MACRO AND MICROSCOPICAL EVALUATION OF ROOT

Transcription

RESEARCH ARTICLE
Department of Botany
Mageswari et.al / IJIPSR / 3 (5), 2015, 551-559
ISSN (online) 2347-2154
International Journal of Innovative
Pharmaceutical Sciences and Research
www.ijipsr.com
MACRO AND MICROSCOPICAL EVALUATION OF ROOT OF
Carmona retusa (Vahl.) MASAM
1
1
S Mageswari*, 2S Karpagam
Research Scholar, Department of Botany, Queen Mary’s College, Chennai 600004, INDIA
2
Associate Professor, Queen Mary’s College, Chennai 600004, INDIA
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.
Corresponding Author
S Mageswari
Research Scholar,
Queen Mary’s College, Chennai, INDIA
E-mail: mageswari0204@gmail.com
Phone: +91 9940401263
Available online: www.ijipsr.com
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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
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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
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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 noncollapsed 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
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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
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1000µ
C - T. S. of Root - Entire view
350µ
D - T. S. of Root – Periderm and
secondary phloem portion enlarged
1000µ
E - T. S. of Bark - Entire view
100µ
G - T.S. of bark non collapsed phloem elements
enlarged
250µ
F - T.S. of bark periderm and cortical zones
enlarged
Pe – Periderm; SPh – Secondary phloem; Sx – Secondary xylem; Co – Cortex; NcPh-Non collapsed phloem
250µ
250µ
I -Tangential longitudinal view of the phloem showing
H - T.S. of secondary phloem with radial
non-storied biseriate, occasionally uniseriate phloem
rows
rays.
of crystal bearing parenchyma cells
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250µ
J - TLS of phloem showing distribution of
styloid type of crystals in the axile
parenchyma
70µ
K & L - Prismatic and Styloid crystals in polarized light
100µ
N - Horizontal procumbent and upright cells
of the phloem rays enlarged.
350µ
M - RLS view of the phloem
showing
horizontal bands of ray cells.
350µ
100µ
O
250µ
P
250µ
Q
Powder
Pe – Periderm;
Pa – Parenchyma:
T – Tail;
VE – Vessel;
PhR – Phloem ray;
Fi – Fibre;
CPh – Collapsed phloem; PhP – Phloem parenchyma;
Cr – Crystals;
St – Styloid crystals;
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.
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ACKNOWLEDGEMENT
The authors are thankful to Prof. Dr. P. Jayaraman, Plant Anatomy Research Centre (PARC)
West Tambaram, Chennai for providing technical facilities.
REFERENCES
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12. Aarthi I, Anbu J K F H Nazeer Ahamed, Ashwini Anjana, V Velpandian, 2012. Effect of
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on Ehretia microphylla Lamk., - Asian Journal of Traditional Medicines, 9(5): 118-129.
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Shahdara, New Delhi,
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MACRO AND MICROSCOPICAL EVALUATION OF ROOT

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