Diversity of Fungal Endophytes in Cymbopogon Caesius

Diversity of Fungal Endophytes in Cymbopogon Caesius Diversity of Fungal Endophytes in Cymbopogon caesius (Hook. Arn.) Stapf. of Kemmannugundi Regions of Karnataka, India Avinash K.S. and Y.L. Krishnamurthy* Abstract: Cymbopogon grass produces many bioactive compounds and used for many medicinal purposes. In this study healthy leaf, stem and root segments were cultured on media like Potato dextrose agar and Malt extract agar medium and pure cultured. Endophytic fungi were isolated from 1200 samples of Cymbopogon caesius grass collected from Kemmannugundi regions of Karnataka. Overall 56% of colonisation rate from surface sterilised tissues were recorded. About 30 different fungal species were isolated and 12 were identified and 18 were grouped as morph taxa. Less isolates were recovered from leaf segments and more isolates were recovered from root segments. Curvularia sp. and Fusarium sp. were frequently isolated endophytes with the high colonisation rate. Key Words: Endophytes, Cymbopogon caesius, Grass, Curvularia, Fusarium. 1. Introduction Endophytes are microorganisms which live inside the host without causing any negative effect by their presence. The plant and endophytic fungi show symbiotic association. Plant protects and feeds the endophyte which in return produces bioactive secondary metabolites to enhance the growth and competitiveness of the host in nature (Carrol, 1988 and Hong Lu et al., 2000). Endophytes are widely investigated for their bioactive metabolites and have proven to produce potential compounds which are applicable in medicine (Clovis and Ewald, 2012). The compounds usually isolated from medicinal plants may not be plant metabolites they may be from fungal endophytes which reside in the host plant. Endophytes of tropical plants are among the groups of fungi that have been studied to arrive at the predicted figure of 1.5 million (Hawksworth, 1991; Subramanian et al 2003) Cymbopogon species are traditional medicinal grasses well recognised for its aromatic oil. Cymbopogon caesius grass was wide spread in areas of Kemmannugundi of Chikmagalur Dist, Karnataka. Despite the wide distribution of Cymbopogon grass along the central Western Ghats of Karnataka only limited work has been done for their association with fungal endophytes. Even though the oil extracted from the grass has been used in medicine currently there is no enough information regarding the diversity of the fungal endophytes with the species. We have selected Cymbopogon caesius (Hook. Arn.) Stapf for the study of diversity of the fungal endophytes. Fungi were isolated from leaves, stem and root followed by the surface sterilisation process. The objectives of the study were to isolate and identify the fungal endophytes from different parts of Cymbopogon caeceous grass of Kemmannugundi regions of Karnataka. 2. Meterials and Methods 2.1 Sampling: C. caeseus (root, stem and leaf samples) was collected from Kemmannugundi (13 º2826 N 75 º4450 E 4275ft) Chikmagalur District. Roots were collected with a ball of soil so that to avoid damage to the roots. Twenty healthy looking culms of grass were collected and transported in a clean polythene bag in closed condition and processed within 24 hour of collection. 2.2 Isolation of fungi The collected samples were washed thoroughly by running tap water to clean soil from roots and were cut into small pieces. The cut segments were surface sterilised using 70% ethanol followed by immersion in 3% Sodium hypochlorite for 4 minutes and again washed twice with distilled water (Maheshwari and Rajagopal, 2013). The efficacy of the surface sterilisation was confirmed by pressing the sterilised segments onto the surface of the medium, the surface sterilised small pieces were cut into .03-.05cm segments (Chuyang et al., 2001). Four hundred segments were inoculated into potato dextrose agar medium which is supplemented with 100mg of Amoxillin to inhibit the growth of bacteria. The inoculated petri plates were wrapped with petriseal and incubated at 27  ±1 degree centigrade. The plates were observed daily after 3 days of inoculation up to one month the emerged fungal endophytes were transferred to new petri plates containing PDA Medium. 