2株根瘤菌的鉴定与共生性能比较  

Verification and comparison of symbiosis performance of 2 strains of rhizobia

作　　者：梁洪睿 荀岚 陆云艳 太若雪 李秋萍 张婷[1,2] 张雅琼 梁泉 LIANG Hong-rui;XUN Lan;LU Yun-yan;TAI Ruo-xue;LI Qiu-ping;ZHANG Ting;ZHANG Ya-qiong;LIANG Quan(College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;Yunnan Province Seed Management Station,Kunming 650031,China;School of Ethnic Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南农业大学农学与生物技术学院,昆明650201 [2]云南省种子管理站,昆明650031 [3]云南中医药大学民族医药学院,昆明650500

出　　处：《西南农业学报》2025年第1期64-71,共8页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(32060723)。

摘　　要：【目的】本研究旨在筛选共生匹配性好的根瘤菌,为更好地发挥根瘤菌株的固氮作用提供理论参考。【方法】采用YMA-刚果红固体培养基分离纯化的2株大豆根瘤菌进行形态学、生理生化和16S rDNA测序鉴定,并以低氮处理[13.24 g/L(NH4)2SO4]不接种根瘤菌为对照,通过回接试验验证和比较其共生性能。【结果】分离纯化得到的2株菌株DD1和YN10为革兰氏阴性菌,溶磷反应、柠檬酸盐反应、淀粉水解反应和泌氢反应均为阴性,无明胶液化能力,不产生生长素,16S rDNA序列测定属于慢生根瘤菌属,综合形态特征、生理生化特性及16S rDNA序列测定,DD1菌株鉴定为大豆慢生根瘤菌(Bradyrhizobium japonicum),YN10鉴定为大庆根瘤菌(B.daqingense);以低氮处理不接种根瘤菌为对照,在接种35 d后DD1和YN10根瘤数分别为57和27个,植株鲜重分别为17.170和15.195 g,植株氮含量分别为44.806和48.664 mg/g,与对照差异达极显著水平;接种2种根瘤菌的大豆新叶、老叶叶绿素含量(SPAD)值在接种前14 d内均处于上升趋势,老叶的SPAD值上升速率更快,SPAD均高于未接种大豆;接种DD1和YN10固氮酶活分别为4027.98和3655.68 mol/(g·h),二者差异达极显著水平(P≤0.01,下同)。【结论】分离纯化得到的DD1菌株为大豆慢生根瘤菌(B.japonicum),YN10属于大庆根瘤菌(B.daqingense),回接根瘤菌的大豆植株光合作用能力与固氮能力显著提高。【Objective】Soybean can engage in symbiotic nitrogen fixation with rhizobia,and the rhizobia with good symbiotic matching can be screened to better exert the nitrogen fixation effect of rhizobia.【Method】In the study,two strains of soybean rhizobia were isolated and purified from YMA-Congo red solid medium for morphological,physiological and biochemical,and 16S rDNA sequencing identification,and the symbiotic properties of two strains of rhizobia were isolated and purified by YMA-Congo red solid medium for identification,and their symbiotic properties were verified and compared by backlash test.【Result】The two strains,DD1 and YN10,were gram-negative,with negative reaction for phosphorus dissolving citrate,starch hydrolysis and hydrogen secretion reaction.They did not liquefy gelatin or pyodua auxin 16S rDNA sequence determination classified as the genus Rhizobia slow-growing rhizobia,and the DD1 strain was identified as Bradyrhizobium japonicum by comprehensive morphological characteristics,physiological and biochemical characteristics and 16S rDNA sequence determination.YN10 was identified as B.liaoningense;After 35 days after inoculation,the number of nodules in DD1 and YN10 were 57 and 27,respectively,the fresh weight of plants were 17.170 and 15.195 g,and the nitrogen content of plants were 44.806 and 48.664 mg/g,respectively,showing very significant compared with the control.The chlorophyll content(SPAD)values of new and old leaves of soybean inoculated with the two rhizobia were on an upward trend in the first 14 days,and the SPAD values of old leaves increased faster,and the SPAD values were higher than those of uninoculated soybeans.The nitrogenase activities of DD1 and YN10 were 4027.98 and 3655.68 mol/(g·h),respectively,and the difference between them was very significant(P≤0.01).【Conclusion】The isolated and purified strains DD1(B.japonicum)and YN10(B.daqingense)significantly enhanced the photosynthesis and nitrogen fixation capacity of soybean plants.

关 键 词：大豆 根瘤菌 鉴定 回接验证 

分 类 号：S565.2[农业科学—作物学]