目录
摘要Ⅱ
关键词Ⅱ
AbstractⅡ
引言
引言
1材料与方法1
1.1研究区域概况 1
1.2采样及实验设计 1
1.3乔木可培养附生微生物计数 2
1.4乔木附生微生物总DNA提取与质量检验 2
1.5细菌16S rRNA基因PCR扩增 2
1.6高通量测序及数据分析 3
1.6.1 PCR产物文库构建及Illumina PE250测序 3
1.6.2数据分析3
2结果与分析3
2.1乔木可培养附生微生物计数（细菌、真菌与放线菌）3
2.2 DNA提取质量分析 4
2.2.1琼脂糖凝胶电泳测定 4
2.2.2 NanoDrop2000紫外分光光度计测定DNA含量及纯度4
2.3五种乔木树干表皮细菌群落多样性分析 5
2.3.1树干表皮细菌α多样性分析 5
2.3.2树干表皮细菌β多样性分析 7
2.3.3微生物群落统计学差异分类水平分析 7
2.4 PICRUSt功能预测9
3讨论 10
致谢12
参考文献13
附录15
五种乔木树干表皮附生细菌多样性研究
摘 要
微生物可以寄生、共生和附生于植物的各个组织器官中。树干表皮作为乔木的一种保护组织，其独特的栖息环境利于微生物特别是细菌生长、繁殖而形成群落，然而人们但对其群落结构和生态系统功能一直认识不足。已有研究表明，植物种类是影响植物附生微生物的关键因素，因此研究不同乔木树干表皮的附生微生物对了解乔木的特点和植物与微生物的相互关系有一定意义。本实验以南京中山陵风景区的5种代表性乔木为研究对象，即重阳木（BP）、香樟（CC）、枫香树（LF）、水杉（MG）和枫杨（PS）树干表皮微生物。采用高通量测序技术，5种乔木树干表皮共鉴定到3237个细菌OTUs，且共有OTUs占10.86%；BP树皮细菌多样性指数最高，PS物种丰度最高，MG两者均最低。在门分类水平上，变形菌门（P *51今日免费论文网|www.51jrft.com +Q: *351916072* 
roteobacteria）和放线菌门（Actinobacteria）在5组乔木树干表皮的附生细菌中均占绝对优势。通过PICRUSt功能预测发现，乔木树皮附生细菌的能量代谢、氮代谢和生物固碳通路的相对丰度较高。这些研究结果揭示了乔木表皮附生着大量的微生物类群，具有重要的生态功能，加深了对乔木附生微生物多样性的理解。
EPIPHYTIC BACTERIA COMMUNITIES AND DIVERSITY ON FIVE DIFFERENT SPECIES OF ARBOR BARK
ABSTRACT
Microorganisms can be parasitic, symbiotic and epiphytic in various organs of plants. As a kind of protective tissue of arbor, bark is a unique habitat beneficial to the growth, propagation and community formation of microbes especially for bacteria. However, the community structure of bacterium epiphytic to arbor bark and its ecosystem function are lack of understanding. It has been shown that plant species are the key factors affecting epiphyte microorganisms. Therefore, it is of importance to study epiphyte microorganisms in bark of different trees for understanding the characteristics of trees and the relationship between plants and microorganisms. In this study, five representative species of arbor were selected, which were the epiphytic microorganism of Bischofia polycarpa (BP), Cinnamomum camphora (CC), Liquidambar formosana Hance (LF), Metasequoia glyptostroboides (MG) and Pterocarya stenoptera (PS). With highthroughput sequencing, there are 3237 bacteria OTUs were identified in five species of tree bark, which accounted for 10.86% in the total OTUs. The bacterial diversity index of BP and species richness of PS bark were the highest, while the figures for MG were the lowest. At the taxonomic level, Proteobacteria and Actinobacteria were dominant in the epiphytic bacteria of five groups tree bark. According to the PICRUSt prediction, the relative abundance of energy metabolism, nitrogen metabolism and carbon fixation pathways of epiphytic bacteria in tree bark were high. These results reveal that a variety of microorganisms which have crucial ecological functions are epiphytic on the arbor bark, deepening the understanding of the diversity of epiphytic microorganisms on arbor.

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