香草兰生防细菌筛选分子鉴定及其抑菌机制初步研究.docVIP

香草兰生防细菌筛选分子鉴定及其抑菌机制初步研究 　　摘 要 通过平板对峙法从香草兰根际微生物中筛选出对香草兰根（茎）腐病菌（Fusarium oxysporum）、香草兰疫病菌（Phytophthora nicotianae）和香草兰细菌性软腐病菌（Erwinia carotovora）均有良好抑制效果的生防菌10株。通过16S rDNA序列比对及系统发育树分析，其中7株为解淀粉芽孢杆菌（Bacillus. amyloliquefaciens），2株为枯草芽孢杆菌（B. subtils），1株为甲基营养型芽孢杆菌（B. methylotrophicus）。提取脂肽类化合物进行抑菌分析，发现10株生防菌脂肽类提取物对香草兰根（茎）腐病菌和香草兰疫病菌均有良好的抑制效果，有4株的脂肽类粗提物对香草兰细菌性软腐病菌有强烈的抑制活性。对其中2个菌株脂肽类粗提物进行MALDI-TOF/TOF-MS检测，发现菌株 VX2R11 产生表面活性素（Surfactin）和伊枯草素A（IturinA）2种脂肽类化合物，菌株 VX2S02 仅产生伊枯草素A，推测产生伊枯草素A是菌株VX2R11和VX2S02拮抗香草兰根（茎）腐病菌和疫病菌重要机制；产生表面活性素是菌株VX2R11拮抗香草兰细菌性软腐病菌的重要机制。 　　关键词 香草兰 ；芽孢杆菌 ；16S rDNA 鉴定 ；脂肽类化合物 　　分类号 S435.73 Doi：10.12008/j.issn.1009-2196.2016.01.009 　　Abstract Ten bio-control Bacillus strains were selected via dual culture method from the isolates obtained from vanilla rhizosphere of Wanning city and Qionghai city in Hainan province. The selected isolates were highly antagonistic against vanilla pathogens：Fusarium oxysporum，Phytophthora nicotianae and Erwinia carotovora. Seven B. amyloliquefaciens strains， two B. subtils strains and one B. methylotrophicus strain were identified by the assays of 16S rDNA sequence and the phylogenefic tree. Lipopeptide compound analysis via dual culture method showed that all 10 lipopeptide extractions from each bio-control strain exhibited highly antagonistic against F. oxysporum and P. nicotianae， while 4 lipopeptide extractions were highly antagonistic against E. carotovora. Lipopeptide compound analysis via MALDI-TOF/TOF -MS showed that strain VX2R11 produced Surfactin and IturinA，while strain VX2S02 produced Iturin A， suggesting that strainVX2R11and VX2S02 might via the production of IturinA to antagonize pathogens F. oxysporum and P. nicotianae， while strainVX2R11 might via the production of Surfactin to antagonize pathogen E. carotovora. 　　Keywords Vanilla fragrans ； Bacillus spp ； 16S rDNA sequence ； lipopeptide 　　香草兰[Vanilla fragrans（salisb.）Ames（V.planifolia）]是一种经济附加值极高的热带经济作物，素有“食品香料之王”的称誉。香草兰根（茎）腐病是一种危害极大的土传病害，其病原物是尖孢镰刀菌（Fu