酸枣愈伤组织转化体系构建及在ZjBRC1调控ZjYUCCA表达中的应用  

Optimization of Jujube Callus Transformation System and Application of ZjBRC1 in Regulating ZjYUCCA Expression

作　　者：李亚梅 马福利 张山奇 黄锦秋 陈梦婷 周军永[1,2] 孙其宝[2] 孙俊 LI Yamei;MA Fuli;ZHANG Shanqi;HUANG Jinqiu;CHEN Mengting;ZHOU Junyong;SUN Qibao;SUN Jun(College of Horticulture,Anhui Agricultural University,Hefei 230036,China;Horticulture Research Institute,Anhui Academy of Agricultural Science,Hefei 230031,China;State Key Laboratory of Tea Plant Biology and Utilization,Hefei 230036,China)

机构地区：[1]安徽农业大学园艺学院,合肥230036 [2]安徽省农业科学院园艺研究所,合肥230031 [3]茶树生物学与资源利用国家重点实验室,合肥230036

出　　处：《园艺学报》2022年第4期749-757,共9页Acta Horticulturae Sinica

基　　金：安徽高校自然科学研究项目(KJ2019A0186);安徽省重点研究与开发计划项目(202004a06020008);国家自然科学基金项目(31971687)。

摘　　要：以来源于同一株酸枣实生苗的茎段、叶片、子叶和胚轴等组织为外植体试材,筛选到适合子叶、胚轴和叶片诱导愈伤组织的培养基为MS+1.0 mg·L^(-1)2,4-D+0.4 mg·L^(-1)TDZ;利用农杆菌介导法,当浸染液浓度OD_(600)为0.6~0.8、浸染时间为30 min时,酸枣叶片、子叶、胚轴和茎段愈伤组织遗传转化率分别为31.8%、25.6%、24.5%和23.8%。利用叶片诱导的愈伤组织转化体系,获得了调控分枝发育关键因子ZjBRC1融合表达嵌合抑制子SRDX和35S::GFP空载体对照的转基因愈伤组织。荧光定量qRT-PCR分析显示,与35S::GFP转基因愈伤组织相比,ZjBRC1-SRDX下调生长素合成基因ZjYUCCA7/10-3/10-4的表达,而上调ZjYUCCA2/4/6表达。综上,本研究中建立和优化了枣愈伤组织诱导和转化体系。The stem segment,leaves,cotyledon and hypocotyls from the same sour jujube(Ziziphus jujuba Mill.var.spinosa)were used as the explants to induce callus.The optimum callus induction was obtained from cotyledon,hypocotyls and leaves in MS media supplemented with 1.0 mg·L^(-1)2,4-dichlorophenoxyacetic acid(2,4-D)and 0.4 mg·L^(-1)thidiazuron(TDZ).Furthermore,the callus transformation efficiency was 31.8%in leaves,25.6%in cotyledons,24.5%in hypocotyls and 23.8%in stem segments,respectively,after inoculating callus in the Agrobacterium infection solution with OD_(600)=0.6-0.8 for 30 min.In addition,ZjBRC1,which was a homologue of CYC/TB1 TCP transcription factor BRC1 and the key repressor to control shoot branching,were fused to the N terminus of SRDX repressor domain.35S::GFP and 35S::ZjBRC1-SRDX transgenic calli were obtained via the leaf-induced callus transformation system.qRT-PCR analysis showed that auxin biosynthetic genes ZjYUCCA7/10-3/10-4 were down-regulated while ZjYUCCA2/4/6 were up-regulated in the 35S::ZjBRC1-SRDX transgenic calli.

关 键 词：枣 愈伤组织 遗传转化 ZjBRC1 基因表达 

分 类 号：S665.1[农业科学—果树学]