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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (07): 1067-1074.doi: 10.3724/SP.J.1006.2010.01067

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Sequence Analysis of KAPP Gene in Brassica oleracea

WU Wui-Ru1,ZHU Li-Quan1*,LI Cheng-Qiong3,YANG Kun1,TANG Zhang-Lin1,REN Xue-Song3,WANG Xiao-Jia3*   

  1. 1 Plant Physiology and Biochemistry Laboratory of Southwest University, Chongqing 400716, China; 2 Chongqing Rapeseed Technology Research Center, Chongqing 400716, China; 3 Key Laboratory in Olericulture of Chongqing, Southwest University, Chongqing 400716, China
  • Received:2010-02-01 Revised:2010-04-19 Online:2010-07-12 Published:2010-05-20
  • Contact: ZHU Li-Quan,E-mail: zhuliquan@swu.edu.cn; wxj@swu.edu.cn E-mail:wwrrb518@hotmail.com; Tel: 13594001094

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Abstract: The gDNA and cdna fragments of KAPP gene were amplified from genomic DNA, bud Rna and leaf Rna in Brassica oleracea by PCR and RT-PCR and other molecular biology methods. We initially obtained a fragment of KAPP gDNA with length of 3 247 bp, a fragment of KAPP cDNA with length of 1 699 bp, as well as a bud cdna with length of 1 578 bp and a leaf cdna with length of 1 581 bp which were called KAPP2 cDNA of bud and KAPP2 cDNA of leaf respectively. Compared gDNA with cDNA of KAPP, we found that there were 11 introns in KAPP gene, and these introns all followed typical GU-AG rule. Between KAPP cDNA which we cloned and KAPP cDNA released there are six nucleotide differences, but they encode a same amino acid sequence. Sequences analysis of the KAPP2 cDNA which we cloned from bud cdna and leaf cdna in Brassica oleracea showed that they share 85.2% and 85.0% identity with the reported KAPP cDNA respectively.We also found that the nonsense mutation in 590 bp of KAPP2 of bud cDNA and 593 bp of KAPP2 of leaf cDNA led to the early appearance of termination codon, who’s Blast indicated that both of them shared more identity in Arabidopsis thaliana than in Brassica oleracea. Based on KAPP cDNA sequence of eight species released by NCBI and two sequences of KAPP2 cDNA cloned in this study, we constructed a phylogenetic tree of KAPP gene, which showed that the two KAPP2 sequences were in the same group with released KAPP cDNA in Brassica oleracea. Based on all the above analysis, as well as some view of the comparative mapping research, we speculated that KAPP gene may have more than one copy in the genome of Brassica oleracea,the KAPP2 sequence cloned in this study may be the other copies of KAPP gene. Moreover, they are likely to be the pseudogene which was inactivated by mutation during evolution process. It has an important significance for the deep research between KAPP and Self-incompatibility. The results will provide some new insights into the research of molecular mechanism in SI and molecular evolution in Brassica oleracea.

Key words: Self-incompatibility, Signal transduction, KAPP gene, Brassica oleracea

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