Molecular Marker Development of Soft Rot Disease Defense-related Genes in Chinese Cabbage (Brassica rapa ssp. pekinensis)
CHEN Chang-Long, ZHAO Xue SUN, Wang-Wang, LI Xiao-Ying, TIAN Yu, LI Wei-Ran, XIE Hua*
Beijing Agro-Biotechnology Research Center/Beijing Academy of Agriculture and Forestry Sciences/Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing 100097, China
Abstract：Bacterial soft rot disease causes severe damage to Chinese cabbage (Brassica rapa ssp. pekinensis), and Pectobacterium carotovorum (Pc) is a major pathogen. Based on defense-related genes in Chinese cabbage to soft rot disease, molecular markers were developed in this study, which would be very important for molecular marker-assisted breeding and studying defense mechanism of Chinese cabbage to soft rot disease. Firstly, the ESTs (expressed sequence tags) database related to soft rot resistance of Chinese cabbage with independent property rights were analyzed by bioinformatics method, and 56 pairs of specific PCR primers were designed and used to test the polymorphism between resistant line (A19-2) and sensitive line (A32-2) of Chinese cabbage to soft rot disease. The genotypes of the F2 population (containing 142 Chinese cabbage individuals) generated by F1 self-breeding were recorded using the markers and the segregation analysis was conducted. All the 56 pairs of specific PCR primers obtained successful PCR-amplification. There were 30 markers which had polymorphism in the resistant and sensitive parents, including 11 EST-PCR markers and 19 EST-CAPS (cleaved amplified polymorphic sequence) markers, and 9 pairs could match with more than one restriction enzymes. There were 14 dominant markers and 16 co-dominant markers. In addition, the genotypes of 142 individuals of the F2 population were statistically analyzed using the above 30 marker loci. The result showed that 24 loci (80%) had the segregation ratio of 3∶1 or 1∶2∶1, and 6 loci (20%) had the genetic distortion (P<0.05) which was inconsistent with Mendelian ratio. This research developed defense-related EST-PCR and EST-CAPS molecular markers of Chinese cabbage to soft rot disease based on the ESTs resources of Chinese cabbage, which have certain significance in the study on utilization of ESTs resources, localization of defense-related genes and molecular marker-assisted breeding.
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