作物学报2016，Vol.42Issue(6)：850-859,10.DOI:10.3724/SP.J.1006.2016.00850
紫色不结球白菜花色苷合酶基因BrcANS的克隆与表达分析
Cloning and Expression Analysis of Anthocyanidin Synthase Gene BrcANS from Purple Non-heading Chinese Cabbage
摘要
Abstract
Purple non-heading Chinese cabbage cultivar NJZX1-3, its green leaf mutant line NJZX1-0, and their progeny F2:NJZX2-1 and NJZX2-2 were used to study the function of anthocyanidin synthase gene in the anthocyanin biosynthesis of non-heading Chinese cabbage leaf. Homology-based cloning was used and anthocyanidin synthase gene was respectively cloned from two cultivars (NJZX1-3 and NJZX1-0). The gene nucleotides and amino acids sequences found in the two materials and Chinese cabbage were exactly the same, with a length of 1077 bp and encoding a peptide with 358 residues. Furthermore, a 2OG-Fe(II) dioxygenase super family domain was found in the amino acid sequence from the 211th to the 307th amino acids and the gene was named as BrcANS. The homology between BrcANS protein and BjANS protein was up to 99%, in accordance with the close relationship between them. Their total anthocyanin content (TAC) was consistent with the degree of purple in fresh leaves of the four materials, of which total anthocyanin content in cultivar NJZX1-3 leaves was the highest, up to 80.15±5.74 mg 100 g–1 FW. Simultaneously, the expression level of BrcANS (NJZX1-0<NJZX2-1<NJZX2-2<NJZX1-3) was positively corre-lated with the increasing trend of TAC. The mRNA of BrcANS exhibited tissue-specific expression in both materials, showing high level in leaves and lower level in other organs. In addition, the expression of two materials was significantly different, indicating that the expression of BrcANS in cultivar NJZX1-3 leaves was obviously higher than that in mutant line NJZX1-0. With the in-creasing of leaf age, the leaf color became shallow and the expression of BrcANS reduced. Meanwhile, the difference of expres-sion between NJZX1-3 and NJZX1-0 decreased significantly. These results indicated that BrcANS gene is one of the key genes in the anthocyanin biosynthesis of non-heading Chinese cabbage leaf, and its expression level is directly related to the purple color of leaves, thus the gene might regulate the formation of the purple color in leaves at transcriptional level.关键词
不结球白菜/花色苷合酶/同源克隆/序列分析/总花色苷含量/基因表达Key words
Non-heading Chinese cabbage/Anthocyanidin synthase/Homology-based cloning/Sequence analysis/Total anthocyanin content/Gene expression引用本文复制引用
许玉超,侯喜林,徐玮玮,沈露露,张仕林,刘世拓,胡春梅..紫色不结球白菜花色苷合酶基因BrcANS的克隆与表达分析[J].作物学报,2016,42(6):850-859,10.基金项目
本研究由江苏省农业科技自主创新项目[CX(15)1015]和江苏省科技支撑计划项目(BE2013429)资助。The study was supported by the Independent Innovation Fund for Agricultural Science and Technology of Jiangsu Province [CX(15)1015], and the Science-technology Support Program of Jiangsu Province (BE2013429) （15）
