摘要
【目的】筛选获得结瘤固氮能力强、促生效果好的土著高效救荒野豌豆根瘤菌菌株,为贵州喀斯特地区土壤改良应用提供优质菌株资源。【方法】采用刚果红YMA平板分离培养法,对贵州省不同生态功能区的救荒野豌豆根瘤进行分离、纯化,结合菌体形态结构和16S r RNA测序方法进行细菌种类鉴定,将筛选获得根瘤菌株与豆科模式植物百脉根进行盆栽回接试验,结合菌株固氮酶活性的测定,筛选土著高效根瘤菌株。【结果】从贵州省境内不同生态功能区的救荒野豌豆根瘤样品中分离获得53株形态、颜色和大小各异的根瘤细菌菌株,革兰氏染色后,共获得30株革兰氏阴性(G-)纯化菌株,经鉴定8株为根瘤内生细菌,22株为根瘤菌分属根瘤菌属(Rhizobium)和假单胞菌属(Pseudomonas),其中Rhizobium的分布频率最高,达91%,豌豆根瘤菌(R. leguminosarum)的分布频率最高,达27%,分别是分离获得的优势属和优势种。将来自不同地域和种类的根瘤菌与百脉根进行盆栽回接试验后,共筛选得到优良固氮促生根瘤菌6株,即XDVR141-4、ZJVR339-3、WEVR362-3、WXVR365-1、WLVR370-5和BQVR415-3,其中,WXVR365-1菌株的酶活力及植物促生效果与其余5株的差异均达显著水平(P<0.05),为最优促生根瘤菌。【结论】从贵州不同生态功能区采集的救荒野豌豆根瘤中分离获得的R. acidisoli属于根瘤菌α-变形菌纲的一类,具有较强的固氮促生效果,可作为开发微生物菌肥的候选菌株资源。
【Objective】To screen and obtain indigenous efficient strains of Vicia sativa L.rhizobia with strong nitrogen fixation ability and growth promoting effect and to provide high quality strain resources for soil improvement in karst area of Guizhou.【Method】Isolation and purification of V.sativa L.nodules in different ecological functional areas of Guizhou were isolated and cultured by Congo red YMA plate method,and bacterial species identification was carried out according to bacterium morphological structure and the 16S rRNA sequencing method.Then rhizobia and legume model plant Lotus corniculatus L.were employed for potted re-inoculation test,and the determination of the nitrogenase activity of the strains were combined to screen indigenous efficient rhizobia.【Result】From different ecological functional areas in Guizhou,53 rhizobia of different shapes,colors and sizes were isolated from V.sativa L.root nodule samples.After Gram staining,a total of 30 Gram-negative(G-)purified strains were obtained,of which 8 strains were identified as nodule endophytic bacteria,22 strains were Rhizobium and Pseudomonas.Rhizobium had the highest distribution frequency,reaching 91%and R.leguminosarum had the highest frequency of distribution,up to 27%,indicating that they were dominant genera and dominant species obtained through isolation,respectively.In potted re-inoculation test of different species of rhizobia from different regions and L.corniculatus L.,6 excellent nitrogen-fixing and growth promoting rhizobial strains were screened:XDVR141-4,ZJVR339-3,WEVR362-3,WXVR365-1,WLVR370-5,BQVR415-3.And enzyme activity and plant probiotic effect of WXVR365-1 strain were significantly different from those of the other 5 strains(P<0.05),proving that it was the optimal growth-promoting rhizobium strain.【Conclusion】R.acidisoli isolated from V.sativa L.root nodules from different ecological functional areas in Guizhou belongs to rhizobial strainα-Proteobacteria with strong nitrogen fixation and growth-promoting effects,which
作者
韦兴迪
曾庆飞
王佳楚函
冉伟男
龙忠富
莫丹禄
余晓玲
WEI Xing-di;ZENG Qing-fei;WANG Jia-chu-han;RANWei-nan;LONG Zhong-fu;MO Dan-lu;YU Xiao-ling(Guizhou Institute of Prataculture,Guizhou Academy of Agricultural Sciences,Guiyang,Guizhou 550006,China;College of Animal Science,Guizhou University,Guiyang,Guizhou 550025,China;Agricultural Comprehensive Service Center of Mawei Town,Dushan County,Qiannan Prefecture,Qiannan,Guizhou 558206,China;Xiaochang Institute of Agricultural Science and Technology,Xiaogan,Hubei 432999,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2023年第1期200-208,共9页
Journal of Southern Agriculture
基金
国家牧草产业技术体系项目(CARS-34)
贵州草地资源管理与高效利用创新能力建设项目(黔科合服企[2022]004)
贵州省农业科技支撑计划项目(黔科合支撑[2019]2365)
贵州省农业科学院青年基金项目(黔农科院青年科技基金[2021]32号)。
关键词
救荒野豌豆
根瘤菌
高效菌株筛选
贵州
Vicia sativa L.
rhizobia
screening of efficient strains
Guizhou
