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甘蓝eSRK重组体的构建及其与SCR的相互作用

Construction of eSRK Chimeras and Interaction between eSRK Chimeras and SCRs from Brassica oleracea L.
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摘要 SRK与SCR是甘蓝自交不亲和雌雄性决定因子,两者间相互作具有单倍型特异性。为了探讨HVI/II区域在SRK单倍型特异性及其与SCR互作中的作用,采用重组技术构建甘蓝不同单倍型eSRK(SRKE与SRKF)间的重组体eSRKE-1、eSRKE-2和eSRKE-3,用酵母双杂交系统3检测各eSRK重组体与SCR之间的相互作用。结果表明:SCRE能与eSRKE作用,而不能与eSRKF作用,说明eSRKE、eSRKF属于不同单倍型;SCRE与重组体eSRKE-1、eSRKE-2、eSRKE-3均不发生作用,HVI和HVII区域内差异的氨基酸位点共同参与了与SCR的作用;SCRF不能与eSRKE-1、eSRKE-2、eSRKE-3作用,替换HVI/II区域后并不能改变SRK的单倍型。 Self-incompatibility in Brassica is mediated by allele-specific interactions between stigma-expressed S-locus receptor kinase （SRK） and pollen coat-localized S-locus cysteinerich （SCR） ligands encoded by the S-locus haplotype. To identify amino acid fragments within the SRK extracellular domain （eSRK） that are required for ligand-selective activation, we constructed chi- meric eSRK between two S-locus haplotypes in Brassica oleracea, and identified the interaction between eSRK chimeras and SCRs by yeast two-hybrid system. The results showed that SRKE （not chimera） could interact with SURE, and SRKF could interact with SCRF. All of eSRK chimeras could not interact with SCRs. The hypervariable regions, HVI and HVII, were essential for specificity in the SRK-SCR interaction. However, eSRK chimeras could not interact with SURF, although they contained hyper- variable regions come from eSRKF, which should be related with the overall sequence or 3D conformation of the segments deter- mining SI specificity.
作者 韦静宜 高启国 任雪松 王小佳 李成琼 宋明
机构地区 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
出处 《作物学报》 CAS CSCD 北大核心 2012年第9期1631-1639,共9页 Acta Agronomica Sinica
基金 国家自然科学基金项目(30900986 31071802) 重庆市自然科学基金项目(2009BB1298) 教育部博士点基金项目(200806350006) 西南大学博士基金项目(SWUB2008042) 中央高校基本科研业务费专项(XDJK2010B010 XDJK2009C126)资助
关键词 自交不亲性 SRK SCR 酵母双杂交 Self-incompatibility SRK SCR Yeast two-hybrid system
分类号 S635 [农业科学—蔬菜学]
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参考文献32

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作物学报
2012年 第9期

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