抗疫病加工型辣椒细胞质雄性不育恢复系的创制  

Development of Cytoplasmic Male Sterility Restorer Line with Phytophthora Blight Resistance in Processing Type Pepper

作　　者：李怡斐[1] 杨小苗 王春萍[2] 段敏杰 黄任中[1] 张世才[1] LI Yifei;YANG Xiaomiao;WANG Chunping;DUAN Minjie;HUANG Renzhong;ZHANG Shicai(Vegetable and Flower Research Institute,Chongqing Academy of Agricultural Sciences,Chongqing 401329,China;Chongqing Key Laboratory of Adversity Agriculture,Biotechnology Research Institute,Chongqing Academy of Agricultural Sciences,Chongqing 401329,China)

机构地区：[1]重庆市农业科学院蔬菜花卉研究所,重庆401329 [2]重庆市农业科学院生物技术研究所,逆境农业研究重庆市市级重点实验室,重庆401329

出　　处：《西北植物学报》2022年第10期1637-1643,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家特色蔬菜产业技术体系(CARS-24-A-04);重庆市川渝联合实施重点研发项目(cstc2021jscx-cylhX0002);重庆市技术创新与应用发展专项面上项目(cstc2020jscx-msxmX0054);2022年重庆市级财政专项基础科研项目(NKY-2022AC005)。

摘　　要：为加快抗疫病加工型辣椒细胞质雄性不育(cytoplasmic male sterility,CMS)恢复系的创制,该试验以单生、长果自交系481-4-10为母本,以抗疫病的恢复系939-1和1021(1)-1为父本配制杂交组合,采用花药培养技术将抗疫病恢复系的Rf基因和抗疫病基因导入单生、长果自交系中,并利用分子标记辅助选择(molecular marker-assisted selection,MAS)技术鉴定DH系(Double Haploid line)的Rf基因以及抗病性,进一步用室内苗期抗性鉴定的方法验证MAS筛选的含Rf DH系对疫病的抗性。结果表明:(1)花药培养的供体亲本(481-4-10×939-1)F_(1)诱导出22个胚状体,成苗后经倍性鉴定获得11个花培DH系;而由供体亲本[481-4-10×1021(1)-1]F_(1)获得9个DH系。(2)分子标记CRF-SCAR对花培DH系进行分子标记辅助选择验证结果表明,来自供体(481-4-10×939-1)F_(1)的DH系中有7个可扩增出870 bp的特异条带,占63.6%;而供体[481-4-10×1021(1)-1]F_(1)获得的DH系中则有8个能扩增出870 bp的特异条带,占88.9%。(3)分子标记FQ01/RQ01对DH系进行分子标记辅助选择筛选结果发现,来自供体(481-4-10×939-1)F_(1)的DH系有5个能扩增出717 bp的特异条带,占45.5%;而来自供体[481-4-10×1021(1)-1]F_(1)的DH系有4个可扩增出717 bp的特异条带,占44.4%。(4)MAS技术初步筛选到7个携带Rf的抗疫病DH系,分别命名为‘渝辣选3-2/3-3/3-5/7-1/7-5/7-6/7-9’;苗期抗性鉴定结果显示,7个DH系中5个DH系抗疫病,2个中抗疫病;农艺性状调查和测交实验表明,‘渝辣选3-2’和‘渝辣选7-1’是单生朝天椒、果实较长,味辣,均为CMS恢复系。该研究创制的2个DH系为利用辣椒CMS三系配套选育抗病新品种奠定了基础。In order to speed up the creation of cytoplasmic male sterility(CMS)restorer line with phytophthora blight resistance in processing type pepper,a hybrid combination was prepared with long fruit inbred line 481-4-10 as female parent and disease resistant restorer lines 939-1 and 1021(1)-1 as male parent.Rf gene and disease resistant gene were introduced into long fruit inbred lines by anther culture technology,molecular marker assisted selection(MAS)was used to preliminarily determine whether DH lines contain Rf gene and disease resistance.The disease resistance of DH lines was verified by seedling resistance identification.The results showed that:(1)twenty-two embryoids were induced by donor parent(481-4-10×939-1)F_(1),and eleven DH lines were obtained.9 DH lines were obtained from donor[481-4-10×1021(1)-1]F_(1).(2)The PCR results of molecular marker CRF-SCAR showed that seven DH lines obtained from donor(481-4-10×939-1)F_(1) could amplify 870 bp specific bands,accounting for 63.6%.However,eight DH lines from donor[481-4-10×1021(1)-1]F_(1) could also amplify 870 bp specific bands,accounting for 88.9%.(3)PCR results of FQ01/RQ01 molecular marker showed that five DH lines from donor(481-4-10×939-1)F_(1) could amplify 717 bp specific bands,accounting for 45.5%.Four DH lines from donor[481-4-10×1021(1)-1]F_(1) could amplify specific bands of 717 bp,accounting for 44.4%.(4)Seven phytophthora blight resistance DH lines carrying Rf gene were preliminarily screened by MAS technology.The identification results of seedling resistance showed that five DH lines were disease resistant and two were moderately disease resistant among the seven DH lines.The investigation of agronomic characters and the test crossing experiment showed that‘Yulaxuan 3-2’and‘Yulaxuan 7-1’were single pepper with longer fruit and spicy taste,and both were cytoplasmic male sterility(CMS)restorers.The two DH lines created in this study will lay a foundation for breeding new disease resistant processing pepper varieties with CMS three line

关 键 词：辣椒 细胞质雄性不育 恢复系 疫病 花药培养 双单倍体 

分 类 号：Q785[生物学—分子生物学] Q789[农业科学—蔬菜学] S641.3[农业科学—园艺学]