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ANATOMICAL INVESTIGATION ON THE LEAVES AND STEM OF PASSIFLORA

INCARNATA (PASSIFLORACEAE)

Article  in  International Journal of Research in Ayurveda and Pharmacy · July 2015

DOI: 10.7897/2277-4343.064101

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Research Article
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ANATOMICAL INVESTIGATION ON THE LEAVES AND STEM OF PASSIFLORA INCARNATA

(PASSIFLORACEAE)
Vijayalakshmi Chinniah*, VenkataRathinaKumar Thiagarajan
Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai, Tamilnadu, India

Received on: 04/03/15 Revised on: 27/04/15 Accepted on: 02/05/15

*Corresponding author
Mrs. Vijayalakshmi Chinniah, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai, Tamilnadu, India
E-mail: [email protected]

DOI: 10.7897/2277-4343.064101

ABSTRACT

The present study was designed to explore the anatomical features of leaf and stem of Passiflora incarnata. Macroscopical, microscopical analysis for
the standardization of Passiflora incarnata (Passifloraceae) which has been widely used to treat various disorders like cancer, insomnia, ulcer and
convulsion etc. Leaves-green colour, palmate shape with 3 lobes, obtuse apex, serrate margin, pinnate venation. Stem- green colour, cylindrical shape.
Microscopic evaluation revealed the presence of anamocytic or anisocytic stomata, unicellular or uniseriate covering trichomes, calcium oxalate
crystals, simple unbranched termination, biforked termination and repeatedly branched dendroid type of termination. The observation of cellular level
morphology or anatomy is a major aid for the authentication of drugs. These characters are especially important for identification of drugs. These
studies will contribute further investigation on this plant.

Keywords: Passiflora incarnata, leaf, stem, insomnia, calcium oxalate crystals

INTRODUCTION Specimen preparation

Passiflora incarnata Linn is dicotyledonous plant Petiole, leaf, stem and tendrils were fixed in FAA solution
belonging to the family Passifloraceae. A perennial, (Formalin-5ml+ Acetic acid-5ml + 70% Ethyl alcohol-
creeping herb, climbing by means of axillary tendrils. 90ml).After 24 hours of fixing, the specimens were
Leaves alternate, palmate three to five serrate lobes. dehydrated with graded series of tertiary –Butyl alcohol
Flowers large, solitary, with long peduncles, whitish, with as per the schedule given by Sass, (1940). Infiltration of
a triple purple and pink crown. Fruits are ovate berries the specimens was carried by gradual addition of paraffin
containing numerous ovoid, flattened seeds covered with wax (melting point 58-60º C) until TBA solution attained
a yellowish or brownish aril1-3. super saturation. The specimens were cast into paraffin
blocks.
It has been reported to contain flavonoids as the major
phytoconstitutents. These include apigenin, luteolin, Sectioning
quercetin, kaempferol etc. It has also been reported to
contain alkaloids like harman, harmol. Carbohydrates The paraffin embedded specimens were sectioned with
such as raffinose, sucrose, d-glucose and d-fructose, the help of Rotary Microtome. The thickness of the
amino acids, cyanogenic glycoside gynocardin and sections was about 10-12 μm. Dewaxing of the sections
Cylcopentenoid cyanogenic glycosides- Passibiflorin, was done by customary procedure7. The sections were
eoipassibiflorin, passitrifasciatin4,5. stained with Toluidine blue as per the method given by
O’Brien8.
Based on the ethano-medicinal information it has been
revealed that Passiflora incarnata has been used as an For studying the stomatal morphology, venation pattern
antispasmodic, sedative, narcotic, neuralgia, insomnia, and trichome distribution, paradermal sections (sections
epilepsy, ulcer, haemorrhoids6. taken parallel to the surface of leaf) as well as clearing of
leafwith 5% sodium hydroxide or epidermal peeling by
MATERIALS AND METHODS partial maceration employing Jeffrey’s maceration fluid
were prepared9. Powdered materials of different parts
Collection of plant material were cleared with NaOH and mounted in glycerine
medium after staining7.
The leaves and stem of Passiflora incarnata were
collected from Salem, Tamilnadu, India. The plant was Photomicrographs
identified and authenticated (PARC/2015/3037) by senior
plant Taxonomist at Plant Anatomy Research Centre Photomicrographs were taken with the help of Nikon lab
(PARC). photo 2 microscopic Unit. For normal observations bright

