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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (12): 2136-2144.doi: 10.3724/SP.J.1006.2011.02136

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Interaction between Two Self-incompatible Signal Elements, EXO70A1 and ARC1

YANG Kun1, ZHANG He-Cui1, Richard CONVERSE2, ZHU Li-Quan1,*, YANG Yong-Jun1, XUE Li-Yan1, LUO Bing1,CHANG Deng-Long1,GAO Qi-Guo2,WANG Xiao-Jia2   

  1. 1 College of Agronomy and Biotechnology, Southwest University, Chongqing 400716, China; 2 Department of Biology, University of Cincinnati, Cincinnati OH 45267-0524, USA; 3 Key Laboratory in Olericulture of Chongqing, Southwest University, Chongqing 400716, China
  • Received:2011-04-23 Revised:2011-07-25 Online:2011-12-12 Published:2011-09-29
  • Contact: 朱利泉, E-mail: zhuliquan@swu.edu.cn, Tel: 023-68250794

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Abstract: ARC1 and EXO70A1 are important signal elements of self-incompatibility in Brassica. To identify the interaction of ARC1-EXO70A1 during the course of SI, we cloned the coding sequences of ARC1 and EXO70A1 from Brassica napus L.and Brassica oleracea L. var. acephala. Sequence analysis showed that ARC1 consisted of 663 amino acids in Brassica oleracea and 661amino acids in Brassica napus, and there existed 45 amino acids difference between them. Sequence alignment showed that similarity positions and identity positions reached 95.9% and 93.9% between BoARC1 and BnARC1, whereas there existed only six-amino-acid difference between BoEXO70A1 and BnEXO70A1. The Similarity positions and identity positions reached 99.4% and 98.9% between BoEXO70A1 and BnEXO70A1 respectively. The homology of EXO70A1 was higher than that of ARC1. Yeast two-hybrid results indicated that the strong interaction existed between ARC1 and EXO70A1, and it could activate the expression of four reporter genes (ADE2, HIS3, AUR1-C,and MEL1) in diploid yeasts. However, low interaction existed between EXO70A1 and ARC1N with 316 amino acids deleted from C-terminal, and it only activated the expression of three reporter genes (ADE2, AUR1-C, and MEL1). This provides an insight that the interface of interaction between ARC1 and EXO70A1 may not consist of the domains of arm repeats in ARC1. N-terminal domains of ARC1 play an essential role in the interaction of ARC1-EXO70A1. The influences of the differences in amino-acid composition between BoARC1 and BnARC1 on the interaction of ARC1-EXO70A1 couldn’t be detected by means of yeast two-hybrid system, which probably resulted from that the binding interface between ARC1 and EXO70A1 was not altered by sequence difference of two proteins in two Brassica species.

Key words: Self-incompatibility, Signal element, Protein interaction, Yeast two-hybrid

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