2321-7871

Weekly Science Research Journal

Vol-1, Issue-10, 26th September 2013

Primary Article

Histochemical Studies O...
2321-7871

Vol-1, Issue-10, 26th September 2013

Histochemical Studies Of Cochlospermum Religiosum (l) Alston

have been r...
2321-7871

Vol-1, Issue-10, 26th September 2013

Histochemical Studies Of Cochlospermum Religiosum (l) Alston

The polyphe...
2321-7871

Vol-1, Issue-10, 26th September 2013

Histochemical Studies Of Cochlospermum Religiosum (l) Alston

polymers in...
2321-7871

Vol-1, Issue-10, 26th September 2013

Histochemical Studies Of Cochlospermum Religiosum (l) Alston

Fig-1: Hist...
2321-7871

Vol-1, Issue-10, 26th September 2013

Histochemical Studies Of Cochlospermum Religiosum (l) Alston

Fig-3: Hist...
2321-7871

Vol-1, Issue-10, 26th September 2013

Histochemical Studies Of Cochlospermum Religiosum (l) Alston

Table-2: Le...
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Histochemical studies of cochlospermum religiosum (l) alston

  1. 1. 2321-7871 Weekly Science Research Journal Vol-1, Issue-10, 26th September 2013 Primary Article Histochemical Studies Of Cochlospermum Religiosum (l) Alston Sasikala A, Linga Rao M And Savithramma N ABSTRACT Traditional medicines are prepared from a single plant or combination of plants. Histochemical studies are helpful in drug adulteration and biosystematics arrangement. The present study deals with the location or identification of phytochemicals like tannins, polyphenols, crystals and starch grains in various regions of leaf, stem and stembark of Cochlospermum religiosum by using different chemicals or reagents (FeCl3, Iodine solution, Toluidine blue reagent and HCl). The results showed that the bluish black, purple or blue, bluish green and dark black colour indicates the presence of tannins, starch grains, polyphenols and crystals in various regions like epidermis, endodermis, midrib, cortex and vascular bundle of leaf, stem and stembark of Cochlospermum religiosum. These observations could be of immense value in the botanical identification and standardization of crude drug. formulations were found to possess KEY WORDS: good antibacterial and antifungal Cochlospermum Religiosum, activity (Cecilie et al., 2005). Stembark, Phenols, Tannins. Sasikala and Savithramma (2012) studied the antimicrobial activity of INTRODUCTION Plants are the great sources biological synthesis of silver o f m e d i c i n e s , e s p e c i a l l y i n nanoparticles from leaves of C. religiosum, preliminary traditional system of medicine, which are useful in the treatment of phytochemical screening (Sasikala various diseases. Indian and Savithramma, 2012), contribution to herbal market and quantification of phytochemical emphasis on novel research is (Sasikala and Savithramma, 2013) c o n t i n u o u s l y i n c r e a s i n g . and also studied the invitro Cochlospermum religiosum (L) p r o p a g a t i o n ( S a s i k a l a a n d Alston is a sparsely branched small Savithramma, 2012). Histochemistry or tree, belonging to the family Cochlospermaceae. It is commonly c y t o c h e m i s t r y d e a l s w i t h called as Yellow Silk Cotton, localization of chemical compounds Buttercup Tree and Torchwood within the cells by means of specific Tree because of flowers are large, colors of the compounds. Staining bright golden yellow and seeds the cells with different stains or covered with silky hairs.C. dyes, which render the compounds religiosum stem bark and root visible under the microscope, powder is traditionally used for makes the specific color reaction fertility and ash of fruit mixed with compounds. The importance of coconut is used for the treatment of histochemistry in solving critical scabies (Goud et al., 2005). The biosystematic problems is as gum of C. religiosum is also found to popular as the use of other be an ingredient of unani medicine markers. According to botanical Qurs-e-Sartaan Kafoori which is l i t e r a t u r e s , t h e u s e o f used for Styptic, Antipyretic, histochemical characters in Phthisis, Tuberculosis, Hectic fever taxonomic conclusions is now a and Qurs-e-Suzak Cicatrizant, common practice. For example, the D i u r e t i c , G o n o r r h e a . T h e s e presences of calcium oxalate crystals in various plant families Page No-1 Sasikala A, Linga Rao M And Savithramma N From 1,2,3 Research Scholar Department of Botany, Sri Venkateswara University, Tirupati – 517 502. Andhra Pradesh, INDIA. Te Article is published on September 2013 issue & available at www.weeklyscience.org DOI : 10.9780/ 2321-7871/1102013/29
