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【目的】分析感柑橘黄龙病长春花植株与健康长春花植株不同部位内生细菌菌群结构变化,为柑橘黄龙病菌与长春花内生细菌的相关性研究提供理论基础。【方法】本研究利用兼性厌氧可培养技术、16S rDNA限制性片段长度多态性分析(Restriction fragment length polymorphism,RFLP)以及16S rDNA序列分析相结合的方法。【结果】分别从感病和健康长春花叶、茎、根的组织中分离获得67株内生细菌,与GenBank中29种细菌的相似性达到97%-100%。其中短小杆菌属(Curtobacterium sp.)、欧文氏菌属(Erwinia sp.)、蜡样芽胞杆菌(Bacillus cereus)为感病长春花内生细菌的优势菌群,鞘胺醇单胞菌属(Brevundimonas sp.)、芽胞杆菌属(Bacillus sp.)为健康长春花内生细菌的优势菌群;马胃葡萄球菌(Staphylococcus equorum)为两者的共同优势菌群。通过RFLP方法分析,感病株得到16个、健株得到23个操作分类单元(Operational TaxonomicUnits,OTUs),感病植株中除柑橘黄龙病菌Candidatus Liberibacter asiaticus外,还有丰富的CandidatusLiberibacter sp.存在。【结论】感病与健康长春花植株中均含有丰富的内生细菌,黄龙病菌的存在改变了长春花原有内生细菌的菌群结构,且菌群多样性下降。可见长春花内生细菌在一定程度上受到柑橘黄龙病菌的抑制。
【Objective】 To analyze the changes of endophytic bacteria flora in different parts of the plant of Citrus Huanglongbing and healthy Catharanthus roseus and provide the theoretical basis for the study of the correlation between Citrus Huanglongye and Catharanthus roseus endophytic bacteria. 【Method】 A facultative anaerobic culturable technique, 16S rDNA restriction fragment length polymorphism (RFLP) and 16S rDNA sequence analysis were used in this study. 【Result】 Seventy - six endophytic bacteria were isolated from the tissues of susceptible and healthy Changchun flowers, stems and roots, respectively. The results showed 97% -100% similarity with 29 kinds of bacteria in GenBank. Among them, Curtobacterium sp., Erwinia sp. And Bacillus cereus are the dominant bacteria of the endophytic bacteria of the susceptible vinca spp., Brevundimonas sp.) and Bacillus sp. were dominant bacteria of endophytic bacteria of healthy vinca. Staphylococcus equorum was the common dominant bacteria. According to RFLP analysis, 16 susceptible plants were obtained, and 23 operating taxonomic units (OTUs) were obtained from healthy plants. In addition to Candidatus Liberibacter asiaticus, Candidatus Liberibacter sp. Also existed in susceptible plants. 【Conclusion】 The susceptible and healthy Catharanthus roseus plants are rich in endophytic bacteria, while the presence of Huanglongbing bacteria changes the flora structure of endophytic bacteria in Catharanthus roseus and the diversity of the flora decreases. Viburnum endophytic bacteria can be seen to some extent, citrus Huanglongbing inhibition.