2.3 Identification of Endophytic fungi For the identification of endophytic fungi, slides were prepared from pure cultures and were stained with Lacto phenol cotton blue stain and observed under Karl Zeiss Primo star microscope. Morphological Characteristics such as growth pattern colour of colony, mycelium texture, spore production type and characters of the spore (Barnet Hunter, Ellis, Subramanian). 3. Statistical analysis The colonisation rate (CR) was determined by total number of segments yielding endophytes divided by the total number of segments inoculated (Petrini et al., 1982). Number of segments yielding endophytes Total number of segments inoculated 4. Results A total of 959 isolates of endophytes were recovered out of 1200 segments inoculated. Total 400 segments each of root, stem and leaf segments were inoculated for the isolation of endophytes. Most frequently found endophytes were Fusarium oxysporum (11.46%), Aspergillus clavatus (8.7%), and Curvularia spp. (7.78%) (Table 1). Table 1: Colonisation Frequency of fungal endophytes of C. caesius. Sl. No. Endophyte Name Colonisation frequency (%) Root Stem Leaf Total Alternaria alternata 4.1 1.09 1.73 Aspergillus clavatus 15.3 6.0 4.8 8.7 Bipolaris Sp. 2.00 0.66 Cephalosporium sp. 0.8 1.9 0.9 Cladosporium sp. 0.6 0.2 0.26 Curvularia andropogonis 16.0 5.2 1.7 7.63 Curvularia lunata 12.2 7.9 3.7 7.93 Fusarium oxysporum 16.1 11.9 6.4 11.46 Hansfordia ovalispora 0.9 0.3 Oidiodendron sp. 1.6 0.9 0.83 Trichoderma sp. 0.4 1.3 0.56 Wardomyces anomala 0.6 0.2 Unidentified Morphotypes 16.4 14.3 14.6 15.1 Table 2: Infection frequency and isolation rate of fungal endophytes of C. caesius. C. caesius plant tissues Leaves Stem Root Total Number of Samples 400 400 400 1200 Number of isolates recovered 126 198 348 672 Colonisation rate 31.5% 49.5% 87% 56% Number of Morph taxa 4 5 9 18 More isolates of endophytes (87%) were recovered from the root segments of the Cymbopogon grass compared to stem (49.49%) and leaf segments (56%). 18 unidentified fungi were numbered and stored as morph taxa. 5. Discussion The Cymbopogon grass is the very popular for its aromatic oil. The purpose of the study was to document endophytic diversity of Cymbopogon grass of the study area which was widely distributed in the area. The endophytic fungi were cultivated on artificial medium (PDA) as pure culture. Some of the fungi exhibited characteristic colony characters and microscopic characters which were helpful in identifying them whereas some of the fungus was produced good mycelia but did not show any sporulation were kept as morph types. All the isolated endophytes belonging to 11 different genera. The endophytes were more resided in root. The endophytes may not show any host specificity as they were recovered from different groups of plants (Petrini 1986). In the present investigation leaves, stems and roots of C. caesius were used for isolation of endophytic fungi. The fungal colonisation was higher in roots as compare to stem and leaves. Higher colonisation of endophytes in leaf and stem tissues, as compared to roots, was reported by Siegel and Latch (1991) and Clay and Schardle (2002) Ajay et al (2012) in grasses and in the study of medicinal plant species leaves colonised greater number of endophytes compared to stem and bark (Raviraja 2005). About 11 genera of fungal species were isolated which indicates the various fungal populations in the grass. The variation in the colonisation rate depends on the host habitat, Soil environment (Shankar Naik et al, 2014). In the study the CR is varied in different parts of the plant. The more number of fungal endophytes isolated from root which similar to the results of Sita et, al., 2011 where thy obtained almost three times more endophytes in roots compared to shoot tissues. Fungi have been widely investigated as a source of bioactive compounds. An excellent example of this is the anticancer drug, taxol, which had been previously supposed to occur only in the plants (Strobel Daisy, 2003) in this aspect present study is very relevant to explore more useful fungal endophytes. 