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field was used. For the study of crystals, starch grains and cells. The ground tissue is parenchymatous, angular and
lignified cells, polarized light was employed. Since these compact. The vascular strand is single, fairly large and
structures have birefringence property, under polarized collateral. The xylem elements are in diffuse masses
light they appear bright against dark background. having thick walled cells with wide lumen. Phloem
Magnifications of the figures are indicated by the scale- elements occur in a few discrete masses Figure (3.2).
bars.
Lamina
RESULTS & DISCUSSION
The lamina is distinctly dorsiventral and amphistomatic.
Macroscopical studies (Table 1 and 2, Figure 1-2.4) The adaxial epidermis consists of thick rectangular cells
with thick cuticle. The abaxial epidermal cells are wide
Table: 1 Organoleptic Evaluation of leaf and rectangular with thin cuticle. The mesophyll tissue
consists of adaxial layer of single vertically elongated thin
Colour Green colour
Shape Palmate with 3 lobes
palisade cells and abaxial zone of spherical and lobed
Odour No distinct odour spongy mesophyll cells. The abaxial layer is undulate
Texture Soft and coriaceous (Leathery) with shallow depressions (Figure 3.3). The lamina is 120
Apex Obtuse mm thick.
Base Symmetrical
Petiole Petiolate
Leaf arrangement Alternate
Leaf margin
Margin Serrate
Venation Pinnate The leaf margin is broadly conical measuring 100 mm in
Leaf also consists of extra floral nectaries and stipules thickness. It includes thick walled, squarish epidermal
Table 2: Organoleptic evaluation of stem
layer with highly prominent cuticle. The inner tissues of
the marginal part include angular and circular thick
Colour Green walled compact mass of cells. Palisade and spongy
Shape Cylindrical mesophyll tissue are not evident in the marginal part.
Odour No distinct odour (Figure 3.4).
Stem bears tendrils in leaf axils

Epidermal cells and stomata

Microscopic studies of the leaves10-13
Paradermal sections were prepared for studying the
Structure of the leaf
epidermal tissues of the lamina. The epidermal cells are
smaller with thin highly wavy anticlinal walls. The
In cross sectional view the leaf exhibits prominent midrib
stomata are abundant and diffuse in distribution. Stomata
and thin, smooth and glabrous lamina (Figure 3.1).The
are either anamocytic (Figure 4.1) or anisocytic. In the
midrib consists of thick and wide adaxial cone and
anamocytic type of stomata no distinct subsidiary cells are
prominent semi-circular abaxial part. The midrib is 900
evident. In the anisocytic type the stomata have one
mm thick. The adaxial cone is 300 mm wide and the smaller and 2 larger subsidiary cells. The adaxial
abaxial part is 900 mm wide. epidermis have fairly wider epidermal cells with wavy
anticlinal walls. The stomata are less in frequency.The
The midrib consists of well-defined epidermal layer of stomata are elliptical measuring 45x60 mm in size. The
cells which are squarish with short, cuticular echinate stomatal pore is narrow slit like in appearance.
outgrowths. The ground tissue in the adaxial cone is
collenchymatous and in the remaining part the ground Trichomes
tissue is parenchymatous. The cells are angular, thin
walled and compact. The vascular system is The lower epidermis has numerous unicellular or
multistranded. There are 3 vascular bundles forming an uniseriate covering trichomes (one to three cells)
abaxial arc and a single adaxial bundle. All the bundles approximately 80–250 μm long, are located
are collateral and are more or less wedge shaped. In all predominantly along veins (Figure 4.3).
the bundles, the xylem elements are directed towards the
centre of the midrib. The bundles have compact masses of Crystals
wide, circular-thick walled xylem elements. Phloem
occurs in large compact mass on the outer part of the Calcium oxalate druses are densely distributed along the
xylem strands. The phloem elements are wide and thick veins of the lamina. The druses are in unseriate strand
walled.
(Figure 4.2). They are about 15 mm in diameter.
Lateral Vein
Venation pattern
The lateral vein consists of a prominent adaxial cone with
The venation is densely reticulate and the veins are
collenchymatous inner cells. The abaxial part is
gradually reduced in thickness. The ultimate veins forms
semicircular and consists of thick walled epidermal cells
the well-defined wide polygonal vein islets. Vein
followed by 2 or 3 layer of similar type of thick walled
terminations are well developed in all islets. They vary

