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International Journal of World Research December 2013, Issue IXI: Volume: I Print-ISSN: ISSN: 2347 – 937X Impact Factor: 0.4327 A STUDY ON PHYLLOSPHERE MYCOFLORA OF TRIBULUS TERRESTRIS, L, WITH SPECIAL REFERENCE TO TRIBULUS TERRESTRIS AS CRUDE DRUG Mary Josephine1 & Kanchana2 1 Associate Professor, Department of Botany,Nirmala college for women, Coimbatore,Tamilnadu,India 2 Scholar, Nirmala college for women, Coimbatore,Tamilnadu,India ABSTRACT Leaf samples of Tribulus terrestris, Linn was collected from the waste lands from ten different locations in valparai, Karanampettai, Maruthamalai, Anaikatti and Siruvani region of Coimbatore District,Tamil Nadu, India.Qualitative studies made on the Aspergillus mycoflora in the phyllosphere of Tribulus terrestris plants using the soil-washing technique. Aspergillus isolates were examined, and the distribution are determined in the phyllosphere .The study revealed, Aspergillus mycoflora was more abundant in all the selected samples.Aspergillus can cause disease in three major ways: through the production of mycotoxins; through induction of allergenic responses; and through localized or systemic infections. Raw part of Tribulus terrestris carries lot of Aspergillus fungus both in root and leaves. In the point of view of using Tribulus terrestris as crude drug, it is an important observation. Key words: phyllosphere, Aspergillus, Tribulus terrestris INTRODUCTION Tribulus weed species are summer growing annuals and have high drought tolerance. with a small yellow flower and spiny fruit, called caltrop (Tribulus terrestris.,L) Plant species are opportunistic weeds of cultivation, waste lands and degraded pastures.Under certain conditions, grazing of Tribulus has been associated with nitrate poisoning, photosensitisation and sheep staggers. Spines on fruit damage the feet of livestock, causing lameness, and fruit can become embedded in sheep fleece, lowering its value and causing discomfort to shearers.Caltrop is also a weed of urban areas and is a nuisance on footpaths and playing fields. Caltrop is now found worldwide, particularly in areas with a Mediterranean climate. It is listed as a weed in 37 countries. Page | 108 International Journal of World Research Print-ISSN: ISSN: 2347 – 937X Tribules terrestris, Linn Classification : Class Sub class Order Family Genus Common Names : Eng Hind Sans Ben Tamil Mal December 2013, Issue IXI: Volume: I Impact Factor: 0.4327 – Magnoliophyla – Dicotyledons – Rosidae – Sapindales – Zygophyllaceal – Creosote – bush family – Tribulus – Punturevine – Small Caltrops – Chota gokhru – Ikshugandha – Gokhuri – Chrunerincle, Nerinji – Nerinnil, Neringil Botanical Description : Annual or bicnnial, prostrate, densely appressed whitish silky pubescent herb. Stem hirsute to sericeous, branches spreading. Leaves paripinnate, 2.5 – 5 cm long; Stipculs lanceolate to falcate, 3-5mm long; leaflets (40) 5-6 (-8) pairs orate to elliptic – ablong, 5-10 (-12) mm long, 3-8mm broad, in-equilateral, acute. Flowers yellow, 1-1.5 cm across; Pedical up to 1.5 (-2) cm long. Sepals orate – lanwdate 5-6 mm long, c3mm broad, acute. Petals obovate, 6-8 mm long, 34 mm broad, obtuse. Stamens 10, filaments c 3-5mm long, another versatile. Ovary ovoid, hirsute; style c 1.5mm long, stigmas decurrent. Fruit up to c 1 cm in diameter, 4-8mm long, mericarps densely crested and tuberculate on dorsal side, densely hairy to glabrescent, with 2 long patent and 2 short downwardly directed spines (rarely all or lower 2 reduced to tubercles) ( Plate 1 ) Uses : Tribulus terrestris is found to be potent diuretic, nephroprotective and entiurolithiatic. It is non-irritant diuretic and urinary antiseptic hence outstanding remedy is urinogeital disease promoting urine flow, soothing the mucosa, and aiding in the excretion of stones and aiding in the excretion of stones and calculi. It is used in case of dysuria, urinary stone, and incontinence of urine. Both the plant and the seeds are used for treatment of phosphaturia, dysuria, gonorrhea, gleets, chronic cystitis, gout, post