6. Acknowledgement The authors are gratefully thankful to Kuvempu University Shankaraghatta Shimoga for constant support throughout the study and Department of Biotechnology, Govt. of India, New Delhi for Financial assistance. 7. References [1] Chunying Zhang, Lijuan Yin, Silan Dai, Diversity of root-associated fungal endophytes in Rhododendron fortune in subtropical forests of China, Mycorrhiza. 19(2009) 417-423. [2] Carroll, G. C, Fungal endophytes in stems and leaves: From latent pathogen to mutualistic symbiont, Ecology. 692 (1988) -9. [3] Hawksworth,D. L, The fungal dimension of biodiversity :magnitude, significance, and conservation, Mycol. Res. 95(1991) 641-655. [4] Petrini, 0. Stone, j. Carroll, F. Endophytic fungi in evergreen shrubs in western Oregon: a preliminary study, Canadian Journal of Botany. 60(1982) 789-796. [5] Petrini, O, Taxonomy of endophytic fungi of aerial plant tissues. In Microbiology of the Phyllosphere (ed. N. j. Fokkema ). van den Heuvel), 1986, pp. 175-187. Cambridge University Press: Cambridge. [6] Pramuan Saithong, Wanchai Panthavee, Siriporn Stonsaovapak and Li Congfa, Isolation and primary identification of endophytic fungi from Cephalotaxus mannii trees, Maejo International journal of Science and Technology. 4(03) (2010) 446-453. [7] Raviraja N S, Fungal endophytes in five medicinal plant species from Kudremukh Range, Western Ghats of India, Journal of Basic Microbiology. 45(3) (2005) 230-235. [8] Shankar Naik B, Krishnappa M, Krishnamurthy Y L. Endophytic assemblage in Strychnous nuxvomica L. and antagonistic activities in vitro, Archives of Phytopathology and Plant Protection. 2014. [9] Sita R Ghimire, Nikki D Charlton, Jeremey D Bell, Yelugere L Krishnamurthy and Kelly D Craven, Biodiversity of fungal endophyte communities inhabiting switch grass (Panicum virgatum L.) growing in the native tall grass prairie of northern Oklahoma, Fungal Diversity. 47(2011)19-27. [10] Suryanarayanan T S and Kumaresan V, Endophytic fungi of some halophytes from an estuarine mangrove forest, Mycological Research. 104(12) (2000) 1465-1467. [11] Hong Lu, Wen Xin Zou, Jun Cai Meng, Jun Hu, Ren Xiang Tan, New bioactive metabolites produced by Colletotrichum sp., an endophytic fungus in Artemisia annua, Plant Science 151 (2000) 67–73. [12] Clovis Douanla-Meli,Ewald Langer, Diversity and molecular phylogeny of fungal endophytes associated with Diospyros crassiflora, Mycology 3(3) (2012):175-187. [13] Maheswari, S., and Rajagopal, K, Biodiversity of endophytic fungi in Kigelia pinnata during two different seasons, Curr. Sci. 104(2013): 515-518. [14] Barnett, H.L., and Hunter, B.B., 1972.Illustrated genera of imperfect fungi, 3rd ed. Burgess publishing company, USA.pp. 218. [15] Ellis M B, More Dematiaceous Hypomycetes, Commonwealth Mycological Institute, Kew, Suirrey, England, 1976. [16] Siegel MR, Latch GCM., Expression of antifungal activity in agar culture by isolates of grass endophytes, Mycologia, 83(1991) 529–537. [17] Clay K, Schardle CL., Evolutionary origin and ecological consequences of endophyte symbiosis with grasses, Am Nat, 160(2002) S99–S127. [18] Ajay Kumar Gautam , Mona Kant Yogita Thakur., Isolation of endophytic fungi from Cannabis sativa and study their antifungal potential, Archives Of Phytopathology And Plant Protection, Vol. 46 (2013), No. 6, 627–635. [19] Gary Strobel and  Bryn Daisy, Bioprospecting for Microbial Endophytes and Their Natural Products, Microbiol Mol Biol Rev. 67(4)2003: 491–502.

