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Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers

  • Cell Biology and Morphogenesis
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Abstract

Two protocols of plant regeneration for cotton were adopted in this study, namely, 2, 4-D and kinetin hormone combination and IBA and kinetin hormone combination. Twenty-eight embryogenic cell lines via somatic embryogenesis and 67 regenerated plants from these embryogenic calli were selected and used for random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), chromosomal number counting, and flow cytometric analysis. The roles of RAPD and SSR markers in detecting somaclonal variation of cotton (Gossypium hirsutum L.) were evaluated. Two cluster analyses were performed to express, in the form of dendrograms, the relationships among the hormone combinations and the genetic variability. Both DNA-based techniques were able to amplify all of the cell clones and regenerated plantlets genomes and relative higher genetic variation could be detected in the culture type with 2, 4-D and kinetin hormone combination. The result suggested that 2, 4-D and kinetin hormone combination could induce relative high somaclonal variation and RAPD and SSR markers are useful in detecting somaclonal variation of regenerated cotton plants via somatic embryogenesis. Chromosome number counting and flow cytometry analysis revealed that the number of chromosomes and ploidy levels were nearly stable in all regenerated plants except two regenerated plantlets (lost 4 and 5 chromosomes, respectively) which meant that cytological changes were not correlated with the frequency of RAPD and SSR polymorphisms. This result also might mean that the cell lines with variation of chromosome numbers were difficult to regenerate plants.

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Acknowledgments

The funds supported by National Natural Science Foundation of China (No. 30771368), National High-Tech program (20060AA00105) and China Postdoctoral Science Foundation (20070410949) were appreciated. We also thank Prof. V. Ravishankar Rai, Department of Microbiology, University of Mysore, India for critically evaluating this manuscript.

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  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, 430070, Wuhan, Hubei, China

    Shuangxia Jin, Ramesh Mushke, Huaguo Zhu, Lili Tu, Zhongxu Lin, Yanxin Zhang & Xianlong Zhang

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  1. Shuangxia Jin
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  2. Ramesh Mushke
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  3. Huaguo Zhu
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Correspondence to Xianlong Zhang.

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Communicated by Y. Lu.

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Jin, S., Mushke, R., Zhu, H. et al. Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers. Plant Cell Rep 27, 1303–1316 (2008). https://doi.org/10.1007/s00299-008-0557-2

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  • DOI: https://doi.org/10.1007/s00299-008-0557-2

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