partum hemorrhage and to ensure fecundity. It is used in various herbal formulas to treat headaches, eye problems such as itching, conjunctivitis and weak vision. It is also used to treat high blood pressure and rib pain. The hormonal balancing effects of the plant for women makes this herb suitable for prementstrual syndrome and menopausal syndrome. Tribulus is used in case of impotence and spermaturia as it helps to increase the level of the luleinizing hormonal in the body and improved the synthesis of the testicular male sex hormone.Saponin present in T.trerrestris dilates the coronary artery and improves coronary circulation. The plant contains harmine alkaloids that are most likely to be responsible for its redative properties. The seeds are strengthening and the ash of plant is good for external application is rheumarthritis. It also has been reported for its anti angina, hepathprotective, and anthelmintie activity. Page | 109 International Journal of World Research Print-ISSN: ISSN: 2347 – 937X December 2013, Issue IXI: Volume: I Impact Factor: 0.4327 Phyllosphere The phyllosphere is a term used in microbiology to refer to leaf surfaces or total aboveground surfaces of a plant as a habitat for microorganisms. The below-ground microbial habitats (i.e. the thin-volume of soil surrounding root or subterranean stem surfaces) are referred to as the rhizosphere and laimosphere, respectively. All plants are host to a numerous and diverse community of microorganisms including bacteria, fungi, and yeasts. Some are beneficial to the plant, others function as plant pathogens and may damage the host plant or even kill it. However, the majority of bacterial colonists on any given plant have no detectable effect on plant growth or function. Research into the characteristics of microbial life in the phyllosphere is of great commercial importance to the agricultural industry for two reasons. First, understanding the survival of plant disease-causing bacteria and fungi is vital for developing new ways to control their spread. Second, there has been a recent rise in the number of food poisoning cases associated with fruit and vegetables contaminated with bacteria, such as Salmonella and E.coli. This is particularly true of fresh fruits and salads which are not cooked prior to consumption. Preventing these outbreaks by developing better decontamination strategies is important to protect public health. Review of Literature The above-ground parts of plants are normally colonized by a variety of bacteria, yeasts, and fungi. While a few microbial species can be isolated from within plant tissues, many more are recovered from the surfaces of healthy plants. The aerial habitat colonized by these microbes is termed the phyllosphere, and the inhabitants are called epiphytes. Bacteria are by far the most numerous colonists of leaves, often being found in numbers averaging 106 to 107 cells/cm2 (up to 108 cells/g) of leaf (Andrews and Harris, 2000; Kim and Sundin, 2001). The microbial communities of leaves are diverse and include many different genera of bacteria, filamentous fungi, yeasts, algae, and, less frequently, protozoa and nematodes. Filamentous fungi are considered transient inhabitants of leaf surfaces, being present predominantly as spores, whereas rapidly sporulating species and yeasts colonize this habitat more actively (Andrews and Harris, 2000). The phyllosphere supports numerous microorganisms, including bacteria, fungi, yeast, and protozoa (Andrews and Harris 2000; Beattie and Lindow 1995; Hirano and Upper 2000; Kinkel 1997; Leveau 2006; Lindow and Leveau 2002; Lindow and Brandl 2003). Bacteria are the most abundant members of the phyllosphere community, and have been shown to colonize leaves at densities of up to 108 cells cm−2 . The description of microbial community structure and the quantification of microbial diversity associated with leaf surfaces has been much improved by the application of culture-independent methods. These have provided many new insights into the microbial species that are common and unique to all plant leaf surfaces, the