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【目的】分析赣南脐橙黄龙病植株和健康植株叶片内生菌,对比不同培养条件下培养出的内生菌,为筛选出对柑橘黄龙病原菌有影响的伴生菌奠定理论基础。【方法】通过PCR方法对脐橙中黄龙病菌进行验证,并基于16S r RNA基因高通量测序技术对患病与健康赣南脐橙叶片内生菌以及不同培养基富集培养后的内生菌进行多样性分析。【结果】所采集样品中有5株患病株,5株健康株。5株病株中共同含有的细菌属有13个,其中7个在5株健株中也共同存在。Defluviicoccus属和Granulicella属在病健株植物中都是优势菌属,且在健株中的平均含量高于病株。病株与健株的样品相似度存在明显界线。富集培养后不同样本和不同培养基中菌属分布不同。肠杆菌属(Enterobacter)、短小杆菌属(Curtobacterium)、假单胞菌属(Pseudomonas)和泛菌属(Pantoea)得到了大量富集,不动杆菌属和沙雷氏菌属等9个菌属富集量较少。另外,培养和未培养各样本间未分类菌(Unclassified)含量差异也较大。【结论】赣南脐橙患病植株和健康植株叶片内生菌有着明显差异,黄龙病菌的存在改变了脐橙叶片原有内生细菌的菌群结构。从活体植物组织内直接检测才能得到真正的植物内生菌群落分布情况。通过分析菌群的差异,有望找到与柑橘黄龙病菌生长相关的伴生菌。
【Objective】 The purpose of this study was to analyze the endophytes in the leaves of Gannan navel orange yellow dragonfly plants and healthy plants and to compare the endophytic bacteria cultured in different culture conditions to lay a theoretical foundation for the screening of the accompanying bacteria that have an impact on the pathogenic bacteria of citrus Huanglong. 【Method】 The results showed that the pathogenic bacteria in the leaves of Navel orange (Caragana anguillicaudatus) were enriched and the endophytes were enriched in different media Diversity analysis. 【Result】 There were 5 diseased plants and 5 healthy plants in the collected samples. Thirteen bacteria were found in the five isolates, of which seven also co-existed in five healthy isolates. Defluviicoccus and Granulicella are dominant species in diseased plants and have a higher average level in healthy plants than in diseased plants. There is a clear boundary between sample similarity of diseased and healthy strains. After enrichment culture in different samples and different media genus distribution. Enterobacter, Curtobacterium, Pseudomonas and Pantoea were abundantly enriched, and nine species of genus Acinetobacter and Serratia were abundant Set a small amount. In addition, there was also a large difference in Unclassified contents between cultured and uncultured samples. 【Conclusion】 There are significant differences in endophytes from the leaves of Navel orange and healthy plants in Gannan. The presence of Huanglongbing changes the original flora of Navel orange leaves. Direct detection from living plant tissue can get the true plant endophyte community distribution. By analyzing the differences in the flora, it is expected to find the companion bacteria associated with the growth of Citrus Huanglongshan.