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cDNA-AFLP analysis on bolting or flowering of flowering Chinese cabbage and molecular characteristics of BrcuDFR-like/BrcuAXS gene

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Abstract

The molecular basis of flower bud differentiation in flowering Chinese cabbage (Brassica rapa L. ssp. Chinensis var. utilis Tsen et Lee) was studied in this work. Samples were taken from two varieties, the early-blooming “Youqin 49” and the late-blooming “Youqingtiancaixin 80”, at five different developmental stages and studied via cDNA-AFLP. Nineteen expression sequence tags (ESTs) associated with bolting or flowering were isolated and cloned. Blast results indicated that 15 ESTs were involved in the synthesis of anthocayanins, photosynthesis, signal transduction, and phytochrome production. Two ESTs had high similarity to hypothetical proteins with unknown function. Two other ESTs shared no similarity to any sequence in the NCBI database and potentially may be newly identified genes. The deduced amino acid sequences of EST amplified by primer A6T4 or A8T4 had high similarity to both dihydroflavonol reductase (DFR) and UDP-d-apiose/UDP-d-xylose synthase (AXS), thus was named BrcuDFR-like/BrcuAXS. Using the cDNA sequence, a putative BrcuDFR-like/BrcuAXS gene was cloned and characterized from flowering Chinese cabbage via rapid amplification of cDNA ends (RACE). The full-length cDNA has 1332 bp with an open frame of 919 bp which codes for a polypeptide of 313 amino acids. The corresponding genome sequence is 2,046 bp. Comparison of cDNA and its corresponding genomic sequence indicates that BrcuDFR-like/BrcuAXS contains 9 exons and 8 introns. The temporal expression patterns indicated the gene is more likely to encode the DFR protein, which catalyzes the synthesis of anthocayanins, than UDP-d-apiose/UDP-d-xylose synthase (AXS), which catalyzes the conversion of UDP-d-glucuronate to a mixture of UDP-d-apiose and UDP-d-xylose. Further work is needed to determine what role BrcuDFR-like/BrcuAXS plays during floral organ development.

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Abbreviations

cDNA-AFLP:

cDNA-amplified fragment length polymorphism

ORF:

Open reading frame

PCR:

Polymerase chain reaction

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcriptase polymerase chain reaction

TDF:

Transcript-derived fragment

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Acknowledgments

This study was supported by the Natural Science Foundation of Jiangxi Province, P. R. China (Grant No. 2010GQN0032) and the Department of Education of Jiangxi Province, P. R. China (Grant No. GJJ11084).

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Authors and Affiliations

  1. College of Agriculture, JiangXi Agricultural University, Nanchang, 330045, People’s Republic of China

    Xufeng Xiao, Jianjun Lei, Bihao Cao, Guoju Chen & Changming Chen

  2. College of Horticulture, South China Agricultural University, Guangzhou, 510642, People’s Republic of China

    Xufeng Xiao, Jianjun Lei, Bihao Cao, Guoju Chen & Changming Chen

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  1. Xufeng Xiao
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  2. Jianjun Lei
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  3. Bihao Cao
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  5. Changming Chen
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Corresponding author

Correspondence to Jianjun Lei.

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Xufeng Xiao and Bihao Cao contributed equally to this study.

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Xiao, X., Lei, J., Cao, B. et al. cDNA-AFLP analysis on bolting or flowering of flowering Chinese cabbage and molecular characteristics of BrcuDFR-like/BrcuAXS gene. Mol Biol Rep 39, 7525–7531 (2012). https://doi.org/10.1007/s11033-012-1586-z

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