利用PCR技术快速检测根际产ACC脱氨酶细菌  

PCR-based Method for the Rapid Detection of ACC Deaminaseproducing Rhizosphere Bacteria

作　　者：秦媛[1] 潘雪玉 袁志林[1] 

机构地区：[1]中国林业科学研究院亚热带林业研究所,杭州311400

出　　处：《生物技术通报》2017年第11期112-122,共11页Biotechnology Bulletin

基　　金：国家自然科学基金面上项目(31370704);2014年度国家林业局引智项目;2016年度留学回国人员科技活动项目择优资助经费

摘　　要：通过产生ACC脱氨酶降低胁迫乙烯水平并缓解盐胁迫危害,是植物根际促生菌(PGPR)促进宿主生长和抗逆的重要机制。本研究提出了利用PCR技术快速检测产ACC脱氨酶细菌的快捷方法。以编码ACC脱氨酶的acd S基因为标记,分别使用acd Sf3/acd Sr4、Deg ACCf/Deg ACCr和F1936f/F1938r三对引物,对多种盐生植物和美洲黑杨(Populus deltoids)的根部及根际土中分离得到的细菌菌株进行检测。结果表明,结合acd Sf3/acd Sr4引物和递减PCR(touchdown-PCR)方法时,能获得单一的特异性扩增条带且扩增成功率高;但Deg ACCf/Deg ACCr和F1936f/F1938r两对引物特异性较差。从247个菌株中检测到25株含有acd S基因,旨为今后研究植物根际细菌acd S基因遗传性及储备丰富的功能性菌株奠定基础。Producing ACC（1-aminocyclopropane-1-carboxylate）deaminase to reduce ethylene levels and eliminate salt stress is thekey mechanism of plant-growth-promoting rhizobacteria（PGPR）in improving plant growth and stress tolerance. This work aims to establishan expeditious approach to screen bacteria containing ACC deaminase based on polymerase chain reaction（PCR）. We set the acd S geneencoding the ACC deaminase as the probe,then selected three pairs of primers（acd Sf3/acd Sr4,Deg ACCf/Deg ACCr and F1936 f/F1938 r）respectively,and detected the various bacterial strains isolated from the root and rhizosphere soil of diverse halophytes and Populus deltoids.The results showed that single specific amplified bands were produced and the rate of amplification was high when using the primers acd Sf3/acd Sr4 combined with touch-down PCR methods;however,Deg ACCf/Deg ACCr or F1936 f/F1938 r was in poor specificity. Finally,weacquired 25 strains containing acds gene from 257 bacterial isolates,aiming at laying a foundation for reserving rich and functional microbialresources and further studying the genetic diversity and function of acd S in PGPR.

关 键 词：盐生植物 杨树 植物促生长根际细菌 内生细菌 产ACC脱氨酶细菌 

分 类 号：Q93[生物学—微生物学]