天麻软腐病相关细菌的分离鉴定

Isolation and Identification of Bacteria Related to Gastrodia elata Soft Rot Disease

分析患软腐病的天麻块茎内的细菌多样性及其组成,为天麻细菌性软腐病的防控研究奠定基础。以健康块茎为对照,利用传统平板法分离培养天麻块茎患软腐病组织内的细菌;通过16S rDNA片段扩增、测序进行鉴定,分析细菌多样性及分布情况。结果表明,从天麻块茎样品中分离获得182株可培养细菌,隶属γ-变型菌纲8个属11个种,包括浅黄色假单胞菌(Pseudomonas lurida)、帕勒隆尼氏假单胞菌(Ps. palleroniana)、拉恩氏菌(Rahnella sp.)、戴氏西地西菌(Cedecea davisae)、解鸟氨酸拉乌尔菌(Raoultella omithinolytica)、变栖克雷伯菌(Klebsiella variicola)和Klebsiella sp.、路德维希肠杆菌(Enterobacter ludwigii)和Enterobacter sp.、泛菌(Pantoea sp.)和塔斯曼尼亚欧文氏菌(Erwinia tasmaniensis)。其中假单胞菌属为优势属,占细菌总数的60%,浅黄色假单胞菌为该属中的主要种类,占该属细菌总数的71%。分析显示,与健康天麻块茎相比,患软腐病的块茎内细菌种类和数量大量增加,破损可能会导致外源性病原细菌的侵入和大量繁殖。假单胞菌属是所有组织中的主要细菌类群,可能在天麻发育和内源性软腐病中发挥着重要作用;欧文氏杆菌属、西地西菌属、克雷白氏杆菌属、泛菌属和拉乌尔菌属可能与天麻软腐病密切相关。

The bacterial diversity and composition in tubers of Gastrodia elata suffering from soft rot disease were analyzed to lay a foundation for further study of prevention and control of G. elata soft rot disease. Taking healthy tubers as control, the traditional plate method was used to obtain the bacteria isolates from soft rot tissue of G. elata tubers. 16S rDNA fragment amplification and sequencing were used for identification, and the bacterial diversity and distribution were analyzed. The results indicated that a total of 182 bacterial isolates belonging to 11 species of 8 genera in γ-Proteobacteria were obtained from tubers of G. elata, including Pseudomonas lurida, Ps. palleroniana, Rahnella sp., Cedecea davisae, Raoultella omithinolytica, Klebsiella variicola, Klebsiella sp., Enterobacter ludwigii, Enterobacter sp., Pantoea sp. And Erwinia tasmaniensis. Pseudomonas was the dominant genus, accounted for 60% of the total bacteria, and P. lurida was the main species in the genus, accounting for 71% of the total number of bacteria in this genus. The analysis showed that compared with the healthy tubers of G. elata, the diversity and quantity of bacteria in the tubers with soft rot disease were greatly increased, and tuber damage may cause the entry and proliferation of exogenous pathogenic bacteria. Pseudomonas is the main bacterial group in all tissues and may play an important role in the development and endogenous soft rot disease of G. elata. The genera Erwinia, Cedecea, Klebsiella, Pantoea and Raoultella may be closely related to G. elata soft rot disease.