红麻细胞质雄性不育系UG93A和保持系UG93B cox1的克隆和表达分析

Cloning and expression analysis of cox1 in cytoplasmic male sterility line UG93A and its maintainer line UG93B of kenaf

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。

Full-length CDS including its flanking sequence of cox1 cloned by hiTAIL-PCR.They were 2 149 and 3 244 bp in cytoplasmic male sterile line UG93 Aand its maintainer line UG93 B,respectively,both contained an 1 602 bp opening reading frame,encoding 533 AA residue with 58.31 ku molecular weight.Four single base variations were found through BLAST analysis,but the deduced amino acid sequence were the same as well as their flanking sequences.RNA blotting results suggested that there was no size difference of mRNA among UG93 A,UG93Band F1（UG93A/992）.qRT-PCR analysis revealed that the expression level of cox1 was reduced significantly in UG93 Acompared to UG93 Band F1（UG93A/992）,indicating that cox1 might play an important role in energetic metabolism during anther development of kenaf.