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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (06): 850-859.doi: 10.3724/SP.J.1006.2016.00850

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

Cloning and Expression Analysis of Anthocyanidin Synthase Gene BrcANS from Purple Non-heading Chinese Cabbage

XU Yu-Chao1,HOU Xi-Lin1,XU Wei-Wei1,SHEN Lu-Lu2,ZHANG Shi-Lin1,LIU Shi-Tuo1,HU Chun-Mei1,*   

  1. 1 State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China; 2 Agriculture Committee of Feixi County, Hefei 230001, China
  • Received:2015-11-11 Revised:2016-03-14 Online:2016-06-12 Published:2016-03-21
  • Contact: 胡春梅, E-mail: jjjhcm@njau.edu.cn
  • Supported by:

    The work was supported by the Independent Innovation Fund for Agricultural Science and Technology of Jiangsu Province [CX (15)1015], the Science-technology Support Program of Jiangsu Province (BE2013429).

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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 correlated 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 increasing of leaf age, the leaf color became shallow and the expression of BrcANS reduced. Meanwhile, the difference of expression 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, so the gene might regulate the formation of the purple color in leaves at transcriptional level.

Key words: Non-heading Chinese cabbage, Anthocyanidin synthase, Homology-based cloning, Sequence analysis, Total anthocyanin content /TAC, Gene expression

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