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园艺学报 ›› 2020, Vol. 47 ›› Issue (1): 63-72.doi: 10.16420/j.issn.0513-353x.2019-0275

• 研究论文 • 上一篇    下一篇

二倍体马铃薯StBRC1a功能缺失突变体的获得及其功能分析

冯爽爽1,2,罗嘉翼1,2,朱曦鉴2,4,蒋继滨2,4,黄三文1,3,*,张金喆2,*   

  1. 1南京农业大学生命科学学院,南京 210095;2中国农业科学院蔬菜花卉研究所,北京 100081;3中国农业科学院深圳农业基因组研究所,广东深圳 518120;4云南师范大学生命科学学院,昆明 650500
  • 出版日期:2020-01-25 发布日期:2020-01-25
  • 基金资助:
    国家自然科学基金项目(31902027);国家高技术研究发展计划项目(2013AA102603)

Homozygous Mutant Construction and Function Analysis of TCP Transcription Factor StBRC1a in Diploid Potato

FENG Shuangshuang1,2,LUO Jiayi1,2,ZHU Xijian2,4,JIANG Jibin2,4,HUANG Sanwen1,3,*,and ZHANG Jinzhe2,*   

  1. 1College of Life Science,Nanjing Agricultural University,Nanjing 210095,China;2Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;3Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen,Guangdong 518120,China;4College of Life Science,Yunnan Normal University,Kunming 650500,China
  • Online:2020-01-25 Published:2020-01-25

摘要: 为研究二倍体马铃薯TCP转录因子基因StBRC1a的功能,在二倍体马铃薯Solanum tuberosum group phureja S15-65中对StBRC1a进行了克隆,发现StBRC1a有两个不同长度的转录本,分别命名为StBRC1aL与StBRC1aS。序列分析后发现,二者均具有TCP转录因子家族特有的由59个氨基酸组成的非典型碱性—螺旋—环—螺旋(bHLH)结构域及1个R结构域。通过构建系统演化树发现,StBRC1aL和StBRC1aS与番茄的SlBRC1a具有最近的亲缘关系,同属于CYC/TB1类TCP转录因子。组织特异性表达分析的结果表明StBRC1aL与StBRC1aS在腋芽中特异性的高表达。通过CRISPR/Cas9系统构建StBRC1a功能缺失的纯合突变体后发现,与野生型相比突变体植株表现为分枝显著增多的表型,说明StBRC1a对于二倍体马铃薯的分枝具有抑制作用。

关键词: 马铃薯, 二倍体, TCP转录因子, CRISPR/Cas9, 分枝

Abstract: Branching is crucial for plant shoot morphogenesis and crop yield. It’s reported that TCP transcription factor play an important role in regulating branching. However, related study in diploid potato remains largely unknown. In order to illustrate the function of TCP transcription factor StBRC1a in diploid potato,we cloned StBRC1a in diploid potato Solanum tuberosum group phureja S15-65 and found that StBRC1a had two transcripts with different length named StBRC1aL and StBRC1aS. Sequence analysis revealed that StBRC1aL and StBRC1aS both had a 59-amino acid basic helix–loop–helix(bHLH)motif and an R domain. By constructing phylogenetic tree,it’s found that StBRC1aL and StBRC1aS had the closest relationship with SlBRC1a,which indicated it belonged to CYC/TB1-like TCP transcription factor. Tissue specific expression analysis showed that StBRC1aL and StBRC1aS were highly expressed in axillary buds. We constructed a StBRC1a homozygous mutant by CRISPR/Cas9 system. Compared with wild type,the mutant showed significantly increased branching,which demonstrated that StBRC1a played an important role in branching for diploid potatoes.

Key words: potato, diploid, TCP transcription factor, CRISPR/Cas9, branching

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