Developing Exon-Targeted Intron-Exon Splicing Junction Marker and Establishing Genetic Map of Upland Cotton

Q3 Agricultural and Biological Sciences Acta Agronomica Sinica Pub Date : 2011-01-01 DOI:10.1016/S1875-2780(11)60002-3
Gang LIN , Ke ZHANG , Jian ZHANG, Zhonghua TENG, Zheng-Sheng ZHANG
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引用次数: 3

Abstract

The objective of this study was to develop new molecular markers and locate them into the existing genetic map of upland cotton (Gossypium hirsutum L.). Exon-targeted intron-exon splice junction (ET-ISJ) maker primers were designed according to the conserved sequences of intron-exon splicing junction. A total of 1280 ET-ISJ primer combinations were used to screen polymorphism between upland cotton cultivars Yumian 1 and T586. Sixty-nine primer combinations were polymorphic, which accounted for 5.4% of the total primer combinations. A mapping population of F2:7 from Yumian 1 × T586 were tested using the 69 ET-ISJ markers, and 70 ET-ISJ loci were obtained. Linkage analysis was conducted using the 70 ET-ISJ loci as well as 523 simple sequence repeat (SSR), 59 intron-tageted intron-exon splice junction (IT-ISJ), 29 sequence-related amplified polymorphism (SRAP), and 8 morphological loci. The linkage map obtained was composed of 59 linkage groups and 673 loci including 68 ET-ISJ, 510 SSR, 58 IT-ISJ, 29 SRAP, and 8 morphological loci. This linkage map covered 3216.7 cM (72.3%) of cotton genome with an average interval of 4.8 cM between 2 adjacent markers. Sixty-eight ET-ISJ loci were located on 20 chromosomes. The present study showed that ET-ISJ markers are relatively stable, highly polymorphic, and effective in constructing genetic map of upland cotton.

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开发外显子定位的内含子-外显子剪接连接标记并建立陆地棉遗传图谱
本研究的目的是开发新的分子标记,并将其定位到陆地棉(Gossypium hirsutum L.)现有遗传图谱中。根据内含子-外显子剪接连接的保守序列设计了外显子靶向内含子-外显子剪接连接(ET-ISJ)引物。利用1280个ET-ISJ引物组合对育棉1号和T586间的多态性进行了筛选。69个引物组合具有多态性,占引物组合总数的5.4%。利用69个ET-ISJ标记对玉棉1 × T586的F2:7定位群体进行了检测,获得了70个ET-ISJ位点。利用70个ET-ISJ位点、523个简单序列重复(SSR)、59个内含子标记的内含子-外显子剪接连接(IT-ISJ)、29个序列相关扩增多态性(SRAP)和8个形态位点进行连锁分析。得到的连锁图谱由59个连锁群和673个位点组成,其中ET-ISJ位点68个,SSR位点510个,IT-ISJ位点58个,SRAP位点29个,形态学位点8个。该图谱覆盖棉花基因组3216.7 cM(72.3%),相邻标记间平均间隔4.8 cM。68个ET-ISJ位点位于20条染色体上。本研究表明,ET-ISJ标记相对稳定,具有高度多态性,是构建陆地棉遗传图谱的有效方法。
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30 weeks
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