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利用hiTAIL-PCR技术扩增红麻cox1全长及侧翼序列,结果表明,在红麻不育系UG93A和保持系UG93B中分别获得长度为2 149和3 244bp的序列,包含cox1基因CDS全长及其5′和3′部分侧翼序列,通过分析得出不育系和保持系中cox1的CDS序列长度为1 602bp,编码533个氨基酸残基,分子量为58.31ku。BLAST序列比对发现,UG93A和UG93Bcox1的CDS序列虽然存在4个碱基差异,但推导的氨基酸序列一样,并且UG93A和UG93Bcox1两端侧翼序列结果一致。RNA Blot分析显示cox1基因在红麻UG93A、UG93B和F1(UG93A/992)的转录本大小基本相同,推测cox1基因不是导致红麻CMS(cytoplasmic male sterility)的直接因子。qRT-PCR结果表明,UG93A中cox1的表达量显著低于UG93B和F1(UG93A/992),推测cox1在红麻花粉发育能量代谢过程中有着重要的作用。
The full length and flanking sequences of cox1 were amplified by hiTAIL-PCR. The results showed that the sequences of 2 149 bp and 3 244 bp in length were obtained in UG93A and maintainer line UG93B, The 5 ’and 3’ flanking sequences of cox1 showed that the CDS sequence of cox1 was 1 602 bp in length, encoding 533 amino acid residues with a molecular weight of 58.31 ku. BLAST sequence alignment found that UG93A and UG93Bcox1 CDS sequence although there are 4 base differences, but the deduced amino acid sequence, UG93A and UG93Bcox1 two flanking sequence results. RNA Blot analysis revealed that the cox1 gene was essentially the same size in the kenaf UG93A, UG93B and F1 (UG93A / 992) transcripts, suggesting that the cox1 gene is not a direct factor in cytoplasmic male sterility. The results of qRT-PCR showed that the expression of cox1 in UG93A was significantly lower than that in UG93B and F1 (UG93A / 992), suggesting that cox1 plays an important role in the energy metabolism of kenaf pollen.