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Cloning and Expression Analysis of Wheat Cytokinin Oxidase/Dehydrogenase Gene TaCKX3

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Abstract

Cytokinin oxidase/dehydrogenase plays an important role in regulating plant growth and development. A novel gene of TaCKX3 was cloned from wheat by specific primers designed according to the Triticeae Full-Length CDS Database and was proved to be located on chromosome 7B by analyzing the nulli-tetrasomic lines of “Chinese Spring.” The genomic coding sequence was interrupted by three introns, which were 103, 421, and 458 bp, successively. The complementary DNA sequence of TaCKX3 is 60% identical to that of TaCKX1, and the homology of their deduced proteins is even low. The putative TaCKX3 protein shows highly homology with ZmCKX10 but not with other known cytokinin oxidase/dehydrogenase. However, it contains conserved motifs as other cytokinin oxidase/dehydrogenase, such as flavin adenosine dinucleotide binding domain and cytokinin binding domain. Consistent with ZmCKX10 and AtCKX7, nor the putative TaCKX3 has signal peptide at N terminus, which means that TaCKX3 functions in cytoplasm. Quantitive polymerase chain reaction analysis indicated that the expression of TaCKX3 gene is significantly up-regulated in germinating embryos treated by 6-BA and slightly up-regulated by NaCl or PEG-6000.

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Abbreviations

BLAST:

Basic Local Alignment Search Tool

cDNA:

complementary DNA

CKs:

cytokinins

CKX:

cytokinin oxidase/dehydrogenase

DNA:

deoxyribonucleic acid

FAD:

flavin adenosine dinucleotide

MW:

molecular weight

NT:

nulli-tetrasomic

PCR:

polymerase chain reaction

pI :

isoelectric point

RNA:

ribonucleic acid

RT-PCR:

reverse transcription polymerase chain reaction

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Acknowledgments

The authors thank Dr. Bin Liu (Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles) for the critical review of the manuscript. They also thank Prof. Bin Wang (Institute of Genetics and Developmental Biology, Chinese Academy of Science) for sending us seeds of nulli-tetrasomic lines of “Chinese Spring.” This project was supported by the Scientific Research Award Foundation for Outstanding Young and Middle-Aged Scientists of Shandong Province of China (BS2009NY037) and the International Cooperation Program for excellent Lecture of 2009 by Shandong Provincial Education Department, P.R. China.

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Authors and Affiliations

  1. State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Daizong Street 61, Taian, 271018, China

    Xin Ma, De-Shun Feng, Hong-Gang Wang, Xing-Feng Li & Ling-Rang Kong

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  1. Xin Ma
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  2. De-Shun Feng
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  3. Hong-Gang Wang
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  4. Xing-Feng Li
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  5. Ling-Rang Kong
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Corresponding author

Correspondence to De-Shun Feng.

Additional information

Sequence data of TaCKX3 from this article have been deposited at GenBank under accession number GQ925404.

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Ma, X., Feng, DS., Wang, HG. et al. Cloning and Expression Analysis of Wheat Cytokinin Oxidase/Dehydrogenase Gene TaCKX3 . Plant Mol Biol Rep 29, 98–105 (2011). https://doi.org/10.1007/s11105-010-0209-x

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  • DOI: https://doi.org/10.1007/s11105-010-0209-x

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