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

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 F_2:7 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.