cDNA-AFLP Analysis on Differentially Expressed Genes Associated with Bolting in Chinese Cabbage (Brassica rapa ssp. pekinensis)-Cabbage (Brassica oleracea var. capitata) Translocation Lines of AT4 Series
Abstract：Bolting and flowering time are important agronomic characters for Chinese cabbage (Brassica rapa ssp. pekinensis), in which the yield and quality of harvested product are influenced by premature bolting. Chinese cabbage-cabbage (Brassica oleracea var. capitata) translocation lines of AT4 series added No. 4 chromosome fragments from cabbage in Chinese cabbage background were used as materials in this experiment. Commercial Chinese cabbage varieties Hanchun58, Yangchun for late bolting and Beijingxiaoza56 were used as control genotypes for bolting identification. All 45 late bolting lines and 15 early bolting lines were identified by phenotypic observation. In late bolting group, all 36 lines were extremely late and 9 lines were late. In early bolting group, 7 lines were early and 8 lines were extremely early. Late bolting lines and early bolting lines with 4 different vernalization controls were displayed by cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis. A total of 126 differential expressed transcript-derived fragments (TDFs) were identified, including the presence of bands in late bolting lines and without bands in early bolting lines, the presence of bands in early bolting lines and without bands in late bolting lines, expression increased or decreased with vernalization time increased in late bolting lines, expression increased or decreased with vernalization time increased in early bolting lines. All 74 TDFs were obtained by sequenced. BLAST and alignments showed that 61 TDFs shared the highest levels of similarity with homologous sequence, including 41 TDFs shared the highest levels of similarity with genes of known function and 20 TDFs with genes of unknown function, and 13 TDFs with novel expressed sequence. The TDFs of known function were involved in genes encoding enzymes working in metabolism, energy metabolism, cellular transport, signal transduction, regulation of transcription, DNA modification, cell cycle, etc. Among the 61 TDFs shared the highest levels of similarity with homologous sequence, 5 of them shared the highest levels of similarity with genes of cabbage, 2 of them shared the highest levels of similarity with genes of Penaeus monodon, 1 of them shared the highest levels of similarity with genes of Anas platyrhynchos, 1 of them shared the highest levels of similarity with genes of uncultured cyanobacterium, and 52 of them shared the highest levels of similarity with genes of Chinese cabbage. The acquisition of late bolting Chinese cabbage-cabbage translocation lines will provide useful materials for breeding new varieties program. The difference in expression of TDFs between late and early bolting lines will lay the foundation of understanding key genes and their regulation mechanism of bolting and flowering in Chinese cabbage.
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