干旱和盐胁迫下棉花根际促生细菌的分离与鉴定

发布时间：2021-10-31 10:56

　　干旱和盐胁迫是影响作物生产布局和产量的重要因素。微生物组被认为是植物体的第二套基因组,对植物的生长和适逆至关重要。本课题主要通过对正常和盐/干旱胁迫下生长的棉花根际微生物群进行分离和测序,研究棉花根际微生物的组成,分离对棉花耐盐和抗旱有利的微生物种类及研究其功能。主要结果如下:研究了盐胁迫条件下棉花植株与PGP根际细菌之间的相互作用。研究首先进行棉花盐胁迫处理下根际微生物的分离和培养,成功获得了枯草芽孢杆菌（Bacillus subtilis,BB）和短小芽孢杆菌（Bacillus pumilus,BC）等细菌菌株。然后将这些菌株施加到盐胁迫处理（200m M Na Cl）的棉花根际,发现BB和BC处理可以增强棉花对盐胁迫的抗性,表现在添加BB或者BC的棉花植株根、茎和叶对盐处理敏感性降低。研究进一步对BB或BC处理的盐胁迫下棉花根系进行转录组测序。结果表明,在有或没有枯草芽孢杆菌（S和BB）的条件下,共有5535个差异表达的基因,上调基因3349个,下调基因2186个;而在有或没有短小芽孢杆菌（S和BC）的条件下,DEG是8452个,其中包括4035个上调基因和4417下调基因。GO... 

【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校

【文章页数】：102 页

【学位级别】：硕士

【文章目录】：
摘要
Abstract
List of abbreviations
Chapter 1
    Introduction
    1.1 Cotton
    1.2 Plants Stress
        1.2.1 Salt Stress
        1.2.2.Drought stress
        1.2.3 Drought stress in cotton
        1.2.4 Effect of salinity on plant growth
        1.2.5 Salt stress in cotton
    1.3 PGP Bacteria and their role in abiotic stress
        1.3.1 Role of PGP in controlling salt stress
    1.4 Aims of the thesis
Chapter 2 Salt stress Tolerance conferred by plant growth promoting rhizosphere bacteria in Cotton
    Introduction
    2.1 Materials and Methods
        2.1.1 Identification and selection of salt stress resistant PGP bacteria in Cotton Rhizosphere
            2.1.1.1 Soil preparation
            2.1.1.2 Cotton seed germination,Salt stress and Harvesting
            2.1.1.3 Isolation of bacteria from Rhizosphere,roots and soil
            2.1.1.4 Screening of Bacteria for Salt stress
            2.1.1.5 16SrRNA sequencing and identification of Bacterial strains
        2.1.2 Cotton plants treatment with salt stress and bacteria
            2.1.2.1 Cotton seeds germination and treatments
            2.1.2.2 Phenotype appearance and selection
            2.1.2.3 Sampling and RNA-extraction
            2.1.2.4 Library construction and sequencing
            2.1.2.5 Illumina sequences analysis
            2.1.2.6 Statistical analysis of differentially expressed genes(DEGs)
            2.1.2.7 Gene ontology and pathway enrichment analysis
            2.1.2.8 Quantitative PCR(q PCR)
    2.2 Results
        2.2.1 Bacillus subtilis(BB)and Bacillus pumilus(BC)promoted Cotton growth under salt stress conditions
        2.2.2 Difference shown in leaves numbers and development
        2.2.3 Height variations of Plants
        2.2.4 Root length variations
        2.2.5 Transcriptomic Analysis
            2.2.5.1 Expression density distribution
            2.2.5.2 Correlation between samples
            2.2.5.3 Effects on the expression patterns of DEGs
            2.2.5.4 Gene Ontology enrichment of DEGs
            2.2.5.5 KEGG pathways Analysis
        2.2.6 Selected Pathways for salt stress resistance and Plant growth promotion
            2.2.6.1 Pentose and Glucuronate Interconversions Pathway
            2.2.6.2 Starch and Sucrose Metabolism Pathway
        2.2.7 Validation of DEGs by q RT-PCR
    2.3 Discussion
    2.4 Summary and future prospects
Chapter 3 Microbiome Diversity in Cotton Rhizosphere Under Normal and Drought Conditions
    Introduction
    3.1 Materials and methods
        3.1.1 Plant material and drought treatment
        3.1.2 Sample collection,DNA extraction,and16S r RNA gene amplification
        3.1.3 Illumina library preparation and High-throughput sequencing
        3.1.4 Primary Data processing and Operational Taxonomic Unit demarcation
        3.1.5 Alpha diversity and Taxonomic analysis
        3.1.6 Beta diversity analysis
        3.1.7 Metagenome analysis and network construction
        3.1.8 Statistical analysis
    3.2 Results
        3.2.1 General analysis of sequencing data
        3.2.2 Alpha diversity analysis and Taxonomic analysis
        3.2.3 Beta diversity
        3.2.4 Metagenome prediction
    3.3 Discussion
    3.4 Conclusions
References
Appendices
    Appendix1
    Appendix2
    Appendix3
    Appendix4
    Appendix5
    Appendix6
    Appendix7
List of publications
Acknowledgement


【参考文献】：
期刊论文
[1]Polyamine Accumulation in Transgenic Tomato Enhances the Tolerance to High Temperature Stress[J]. Lin Cheng,Yijing Zou,Shuli Ding,Jiajing Zhang,Xiaolin Yu,Jiashu Cao and Gang Lu(Key Laboratory of Horticultural Plant Growth,Development and Biotechnology,Ministry of Agriculture,Department of Horticulture,Zhejiang University,Hangzhou 310029,China).  Journal of Integrative Plant Biology. 2009(05)



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