  2. 2. 2321-7871 Vol-1, Issue-10, 26th September 2013 Histochemical Studies Of Cochlospermum Religiosum (l) Alston have been reported by various scientists (Mbagwu et al., 2009) reported that the size and shape of calcium oxalate crystals though variable in each species showed enough interspecific differences that may be used for taxonomic references in Vigna species. This has been done in other groups of plants such as Dioscoreaceae, (Edeoga and Okoli, 1995). Icacinaceae, (Heintzelman and Howard, 1948) Nyctaginaceae, (Edeoga and Ikem, 2002) and Verbenaceae (Mathew and Shah, 2006). The biosystematic importance and implications of histochemical features of ergastics, calcium oxalate crystals, nature of tannins and saponins have been investigated in various plants families such as Dioscoreaceae (Edeoga and Ogbegbor, 1999), Leguminosaepapilionoideae (Mbagwu and Edeoga, 2006). Identification of localization of secondary metabolites in plant parts which are using in the preparation of drug is an immense importance to prevent adulteration and also helpful in taxonomic hierarchy. Hence in the present study an attempt has been made to identification and localization of secondary metabolites in the medicinal plants. MATERIAL AND METHODS Fresh and healthy leaf, stem and sembark of Cochlospermum religiosum were collected from Tirumal hills of Chittoor district, Andhra Pradesh, India during the year May, 2012. These specimens were initially fixed in FAA (1:1:18) glacial acetic: 40% formaldehyde: 70% ethanol (v/v) for 48-72 h. after 72 h the specimens were transverse (T.S) section were taken using a rotary microtome (RMT-30). Anatomical staining was done by initially staining with few drops of alcian blue for 5 min and counter stained with safranin solution for 2 min. The specimens were treated with FeCl3, Iodine solution, Toluidine blue reagent and HCl for identification of polyphenols, tannins, crystals and starch grains. Photomicrographs of the anatomical features were then taken from the slides using Nikhon Labhot 2 microscopic unit (Asokan, 2006). Fluorescence Studies The fluorescence studies were carried out as per the method of Bhattacharya and Zaman (2009). A small quantity of the leaf powder was placed on a grease free clean microscopic slide and added 1-2 drops of the freshly prepared reagent solution, mixed by gentle tilting the slide and waited for 1-2 min. Then the slides were placed inside the UV-viewer chamber and viewed in day light short (254 nm) and long (365 nm) ultraviolet radiations. The colours observed by application of different reagents in different radiations were recorded. Behavior of the leaf powder with different chemical reagents was studied to detect the presence of major phytoconstituents with color changes under day light by reported method of Pratt and Chase (1949) RESULTS AND DISCUSSION Histochemical Studies Present day's adulteration of the drugs is a common phenomenon. It is inevitable to study the pharmacognostic studies to avoid adulteration.Hence the following experiments were carried out. Histochemical color reactions were carried out through transverse sections of leaf, stem and bark of Cochlospermum religiosum. The results were showed in Table-1. Fig-1 showed various secondary metabolites present in leaf with treatment of different reagents. The presence of tannins was indicated by the development of bluish black colour, when treated with ferric chloride (FeCl3). The tannins were found mainly in the parenchyma tissue of the midrib region, whereas the starch grains were indicated by the development of blue color or purple, when treated with iodine solution. The starch grains were located in the mesophyll and parenchymatous region of midrib. The presence of polyphenols was indicated by the development of bluish green color, when treated with toluidine blue reagent. The polyphenols were found surrounding the vascular bundle sheath. The presences of crystals were indicated by the development of dark black color, when treated with HCl, the crystals were present in the midrib region and vascular bundles. Fig-2 showed the histochemical studies of stem, the presence of tannins were indicated by the development of bluish black color, when treated with ferric chloride (FeCl3). The tannins were present in endodermis and cortex region. The starch grains were indicated by the development of blue color or purple, when treated with iodine solution found in epidermis, cortex and vascular bundles. Page No-2
