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首页> 外文期刊>Plant Cell Reports >Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers.
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Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers.

机译：使用细胞遗传学，流式细胞仪和分子标记物检测棉花的体细胞克隆变异。

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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.
机译：本研究采用了两种棉花植株再生方案，即2，4-D和动蛋白激素组合以及IBA和动蛋白激素组合。选择了28种通过体细胞胚发生的胚发生细胞系和67种来自这些胚发生愈伤组织的再生植物，并将其用于随机扩增多态性DNA（RAPD），简单序列重复（SSR），染色体数目计数和流式细胞仪分析。评估了RAPD和SSR标记在检测棉花体细胞无性变异中的作用。进行了两项聚类分析，以树状图的形式表示激素组合与遗传变异之间的关系。两种基于DNA的技术都能够扩增所有细胞克隆和再生的小植株基因组，并且在含有2、4-D和激动素激素组合的培养类型中可以检测到相对较高的遗传变异。结果表明，2-4-D和动蛋白激素组合可以诱导较高的体细胞克隆变异，RAPD和SSR标记可用于通过体细胞胚发生检测再生棉花植株的体细胞克隆变异。染色体计数和流式细胞仪分析表明，除两个再生小植株（分别丢失了4和5个染色体）外，所有再生植物中的染色体数目和倍性水平几乎稳定，这意味着细胞学变化与RAPD和RAPD的发生频率无关。 SSR多态性。该结果也可能意味着具有染色体数目变化的细胞系难以再生。

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