Levi and Strauss & Co

Levi Strauss & Co. is a flourishing business. Since the early days, it has been a leader in the garment industry. The original and most famous Levi Strauss product is blue jeans. Jeans have become desirable and even fashionable clothing for not only miners, farmers, and cowboys, but also for movie stars, executives, women, children, and teenagers from all over the world. Throughout its history, however, the company has researched and developed a number of other products. The company now markets a wide range of clothing and accessories, all under the brand name Levi’s.Many new Levi products have been launched over the years. Some of these have succeeded beautifully, but others have flopped completely. The company is still best known, however, as the maker of Levi jeans, the pants that are guaranteed to shrink1, wrinkle, and fade2. In 1954, flushed with the success of the cotton twill pants it had introduced a few years earlier, Levi brought out a line of permanent press (no-iro n) slacks. Within six months, 5 out of every 100 pairs sold had been returned, and Levi had to admit it didn’t have the right fabric for permanent press.Fifteen years later, as the company was planning its major expansion, it hit on a couple of equally dramatic flops. First was the denim bathing suit—which, when wet, weighed the wearer down to the point of imminent drowning. Next was a line of disposable (throw-away) sheets and towels. These, Levi discovered, were not high on the consumer’s list of priorities. Unable to interest hotels in the product, the company was saved when the factory that made the sheets burned down. Levi absorbed the $250,000 loss.Eventually Levi created six new divisions, ranging from jeans to accessories and including a sizable effort in women’s sportswear, Levi’s for Gals. The diversification3 worked. In the mid-1970s Levi’s sales hit the billion-dollar mark, having taken 125 years to reach that milestone. Four yea rs later sales hit $2 billion. In 1979 the company ranked 167 on Fortune’s4 list of the 500 largest industrial corporations, and 20 in net profits. 5 Between 1970 and 1980 Levi had grown an average of 23 percent a year.In 1979 alone it sold 143 million garments. In menswear, though, all Levi products had been aimed at the middle of the market. The company had brought out a line of moderately dressy slacks and polyester leisure suits—the Action Slack and Action Suit—and was doing a brisk business with them. But the tempting upper end of the market remained untouched. â€Å"If we want to grow we’re probably going to have to go to upper moderate price points,† one Levi official explained, â€Å"and somewhat higher taste level for our products. In short, they needed to sell more expensive clothes—like the Tailored Classic. If Levi could sell sport coats, dress slacks, and, above all, suits, a whole new market would open up. The Tailored Classic might make money all by itself. But even more important, it would get Levi into the business of producing fancier and costlier clothing. The consumer would come to think of it as a manufacturer of dress apparel and it could spin off6 many more such lines in the future. Why, with such a record of success would any company be worrying about making new products?Part of the answer, obviously, is the sheer riskiness of depending so heavily on a single item. The boom in jeans was in many ways a historical accident, and what history has given it can also take away. Then, too, the demographics of the marketplace were already beginning to change. Jeans, to be sure, were no longer the exclusive province of youth: baby boomers7 who had grown up on Levi’s kept on wearing them into their twenties. But they would no longer be wearing them everywhere and all the time, as they did when they were teenagers.And the next generation of adolescents was not so numerous. The birthrate had peaked i n 1957; by 1964, demographers agreed, the baby boom in the United Stated was over. The bulge in the population that the boom had created would soon be moving into a world of casual slacks, leisure suits, and coats and ties. From a marketing point of view, that’s where the action would be. Finally, the competition had been gearing up. Levi’s had always shared the market with Blue Bell’s Wranglers and other national brands like Lee. But now everyone seemed to be selling jeans.Back in 1970 Levi probably couldn’t have foreseen the popularity of â€Å"designer† jeans skimming off8 the upper end of the market. But they could certainly anticipate cut-rate models gnawing away at the low end. To sell their wares, Levi knew, retailers would have to slash prices. The profit outlook in a saturated, competitive marketplace like this was bleak. Adapted from: Boyd, F. (1994). Making Business Decisions: Real Cases from Real Companies. USA: Addison-Wesley Publishin g Company. _________________________________________________________________________________________

The Fundamentals of Thematic Essay Topics for Us History Regents Revealed

