Knowledge Resource Center for Ecological Environment in Arid Area
| 不同草原类型针茅根部内生菌群落结构研究 | |
| 其他题名 | Endophyte communities of Stipa sp. root cross differrent types of steppes in Northern China |
| 张少鹏 | |
| 出版年 | 2014 |
| 学位类型 | 硕士 |
| 导师 | 张惠文 |
| 学位授予单位 | 中国科学院大学 |
| 中文摘要 | 内生菌是植物微生态的重要组成部分。针茅作为北方草原的优势种或建群种，分布非常广泛。开展针茅根部内生菌研究，不但有助于揭示内生菌与宿主之间的共生关系，亦可丰富内生菌资源。 从我国新疆、甘肃、内蒙古三省选择了六种不同草原类型（亚高山草甸，高山草甸，戈壁，荒漠草原，典型草原和草甸草原）并采集针茅根部材料，采用培养法和非培养法对针茅根部内生菌的群落结构进行了研究，并获得了以下主要结果。 从针茅根部共分离出可培养内生真菌213株，根据序列的相似性（以97%为阈值），共获得51个真菌分类操作单元（OTUs），覆盖了4门7纲23科27属，子囊菌门真菌为绝对优势菌群，占分离真菌总数的93.42%，镰孢菌属（Fusarium，41.31%）为优势属真菌，在各草原类型中均有分布；六种草原类型针茅根部内生真菌的优势属差别较大，仅Fusarium为各草原类型共有优势菌属,亚高山草甸的Microdochium，高山草甸的Saccharicola和Aureobasidium，戈壁草原的Curvularia和Rhizopus，草甸草原的Trichoderma和Cochliobolus，则为各草原类型针茅根部内生真菌的优势属；研究结果表明，各草原类型中针茅根部可培养内生真菌群落之间的共有属较少，群落结构相似性较低。 从针茅根部共分离到可培养内生细菌200株，按照97%的序列相似性进行聚类，共获得37个细菌OTUs，覆盖了4门7纲19科26属，其中芽孢杆菌属（Bacillus，48.50%）和假单胞菌属（Pseudomonas，17.50%）为内生细菌优势属，在各草原类型中均有分布，六种草原类型针茅根部内生细菌优势属不尽相同，仅Bacillus为各草原类型共有优势属，Pseudomonas为高山草甸、戈壁、草甸草原的共有优势属，亚高山草甸的Promicromonospora，戈壁草原的Paenibacillus和Achromobacter，则为各自的内生细菌优势属；研究结果表明，除高山草甸外，其他五种草原类型间针茅根部可培养内生细菌群落结构相似程度较高。 应用Illumina Miseq高通量测序研究了不同草原类型中针茅根部的内生细菌群落，在97%的相似度水平共获得1689个OTUs，覆盖了36门257属。高山草甸中比例最高的是变形菌门细菌（87.75%），而其他五种草原类型中比例最高的均为放线菌门细菌。不同草原类型中针茅根部内生细菌群落结构存在明显差异，戈壁与荒漠草原的内生细菌群落结构最为相近，高山草甸与其他草原类型间相似性最低。RDA分析表明降雨量、年平均气温、土壤pH是引起不同草原类型中针茅根部内生细菌群落结构变化的主要环境因子。 应用PCR-DGGE和Illumina Miseq高通量测序研究针茅根部内生菌的初步结果表明，选择ITS1区对针茅根部内生真菌群落进行研究可获得很高的多样性。 本研究分别采用传统培养方法和非培养方法对不同草原类型中针茅根部内生菌进行了研究，获得了大量的内生细菌、内生真菌菌株资源，通过高通量测序技术揭示了针茅根部内生细菌的群落结构变化；采用非培养方法（PCR-DGGE，高通量测序）研究针茅根部内生真菌群落结构的初步探索为后期对全部样品进行高通量测序提供了方法学基础。 |
| 英文摘要 | Endophytes are microorganisms that live in plant tissues without causing symptoms of disease and important components of plant microbiomes. As the dominant specie or constructive specie, Stipa sp. is widely distributed in the steppes of northern China. Besides to enrich the endophyte resource, researching on endophyte communities of Stipa sp. could help to reveal the symbiotic relationships between endophytes and their host plant. This study was conducted to explore endophytes communities of Stipa sp. in different types of steppes in the north of China. Roots of Stipa sp. were collected in six different types of steppes, which included subalpine meadow, alpine meadow, Gobi desert, desert steppe, typical steppe, and meadow steppe, in the province of Xinjiang, Gansu and Inner Mongolia. Endophyte communities were investigated by culture-dependent and culture-independent approaches, respectively. The main results are as follows: In total, 213 endophytic fungal isolates were obtained. Fungal isolates were identified based on comparative analysis of ITS rDNA gene sequence and were divided into 51 operational taxonomic units (OTUs) at the 97% similarity threshold and classified to 4 phyla, 7 classes, 23 families and 27 genera. Most of the isolated fungi were members of Ascomycota (93.42% of the total isolates). And the most abundant genus was Fusarium (41.31% of the total isolates), which also was dominant genus in all six steppe types. Besides Fusarium, other dominant genera in six steppe types were different, such as Microdochium in subalpine meadow, Saccharicola and Aureobasidium in alpine meadow, Curvularia and Rhizopus in Gobi desert and Trichoderma in typical meadow. Results indicated that there are few common genera in the six steppe types and the structure of culturable endophytic fungal communities varied in different steppe types. 200 endophytic bacterial isolates were obtained in the six different types of steppes. These isolates were identified based on comparative analysis of 16S rDNA sequence and were divided into 37 operational taxonomic units (OTUs) at the 97% similarity threshold and classified to 4 phyla, 7 classes, 19 families and 26 genera.The predominant genera were Bacillus (48.50% of the total isolates) and Pseudomonas (17.50% of the total isolates), which were distributed in all six steppe types. As to different steppe types, Bacillus was predominant bacteria in the roots of Stipa sp. in all the six steppe types and Pseudomonas was predominant genera in alpine meadow, Gobi desert and meadow steppes. Besides, other dominant genera in the six steppe types were different, such as Promicromonospora in subalpine meadow, Paenibacillus and Achromobacter in Gobi desert. The results indicated that，with the exception of alpine meadow， the structure of endophytic bacterial communities in the roots of Stipa sp. in different steppe types were similar to each other. Endophytic bacterial community in the roots of Stipa sp. of six different steppe types were investigated untilized Illumina Miseq high-throughput sequencing. All sequence were divided into 1689 OTUs at the 97% similarity threshold and classified to 36phyla and 257genera besides unclassified sequence. Endophytic bacetria in the roots of Stipa sp. in alpine meadow were dominated by Proteobacteria(87.75%), while most endophytes were from the Actinobacteria in other five steppe types. Our research indicated thatthere were significant difference among the endophytic bacterial communities of different steppe types, the endophtic bacterial community in the roots of Stipa sp. of alpine had low similarities with that of other five steppe types while there were relatively high similarity between that of Gobi desert and desert steppe. Redundancy analysis (RDA) revealed that annual precipitation, mean annual temperature and soil pH were the main environmental factors affecting endophytic bacterial community distribution in the roots of Stipa sp. in six steppe types. |
| 中文关键词 | 针茅 ; 内生真菌 ; 内生细菌 ; 高通量测序 |
| 英文关键词 | Endophytic fungi Endophytic bacteria Stipa sp. High-throughput sequencing |
| 语种 | 中文 |
| 国家 | 中国 |
| 来源学科分类 | 微生物学 |
| 来源机构 | 中国科学院沈阳应用生态研究所 |
| 资源类型 | 学位论文 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/287414 |
| 推荐引用方式 GB/T 7714 | 张少鹏. 不同草原类型针茅根部内生菌群落结构研究[D]. 中国科学院大学,2014. |
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