关键词: 甘蓝型油菜, cDNA-SRAP, 群体分离分析法, 差异显示

Abstract: The yellow-seed and black-seed in Brassica napus L. have different characters and different mechanisms in the seed color formation. The objective of this research was to explore the molecular mechanism of the differential expression between yellow and black immature seeds and demonstrate the feasibility of cDNA-SRAP for differential display. On the basis of the different lines with yellow and black seed-coat from recombinant inbred lines (RILs) which had self-pollinated for 7 generations, the yellow and black immature seed RNA bulked pools were constructed. SRAP was used to analysis the different cDNA pools transcribed from RNA pools. Totally, 2 100 amplifiedbands were obtained with 996 pairs of SRAP primer and the repeated bands were 65.2%. Twelve steady differential fragments with length of 100–300 bp between yellow and black immature seeds were acquired. Seven stable differential fragments were sorted out, cloned and sequenced. Sequence analysis revealed that only two of the fragments had significant homologous nucleotide sequence with Arabidopsis thaliana, one was similar to NAD+ADP-ribosyl transferase and the other was peptide transporter protein 3. The results indicated that cDNA pools with SRAP marker could be used for differential display on Brassica napus, and the two fragments may be related to the seed coat color expression of Brassica napus.

Key words: Brassica napus L., cDNA-SRAP, Bulked segregant analysis (BSA), Differential display