specific adaptations Page | 110 International Journal of World Research Print-ISSN: ISSN: 2347 – 937X December 2013, Issue IXI: Volume: I Impact Factor: 0.4327 that bacteria and fungi possess and express to meet the chemical and physical challenges of the phyllosphere environment, and the factors that determine community composition, such as the plant species and weather conditions. Materials and Methods Leaf Sampling for Phyllosphere study Leaf samples of Tribulus terrestris, Linn was collected from the waste lands from ten different locations in valparai, Karanampettai, Maruthamalai, Anaikatti and Siruvani region of Coimbatore District,Tamil Nadu, India. Leaf samples were collected from the month of December to February,2012-2013.The leaves were selected randomly and directly extracted from the leaf surface by washing the leaf surface. The leaves were homogenized with sterile quartz sand and 9 mi of sterile saline (9 %.), one gram of fresh weight of leaves were taken from the collected samples. Samples were replicated three times. The dilutions were plated on Potato Dextrose Agar media suitable for the fungus group. Developed colonies were observed by using Compound microscope to find out the occurrence of Aspergillus fungus. Results and Discussion The phyllosphere study showed the colonies of Aspergillus were found to be occur in all the samples collected(Plate 2). Similar results were obtained (Abdel-Hafez.,1984). Qualitative studies made on the Aspergillus mycoflora in the phyllosphere of Tribulus terrestris plants using the soil-washing technique. Aspergillus isolates were examined, and the distributions were determined in the phyllosphere . Aerosolized Aspergillus spores are found nearly everywhere. So we are routinely and almost constantly exposed to them. Such exposure is a normal part of the human condition and generally poses no adverse health effects. Aspergillus can and does cause disease in three major ways: through the production of mycotoxins; through induction of allergenic responses; and through localized or systemic infections. The most common pathogenic species are Aspergillus fumigatus and Aspergillus flavus. Aspergillus flavus produces aflatoxin which is both a toxin and a carcinogen and which can potentially contaminate foods such as nuts. Aspergillosis is an infection or allergic reaction caused by various kinds of mold (a type of fungus). In traditional medicines the various parts of the plant materials are used as raw. These raw extracts contain Aspergillus. Raw part of Tribulus terrestris carries lot of Aspergillus fungus in the leaves. In the point of view of using Tribulus terrestris as crude drug, it is an important observation. References 1. Andrews, J. H., and R. F. Harris. 2000. The ecology and biogeography of microorganisms on plant surfaces. Annu. Rev. Phytopathol.38:145-180. 2. Beattie, G. A., and S. E. Lindow. 1995. The secret life of foliar bacterial pathogens on leaves. Annu. Rev. Phytopathol. 33:145-172. Page | 111 International Journal of World Research Print-ISSN: ISSN: 2347 – 937X December 2013, Issue IXI: Volume: I Impact Factor: 0.4327 3. Hirano, S. S., and C. D. Upper. 2000. Bacteria in the leaf ecosystem with emphasis on Pseudomonas syringae—a pathogen, ice nucleus, and epiphyte. Microbiol. Mol. Biol. Rev. 64:624-653. 4. Kim, J. J., and G. W. Sundin. 2001. Construction and analysis of photolyase mutants of Pseudomonas aeruginosa and Pseudomonas syringae: contribution of photoreactivation, nucleotide excision repair, and mutagenic DNA repair to cell survival and mutability following exposure to UV-B radiation. Appl. Environ. Microbiol. 67:1405-1411. 5. Kinkel, L. L., M. Wilson, and S. E. Lindow. 2000. Plant species and plant incubation conditions influence variability in epiphytic bacterial population size. Microb. Ecol. 39:111. 6. Lindow, S. E., and J. H. J. Leveau. 2002. Phyllosphere microbiology. Curr. Opin. Biotechnol. 13:238-243. Plate 1 Plate 2 Page | 112 International Journal of World Research Print-ISSN: ISSN: 2347 – 937X December 2013, Issue IXI: Volume: I Impact Factor: 0.4327 Page | 113