prismatic and Styloid crystals in polarized light

J - TLS of phloem showing distribution of

styloid type of crystals in the axile

parenchyma

250µ 70µ 100µ




ACKNOWLEDGEMENT

The authors are thankful to Prof. Dr. P. Jayaraman, Plant Anatomy Research Centre (PARC)

West Tambaram, Chennai for providing technical facilities.

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