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Ethnobotanical Leaflets 14: 826-35. 2010. Anatomical and
Phytochemical Studies of the Leaves and Roots of Urginea
grandiflora Bak. and Pancratium tortuosum Herbert H. A. S. Sultan, B. I. Abu Elreish and S. M.
Yagi* Department of Botany, Faculty of Science, *Corresponding author E-mail address: sakinayagi@yahoo.com Issued: July 1, 2010 Abstract Urginea grandiflora Bak. and Pancratium tortuosum Herbert are bulbous, medicinal plants endemic to the Key words: bulbous
plants; Urginea grandiflora; Pancratium tortuosum anatomy; phytochemistry. Introduction To ensure
reproducible quality of herbal products, proper control of starting material
is utmost essential. Thus in recent years there have been an emphasis in
standardization of medicinal plants of therapeutic potential. According to
World Health Organization (WHO) the macroscopic and microscopic description
of a medicinal plant is the first step towards establishing its identity and
purity and should be carried out before any tests are undertaken (Anonymous.
1996). Correct botanical identity based on
the external morphology is possible when a complete plant specimen is
available. Anatomical characters can also help the identification when
morphological features are indistinct (David et al., 2008). Urginea grandiflora Bak. (Hyacinthaceae) and Pancratium tortuosum Herbert (Amaryllidaceae)
are perennial,
herbaceous
and bulbous plants, distributed in the Red Sea Hills in Eastern Sudan (Andrews,
1956). The latex of U. grandiflora obtained directly from the bulb of the plant can be
applied directly to cuts and wounds. The bulb of P. tortuosum is used
as malaria therapy. Few studies have been devoted to these two
species, therefore this work has aimed to investigate the leaf and root
anatomy in order to contribute to the medicinal plant identification and to
investigate their secondary metabolites content. Materials and methods Bulbs of the two plants were collected from Erkawit area in Eastern Sudan. The taxonomic identification of the two plants was carried out at the Herbarium of Botany Department, Faculty of Science, and University of Khartoum. Voucher specimens were deposited at the herbarium as reference materials. For anatomical analysis, fresh
mature Leaves and roots were fixed in formalin: glacial acetic acid: 70% alcohol
(5:5:90). Sections were prepared according to the method described by Alexander
(1940). Transverse sections were prepared by using a rotatory microtome
(Leitz 1512-West Germany) and stained with safranin. A light microscope was
used to view the slides and adjusted to finest resolution. Photographs were
obtained using a Nickkon digital camera focused through the microscope
eyepiece. Qualitative analysis of chemical constituents was carried out according to the method described by Sofowara (1993. Results and discussion Anatomy of
Urginea grandiflora The root: Transverse section of the root appears as a complete circle Fig. (1a). The outer region is the bark covered with suberin and tannins. The epidermis consists of a single layer, followed by the hypodermis which consists of one raw of large cells. Inner to the hypodermis is the cortex which consists of 22-25 layers of parenchyma cells. The endodermis is the last layer of the cortex, formed from elongated very thin- walled cells. One layer of the pericycle is found inner to the endodermis, followed by the typically alternating radial vascular bundles (15 – 17) arranged in a circle. The vascular bundles are formed from phloem and xylem. Phloem consists of sieve tube cells, companion cells and phloem parenchyma. Xylem consists of protoxylem vessels to the outside and metaxylem vessels to the inside connected by conjunctive tissue with the phloem. Although roots of monocotyledons commonly, lack normal secondary growth (Esau, 1977), abnormal secondary thickening is observed in U. grandiflora root. Large secondary vessels in a circle are observed inner to the primary xylem vessels Fig. (1b). Also abnormal secondary phloem is found alternating with the secondary vessels. Fibers are observed around the large secondary vessels. Calcium oxalate crystals appear in the cortex as clusters and single raphides (Fig.1c). Appearance and location of the crystals may be specific and useful in taxonomic classification (Kuster, 1956). Few parenchyma cells form the pith at the centre. The leaf: Transverse section showed that, the leave is isobilateral, broad and dark green Fig (2a). The abaxial and adaxial epidermis consists of one raw of elongated cells