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Molecular cloning and expression analysis of a cytokinin oxidase (DhCKX) gene in Dendrobium huoshanense

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Abstract

Cytokinin oxidase plays an important role in the cytokinins regulatory processes. A putative cytokinin oxidase gene, D endrobium h uoshanense cytokinin oxidase (DhCKX), was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) of total RNA isolated from shoot apices of D. huoshanense plantlets in the presence of 20 μmol l−1 6-benzylamino-purine (BAP). The full-length cDNA contains an open reading frame (ORF) encoding a putative protein of 537 amino acids with high homology to known CKX genes and with a predicted molecular mass of 60.383 kDa. Compared with the CKX sequences from other species, the DhCKX shows high identity with Dendrobium hybrid “Sonia”, 92.2% in nucleotide sequence and 89.9% in amino acid sequence, respectively. Based on the alignment of amino acid sequences, the phylogenetic tree of 15 CKX genes is constructed. Semi-quantitative RT-PCR analysis showed that the expression of the DhCKX gene was induced in roots, shoot apices and leaves, but not in stems, after treatment with 20 μmol l−1 BAP. The DhCKX gene showed maximum expression in shoot apices at day 12 during continuous treatment with 20 μmol l−1 BAP. Furthermore, CKX activity appears to play an important role in controlling the cytokinin levels and growth during shoot morphogenesis in D. huoshanense.

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Acknowledgements

The supports from Specialized Research Fund for the Doctoral Program of Higher Education of China (No. 20060359006) and the Research Fund for the Outstanding Doctor’s paper Program of Hefei University of Technology are gratefully acknowledged.

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Authors and Affiliations

  1. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei, 230009, People’s Republic of China

    Ying Wang, Jian-Ping Luo, Zhao-Jun Wei & Jing-Cheng Zhang

  2. College of Life Science, Hebei University, Baoding, 071002, People’s Republic of China

    Ying Wang

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  1. Ying Wang
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Correspondence to Jian-Ping Luo.

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Wang, Y., Luo, JP., Wei, ZJ. et al. Molecular cloning and expression analysis of a cytokinin oxidase (DhCKX) gene in Dendrobium huoshanense . Mol Biol Rep 36, 1331–1338 (2009). https://doi.org/10.1007/s11033-008-9316-2

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  • DOI: https://doi.org/10.1007/s11033-008-9316-2

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