Kim Hyoun-Joung, Lee Heung-Ryul, Han Jung-Heon, Yeom Seon-In, Harn Chee-Hark, Kim Byung-Dong
Department of Plant Science, Seoul National University, Seoul 151-921, Korea.
Mol Cells. 2008 Apr 30;25(2):196-204. Epub 2008 Mar 28.
Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsungcho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to F2 genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.
尽管人们越来越意识到WRKY基因在植物防御信号传导中的重要性，但这些基因在辣椒基因组中的位置尚未确定。为了开发基于WRKY的标记，从番茄和辣椒基因WRKY结构域内DNA结合基序的保守序列推导引物序列。这些引物来自三个WRKY组保守序列的上游和下游部分。测试了每个WRKY组的六种引物组合在作图亲本辣椒“CM334”和辣椒“Chilsungcho”之间的多态性。由WRKY第II组基因推导的引物对扩增的DNA片段显示出高水平的多态性。使用32对引物扩增WRKY第II组基因WRKY结构域的上游和下游部分，检测到60条多态性条带。从WRKY第II组基因下游部分的引物对未检测到多态性。这些引物中的一半用于F2基因分型以构建连锁图谱。41个标记中的30个均匀分布在28个连锁群中的20个上，没有聚类。该连锁图谱还由199个AFLP和26个SSR标记组成。这种基于WRKY的标记系统是一种为植物基因家族生成序列特异性标记的快速简单方法。
