紫云英优势根瘤菌的发掘与鉴定OA
Isolation and identification of dominant rhizobia associated with Astragalus sinicus
[目的]从不同地区紫云英根瘤菌资源中筛选优势菌株,为后续菌剂开发和应用奠定基础.[方法]采用人工气候室"双层钵"盆栽法,基于株高、结瘤数、叶片叶绿素含量和干质量等指标,对前期分离自河南、四川、广西、湖北和湖南5地的37株紫云英根瘤菌代表菌株开展共生效率测定和分析.挑选出共生固氮效率较好的菌株进行耐酸碱、耐盐和耐干旱的抗逆性检测,并进行基于16S rRNA、持家基因atpD和结瘤基因nodC的系统发育分析,明确其分类地位.[结果]根瘤菌WYCCWR 10750和WYCCWR 10704为河南省与紫云英共生效果最佳菌株,能够耐受pH值范围4～11、质量分数2%的NaCl及质量分数7%的聚乙二醇;根瘤菌WYCCWR 12312和WYCCWR 12365为四川省与紫云英共生效果最佳菌株,能够耐受pH值范围4～11、质量分数2%的NaCl及质量分数5%的聚乙二醇;根瘤菌WYCCWR 12508和WYCCWR 12418为广西壮族自治区与紫云英共生效果最佳菌株,能够耐受pH值范围5～10、质量分数2%的NaCl及质量分数7%的聚乙二醇;根瘤菌WYCCWR 12656和WYCCWR 12542为湖北省与紫云英共生效果最佳菌株,能够耐受pH值范围5～10、质量分数2%的NaCl及质量分数7%的聚乙二醇;根瘤菌WYCCWR 12123和WYCCWR 12091为湖南省与紫云英共生效果最佳菌株,能够耐受pH值范围5～10、质量分数2%的NaCl及质量分数7%的聚乙二醇.上述10株菌为最终发掘的不同地区的紫云英优势根瘤菌.系统发育分析结果表明,10株优势菌均属于中慢生根瘤菌属,被鉴定为Mesorhizobium huakuii、Mesorhizobium qingshengii和Mesorhizobium jarvisii 3个种群,且结瘤基因型单一.[结论]从河南、四川、广西、湖北和湖南5地发掘鉴定紫云英优势根瘤菌10株,均可耐受质量分数2%的NaCl,酸碱耐受性较广,对聚乙二醇具有一定耐受能力,同时含有保守的结瘤基因.
[Objective]This study aimed to screen out the dominant strains from the Astragalus sinicus rhizobium resources in different region of China,so as to lay a solid foundation for the subsequent development and application of rhizobial inoculants.[Method]The symbiotic efficiency of 37 represen-tative Astragalus sinicus rhizobial strains previously isolated from Henan,Sichuan,Guangxi,Hubei and Hunan Provinces was determined and analyzed by the"double-pot"pot culture method in an artifi-cial climate chamber,with plant height,nodule number,leaf chlorophyll content and dry biomass as the key evaluation indicators.The strains with excellent symbiotic nitrogen fixation efficiency were selected for stress tolerance detection including acid-alkali resistance,salt tolerance and drought resis-tance.Furthermore,phylogenetic analysis based on the 16S rRNA gene,housekeeping gene atpD and nodulation gene nodC was conducted to clarify their taxonomic status.[Result]The optimal rhizobial strains for symbiosis with Astragalus sinicus in each province were identified as follows:WYCCWR 10750 and WYCCWR 10704 in Henan Province,which could tolerate a pH value range of 4-11,2%(mass fraction)NaCl and 7%(mass fraction)polyethylene glycol(PEG);WYCCWR 12312 and WYCCWR 12365 in Sichuan Province,which could tolerate a pH value range of 4-11,2%(mass frac-tion)NaCl and 5%(mass fraction)PEG;WYCCWR 12508 and WYCCWR 12418 in the Guangxi Zhuang Autonomous Region,which could tolerate a pH value range of 5-10,2%(mass fraction)NaCl and 7%(mass fraction)PEG;WYCCWR 12656 and WYCCWR 12542 in Hubei Province,which could tolerate a pH value range of 5-10,2%(mass fraction)NaCl and 7%(mass fraction)PEG;WYCCWR 12123 and WYCCWR 12091 in Hunan Province,which could tolerate a pH value range of 5-10,2%(mass fraction)NaCl and 7%(mass fraction)PEG.The above 10 strains were the domi-nant Astragalus sinicus rhizobia identified from different provinces in this study.Phylogenetic analysis results showed that all the 10 dominant strains belonged to the genus Mesorhizobium,and were identi-fied as three species including Mesorhizobium huakuii,Mesorhizobium qingshengii and Mesorhizobium jarvisii,with a single nodulation genotype.[Conclusion]A total of 10 dominant Astragalus sinicus rhizo-bial strains were excavated and identified from Henan,Sichuan,Guangxi,Hubei and Hunan Pro-vinces.All these strains could tolerate 2%(mass fraction)NaCl,had a wide acid-alkali tolerance range and a certain tolerance to polyethylene glycol,and contained conserved nodulation genes,which could provide excellent germplasm resources for the development of Astragalus sinicus rhizobial inoculants.
李可;赵泽洋;杨柳;杨国平;张俊杰
郑州轻工业大学食品与生物工程学院,河南 郑州 450000郑州轻工业大学食品与生物工程学院,河南 郑州 450000郑州轻工业大学食品与生物工程学院,河南 郑州 450000宁夏五丰农业科技有限公司,宁夏 银川 750021郑州轻工业大学食品与生物工程学院,河南 郑州 450000
农业科技
紫云英根瘤菌高效固氮抗逆性系统发育分析
Astragalus sinicusRhizobiumhigh efficient nitrogen fixationstress resistancephylo-genetic analysis
《河南农业大学学报》 2026 (2)
284-296,13
国家重点研发计划项目(2021YFD1700200)
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