柑橘黃龍�。℉LB）是影響全球柑橘產業(yè)發(fā)展的最嚴重病害之一。目前該細菌尚無法獲得離體純培養(yǎng),嚴重阻礙了其生物學特性、遺傳變異和致病分子機制的研究。HLB菌全基因組序列的公布,為其致病機制研究提供了理論參考。在本研究中,為了探討CLas致病的可能分子機制,我們對假定致病相關效應子基因在不同癥狀樣本中的表達水平,以及柑橘中部分防衛(wèi)相關基因進行了表達特性分析。另外對巴基斯坦樣品中的黃龍病菌進行基于Cthr基因的遺傳多樣性分析。同時對利用菟絲子從柑橘向長春花和煙草傳播CLas進行了研究,獲得的主要結果如下:1.致病相關效應子基因表達分析:我們從中國主要的柑橘產區(qū),包括江西、云南、海南等地,收集了表現(xiàn)不同癥狀的樣品。運用q RT-PCR分析了9個效應子基因在均勻黃化（Y）、斑駁黃化（BM）、脈凸（VK）和缺鋅黃化（ZD）癥狀中的表達特性。結果如下:CLas₀3230、CLas₀4025和CLas₀4560在VK癥狀中表達水平較高,而CLas₀2075、CLas₀2145、CLas

【文章來源】：華中農業(yè)大學湖北省 211工程院校 教育部直屬院校

【文章頁數(shù)】：87 頁

【學位級別】：碩士

【文章目錄】：
Abstract
摘要
Abbreviation
Chapter1 Literature Review
    1.1 Introduction
    1.2 Symptoms of Huanglongbing and its Impact on Orange Trees
    1.3 Classification and distribution of pathogens
    1.4 Host plant-pathogen interactions
    1.5 Genomics of HLB bacteria
    1.6 Techniques for the detection of HLB bacteria
        1.6.1 Biological assay
        1.6.2 Microscopic techniques
        1.6.3 Molecular techniques
        1.6.4 Spectroscopic and imaging technique
        1.6.5 Profiling of plant volatile organic compounds for disease identification
        1.6.6 Isothermal amplification combined with a lateral flow dipstick for rapidand sensitive detection of Candidatus Liberibacter
    1.7 Transient expression of Candidatus Liberibacter asiaticus effectors
    1.8 Collagen-triple helix gene
        1.8.1 Triple-helix structure and stability
        1.8.2 Formation of higher order structures
        1.8.3 Bacterial collagens that are known to form a triple helix structure
    1.9 Aim and scope of this study
Chapter2 Gene expression analysis of putative pathogenesis-related genes of CandidatusLiberibacter asiaticus
    2.1 Introduction
    2.2 Material and methods
        2.2.1 Plant materials
        2.2.2 Primers used in this study
        2.2.3 Plant total genomic DNA extraction
        2.2.4 Plant total RNA extraction
        2.2.5 Preparation of c DNA
        2.2.6 RT-PCR reaction
        2.2.7 q RT-PCR reaction
    2.3 Result and Analysis
        2.3.1 Gene expression analysis by RT-PCR
        2.3.2 Gene expression analysis of effectors of CLas by q RT-PCR
    2.4 Discussion
    2.5 Conclusion
Chapter3 Genetic diversity analysis of Pakistan samples based on Cthr gene
    3.1 Introduction
    3.2 Material and methods
        3.2.1 Plant samples
        3.2.2 Plant total genomic DNA extraction
        3.2.3 Primers and PCR assays
        3.2.4 Recovery and purification of PCR products
        3.2.5 Ligation of target DNA fragments
        3.2.6 Recombinant heat shock transformation
        3.2.7 Phylogenetic analysis
    3.3 Results and analysis
        3.3.1 Identification of Cthr gene in CLas Psy62 genome
        3.3.2 PCR identification of Cthr gene
        3.3.3 Cthr gene sequence analysis
        3.3.4 Comparison of Cthr gene between Pakistan and representative isolates
        3.3.5 Phylogenetic analysis of CLas from Pakistan isolates based on Cthr gene
    3.4 Discussion
    3.5 Conclusion
Chapter4 CLas transmission from citrus to periwinkle and tobacco via dodder
    4.1 Introduction
    4.2 Materials and methods
        4.2.1 Preparation of plant materials
        4.2.2 Sample collection
        4.2.3 Plant total genomic DNA extraction
        4.2.4 PCR amplification
    4.3 Result
        4.3.1 CLas transmission via dodder from citrus to periwinkle
        4.3.2 CLas transmission via dodder from citrus to tobacco
        4.3.3 Identification of CLas in periwinkle and tobacco after transmission via dodder
    4.4 Discussion
    4.5 Conclusion
References
Acknowledgement


【參考文獻】：
期刊論文
[1]應用PCR及Nested-PCR技術檢測柑橘黃龍病病原研究[J]. 丁芳,易干軍,王國平.  園藝學報. 2004(06)



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