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Changes in Bacterial and Actinomycetes Diversity of Groundnut Phyllosphere with reference to Plant age, Kind of leaves and Season Adopting Culture Dependent Methods

机译:花生毛球菌和放线菌多样性的变化,取决于植物年龄,叶片种类和采用培养依赖方法的季节

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Objective; To study the changes in bacterial and actinomycetes diversity of groundnut phyllosphere of different age of the plant, kind of leaves and season adopting culture dependent methods.Methods Leaf washing method was used to isolate bacteria and actinomycetes from groundnut leaves of 20, 40, 60 and 80 days old with different kinds viz healthy, aphids lepidopteron infested and diseased sampled during summer and khariff season.Results. The bacterial and actinomycetes diversity showed variation with respect to the age, kind of leaves and season… Bacterial count was found to be maximum in summer. Irrespective of season aphids infested leaves recorded highest bacterial count and diseased leaves recorded least population of both.367 bacterial isolates belonging to 9 bacterial species and 127 actinomycetes isolates belonging to 3 genera were recovered from different kind of leaves, ages and season. But their composition showed variation with respect to age, season and kind of leaves. Introduction Phyllosphere or plant leaf surface is a habitat for many microorganisms 1 (Raimen, 1961). The leaf surface microbes are important in several ways. Some of them are known to fix atmospheric nitrogen 2,3 (Murty, 1983; Favilli and Messini, 1990), produce plant growth regulators 4 (Buckley and Pugh, 1971) and can control plant parasites either by stimulating plants to synthesize phytoalexins 5 (Last and Warren, 1972) or by producing antibacterial (Mc Cormack et al., 1994) 6 and antifungal compounds (Starmer et al., 1987). 7 The epiphytic microflora are also known to produce sugars, aminoacids, peptides, enzymes, vitamins, organic acids and nucleotides which exert influence on the plant growth (Chandramohan and Mahadevan, 1968 ). 8 The microbial communities on leaves vary drastically from one leaf to another and undergo constant change in size and composition (Hirane and Upper, 2000). 9 The microflora are challenged with the tharsh condition of the leaf environment (Beattie and Lindow, 1995) including highly fluctuating water availability, exposure to UV radiation from sunlight and limited access to nutrients. 10 The present study was undertaken to evaluate the bacterial including actinomycetes diversity of groundnut phyllosphere of different ages, seasons and kind of levees adopting culture dependent method. Materials & Methods Sample collection12 different Groundnut fields in an around Shencottai Taluk (77 0 25 1 E LONG, 8 0 97 1 N ALT), Tirunelvli district, Tamil Nadu, India were selected in this study. Sampling was done at the age of 20, 40, 60 and 80 DASE (Days After Seedlings Emergence) in both summer (February to May) and khariff (June to August). The leaves of healthy, pest infested aphids and lepidopteron) and diseased leaves were collected in sterile polythene bags kept in icebox, brought to the laboratory and identification of the leaves was done immediately. Pests' infestation and diseased leaves (Leaf spot) were identified by the criteria suggested by Wightman and Rao (1993) 11 and Sokhi (1983) respectively. 12 Microbiological analysisFive gram of (each kind) leaves were suspended separately in 100 ml of sterile distilled water with 0.01% Tween 80 and shaken in rotatory shaker at 100 rpm for 30 minutes. The suspension was serially diluted and 0.1 ml of aliquote was spread plated on standard plate count agar (Hi media,India ). and starch casein agar(Hi media,India) The seeded plates were incubated at 37 0 C for 48 hours and 37 0 C for 14 days (actinomycetes) Developed colonies were counted and then randomly chosen and purified for identification. The purified colonies were stored on respective agar slants at 4 0 C in refrigerator. All the bacterial including actionmycetes by Bergey's manual of systemic bacteriology (Buchanon and Gibbons, 1979; 13 Kirieg and Holt 1984) 14 Statistical analysisThe microbial populations were correlated with the season and kind of leaves using STATISTICA statistical package. Similar statistical packa
机译:目的;采用培养依赖方法研究不同植物年龄,叶片种类和季节的花生叶球体细菌和放线菌多样性的变化。方法采用叶片洗涤法从20、40、60和20的花生叶片中分离细菌和放线菌。在夏季和Khariff季节采集了80天大的不同种类的样本,即健康,蚜虫鳞翅目被侵染和患病。细菌和放线菌的多样性随年龄,叶片种类和季节的变化而变化……在夏季,细菌计数最高。不论季节蚜虫侵染的叶片,细菌计数均最高,病叶叶片的细菌计数均最低。分别从不同种类,年龄和季节的叶片中回收了属于9种细菌的367种细菌和属于3个属的127种放线菌。但是它们的成分随年龄,季节和叶片种类而变化。引言毛层或植物叶片表面是许多微生物的栖息地1(Raimen,1961)。叶表面微生物在几种方面都很重要。已知其中一些可以固定大气中的氮2,3(Murty,1983; Favilli和Messini,1990),产生植物生长调节剂4(Buckley和Pugh,1971)并可以通过刺激植物合成植物抗毒素5(5( Last and Warren,1972)或通过产生抗菌剂(Mc Cormack等,1994)6和抗真菌化合物(Starmer等,1987)。 7附生微生物区系还已知会产生糖,氨基酸,肽,酶,维生素,有机酸和核苷酸,从而影响植物的生长(Chandramohan和Mahadevan,1968)。 8叶片上的微生物群落从一片叶子到另一片叶子变化很大,并且在大小和组成上不断变化(Hirane和Upper,2000年)。 9微生物区系面临着叶子环境的干旱条件的挑战(Beattie和Lindow,1995年),其中包括水分的高度波动,阳光照射下的紫外线辐射以及有限的养分获取。 10本研究旨在采用培养依赖性方法评估不同年龄,不同季节和不同堤防类型的花生叶球中包括放线菌的细菌的多样性。材料与方法样品收集本研究选择了印度泰米尔纳德邦Tirunelvli区Shencottai Taluk(77 0 25 1 E LONG,8 0 97 1 N ALT)周围的12个不​​同的花生田。在夏季(2月至5月)和卡里夫(6月至8月)分别于20、40、60和80 DASE(出苗后几天)进行采样。将健康的,有害生物侵染的蚜虫和鳞翅目的叶片和患病的叶片收集在保存在冰箱中的无菌聚乙烯袋中,送至实验室,并立即进行叶片鉴定。分别根据Wightman和Rao(1993)11和Sokhi(1983)提出的标准确定了害虫的侵染和病叶(叶斑)。 12微生物学分析将5克(每种)叶片分别悬浮在100毫升含0.01%Tween 80的无菌蒸馏水中,并在100 rpm的旋转摇床上摇动30分钟。将悬浮液连续稀释,并将0.1ml等分试样铺展在标准平板计数琼脂(Himedia,India)上。接种板在37 0 C下孵育48小时,在37 0 C下孵育14天(放线菌),对发育的菌落进行计数,然后随机选择并纯化以鉴定。将纯化的菌落分别保存在冰箱中4 0 C的琼脂斜面上。 Bergey的系统细菌学手册(Buchanon和Gibbons,1979年; 13 Kirieg和Holt 1984年)中的所有细菌(包括行动真菌)14统计分析使用STATISTICA统计软件包,微生物种群与季节和叶片种类相关。相似的统计数据包

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