芍药内生细菌的分离及产酶菌株的初筛选.doc

芍药内生细菌的分离及产酶菌株的初筛选 摘 要 采用表面消毒法从芍药茎叶组织中共分离得到42株内生细菌，其种群数量可达到1.33×103cfu/g鲜重，颜色以白色为主，细胞形态多为杆状、球状。革兰氏染色结果表明，其中革兰氏阳性菌35株，革兰氏阴性菌7株，结果表明芍药茎叶组织中具有丰富的内生细菌种群多样性。对分离得到的内生细菌进行产功能酶活性菌株的筛选，结果表明淀粉酶产生菌有26株，纤维素酶产生菌有9株，蛋白酶产生菌有16株，脂肪酶产生菌有6株。本研究结果表明，芍药茎叶组织中不仅有丰富的微生物种群多样性，同时是筛选酶资源菌株的资源库，为进一步开发利用芍药内生细菌中的功能酶菌株奠定了理论基础。 关键词：芍药，内生菌，多样性，产酶菌株 ISOLATION AND PRELIMINARY SCREENING ENZYME?PRODUCTION STRAINS?OF ENDOPHYTIC BACTERIA FROM?CHINESE HERBACEOUS PEONY Abstract The 42 endopytic bacterial isolates were isolated from?from the stems and leaves of Chinese herbaceous peony (Paeonia lactiflora?Pall.) by the surface sterilization method.?The population densities?can reach?1.33×103cfu/g?fresh weight.?The color of the colony of mostly endophytic bacteria was white.?The shape of mostly bacterial cells were rod-shaped. A Gram stain’s result showed that 35 strains showed gram positive reaction and 7 strains showed gram negative?reaction.?The results indicated that many endopytic bacteria existed in herbaceous peony leaf and stems tissue. The endopytic bacterial populations had abundant diversity.?The enzyme- producing strains from iaolated endophytic bacteria were screened. The result showed that 26 strains produced amylase, 9 strains produced cellulase, 16 strains produced protease, and 6 strains produced lipase. The results of this study suggested that the endophytic bacteria ?was not only a rich population diversity in herbaceous peony leaf and stems tissue.The enzyme-producing strains resources? laid a theoretical foundation for further development and utilization of endophytic bacteria. Keywords:?Chinese herbaceous peony,?endophytic bacteria, diversity, Enzyme -producing strains 目 录 摘 要 1 Abstract 1 前 言 4 第一章 绪论 4 1.1 内生细菌的概念 4 1.2 内生细菌的种群多样性 5 1.3 内生细菌的作用 5 1.3.1 固氮作用 5 1.3.2 促植物生长作用 6 1.3.3 增强宿主植物抗逆境能力 6 1.3.4 抗病虫害 6 1.4 内生细菌产酶菌株的研究 6 第二章 芍药茎叶内生细菌的分离、纯化与保存 8 2.1 材料和方法 8 2.1.1 样品来源 8 2.1.2 主要试剂及仪器设备 8 2.1.3培养基 8 2.1.4 内生细菌的分离 8 2.1.5 内生细菌的分类 9 2.1.6菌株的保藏 10 2.2 结果与分析 10 2.3 讨论 14 第三章 功能酶