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作物学报 ›› 2015, Vol. 41 ›› Issue (06): 972-978.doi: 10.3724/SP.J.1006.2015.00972

• 研究简报 • 上一篇    下一篇

甘蓝BoExo70A1与BoSEC3、BoExo84蛋白相互作用的酵母双杂交检测

高启国,刘豫东,蒲全明,张林成,朱利泉,王小佳   

  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 400716; 2 西南大学农学与生物科技学院, 重庆 400716
  • 收稿日期:2014-11-12 修回日期:2015-03-19 出版日期:2015-06-12 网络出版日期:2015-04-07
  • 基金资助:

    本研究由国家重点基础研究发展计划(973计划)项目(2012CB113900), 国家自然科学基金项目(30900986), 中央高校基本科研业务费专项资金项目(XDJK2010B010)和重庆市自然基金重点项目(cstc2012jjB80010)资助。

Interaction Analysis of Brassica oleracea L. BoExo70A1 with BoSEC3 and BoExo84 Proteins by Using Yeast Two-hybird System

GAO Qi-Guo1**,LIU Yu-Dong1,**,PU Quan-Ming1,ZHANG Lin-Cheng1,ZHU Li-Quan2,WANG Xiao-Jia1   

  1. 1 College of Horticulture and Landscape Architecture, Southwest University / Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Chongqing 400716, China; 2 College of Agronomy and Biotechnology, Southwest University, Chongqing 400716, China
  • Received:2014-11-12 Revised:2015-03-19 Published:2015-06-12 Published online:2015-04-07

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摘要：

Exo70A1是芸薹属作物柱头接受亲和花粉的必需因子, 可能参与了干性柱头水分分泌和花粉水合过程。本文从自交不亲和甘蓝A1柱头cDNA中获取了BoExo70A1、BoSEC3、BoSEC10、BoSEC15和BoExo84基因的编码序列, 序列分析表明BoExo70A1、BoSEC3、BoSEC10、BoSEC15和BoExo84基因分别与AtExo70A1、AtSEC3A、AtSEC10、AtSEC15B和AtExo84B基因的cDNA序列高度同源, 其编码的5个蛋白均没有信号肽, BoSEC3蛋白含有一个典型的EF-hand钙离子结合结构域。将BoExo84蛋白拆分为2个片段(BoExo84-N和BoExo84-C), 并与BoSEC3、BoSEC10和BoSEC15蛋白编码序列一起分别亚克隆至pGADT7质粒, BoExo70A1蛋白编码序列亚克隆至pGBKT7质粒, 酵母双杂交检测结果表明BoExo70A1蛋白能与BoSEC3、BoExo84-N蛋白互作, 但不能与BoSEC10、BoSEC15蛋白互作, 这为深入探讨BoExo70A1蛋白乃至整个胞泌复合体在甘蓝干性柱头接受自花花粉排斥异花花粉过程中的作用机制提供了依据。

关键词: 甘蓝自交不亲和, BoExo70A1, 胞泌复合体, 相互作用

Abstract:

Exo70A1 is necessary for Brassica stigma accepting the compatible pollen, and may play an important role in regulating the movement of water from the dry stigma to the pollen grain and compatible pollen hydration. Here, the coding sequences of BoExo70A1, BoSEC3, BoSEC10, BoSEC15, and BoExo84 genes were amplified from the stigma cDNA of the highly self-incompatible Brassica oleracea L. line A1. Sequence analysis showed that the cDNA sequences of BoExo70A1, BoSEC3, BoSEC10, BoSEC15, and BoExo84 genes were highly homologous to those of AtExo70A1, AtSEC3A, AtSEC10, AtSEC15B, and AtExo84B genes respectively. No signal peptide was found in their deduced protein sequences. BoSEC3 protein contained an EF-hand calcium-binding domain. The BoExo84 protein was divided into two fragments, BoExo84-N and BoExo84-C, and then their encoding sequences were subcloned into pGADT7 vector together with those of BoSEC3, BoSEC10, and BoSEC15 proteins respectively, whereas the encoding sequence of BoExo70A1 protein was subcloned into pGBKT7 vector for analysis of the proteins interaction by yeast two-hybid system. The results showed that BoExo70A1 protein interacted with BoSEC3 or BoExo84-N protein, but not with BoSEC10 or BoSEC15 protein.

Key words: Brassica oleracea L. self-incompatibility, BoExo70A1, Exocyst complex, Interaction

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