微波诱变选育耐药高效溶磷苜蓿根瘤菌

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1、微波诱变选育耐药高效溶磷苜蓿根瘤菌第43卷第12期2009年12月原Atomic子能科EnergyScience学技术andTechnologyVo1.43,NO.12Dec.2009微波诱变选育耐药高效溶磷苜蓿根瘤茵李剑峰,张淑卿,师尚礼一,霍平慧(1.甘肃农业大学草业学院,甘肃兰州730070;2.草业生态系统教育部重点实验室/中一美草地畜牧业可持续发展研究中心,甘肃兰州730070)摘要:为获得高效溶磷,同时具备卡那霉素和青霉素双抗药性的根瘤菌突变株,以苜蓿根瘤菌L一5为原始菌株进行微波诱变处理,对微波诱变参数进行优化

2、,考察突变株的遗传稳定性,溶磷量和结瘤特性.经微波(600W,30s)诱变,得到5株具备较高溶磷能力的双耐药突变株LW59,LW81,IWl04,LW107,LW135.多次传代试验证明,LW107是稳定的高效溶磷双耐药突变株.突变株LW107具备对卡那霉素100mg/I和青霉素300mg/I的抗性,14d的溶磷量达到16.67mg/L,较原始菌株提高112.59,固氮酶活性高于原始菌株l1.7.经植株回接试验证明,突变株结瘤能力与原始菌株无显着差异.关键词:根瘤菌;微波诱导;溶磷量中图分类号:Q936文献标志码:A文章编号

3、:l0006931(2009)12—1071一O6ScreeningofDissolvePhosphorusRhizobiummelilotiAntibiotic—ResistantStrainUsingMicrowaveMutagenesisLIJian—feng,ZHANGShu—qingt,SHIShang—li,.,,HUOPing—huit(1.CollegeofGrasslandScience,GansuAgriculturalUniversity,Lanzhou730070,China2.KeyLaborato

4、ryofGrasslandEcosystemofMinistryofEducation/Sino-U.S.Centers/_0rGrazingLandEcosystemS"5￡以n6f如,Lanzhou730070,China)Abstract:InordertOobtainhighlyefficientdissolvephosphorusRhizobiummelilotistrainwithtwoantibiotic—resistant.adissolvephosphorusrhizobiumstrainL-5wasirr

5、adiatedbymicrowave.Themicrowavemutagenesisparameterswereoptimized,andthegeneticstability,phosphatesolubilizingpowerandantibioticresistanceofthemu—tantwereinvestigated.Fivehighlyefficientdissolvephosphorusstrainswithtwoantibi—otic～resistantnamedLW59,LW81,LW104,LW107

6、andLW135wereobtainedthroughthemicrowaveirradiation(600W,30s).Inseveralpassages,amutantnamedLW107withsteadyphosphorussolubilizingcapabilitywasobtainedanditexhibitsresistanceto300mg/Iampicillinand100mg/Ikanamycin.Thephosphatesolubilizingpowerreaching16.67mg/L(14d)and

7、nitrogenaseactivityare112.59%and11.7higher收稿日期:2009—0513;修回日期:20090618基金项目:国家科技支撑项目资助(2007BAD52B06,2006BAD04A04,2006BAD01A19);农业部行业专项资助项目(nyhyzx07022);现代农业产业技术体系建设专项资助项目作者简介:李剑峰(1979),男,甘肃天水人,博士研究生,牧草及根瘤菌种质资源研究专业*通信作者:师尚礼,教授,博士生导师,从事牧草种质资源研究,Email:shishl@gsau.edu.c

8、a1O72原子能科学技术第43卷thanthoseofinitialstrains,respectively.TwostrainsoftheLW107andL-5areinocu—latedontotheliveplantsanditistestifiedthatnodulationabi