与黑龙江大豆主栽品种匹配的优良根瘤菌筛选与鉴定  

Screening and identification of elite rhizobia matched with main cultivars of soybean in Heilongjiang Province

作　　者：伍惠[1] 钟喆栋 王学路[1] 陈大松[1] 李友国[1] 

机构地区：[1]华中农业大学生命科学技术学院农业微生物学国家重点实验室

出　　处：《应用与环境生物学报》2018年第1期39-46,共8页Chinese Journal of Applied and Environmental Biology

基　　金：国家高技术研究发展计划(863)项目(2013AA102802-04);国家重点研发计划项目(2016YFD0100702)资助~~

摘　　要：根瘤菌剂在大豆上能否充分发挥共生固氮效应涉及到根瘤菌的结瘤固氮能力、与大豆品种的匹配性、土壤特性等多种因素.黑龙江省是我国大豆的主产区之一,为筛选得到适于在黑龙江推广应用的根瘤菌资源,从黑龙江、河北和湖南等地区采集土壤样品,采用大豆结瘤法,从10个土样中分离获得59株大豆根瘤菌株,再依据BTB显色反应和16S r DNA分析,鉴定出36个慢生型大豆根瘤菌.沙培结果表明,有34株根瘤菌能够在大豆品种黑农61上有效结瘤;再从中选出4株共生效应较好的菌株做进一步实验.匹配实验表明,这4株菌SN2-5、NW5-3、SN7-2和SN10-3同样能与黑龙江另外16个主栽大豆品种结瘤;与不接根瘤菌的对照组相比,SN2-5、NW5-3、SN7-2和SN10-3处理的大豆的根瘤固氮酶活和生物量显著提高,且整体表型优于HN01和USDA110.土壤盆栽实验结果表明,NW5-3、SN10-3、SN7-2在黑土中具有显著的接种效果,且在10 mmol/L NH_4NO_3高氮处理下,4个优良供试菌均能够与黑农61结瘤.与不加氮素的处理相比,其中NW5-3和SN10-3形成根瘤具有近50%的固氮酶活性.本研究筛选的4个慢生型大豆根瘤菌SN2-5、NW5-3、SN7-2、SN10-3与黑龙江的主栽大豆品种具有良好匹配性能,可进一步用于黑龙江大豆产区的田间实验.The nitrogen-fixing efficiency of rhizobia inoculated on host legume is related to the nitrogen fixation ability of rhizobia, the matching performance between bacteria and host plant, and soil characteristics. Heilongjiang Province is one of the main soybean cultivation areas in China. In order to screen elite rhizobia that can be applied in Heilongjiang, 10 soil samples from Heilongjiang, Hebei, and Hunan were collected. In all, 59 rhizobial strains were isolated from soil samples using soybean nodulation capturing method; of which, 36 Bradyrhizobium japonicum strains were identified using bromothymol blue（BTB） reaction and 16 S rDNA analysis. Sand pot test showed that 34 of the 36 B. japonicum strains could effectively nodulate on soybean cultivar Hei Long 61. Among them, the SN2-5, NW5-3, SN7-2, and SN10-3 strains could also effectively nodulate on other 16 soybean varieties from Heilongjiang; further, the nitrogenase activity of nodule and biomass of soybean inoculated by these 4 strains respectively were significantly higher than those of the negative control. The symbiosis phenotype of these 4 rhizobial strains was superior to those of Sinorhizobium fredii HN01 and B. diazoefficiens USDA110. The NW5-3, SN10-3, and SN7-2 strains also showed significant inoculation efficiency on soybean planted in black soil. Under 10 mmol/L NH_4NO_3 stress, the SN2-5, NW5-3, SN7-2, and SN10-3 strains could still nodulate on soybean, and, compared with the nodules without nitrogen stress, the nodules formed by NW5-3 and SN10-3 still retained 50% nitrogenase activity. Above results showed that the rhizobia isolated in this study had high nodulating and nitrogen fixing efficiency on soybean cultivars from Heilongjiang and could be applied in Heilongjiang soybean cultivation area, because they are suitable for soybean varieties and soil characteristics.

关 键 词：根瘤菌接种剂 大豆品种 结瘤效能 固氮效能 匹配性 

分 类 号：Q939.114[生物学—微生物学]