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from simple unbranched termination, biforked termination The cortical zone includes a small rectangular outer layer
and repeatedly branched dendroid type of termination. of thick walled cells and 2 or 3 inner layer of wide thin
The terminations are uniformly thin, straight or curved walled parenchyma cells. The vascular cylinder consists
(Figure 5). of an outer circle of thick curved segments of fibres and a
thin layer of phloem encircling a thick cylinder of xylem.
Petiole The xylem includes mostly thick walled wide lignified
fibres and a few vessels which are not well distinct from
The petiole is horizontally flat with adaxial shallow xylem fibres.
groove (Figure 6). It is multistranded having about 8
vascular bundles organized into distinct pattern, amidst The central part of the tendril consists of circular, fairly
the 8 vascular bundles, 7 bundles are arranged in a thick walled and lignified compact cells
shallow arc with wide gaps in between them. All the eight
bundles are located in the adaxial median part. Powder microscopy

The petiole consists of thin layer of epidermis and 3 or 4 The powder microscopical study showed the presence of
layers of thick walled cells resembling the epidermal the following microscopical features (Figure 9)
cells. The remaining ground tissue consists of large,
angular or circular thin walled parenchyma cells. The Druses
vascular bundles are collateral and the xylem elements are
facing towards the centre. The xylem elements are short, Calcium oxalate druses were located in the vertical
parallel, compact and they are circular and thick walled. strands along the veins. The druses are quite wide and
Phloem occurs in thick mass on the outer part of each thick measuring 20 mm in diameter.
vascular bundle.
Trichomes
Stem
Uniseriate trichomes with one to three thin-walled cells,
The stem is circular in sectional view measuring 2.4 mm straight or slightly curved, ending in a point or sometimes
in diameter. It consists of a continuous epidermal layer, as a hook.
narrow cortex with a circle of several discrete fibre
strands and a wide hollow cylinder of vascular elements. Vein termination
There is a wide pith with central empty cavity (Figure 7).
The epidermal layer consists of small, thin walled Fragments of lamina showed the presence of branched
elliptical cells. The cortical zone includes 6 or 7 layers of dendroid type of vein terminations.
rectangular parenchyma cells. Along the inner border of
the cortex occur thick segments of thick walled lignified Stomata
fibres. 14-15
Abaxial epidermal peelings showed the presence of
The vascular cylinder consists of outer thin continuous epidermal cells and stomata. The epidermal cells are
layer of secondary phloem and inner cylinder of amoeboid in outline with wavy anticlinal walls. Stomata
secondary xylem and primary xylem. were densely distributed and they are anamocytic type.
The guard cells are elliptical and thin walled.
The secondary phloem includes wide, angular and thick
walled sieve elements with well-preserved companion Fibres
cells. The phloem parenchyma has small, thick walled
cells. The secondary xylem consists of compact radial Xylem fibres are predominantly seen in the powder. They
files of thick walled and lignified xylem fibres and a few are of 2 types, some are wide and others are narrow. The
wide, circular thick walled cells. wide fibres are thin walled, short with wide lumen. They
are 400 mm long and 20 mm wide. The narrow fibres are
The inner boundary of the xylem cylinder exhibits the comparatively thick walled with reduced lumen. They are
primary xylem strands. The primary xylem consists of 1 also 400 mm wide but 10 mm thick.
or 2 wide meta xylem elements and a group of small proto
xylem elements.The secondary xylem vessels are up to 80 Parenchyma cells
mm in diameter.
Parenchyma cells are abundant and scattered in the
Tendril powder. They vary in shape and size. They may be
narrow and rectangular or wide and elongated. Some of
The tendril is circular in sectional view with slightly the parenchyma cells are squarish and thin walled. The
undulate outline (Figure 8). It is 1.25 mm thick. The parenchyma cells are up to 140 mm long and 20 mm thick.
tendril consists of fairly large radially oblong thin walled Some of these cells have granular inclusions. These cells
epidermal cells with thick cuticle. are also attached laterally with the vessel elements.

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Tracheids Vessel elements

Occasionally tracheids are seen in the powder. The Vessel elements are more common in the powder. They
tracheids are long, narrow with dense pits on the lateral are mostly long narrow cylinders. Some vessel elements
wall. The tracheids do not have end wall perforation. are comparatively shorter and wider.The vessel elements
have wide, circular, horizontal end wall perforations on
the lateral walls are seen dense multiseriate and circular
bordered pits. The narrow vessel elements are 320 mm
long and 50 mm wide. The short wide vessel elements are
270 mm long and 80 mm wide.

