摘要
【目的】对柑橘健康植株和感染黄龙病植株的韧皮部内生细菌的多样性进行比较分析,为研究柑橘植株韧皮部内生细菌与黄龙病菌之间的相互关系奠定基础。【方法】采用常规分离培养与分子鉴定的方法和基于16S rDNA基因的限制性酶切长度多态性(restriction fragment length polymorphism,RFLP)分析的方法。【结果】用常规分离与分子鉴定方法获得10个属细菌类群,其中芽孢杆菌属(Bacillus sp.)、假单胞菌属(Pseudomonas sp.)、考克氏菌属(Kocuria sp.)为无症健株的优势菌群;微杆菌属(Microbacterium sp.)、芽孢杆菌属、假单胞菌属和考克氏菌属.为黄龙病罹病植株的优势菌群。PCR-RFLP方法得到29种酶切图谱,共鉴定出10属可培养细菌以及11种不可培养细菌。健株与病株中Dyella sp.、Pseudomonas sp.、Serratia sp.和未培养细菌所占比例均大于5%。柑橘健株与病株中细菌的种群密度和菌群种类存在明显差异。【结论】研究结果表明柑橘健株和病株韧皮部组织中的内生细菌优势菌群存在明显差异,而伴生细菌的优势菌群与黄龙病菌的相互关系正在深入分析研究。
【Objective】 The endophytic bacterial diversity in the Huanglongbing pathogen-infected and healthy citrus phloem tissue in Guangxi were investigated in order to decipher the interactions of Huanglongbing pathogen with companion microbe.【Method】Both classical morphological,physiological,chemotaxonomic characteristics methodology and 16S rRNA,and PCR-RFLP analysis methods were applied.【Result】 By the traditional isolation and identification and 16S rDNA amplificons methods,10 genera of bacteria were identified from 21 culturable bacterial populations.The sequences aligned with NCBI GenBank database and showed that they belonged to 10 different genera of bacterium.The dominant bacterial population in healthy citrus belonged to Bacillus sp.,Pseudomonas sp.,and Kocuria sp..While in phloem tissue of infected citrus plant over ten years were Bacillus sp.,Pseudomonas sp.,Kocuria halotolerans,and Microbacterium sp..By the PCR-RFLP methods,two 16S rDNA clone libraries of endophytic bacteria were constructed and 29 restriction endonuclease types were detected,10 genera of bacteria and 11 genera of uncultured bacteria were identified.The dominant bacterium population in libraries belonged to Dyella sp.,Pseudomonas sp.,Serratia sp.and uncultured bacterium.The density and species of endophytic bacteria were observed in remarkable difference compared disease tree to healthy one.【Conclusion】 By traditional cultivation and 16S rDNA sequencing,PCR-RFLP analysis,the diversity of population and species of endophytic bacteria mainly in citrus phloem tissue has difference among infected and healthy citrus plant,while those dominant bacterial populations if related to Huanglongbing pathogen is currently still under investigation.
出处
《中国农业科学》
CAS
CSCD
北大核心
2010年第23期4823-4833,共11页
Scientia Agricultura Sinica
基金
国家自然科学基金项目(30971875)
