Expression of Genes Associated with Bolting in Chinese Cabbage (Brassica rapa ssp. pekinensis)-Cabbage (B. oleracea var. capitata) Translocation Lines AT4 Series
Abstract：Premature bolting will cause serious losses to the production of Chinese cabbage (Brassica rapa ssp. pekinensis). The molecular mechanism involved in bolting and flowering is important for the improvement of late bolting Chinese cabbage varieties. The late and early Chinese cabbage-cabbage translocation lines of AT4 series added No. 4 chromosome fragments from cabbage (Brassica oleracea var. capitata) under Chinese cabbage background were used as materials in this experiment, in which a total of 8 differentially expressed transcript-derived fragments (TDF) between late and early bolting translocation lines were screened by cDNA-amplified fragment length polymorphism (cDNA-AFLP). After being sequenced and BLAST, homologous alignment and function analysis indicated that 8 TDFs included 3 types: transcription translation, signal transduction and membrane transport category. In function prediction of TDFs, cellular signal transduction and material transport, material and energy metabolism, stress response, gene regulatory and expression and cell cycle were involved. In order to confirm whether and/or how genes expression changed and to evaluate the relationship between time of bolting and TDFs expression level, the 8 TDFs were further analyzed by qRT-PCR for 3 extra-late bolting Chinese cabbage-cabbage translocation lines, 3 extra-early bolting Chinese cabbage-cabbage translocation lines, one extra- late bolting inbred lines, one extra- early bolting inbred lines, one bolting resistance Chinese cabbage variety and one extra-early bolting Chinese cabbage variety. These materials by vernalization treatment at 0, 7, 14 and 21 d. The results showed that the expression peaks of extra-late bolting lines were significantly higher than that of the extra-early bolting lines in fragments of P65M58-11, P65M58-12 and P75M47-9, respectively. The expression level of P65M58-11 and P65M58-12 were the highest after vernalization treatment for 14 d. In extra-late bolting lines, the expression level of P75M47-9 were the highest in the 7 d of vernalization treatment. But the expression in extra-early bolting lines presented down-regulated trend from vernalization treatment initiation. The expression peaks of 4 TDFs, such as P77M58-9, P77M58-12, P63M49-8-1 and P63M61-4-2, were significantly lower in the extra-late bolting lines than that in extra-early bolting lines. The expression peak of P63M49-8-1 was at 7 d of vernalization treatments in the extra-early bolting lines. However, the expression of extra-late bolting lines showed gradually increased of 7~21 d vernalization treatments. The expression peaks of P77M58-9, P77M58-12 and P63M61-4-2 were at 14th d of vernalization treatments. The expression of P63M49-1-2 had no significant differences between the extra-late and extra-early bolting lines. In the extra-early bolting lines, the expression peak of P63M49-1-2 appeared at 7th d of vernalization treatments. However, in extra-late bolting lines the expression peak appeared at 21th d of vernalization treatments. In this research, we identified 8 genes with different expression level and expression patterns between extra-late bolting lines and extra-early bolting lines for 6 Chinese cabbage-cabbage translocation lines, 2 inbred lines and 2 Chinese cabbage varieties. These results will lay the foundation to further discover genes related to bolting and flowering, validate genes function, and obtain late bolting materials in Chinese cabbage.
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