Figure 1: Aerial parts of P. incarnata

Figure 2.1: Serrate margin Figure 2.2: Extra floral nectaries

Figure 2.3: Tendrils Figure 2.4: Stipules

Macroscopical (Features of P. incarnata)

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Figure 3.1: T.S of midrib enlarged Figure 3.2: T.S of lateral vein

Figure 3.4: T.S through margin

Figure 3.3: T.S of lamina

Figure 4.3: Trichomes

Figure 4.1: Stomata enlarged Figure 4.2: Vertical row of druses

Figure 5: Venation pattern

Figure 6: T.S of petiole

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Figure 7: T.S of stem Figure 8: T.S of tendril

Druses Trichomes Venation

Stomata

Fibers Parenchyma Cells Tracheids Vessel Elements

Figure 9: Powder Microscopy

CONCLUSION REFERENCES

1. Gamble JS. Flora of the Presidency of Madras. Vol 3. Calcutta,

The observation of cellular level morphology or anatomy India: Botanical Survey of India; 1935.p.235.
is a major aid for the authentication of drugs. These 2. Henry AN, Kumari GR, and Chitra V. Flora of Tamilnadu. Vol.3.
characters are especially important for identification of Southern Circle, Coimbatore, India, Botanical Survey of India;
powdered drugs, because in these cases most of the 1987.p.258.
3. Mathew KM, The flora of Tamil Nadu Karnatic, Vol. 3.
morphological diagnostic features are lost. Microscopic Tiruchirappalli, The Rapinat Herbarium, St.John's College;
evaluation is one of the simplest and cheapest methods for 1983.p.689-1540.
the correct identification of the source of the materials. In 4. Borrelli F, Pinto L, Izzo AA, Mascolo N, Capasso F, Mercati V,
our present work we selected the plant P.incarnata L Toja E, Autore G, Anti-inflammatory activity of Passiflora
incarnata L. in rats. Phytother. Res, 1996; 10: S104–S106.
(Passifloraceae). The macroscopic and organoleptic 5. Chakravarty HL, Excerpt from the Bulletin of the Botanical Society
characters of the plant can serve as diagnostic parameters. of Bengal, A Revision of the Indian Passifloraceae. The Botanical
The microscopic study revealed the presence of Society of Bengal, Calcutta, India, 1949; 61–62.
anamocytic or anisocytic stomata, unicellular or uniseriate 6. Anonymous, Quality control Methods for Medicinal Plant
Materials, WHO; 1998.p. 28-35.
covering trichomes, calcium oxalate crystals, simple 7. Johansen DA, Plant Micro technique, McGraw Hill Book Company;
unbranched termination, biforked termination and 1940.p. 27-93,126-154.
repeatedly branched dendroid type of termination. In 8. O'Brien, TP, Feder N and Mc Cull ME, Polychromatic Staining of
conclusion, the present work was undertaken with a view Plant Cell walls by toluidine blue-O, Protoplasma;1964, 59,364-
373. http://dx.doi.org/10.1007/BF01248568
to lay down standards which could be useful to detect the 9. Sass JE, Elements of Botanical Microtechnique, McGraw Hill Book
authenticity of this medicinally useful plant. Co, New York; 1940. p. 222.
10. Easu K, Plant Anatomy, John Wiley and sons,New York; 1964.p.
ACKNOWLEDGEMENT 767.
11. Easu K, Anatomy of seed Plants, John Wiley and sons, New York;
Dr. P. Jayaraman, Director of Plant Anatomy Research Institute, 1979.p. 550.
Tambaram, Chennai, TamilNadu, India for the histochemical studies.

542
 Vijayalakshmi Chinniah et al / Int. J. Res. Ayurveda Pharm. 6(4), July – August 2015

12. Metcalfe CR and Chalk L, Anatomy of the Dicotyledons, Vol. I, 15. Kishore Kumar V, Lalitha KG. Pharmacognostical studies on the
Clarendon Press, Oxford; 1950.p. 289,458. root of Anacyclus pyrethrum DC. Indian J Nat Pro
13. Giby Abraham. Pharmacognostical and phytochemical studies of Res.2012;3(4):518-526.
the plant Sahadevi [Vernonia cinerea (L) Less]. Int. J. Res.
Ayurveda Pharm. 2015; 6(1):47-54. http://dx.doi.org/10.7897/2277- Cite this article as:
4343.06112
14. Ashish S. Zalke, B. Duraiswamy, Upendra B. Gandagule, and Nidhi Vijayalakshmi Chinniah, VenkataRathinaKumar Thiagarajan.
Singh. Pharmacognostical evaluation of Cardiospermum Anatomical investigation on the leaves and stem of Passiflora incarnata
halicacabum Linn. leaf and stem. Anc Sci Life.2013; 33(1):15-21. (Passifloraceae). Int. J. Res. Ayurveda Pharm. 2015;6(4):537-543
http://dx.doi.org/10.4103/0257-7941.134561 http://dx.doi.org/10.7897/2277-4343.064101

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