以ems诱变创制软质小麦宁麦9号高分子量谷蛋白亚基突变体

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1、作物学报ACTAAGRONOMICASINICA2014,40(9):15791584http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2014.01579以EMS诱变创制软质小麦宁麦9号高分子量谷蛋白亚基突变体*张纪元张平平姚金保杨丹杨学明马鸿翔江苏省农业科学院/江苏省农业生物学重点实验室,江苏南京210014摘要:本研究旨在创制遗传背景一致的不同高分子量谷蛋白亚基(H

2、MW-GS)缺失系,为小麦品质研究和育种提供材料。将软质小麦品种宁麦9号,用0.4%EMS溶液处理10000粒种子,获得3781个M1代单株,采用“半粒法”对每个株系进行SDS-PAGE鉴定,从中筛选出299个(7.91%)HMW-GS突变株,包括HMW-GS缺失和分子量突变2种类型。其中,HMW-GS缺失突变176株,突变频率为4.65%,缺失亚基涉及Ax1、Bx7、By8、Dx2和Dy12,突变频率为0.24%~3.28%;分子量突变130株,突变频率为3.44%。将突变体的具胚端种子于温室中繁殖获得M2代,再次鉴定各株系的H

3、MW-GS,并经M3代验证,最终获得Ax1、Dx2、Bx7、By8和Dy12缺失突变体,及Ax1+By8双缺失突变体。用高效液相色谱(HPLC)分析这些突变体的谷蛋白大聚体(GMP)和谷蛋白/醇溶蛋白(GLU/GLI)比值,发现不同缺失突变体的GMP含量都有不同程度的降低,尤以Bx7亚基缺失突变体中GMP含量降幅最大,高达42%。另外,不同缺失突变体的谷蛋白总量和GLU/GLI比值也低于对照,Ax1+By8双缺失突变体的GLU/GLI比值最小。关键词:小麦;甲基磺酸乙酯;高分子量谷蛋白亚基;谷蛋白大聚体EMSInducedHMW-

4、GSMutantsfromSoftWheatNingmai9*ZHANGJi-Yuan,ZHANGPing-Ping,YAOJin-Bao,YANGDan,YANGXue-Ming,andMAHong-XiangJiangsuProvincialKeyLaboratoryofAgrobiology/JiangsuAcademyofAgriculturalSciences,Nanjing210014,ChinaAbstract:Theobjectiveofthisstudywastocreateaseriesofdeletionli

5、nesofhighmolecularweight-gluteninsubunit(HMW-GS)insimilarbackgroundsforwheatqualityresearchandbreeding.Atotalof3781M1plantsofNingmai9(asoftwheatcultivar)wereobtainedfrom10000seedsinducedby0.4%ethylmethanesulfonate(EMS).TheHMW-GScompositionsoftheseplantswereidentifiedv

6、iaSDS-PAGEusinghalfseed.Twohundredandninety-ninelineswithamutationpercentageof7.91%werefoundtobeHMW-GSmutants,includingHMW-GSdeletionandmolecularweightmutation.Amongthem,176lineswereHMW-GSdeletionmutants,withamutationpercentageof4.65%containingsubunitsAx1,Bx7,By8,Dx2,

7、andDy12,withthemutationpercentagesrangingfrom0.24%to3.28%.Themutantsofmolecularweightwere130lineswithamutationpercentageof3.44%.EachofthehalfM1seedwithembryowasusedtoproduceM2generationingreenhouse.HomozygousdeletionlinesofHMW-GSAx1,Dx2,Bx7,By8,Dy12,andAx1+By8weredete

8、ctedusingSDS-PAGEandconfirmedinM3generation.Thecon-tentofgluteninmacropolymer(GMP)andtheratioofglutenin-to-gliadin(GLU/GLI)r