  3. 3. 2321-7871 Vol-1, Issue-10, 26th September 2013 Histochemical Studies Of Cochlospermum Religiosum (l) Alston The polyphenols were found mainly in the endodermis and cortex region, which were indicated by the development of bluish green colour, when treated with toluidine blue reagent. The presences of crystals were indicated by the development of dark black color, when treated with HCl the crystals were present mainly in the cortex. The histochemical analysis of bark showed in Fig-3 the presence of tannins were indicated by the development of bluish black color, when treated with ferric chloride (FeCl3) and these were found in cortex and vascular bundle region. The polyphenols were found mainly in the cortex region which was indicated by the development of bluish green color, when treated with toluidine blue reagent. The presence of crystal indicated by the development of dark black color, when treated with HCl the crystals were present mainly in the cortex. The starch grains were indicated by the development of blue or purple colour, when treated with iodine solution the starch grains were present mainly in the cortex and vascular bundle region. Toluidine blue is a cationic dye that binds to negatively charged groups. An aqueous solution of this dye is blue, but different colors are generated when the dye binds with different anionic groups in the cell for example, a pinkish purple colour will appear when the dye reacts with carboxylated polysaccharides such as pectic acid; green, greenish blue or bright blue with polyphenolic substances such as ligning and tannins; and purplish or greenish blue with nucleic acids (Baker, 1966). Plants store glucose as the polysaccharide starch, it can be separated into two fractionsamylose and amylopectin. Amylose forms a colloidal dispersion in hot water, whereas amylopectin is completely insoluble. The structure of amylose consists of long polymer chains of glucose units connected by an alpha acetyl linkage. Amylose in starch is responsible for the formation of a deep blue colour in the presence of iodine. The iodine molecule slips inside of the amylose coil. When added the Iodine-KI reagent to a solution or other materials blue colour is present. If starch amylose is not present, then the colour will stay orange or yellow. Leaf Powder Behavior Behavior of the leaf powder with different chemical reagents was studied Page No-3 to detect the presence of major phytoconstituents with color changes under day light by reported method (Pratt and Chase, 1949). And the results were showed in Table-2. Fluorescence Analysis The fluorescence characteristics of leaf powder with different chemical reagents were summarized in Table-3. Although a change in color was observed by the addition of various reagents under day light, none of the reagents induced any fluorescence to the leaf powder under both short and long UV radiations. Under UV light dark brown and black colors were prominent. Fluorescence is the phenomenon exhibited by various chemical constituents present in the plant material. Some constituents showed fluorescence in the visible range in the day light. The natural products (Alkaloids) produce fluorescence in UV light but do not produce fluorescence in visible day light. If the substances themselves are not fluorescent, they often be converted fluorescent derivatives or decomposition products by applying different reagents (Mukherjee, 2002). The pharmacognostical studies and histochemical studies and behavior analysis of fluorescence studies of Cochlospermum religiosum are useful to supplement the information with regard to its botanical identification and drug standardization. Moreover, it also helps in distinction from other allied species and adulteration. REFERENCES Asokan J. (2006) Botanicalmicrotechnique principles and practice. Sky Graphics, Chennai, Pp 1420. Baker JR. (1966) Cytological technique 5th Eds, Methven, London Pp 125. Bhattacharya S and Zaman MK. (2009) Pharmacognostical evaluation of Zanthoxylum nitidum bark. Int J Pharm Tech Res 1:292-298. Bowes BG. (1996) A colour atlas of plant structure. Manson, London, UK Cecilie SN, Drissa D, Kari I, Terje EM, Tsukasa M, Hiroaki K. (2005) Medicinal use of Cochlospermum tinctorium in Mali Anti-ulcer, radical scavenging and immunomodulating activities of
