李梦娇^1，， 高泽文^1，， 何甜^1，， 赵明富^1，2，*

（  1、云南农业大学植物保护学院，云南 昆明 650021；       2、云南农业大学特色资源植物改良与利用研究所，云南 昆明 650021；  ）

摘要： 以新鲜健康的铁皮石斛为实验材料，采用组织分离法分别从根和茎中分离内生细菌，并结合形态学和分子生物学方法（PCR扩增菌株的16S rDNA）对分离到的细菌进行鉴定。结果表明，从铁皮石斛组织中共分离到47株内生细菌，通过测序及比对得知内生细菌中伯克氏菌属(Burkholderia)10株、肠杆菌科(Enterobacter)3株、假单胞菌(Pseudomonas)4 株、芽孢杆菌属(Bacillus)23株、贪铜菌属(Cupriavidus)7株。茎和根分别分离得31株和16株内生细菌，数量和种类均是茎>根。并利用内生细菌对铁皮石斛疫病病原菌棕榈疫霉(Phytophthora palmivora)进行室内抑菌试验，结果表明四株芽孢杆菌对棕榈疫霉有明显的抑菌作用，其中巨大芽孢杆菌(Bacillus megaterium)的抑制率最高，达到78.13%。
关键词： 铁皮石斛；内生细菌；芽孢杆菌；生物防治

LI Mengjiao^1,， GAO Zewen^1,， HE Tian^1,， ZHAO Mingfu^1,2,*

（  1、Plant Protection College, Yunnan Agricultural University, Kunming 650021, China；       2、Institute of improvement and application of featured resources plants, Yunnan Agricultural University, Kunming 650021, China；  ）

Abstract： Endophytic bacteria were isolated from the fresh root and stem of Dendrobium officinale by tissue culture. The bacteria were identified based on morphological observation and their 16S rDNA sequence by PCR amplification.The results showed that a total of 47 endophytic bacteria were isolated, which 31 strains from stem,16 from root.Among them, though sequencing and alignment 10 strains belonged to Burkholderia, 3 strains to Enterobacter, 4 strains to Pseudomonas, 23 strains to Bacillus, 23 strains to Cupriavidus. The strains were obtained with the number and type order as: stem＞root. The inhibition effects of an antagonistic endophytic bacteria on Phytophthora palmivora (the pathogen of Dendrobium Officinale Blight) were investigated under laboratory conditions. The result demonstrate that four Bacillus effectively inhibits the pathogen, and the highest rate of inhabitation on P.palmivora is Bacillus megaterium which reached 78.13%. 
Keywords： Dendrobium officinale; Endophytic Bacteria; Bacillus; Biocontrol

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