The Fundamentals of Thematic Essay Topics for Us History Regents Revealed Though it has several similarities with other essay types, there are a few significant differences too. Consistency can't be mentioned enough when it regards a great thematic essay. Conduct some analysis to demonstrate your theory and explain your chief claims. There are different types of essays. Creating such a literary work is no simple feat. You ought to be quite careful when selecting your themes. Students lead busy lives and frequently forget about an approaching deadline. Thematic Essay Topics for Us History Regents - Dead or Alive? Being a complicated job, essay writing is related to many mistakes students have a tendency to make. It's probably clear to each student that each and every bit of academic writing has a goal. In case, you feel that it's impossible to compose your thematic essay in time or you're not sure you're writing it properly use our help. Nowadays it's very difficult to locate a trustworthy essay writing service. As soon as you inform us about all of the paper information, we'll begin trying to find an ideal writer for your paper. In such way, you will find some brilliant suggestions to introduce in your paper and will determine the most suitable theme. It is essential to opt for the most crucial idea dependent on the prompt. Be logical and coherent whilst giving your major suggestions and concepts. A paper proposal is the initial step to writing a research paper the intention behind the proposal is to gain clearance from the instructor that the topic you've chosen. Your very first draft will incorporate all the essay's vital info but might need minor revisions to guarantee the maximum quality of writing. Following that, choose a strong thesis statement. Compose extra paragraphs if necessary to be able to completely answer the essay question. If you don't understand how to plan your work, consider writing a thematic essay that's easy about the outline. An essay should tie all paragraphs back to your theme and offer interesting information that could easily educate readers. Writing an effective essay demands careful attention to detail and capacity to organize your thoughts about the most important subject of your research. A thematic essay is a significant portion of the education approach. As the chief aim of a thematic essay prompt is to produce the student respond to the particular questions, so give your solution at the end! Your goal is to compose a succinct review of the job you've already done to produce clear each point that is quite significant for your paper. The intent of this essay type is to earn a point. Commonly, the thematic essay is connected with social problems in various periods of human civilization. The significant challenge comes from how the search for the appropriate and intriguing subject is truly hard. You may select a subject that appears to be relevant. It is crucial to identify and disclose the principal subject of the studied work. The task creates an individual actively reflect on the problem (subject, question) and search for optimal kinds of expression, which is a significant factor in mental development of somebody. To make certain you will locate a complete answer to every question, we've got a support team that's always online. It's important to keep in mind that the clearer the dilemma is, the simpler it is going to be for you to locate the appropriate material, the more dynamic and interesting your work will be. The only issue is that you ought to find out how to compose a thematic essay.

The Jilting of Granny Weatherall Essay - 786 Words

In this short story of The Jilting of Granny Weatherall by Katherine Anne Porter, there is a powerful symbolic meaning through out the entire narrative. Although the symbols are not obvious in some paragraphs, they are in hidden text in others, which has to be, examined thoroughly by the reader. Granny is an eighty-year-old woman on her deathbed. She is in a state of confusion drifting in and out of consciences; she is reminiscing and blurring the past with the present. Although she comes to her senses every now and then, she is still perplexing some important details of her life. Through this story, all the symbols help define Grannys character, provide greater understanding of her life, and reinforces the important parts of†¦show more content†¦Granny weathered through all the storms in her life and had conquered many obstacles except her jiltings. While on her deathbed, she wanted her daughter Cornelia to find George after sixty years she wanted him to know that she had her husband just the same, and her children, and her house like any other woman (page 99). She also wanted him to know that every thing that he had taken away from her, had been given back to her and even more, than she had hoped for. The text implies that she had been pregnant by George perhaps with Hapsy and that is why they were getting married. When George had left her with her white veil on and the white cake set out for a man (page 98), she compared that feeling to being in deepest pit of hell. Now that she was diminishing, that same feeling of hell returned to haunt her. Her husband John and Hapsy had died and left her, which also made her feel forsaken by the two men she had truly loved and the child she had truly wanted. All of the abandonment that occurred in her life in the past six decades had just made her a stronger woman, who was straightf orward, stern, and meticulous in the way she lived her life. Granny wanted a sign from God (page 101), so she could let go and release everything that had been building up inside of her all for sixty years, theShow MoreRelatedThe Jilting of Granny Weatherall700 Words   |  3 PagesIn â€Å"The Jilting of Granny Weatherall† by Katherine Anne Porter, we learn of an elderly woman who is lying on her death bed watching her life pass before her eyes. We learn, from these flashbacks, how much she has overcome and endured, and how shes put her whole heart into being a mother and wife up until her last breath, when she blew out the candle and rode with her Father in a cart to heaven. 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