野大豆多功能根际促生菌的筛选鉴定和促生效果研究  

Screening,Identification and Growth-promoting Effect of Multifunction Rhizosphere Growth-promoting Strain of Wild Soybean

作　　者：郭英 杨萍 张丹雨 刘莹莹 马莲菊 卜宁 GUO Ying;YANG Ping;ZHANG Dan-yu;LIU Ying-ying;MA Lian-ju;BU Ning(School of Life Science,Shenyang Normal University,Shenyang 110034)

机构地区：[1]沈阳师范大学生命科学学院,沈阳110034

出　　处：《生物技术通报》2018年第10期108-115,共8页Biotechnology Bulletin

基　　金：国家自然科学基金面上资助项目(31270369);国家重点研发计划项目子课题(2016YFC0500707);沈阳市科学技术计划项目(F16-205-1-50)

摘　　要：为获得野大豆根际促生菌(Plant growth promoting rhizobacteria,PGPR)并明确其促生特性,采用选择培养方法从沈阳沈北新区蒲河沿岸生长的野大豆根瘤及根际土壤中分离筛选固氮菌与解磷菌,进行形态学、生理生化指标和16S rDNA分子生物学鉴定。采用Touch-Up PCR(上升PCR)的方法扩增固氮酶nifH结构基因,对菌株的解磷、分泌3-吲哚乙酸(IAA)的能力及ACC脱氨酶的活性进行测定分析,筛选出优良菌株进行盆栽大豆的促生作用研究。结果得到12株野大豆PGPR菌,经鉴定分别归属于芽孢杆菌属(Bacillus)、伯克霍尔德菌属(Burkholderia)、根瘤菌属(Rhizobium)、詹森氏菌属(Janthinobacterium)、鞘氨醇单胞菌(Phingomonas)、分枝杆菌属(Mycobacterium)、假单胞菌属(Pseudomonas)和根癌农杆菌属(Agrobacterium);其中2株可解有机磷,11株可解无机磷,3株具分泌IAA能力,5株具ACC脱氨酶活性。盆栽试验表明:3株菌(GD11、GD17、GD58)可使大豆幼苗株高增长,5株菌(GD9、GD11、GD17、GD30、GD58)可使根长增长;6株菌可提高大豆幼苗的茎干重、叶干重、根干重。野大豆根瘤和根际土壤含有多功能根际促生菌,其在农业生产上具有潜在的应用价值。In order to obtain plant growth promoting rhizobacteria(PGPR)of wild soybean and clarify its growth-promoting characteristics,a selective culture method was used to isolate and screen nitrogen-fixing bacteria and phosphorus-solubilizing bacteria from wild soybean nodules and rhizosphere soils growing along Puhe River in Shenbei New District,Shenyang.Morphological,physiological and biochemical indicators and molecular biological identification of 16S rDNA were adapted to identify their biology,and touch-Up PCR(Rising PCR)method was employed to amplify the structural gene nifH of nitrogenase.Their phosphorus release,IAA secretion,and ACC deaminase activity were determined and analyzed for screening fine strains that then were used to study their growth-promoting effect on potted soybean.As a result,12 PGPR strains of wild soybean were obtained and identified as Bacillus,Burkholderia,Rhizobium,Janthinobacterium,Sphingomyces,Phingomonas,Mycobacterium,Pseudomonas,and Agrobacterium;2 of them dissolved organic phosphorus,11 solubilized inorganic phosphorus,3 secreted IAA,and 5 presented ACC deaminase activity.Pot experiments showed that 3 strains(GD11,GD17,and GD58)resulted in the plant height increase of soybean seedlings,5 strains(GD9,GD11,GD17,GD30,and GD58)promoted root length,and 6 strains led to the increment of stem weight,leaf dry weight,and root dry weight of soybean seedlings.Soybean root nodule and rhizosphere soil contain multi-functional rhizosphere growth-promoting bacteria,thus it has potential application value in agricultural production.

关 键 词：野大豆 根际促生菌 NIFH基因 解磷 

分 类 号：S154.3[农业科学—土壤学]