The Adaptive AFLP-Pst I/Mse I Primer Combinations of Chinese Cabbage

doi:10.7668/hbnxb.2009.04.020

Abstract

Abstract: The PCR-based DNA fingerprinting technique AFLP is widely used in analysis of genetic diversity, the construction of linkage maps, gene mapping, molecular phylogeny and cultivar identification. In this study the clubroot resistant CR DH line, suscept ible inbred line 94SK of Chinese cabbage, the resistant and susceptible DNA bulks constructed from F2 individuals of CR DH line and 94SK were used to select the adaptive AFLP primer combinat ion. AFLP analysis was performed with 256 Pst I/ Mse I primer combinat ions.The result indicated that 237 combinations amplif ied a total of 13 079 AFLP fragments with an average of 55 fragments, the average of 1. 30 markers per 1 000 AFLP fragments were found to be linked to clubroot resistant gene; the amplified polymorphic fragments were 3 167 between the two parents, account for 24 percent of polymorphic frequency; the number of amplified fragments and the number of polymorphic fragments was in pos-i tive linear correlation; GC content of AFLP primer combinat ions significantly affects the number of amplified fragments and the number of polymorphic fragments, indicating negative linear correlat ion between them. Fifty-three primer combinations were found to amplify nuclear DNA of Chinese cabbage, properly. They amplified the fragments up to 4 381 with an average of the 83.Among them, 9 markers derived from 9 primer combinations were linked to the clubroot resistance gene.

Key words: Chinese cabbage, AFLP, Primer combination, GC content

CLC Number:

XU Lei, LI Hong-bo, LIM Yong-pyo, PIAO Zhong-yun. The Adaptive AFLP-Pst I/Mse I Primer Combinations of Chinese Cabbage[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(4): 96-101. doi: 10.7668/hbnxb.2009.04.020.

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