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新疆两种药用阿魏植物内生细菌多样性及抗病促生菌株筛选
刘永红
学位类型硕士
导师李文均
2017-05-01
学位授予单位中国科学院大学
学位授予地点新疆乌鲁木齐
学位专业工程硕士
关键词阿魏 内生菌 多样性 抗病 促生 Ferula endophytes diversity disease resistance growth promotion
摘要Xinjiang, a region of high salinity and drought, is a host to many arid and semi-arid plants. Endophytes live within a plant for at least part of its life cycle without causing apparent disease. Many of these plants including Ferula spp. have indigenous pharmaceutical histories. As many of the medicinal properties of plants are in tandem with the associated microorganisms residing within the plant tissues, it is advisable to explore the endophytic potential of such plants. The present study involved the isolation of endophytes from different tissues of F. sinkiangensis K. M. Shen and F. songorica Pall. ex Spreng and analyzed the distribution pattern among the plant tissues. In addition, the plant growth promoting traits and their biocontrol potential ontaining proction of protease and cellulose, posphate solubilization, ntrogen fixation, sderophore production, indole acetic acid (IAA) synthesis and atagonism assay were studied. During the study, a total of 170 endophytic bacteria belonging to 3 phyla, 15 orders, 20 families, 27 genera and 58 species were isolated from F. songorica. 125 endophytic bacteria belonging to 3 phyla, 13 orders, 23 families and 29 genera were isolated from F. sinkiangensis. The genera Bacillus and Streptomyces constituted a major portion of the endophytic microbes associated with F. songorica and also F. sinkiangensis. The index values according to Shannon and Margalef were 4.02 and 16.55 respectively. Comparing the endophytes from different habitats, it was found that endophytes from Tacheng exhibited more abundant, but those from Shihezi showed more diverse. The 170 endophytes strains in F. songorica were distributed in 27 genera, but 125 endophytes strains in F. sinkiangensis were distributed 29 genera. During this study, root tissues were found to be a more suitable host for endophytic bacteria as compared to stem.Growth promotion assays indicate that most of the endophytes were able to promote plant growth. In this study, 88.0% of the total isolates from F. songorica were capable of grow in nitrogen-free medium, 79.4% and 75.8% of the isolates from F. sinkiangensis were able to produce IAA and fix nitrogen respectively, and 57.1 % of the strains were positive in siderophore production. Salt tolerant experiment of endophytes showed that 5 strains of endophytes grew normally under the condition of 7 % NaCl. Further research about growth promotion ability was carried out by seed germination of wheat.It demonstrated that endophytes bacteria could improve the germination rate of wheat seeds, increase the length and biomass accumulation under salt stress.During This study, there are some potential novel specises belonging to the genera Nocardioides, Paracoccus, Amycolatopsis and Porphyrobacter discovered. This research showed that the protcection of Ferula resource and endophytes in plant tissue were urgently required. This study revealed the great significant ecological adaptability of Ferula, and also the function of endophyes in improving plant disease resistance and stress toleration.
其他摘要新疆位于欧亚大陆中心,降水量少、蒸发强烈,盐碱和荒漠化严重,许多植物不能在正常生长。盐碱和荒漠化是目前影响生态环境,限制农业经济发展的重要原因之一。植物内生菌是植物体微生态系统中的一部分,具有固氮、溶磷、产生长素和产铁载体等能力,能够促进植物生长、提高抗逆性,产生活性与宿主相同的次生代谢物,可作为接种剂,改善农作物在不利环境条件的生长。尤其是生长在盐碱和干旱等逆境胁迫下的植物,其内生菌可能已经适应了逆境,并且能够使宿主植物受益。关于内生菌促进植物生长和提高植物抗胁迫能力的研究已有诸多报道,但关于新疆地区药用植物内生菌的报道并不多。新疆是我国重要的中药材产区,在新疆的盐碱或荒漠环境中,药用植物丰富。阿魏就是新疆特有的珍稀药材资源之一,其历史悠久,药用价值和经济价值高,是我国野生药材二级保护物种。本研究选取来自于新疆塔城地区和布克赛尔县的准噶尔阿魏和来自于石河子地区的新疆阿魏为研究对象,用9种选择性培养基分离其内生菌,并结合其系统发育分析手段探讨内生菌多样性分布特点,同时比较了不同生境和不同组织部位中阿魏内生菌的群落结构变化;另外还对内生菌的固氮、溶磷、产生长素(IAA),产铁载体及产纤维素酶和蛋白酶能力以及抗病能力进行了检测。研究结果表明,阿魏植物均具有丰富的内生菌资源,仅从准噶尔阿魏就分离到三个细菌门中的15个目,20个科,27个属和58个种;新疆阿魏中共分离到13个目,23个科,29个属,59个种。其中准噶尔阿魏内生菌的优势菌为芽孢杆菌属(Bacillus),新疆阿魏内生菌的优势菌为芽孢杆菌属(Bacillus)和链霉菌属(Streptomyces)。多样性指数同样表明阿魏内生菌具有丰富的多样性,如准噶尔阿魏的多样性指数为4.02,丰富度指数为16.55。通过比较不同生境中的阿魏内生菌发现,生长在塔城地区的准噶尔阿魏内生菌资源更丰富,而生长在石河子地区的新疆阿魏内生菌多样性更丰富。其中准噶尔阿魏共分离到170株内生菌,分布于27个属,而新疆阿魏共分离到125株内生菌,分布于29个属。通过分析和比较不同组织部位的内生菌群落后发现,两种阿魏的地下组织根部具有更丰富的内生菌资源,且较茎叶等地上组织,根部也具有更明显的多样。促生实验表明,实验所分离到的准噶尔阿魏和新疆阿魏内生菌资源中大部分具固氮、产生长素和产铁载体等能力,准噶尔阿魏内生菌中88%有固氮能力,新疆阿魏中79.4%能产生生长素,75.8%具有固氮潜力,还有57.1% 能够产生铁载体。本实验对内生菌的促生属性进行了进一步的研究,对同时具有多种促生属性的菌株进行盐胁迫试验,结果发现有5株菌能够在NaCl浓度为7%的条件下正常生长,因此进一步检测这5株菌在盐胁迫条件下对小麦种子萌发的影响,结果表明,内生菌能够提高盐胁迫下小麦种子的发芽率,增加根茎长度和生物量的累积。本研究实施过程中,我们还发现了类诺卡氏菌(Nocardioides),副球菌属(Paracoccus),拟无枝菌酸菌属(Amycolatopsis),产卟啉杆菌属(Porphyrobacter)等类群的潜在新物种。这进一步说明,不仅保护阿魏植物资源具有紧迫性,生长在阿魏植物组织中的内生菌资源更具有研究和应用的价值。本研究成果为揭示阿魏的生态适应性,利用内生菌资源提高植物抗病性和抗逆性提供了可能,对于从微生物角度研究干旱区植物抗病和抗逆等机制具有重要意义。
学科领域生物工程
语种中文
文献类型学位论文
条目标识符http://ir.xjlas.org/handle/365004/14887
专题研究系统_荒漠环境研究室
作者单位中国科学院新疆生态与地理研究所
第一作者单位中国科学院新疆生态与地理研究所
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刘永红. 新疆两种药用阿魏植物内生细菌多样性及抗病促生菌株筛选[D]. 新疆乌鲁木齐. 中国科学院大学,2017.
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