  4. 4. 2321-7871 Vol-1, Issue-10, 26th September 2013 Histochemical Studies Of Cochlospermum Religiosum (l) Alston polymers in the aqueous extract of the roots. J Ethnopharmacol 255-269. – type ecosystems of S.W. Australia. Ann Bot 65: 585-601. Edeoga HO and Ikem IC. (2002) Tannins, saponnins and calcium oxalate crystals from Nigerian species of Boerhaavia L. (Nyctaginaceae). South Afr J Bot 68: 386-388. Pratt RT and Chase ER. (1949) Fluorescence powder vegetable drugs in particular to development systems of identification. J Am Pharm Assoc 38: 324-331. Edeoga HO and Ogbegbor NO. (1999) Distribution of calcium oxalate crystals in some Nigerian species of Aneilema R. Br. (Commelinaceae). Plant Biosys 133: 193-198. Sasikala A and Savithramma N.(2012) Biological synthesis of silver nanoparticles from Cochlospermum Religiosum and their antibacterial efficacy. J Pharm Sci Res 4: 1836-1839. Edeoga HO and Okoli BE. (1995) Histochemical studies in the leaves of some Dioscorea L. (Dioscoreaceae) and the taxonomic importance. Feddes Report 106:113-120. Sasikala A and Savithramma N.(2012) Phytochemical diversity of Cochlospermum religiosum (L.) Alsten – A medicinal tree species. Threats and Concerns to Biodiversity edited by N. Savithramma, Pp 50-59. Goud PS, Murthy KS, Pullaiah T, Babu GV. (2005) Screening for antibacterial and antifungal activity of some medicinal plants of Nallamalais-Andhra Pradesh, India. J Econo Taxono Bot 29: 704-08. Heintzelman CE and Howard RA. (1948) The comparative morphology of the Icaciraceae and the pubescence crystals. Am J Bot 35: 45-52. Sasikala A and Savithramma N. (2012) Invitro plant regeneration of Cochlospermum rligiosum through Shoot Tip Culture .Res J Biot 7 (4): 78-81 Sasikala A, Linga Rao M and Savithramma N. (2013) Quantification of Primary And secondary metabolites From leaves and stem bark of Cochlospermum religiosum (L.) Alsten 2013; 4: Kokashi CJ, Kokasha RJ and Sharma M. (1958) Fluorescence of powdered vegetable drugs in Ultra-violet radiation. J Am Pharm Assoc 47: 715-717. Mathew L and Shah GL. (2006) Crystals and their taxonomic significance in some Verbenaceae. Bot J Linn Soc 88: 279289. Mbagwu FN and Edeoga HO. (2006) Histo-chemical studies on some Nigerian species of Vigna savi (LeguminosaePapilionoideae). J Agron 5: 605-608. Mbagwu FN, Unamb CIN, Onuoha CI and Ezeibekwe IO. (2009) Histochemical studies on fine variants of Viscum L. (Loranthaceae). Res J Biol Sic 4: 254257. Mukherjee PK. (2002) Quality control of herbal drugs: An approach to evaluation of botanicals. 1st Eds, New Delhi: Business Horizons. Pp 114-124. Pate JS, Froend RH, Bowne BJ, Hansen A and Kuo J. (1990) Seedling growth and storage charge characteristics of seeder and resprouter species of Mediterranean Page No-4
  5. 5. 2321-7871 Vol-1, Issue-10, 26th September 2013 Histochemical Studies Of Cochlospermum Religiosum (l) Alston Fig-1: Histochemical studies of leaf of Cochlospermum religiosum (a) Tannins, (b) Starch, (c) Poly phenols and (d) Crystals Fig-2: Histochemical studies of stem of Cochlospermum religiosum (a) Polyphenols, (b) Crystals, (c), (d) Starch and (e) (f) Tannins Page No-5
  6. 6. 2321-7871 Vol-1, Issue-10, 26th September 2013 Histochemical Studies Of Cochlospermum Religiosum (l) Alston Fig-3: Histochemical studies of stembark of Cochlospermum religiosum (a) Tannins, (b) Crystals, (c) Poly phenols and (d) Starch Table-1: Histochemical analysis of leaf, stem and bark of Cochlospermum religiosum Page No-6
  7. 7. 2321-7871 Vol-1, Issue-10, 26th September 2013 Histochemical Studies Of Cochlospermum Religiosum (l) Alston Table-2: Leaf powder behavior of the Cochlospermum religiosum with different chemical reagents Table-3: Fluorescence analysis of leaf powder of Cochlospermum religiosum Page No-7

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