Isolation and Molecular Identification of Soybean Rhizobium in Zhoukou

doi:10.7668/hbnxb.2017.04.016

Abstract

Abstract: In order to explore new rhizobium resource,the method of separating and purifying the root nodule bacteria from soybean root nodules was studied,and a pot experiment was carried out. The results showed that the isolated rhizobium of soybean could form nodules in soybean and alfalfa. According to the published soybean rhizobium common node tumor genes of nodA, nifH, 16S rDNA,all the three genes were amplified. Phylogenetic analysis demonstrated that the strain isolated from Zhoukou was highly conserved with Sinorhizabium fredii of Germany,which convinced that rhizobium strain was S. fredii. The research provided the quick and accurate method for identification of soybean rhizobium at the molecular level.

Key words: Soybean, Rhizobium, Re-inoculation and identification, Sinorhizabium fredii

CLC Number:

S432.4

ZHANG Yi, ZHANG Chao, YANG Yanjie, YU Jie, JI Huijia, CHEN Hong, LIU Wenting, LI Xiaobo, LI Meng, LI Chengwei. Isolation and Molecular Identification of Soybean Rhizobium in Zhoukou[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(4): 98-102. doi: 10.7668/hbnxb.2017.04.016.

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References

[1] Haichar F E, Santaella C, Heulin T A. Root exudates mediated interactions belowground[J]. Soil Biology & Biochemistry, 2014, 77(7):69-80.
[2] 李欣欣, 许锐能, 廖红.大豆共生固氮在农业减肥增效中的贡献及应用潜力[J]. 大豆科学, 2016, 35(4):531-535.
[3] 赵叶舟, 王浩铭, 汪自强.豆科植物和根瘤菌在生态环境中的地位和作用[J]. 农业环境与发展, 2013, 30(4):7-12.
[4] 刘亚柏, 王润芳.氮磷钾对红花草固氮根瘤菌生长及种植后土壤肥力的影响[J]. 中国农学通报, 2015, 31(36):220-225.
[5] 韦革宏, 马占强.根瘤菌-豆科植物共生体系在重金属污染环境修复中的地位、应用及潜力[J]. 微生物学报, 2010, 50(11):1421-1430.
[6] 张爱媛. 根瘤菌与钼肥对大豆养分吸收和产量影响的研究[D]. 哈尔滨:东北农业大学, 2015.
[7] 高俊莲, 孙建光, 陈文新. 斜茎黄芪根瘤菌结瘤基因nodA PCR扩增及PCR-RFLP分析[J]. 微生物学杂志, 2006, 26(4):1-5.
[8] Roche P, Maillet F, Plazanet C, et al. The common nodABC genes of Rhizobium meliloti are host-range determinants[J]. Proceedings of the National Academy of Sciences of the United States of America, 1996, 93(26):15305-15310.
[9] Okazaki S, Tittabutr P, Teulet A, et al. Rhizobium-legume symbiosis in the absence of Nod factors:two possible scenarios with or without the T3SS[J]. ISME Journal, 2016, 10(1):64-74.
[10] Dhole A, Shelat H, Vyas R, et al. Endophytic occupation of legume root nodules by nifH-positive non-rhizobial bacteria, and their efficacy in the groundnut (Arachis hypogaea)[J]. Annals of Microbiology, 2016, 66(4):1397-1407.
[11] Alvarez-Morales A, Hennecke H. Expression of Rhizobium japonicum nifH and nifDK operons can be activated by the klebsiella pneumonia nifA protein but not by the product of ntrC[J]. Molecular & General Genetics:MGG, 1985, 199(2):306-314.
[12] Terakado-Tonooka J, Ohwaki Y, Yamakawa H, et al. Expressed nifH genes of endophytic bacteria detected in Field-Grown sweet potatoes (Ipomoea batatas L.)[J]. Microbes and Environments, 2008, 23(1):89-93.
[13] Chun J S, Goodfellow M. A phylogenetic analysis of the genus Nocardia with 16S rRNA gene sequences[J]. International Journal of Systematic Bacteriology, 1995, 45(2):240-245.
[14] Ntushelo K. Identifying bacteria and studying bacterial diversity using the 16S ribosomal RNA gene-based sequencing techniques:A review[J]. African Journal of Microbiology Research, 2013, 7(49):5533-5540.
[15] Peter M G, Barry G R. Viability of rhizobium bacteroids isolated from soybean nodule protoplasts[J]. Planta, 1978, 142:329-333.
[16] 陈强, 张小平, 李登煜, 等. 从豆科植物的根瘤中直接提取根瘤菌DNA的方法[J]. 微生物学通报, 2002, 29(6):63-67.
[17] Yanagi M, Yamasato K. Phylogenetic analysis of the family Rhizobiaceae and related bacteria by sequencing of 16S rRNA gene using PCR and DNA sequencer[J]. FEMS Microbiology Letters, 1993, 107(1):115-120.
[18] Cardenas L, Feijo J A, Kunkel J G, et al. Rhizobium nod factors induce increases in intracellular free Calcium and extracellular Calcium influxes in bean root hairs[J]. The Plant Journal, 1999, 19(3):347-352.
[19] Azcón R, Rubio R, Barea J M. Selective interactions between different species of mycorrhizal fungi and rhizobium-meliloti strains, and their effects on growth, N2-fixation (N-15) and nutrition of Medicago sativa L.[J]. New Phytologist, 1991, 117(3):399-404.
[20] 高萍,李芳,郭艳娥,等.丛枝菌根真菌和根瘤菌防控植物真菌病害的研究进展[J].草地学报,2017,25(2):236-242.

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