2018年8月17日 星期五
金钗石斛内生细菌的组成及多样性分析
Analyses on composition and diversity of endophytic bacteria in Dendrobium nobile
2018年 第27卷 第1期 页码[79-90]
摘要

采用9种培养基对贵州省的赤水市旺隆镇(P1)、贵阳市贵州师范大学生命科学学院大棚(P2)和金沙县台金生态农业观光园(P3)内栽培的金钗石斛(Dendrobium nobile Lindl.)根、茎和叶中的内生细菌进行了分离,并采用扩增核糖体DNA限制性分析(ARDRA)法对分离的内生细菌进行了多样性分析。结果表明:从3个样地金钗石斛的根、茎和叶中共分离出1 081株内生细菌,隶属2门5科12属40种1亚种,可分成41个OTUs;其中,芽孢杆菌属(Bacillus)和短芽孢杆菌属(Brevibacillus)为优势属,Bacillus subtilis subsp. subtilis和Brevibacillus invocatus为优势种(亚种)。金钗石斛的内生细菌在不同器官中的分布存在差异,根中内生细菌的菌株数量最多(695),且根中内生细菌的Shannon-Wiener多样性指数(2.20)和Margalef丰富度指数(5.81)均高于茎和叶中内生细菌,而叶中内生细菌的Pielou均匀度指数却最高(0.69);并且,茎和叶间内生细菌的Srenson相似性系数最高(0.82),显著高于根和茎间以及根和叶间内生细菌的Srenson相似性系数。3个样地金钗石斛内生细菌的共有OTUs有14个,分离菌株数占金钗石斛内生细菌分离菌株总数的96.5%;并且,3个样地金钗石斛内生细菌的Shannon-Wiener多样性指数和Pielou均匀度指数差异较小,仅P1样地金钗石斛内生细菌的Margalef丰富度指数明显高于P2和P3样地。NA、TSA、LB、KB、R2A和TB培养基从金钗石斛中分离出的内生细菌菌落数和OTU数量均高于YPD、YT和YG培养基。研究结果显示:金钗石斛体内含有丰富的内生细菌资源,并且,其内生细菌的组成和多样性与分离器官、种植地和培养基密切相关。

 

Abstract

Endophytic bacteria in root, stem, and leaf of Dendrobium nobile Lindl. cultivated in Wanglong Town of Chishui City (P1), greenhouse of School of Life Sciences of Guizhou Normal University of Guiyang City (P2), and Taijin Ecological Agricultural Sightseeing Garden of Jinsha County (P3) in Guizhou Province were isolated by using nine kinds of media, and diversity analysis of isolated endophytic bacteria was conducted by using amplified ribosomal DNA restriction analysis (ARDRA). The results show that 1 081 endophytic bacteria are isolated from root, stem, and leaf of D. nobile in three plots, which are belonging to 40 species and 1 subspecies in 12 genera of 5 families in 2 phyla, and can be divided into 41 OTUs; in which, Bacillus and Brevibacillus are dominant genera, and Bacillus subtilis subsp. subtilis and Brevibacillus invocatus are dominant species (subspecies). There is difference in distribution of endophytic bacteria in different organs of D. nobile. Strain number of endophytic bacteria in root is the largest (695), and Shannon-Wiener diversity index (2.20) and Margalef richness index (5.81) of endophytic bacteria in root are both higher than those of endophytic bacteria in stem and leaf, while Pielou evenness index of endophytic bacteria in leaf is the highest (0.69). In addition, Srenson similarity index of endophytic bacteria between stem and leaf is the highest (0.82), and is significantly higher than that between root and stem and between root and leaf. There are 14 common OTUs in endophytic bacteria of D. nobile in three plots, its isolated strain number accounts for 96.5% of total isolated strain number of endophytic bacterium resources from D. nobile. In addition, there are relatively small differences in Shannon-Wiener diversity index and Pielou evenness index of endophytic bacteria of D. nobile in three plots, and only Margalef richness index of endophytic bacteria of D. nobile in P1 plot is obviously higher than that in P2 and P3 plots. Colony number and OTU number of endophytic bacteria isolated from D. nobile in NA, TSA, LB, KB, R2A, and TB media are higher than those in YPD, YT, and YG media. It is suggested that there are abundant endophytic bacterium resources in D. nobile, and the composition and diversity of its endophytic bacteria are closely related with isolated organs, cultivated fields, and media.

 

关键词金钗石斛; 内生细菌; 扩增核糖体DNA限制性分析(ARDRA); 组成分析; 多样性分析
Key wordsDendrobium nobile Lindl.; endophytic bacteria; amplified ribosomal DNA restriction analysis (ARDRA); composition analysis; diversity analysis
作者吴庆珊, 雷珣, 雷友梅, 蒲星, 刘颖, 翁庆北
所在单位贵州师范大学生命科学学院
点击量120
基金项目贵州师范大学研究生创新基金(研创[2016]22); 贵州省优秀科技教育人才省长专项资金([2011]47); 国家大学生创新创业训练计划项目(20131001); 贵州省大学生创新创业训练计划项目(201410663004)