转badh基因玉米植株的获得及其耐盐性分析

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1、作物学报ACTAAGRONOMICASINICA2014,40(11):19731979http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2014.01973转BADH基因玉米植株的获得及其耐盐性分析1,21111,211,2,*王小丽杜建中郝曜山张丽君赵欣梅王亦学孙毅12山西省农业科学院生物技术研究中心/农业部黄土高原作物基因资源与种质创制重点实验室,山西

2、太原030031;山西大学生物工程学院,山西太原030006摘要:采用超声波辅助花粉介导植物转基因方法,将甜菜碱醛脱氢酶(BADH)基因导入玉米自交系郑58,获得了耐盐性强的转基因玉米植株。经卡那霉素抗性初筛、PCR扩增、Southernblot分析,证明BADH基因已导入转化植株并整合到其基因组中。用不同浓度的NaCl溶液对T2代转基因玉米植株与对照进行盐胁迫处理,结果表明,转BADH基因玉米植株表现出一定的抗逆性,生长状况明显优于对照;根据非转化苗对NaCl的反应以及生长状况,确定250–1mmolLNaCl溶液

3、为玉米幼苗耐盐性筛选的适宜浓度;依据此临界浓度下形态指标和生理生化指标的测定结果,与对照相比,转基因植株的株高提高10.94%~25.70%,鲜重增加8.62%~18.20%,干重增加9.00%~18.18%,相对电导率降低37.21%~58.14%,叶绿素含量增加15.89%~90.65%,超氧化物歧化酶(SOD)活性提高64.92%~148.29%,丙二醛(MDA)含量减少26.97%~48.05%。综上所述,转入甜菜碱醛脱氢酶(BADH)基因提高了玉米的耐盐性。这是首例将BADH基因导入优良玉米自交系郑58的报

4、道。超声波辅助花粉介导法是一种经济、高效、实用和无基因型依赖性的植物基因转化方法。关键词:玉米;BADH基因;转基因;耐盐性;形态指标;生理指标TransformationofBADHGeneintoMaizeandSaltTolerenceofTransgenicPlant1,21111,21WANGXiao-Li,DUJian-Zhong,HAOYao-Shan,ZHANGLi-Jun,ZHAOXin-Mei,WANGYi-Xue,1,2,*andSUNYi1BiotechnologyResearchCenter

5、,ShanxiAcademyofAgriculturalSciences/KeyLaboratoryofCropGeneResourcesandGermplasmEnhancement2onLoessPlateau,MinistryofAgriculture,Taiyuan030031,China;BiologicalEngineeringInstitute,ShanxiUniversity,Taiyuan030006,ChinaAbstract:Inordertoobtaintransgenicmaizeplant

6、stoleranttosaltstress,maizeinbredZheng58wastransformedwithBADHgeneusingpollen-mediatedmethod.TheresultsofKm-resistantscreening,PCRdetectionandSouthernblotanalysisprovedthatBADHgenewasintroducedintomaizeplantsandintegratedintothemaizegenome.Effectsofvariousconce

7、ntrationsofNaClsolutiononthegrowthofT2transgenicandnon-transgenicmaizeplantswereinvestigated．Theresultsindicatedthattransgenicmaizeseedlingshadanimprovedresistancetosaltstress,andtheirgrowthperformancewassuperiortothatofnon-transgenic–1maizeseedlings.Onthebasis

8、ofthegrowthstatusofnon-transgenicplants,250mmolLofNaClsolutionwasusedtoscreen–1transgenicplants.Ananalysisonmorphologicalandphysiologicalindexesunderthestressof250mmolLNaCls