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园艺学报 ›› 2017, Vol. 44 ›› Issue (3): 463-474.doi: 10.16420/j.issn.0513-353x.2016-0867

• 研究论文 • 上一篇    下一篇

芥菜AGL18家族成员与开花整合子SOC1的互作分析

李朝闯1,马关鹏1,2,谢 婷1,3,陈 娇1,王志敏1,宋 明1,汤青林1,*   

  1. 1西南大学园艺园林学院,南方山地园艺学教育部重点实验室,重庆市蔬菜学重点实验室,重庆 400715;2贵州省园艺研究所,贵阳 550006;3毕节市农业技术推广站,贵州毕节 551700
  • 出版日期:2017-03-25 发布日期:2017-03-25

Analysis of Interactions Between AGL18 Family Members and Flowering Time Integrator Factor SOC1in Brassica juncea

LI Chaochuang1,MA Guanpeng1,2,XIE Ting1,3,CHEN Jiao1,WANG Zhimin1,SONG Ming1,and TANG Qinglin1,*   

  1. 1College of Horticulture and Landscape Architecture,Southwest University,Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education,Key Laboratory of Olericulture,Chongqing 400715,China;2Guizhou Province Horticulture Research Institute,Guiyang 550006,China;3Bijie Station for Popularizing Agricultural Technique,Bijie,Guizhou 551700,China
  • Online:2017-03-25 Published:2017-03-25

摘要:

为阐明芥菜开花抑制因子AGL18与开花整合子SOC1间的互作调控机制,在芥菜‘QJ’的开花期克隆了AGL18-1,幼苗期克隆了AGL18-2和AGL18-3,它们分别编码257、257和258个氨基酸,为AGL18家族的3个成员。序列比对表明,芥菜AGL18家族成员与十字花科芜菁和油菜同源性均高达90%。酵母双杂交和BiFC试验表明:芥菜AGL18-1、AGL18-2和AGL18-3蛋白与SOC1不会发生蛋白相互作用。酵母单杂交和Dual-Glo® Luciferase试验表明:AGL18-1、AGL18-2和AGL18-3中仅有花期AGL18-1蛋白与SOC1启动子间存在互作。为进一步筛选AGL18-1/SOC1的互作区域,分别截取了AGL18-1蛋白的M域和IKC域,发现仅M域与SOC1启动子存在相互作用,说明M域是介导花期AGL18-1蛋白与SOC1启动子互作的关键区域。这为深入研究AGL18与SOC1互作的分子机制及其对开花时间的调控奠定了基础。

关键词: 芥菜, AGL18, SOC1, 开花调控, 相互作用

Abstract:

AGAMOUS-LIKE 18(AGL18)is a key regulatory factor in flowering time control of Brassica juncea. In order to clarify the regulatory mechanism of AGL18 protein interact with SOC1 in flowering pathways,AGL18-1 gene from reproductive shoots as well as AGL18-2 and AGL18-3 from vegetative shoots in‘QJ’germplasm of Brassica juncea,were cloned respcetively. AGL18-1,AGL18-2 and AGL18-3were members of AGL18 family and respectively encoded 257,257 and 258 amino acids. Sequences analysis showed that they were more than 90% similar to Brassica rapa and Brassica napus. Both Yeast two hybrid and BiFC showed that SOC1 protein could not interact with AGL18-1,AGL18-2 or AGL18-3. Yeast one hybrid and Dual-Glo® Luciferase assays indicated that SOC1 promoter could interact with AGL18-1 but not AGL18-2 and AGL18-3. To screen the interaction regions of AGL18-1/SOC1,the M-domain and IKC-domain truncations of AGL18-1 protein were separated and found that only the M-domain truncation could interact with SOC1 promoter. It suggested that M-domain was the key region that mediated the interaction between AGL18-1 and SOC1 promoter. The work provided valuable information for in-depth studies on the molecular mechanisms of AGL18 protein with SOC1 and the flowering time in Brassica juncea.

Key words: Brassica juncea, AGL18, SOC1, flowering regulation, interaction