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1、作物学报ACTAAGRONOMICASINICA2014,40(6):1132−1139http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2014.01132绿色荧光蛋白基因标记的固氮菌DX120E在甘蔗植株内的定殖11,2,*121,2,331,2,3,*魏春燕邢永秀莫遥林丽杨丽涛胡春锦李杨瑞12广西大学农学院/亚热带农业生物资源保护与利用
2、国家重点实验室,广西南宁530004;广西农业科学院/中国农业科学院甘蔗3研究中心/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良生物技术重点开放实验室,广西南宁530007;广西农业科学院微生物研究所,广西南宁530007摘要:为了探索甘蔗固氮菌DX120E菌株侵入根的方式以及菌株在甘蔗植株内的定殖规律,确定人工接种固氮菌适宜的侵染浓度和定殖特点,采用绿色荧光蛋白(GFP)标记的固氮菌DX120E接种甘蔗组培苗,以荧光显微镜观察其侵染和定殖情况,以抗生素抗性标记法和平板分离计数法测定
3、DX120E菌株的种群动态。结果表明,菌株DX120E在2个甘2蔗品种的根、叶鞘和叶内均能定殖,定殖菌量依次均为根>叶鞘>叶;不同接种浓度下,最大定殖数量无显著差异,1×10–1CFUmL的接种量足够侵入甘蔗并积累定殖。DX120E从甘蔗根表面的裂隙、主根和侧根发生处及根的断裂处均可侵入,主要在根表面细胞间隙和细胞内大量定殖,同时也可迁移到叶片的叶肉细胞和维管束细胞中定殖。关键词:固氮菌(Klebsiellasp.);甘蔗;接种浓度;细菌数量;定殖ColonizationofNitrogenFixi
4、ngBacterialStrainKlebsiellasp.DX120ELabeledwithGreenFluorescentProtein(GFP)GenewithinSugarcanePlants11,2,*121,2,33WEIChun-Yan,XINGYong-Xiu,MOYao,LINLi,YANGLi-Tao,HUChun-Jin,andLI1,2,3,*Yang-Rui1StateKeyLaboratoryforConservationandUtilizationofSubtropic
5、alAgro-bioresources/CollegeofAgriculture,GuangxiUniversity,Nanning2530004,China;GuangxiAcademyofAgriculturalSciences/SugarcaneResearchCenter,ChineseAcademyofAgriculturalSciences/KeyLabora-toryofSugarcaneBiotechnologyandGeneticImprovement(Guangxi),Minis
6、tryofAgriculture/GuangxiKeyLaboratoryofSugarcaneGeneticIm-3provement,Nanning530007,China;MicrobiologyResearchInstitute,GuangxiAcademyofAgriculturalSciences,Nanning530007,ChinaAbstract:ToexploretheinvasionpatternsofN2-fixingbacteriastrainKlebsiellasp.DX
7、120Eanditscolonizationinsugarcane,andtodeterminetheoptimumconcentrationsofinoculum,weinoculatedtissuecultureseedlingsoftwosugarcanevarieties(GT21andB8)withdifferentconcentrationsofKlebsiellasp.bacteriastrainDX120Elabeledbygreenfluorescentprotein(GFP)ge
8、ne,andwithddH2Oascontrol.TheresultsshowedthatthestrainDX120Epropagatedinroot,leafsheathandleafofsugarcaneseedlings,andbacteriaquantitiesindifferentpartswereroot>leafsheath>leaf.Therewerenodifferencesinthemaximum2–1propagationquantityund