covered with zigzagging (toothed) cuticle Fig. (2b), the cuticle is absent at the stomatal area. Sunken stomata are observed which is kidney-shaped in surface view. The mesophyll between upper and lower epidermis is not differentiated into palisade and spongy parenchyma. The cells inner to both epidermal layers are dense with chloroplasts (chlorenchyma cells). They are more regularly arranged than the central cells of the mesophyll. The central part consists of very large parenchyma cells with no chloroplast Fig (3a). Parallel vascular bundles (10 – 12) are distributed. Each collateral bundle consists of xylem and phloem. The xylem vessels towards the upper abaxial surface are found in two rows connected with parenchyma cells. The phloem consists of sieve tube cells and companion cells with few parenchymas Fig (2c). Starch granules are distributed in the mesophyll. Fibers are observed around the xylem vessels. Figure 1: Transverse section of U. grandiflora root: (a): Whole view X10; (b): Vascular cylinder X40; (c): raphides in the cortex X40 a
b
c
Figure 2: transverse section of U. grandiflora leaves (a): Whole view X10 (b):
Epidermis and stomata X40. (c): Vascular bundle X40 a
b
c
Anatomy
of Pancratium tortuosum The root: Transverse section of the root appears as complete circle Fig. (3a). Suberized cells are found to the outside forming the bark, inner to the bark a single layer of epidermis followed by one layer of hypodermis. Inner to this 12 – 14 raws of large parenchyma cells of the cortex ended with endodermis. The endodermis is a single layer with very thin-walled cells. The pericycle is found as one layer in between the endodermis and the first protoxylem. Seven to eight radial vascular bundles are found in a circle. The xylem and the phloem are radially alternate. The xylem consists of protoxylem to the outside and metaxylem to the inside, phloem consists of sieve tube cells and companion cells. Formation of abnormal secondary thickening is observed with secondary vessels occupying all the central area. No pith is found Fig. (3b). The leaf: The leaf is isobilateral
(Fig. 4a). The interesting character of this
leaf is the twisting behavior. The twisting behavior may be a xerophytic
character to reduce water loses by transpiration (Esau, 1977). The upper and
lower epidermis each consist of one raw of elongated cells covered with
toothed cuticle interrupted by the sunken stomata Fig. (4b). Surface view showed kidney-shaped stomata.
The mesophyll consists of 3-5 rows of irregular chlorenchyma cells dense with
chloroplasts. The central cells of the mesophyll are large parenchyma cells
without chloroplasts. Very large thin-walled water storing parenchyma cells
extending in the centre of the section alternating with the vascular bundles
are observed Fig. (4a & c). Vascular
bundles are Anatomically the presence of large water storing parenchyma cells in the mesophyll together with the very weak narrow vascular bundles and the absence of supportive tissues may facilitate the twisting behavior of the leaf to adapt the arid environment. The observed thick cuticle and sunken stomata are also xerophytic characters help to decrease the rate of transpiration (Esau, 1977). Figure 3: transverse section of P. tortuosum root (a):
Whole view X10; (b): Vasclar bundle X40 a
b
Figure 4: Transverse section of P. tortuosum leaf. (a): Whole
view X40; (b): Epidermis and stomata X40; (c): Vascular bundle X40 a
b
c
Phytochemical screening Phytochemical
screening of secondary metabolites constituents of U. grandiflora and P. tortuosum bulbs was carried out.
Results were summarized in Table (1). U. grandiflora is rich in alkaloids
and cardiac glycosides. Also it revealed the presence of flavonoids,
saponins, terpenes and steroids. P. tortuosum is also rich in
alkaloids and showed the presence of cardiac glycosides, flavonoids,
saponins, terpenes and steroids. Both bulbs were devoid of
anthraquinones.
Several studies showed that alkaloids (Fennel and van Staden, (2001); Berkov et
al. (2003); Cedron et
al. (2008)), cardiac glycosides
(Kellerman et al. (1988); Kopp et al. (1996); Crouch et
al. (2006)), terpenes (Chantal et al. (2005)) steroids and
saponins (Burkill, 1985) are wildly spread in
Urginea and Pancratium
genera. Table (1): Phytochemical
constituents of U. grandiflora and P. tortuosum
++: high
content; +: present;-: absent. Conclusion This study could provide a good basis for the
selection of Urginea grandiflora
and Pancratium tortousum for further investigation in the
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