摘要
为明确由豇豆叶片分离出的20株生防内生菌的生理生化性质及其潜在生防价值,本研究通过形态学、平板对峙、PCR技术、生物信息学等方法对20株生防细菌进行鉴定研究,并以4株病原真菌为靶标病菌,测定20株生防菌对病原真菌的抑制效果。结果显示,20株内生细菌中有3株属于假单胞菌属(Pseudomonas spp.);12株为枯草芽孢杆菌(Bacillus subtilis);剩余5株为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。经过革兰氏染色得知2号、16号、19号三株假单胞菌株为革兰氏阴性菌株,其余为阳性菌;通过解淀粉测定,2、16、19三株菌株没有形成不变色透明圈,无解淀粉功能,其余中间形成明显的透明圈,有解淀粉功能;经过V-P测定,发现1号、2号、11号、15号、16号、18号共6株菌株呈阴性,其余呈阳性;在MR甲基红测定中,1号、6号、7号、10号、11号共5株菌株测定呈阳性,其余呈阴性;由氧化酶测定可知2号、16号、17号、19号4株菌株氧化酶测定为阳性,其余为阴性;由接触酶测定得知20株细菌均为阳性;在抑菌试验中20株生防细菌对4株供试的植物病原真菌均产生了拮抗作用,其抑菌率为29.58%~80.28%,其中抑菌率最高的是18号菌株枯草芽孢杆菌对真菌橡胶树炭疽病菌(Colletotrichum gloeosporioides),抑菌率为80.28%,抑菌率最低的是2号菌株假单胞菌对芒果炭疽病菌(Colletotrichum gloeosporioiles),抑菌率为29.58%。不同菌株之间抑制效果差异较大,大部分菌株对供试的病原真菌具有良好的抑制效果。本研究结果为后续生防机理的研究、植物病害的防治和生物药剂的开发提供理论依据。
In order to clarify the potential bio-control value of 20 strains of endophytic bacteria isolated from cowpea leaves and the evolutionary relationship between them,twenty strains of biocontrol bacteria were studied by morphological method,plate confrontation method,PCR technique and bioinformatics method.The pathogenic fungi were tested for their inhibitory effects against various pathogenic fungi.The results showed that 20 strains of endophytic bacteria were identified as Bacillus subtilis,Pseudomonas ssp.and Bacillus amyloliquefaciens,12 strains of 20 endophytic bacteria were identified as Bacillus subtilis,3 strains were Pseudomonas ssp.,5 strains were Bacillus amyloliquefaciens.All of the biocontrol bacteria were found to inhibit the growth of 4 tested plantpathogens antagonism produced by fungi.Among all of them,the highest inhibition rate was No.18 Bacillus subtilis against No.8 Colletotrichum gloeosporioides,the antibacterial rate was 80.28%,and the worst effect was Bacteria No.2 pseudomonas against the fungus No.4 Colletotrichum gloeosporioiles,had an antibacterial rate of 29.58%.Bio-controI effects vary greatly among different strains,and some strains have significant biocontrol effects.The results of this study provide a theoretical basis for the study of the subsequent biocontrol mechanism, prevention of plant diseases,and the development ofbiopharmaceuticals.
作者
张骕
缪卫国
韦明德
孙亮
刘文波
Zhang Su;Miao Weiguo;Wei Mingde;Sun Liang;Liu Wenbo(Hainan Key Laboratory for Sustainable Utilization of Tropical B ioresource,Institute of Tropical Agriculture and Forestry,Hainan University,Haikou,570228;Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Danzhou,571737)
出处
《分子植物育种》
CAS
CSCD
北大核心
2018年第23期7760-7774,共15页
Molecular Plant Breeding
基金
海南大学青年教师基金项目(hdkyxj201708)
海南省自然科学基金项目(20153131
2063140)
中国热带农科院基本科研业务费专项(1630022015016)
海南茄果类蔬菜化肥农药减施增效技术研究项目(1630052016007)共同资助
关键词
生防细菌
鉴定
病原真菌
拮抗作用
Biocontrol bacteria
Identify
Pathogenic fungus
Antagonistic action
