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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (06): 972-978.doi: 10.3724/SP.J.1006.2015.00972

• RESEARCH NOTES • Previous Articles     Next Articles

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 Online:2015-06-12 Published:2015-04-07

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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