论文部分内容阅读
[目的]WOX9基因在植物的胚胎发育过程中发挥一定作用。[方法]根据已公布的雷蒙德氏棉WOX9基因序列的CDS设计一对引物,从海岛棉‘新海16’中克隆一个同源基因GbWOX9。通过实时荧光定量分析其在棉花体细胞胚不同阶段的表达模式。[结果]获得了1 038 bp的GbWOX9基因的开放阅读框(ORF),编码345个氨基酸,预测蛋白分子量约为38 264.54 k Da,等电点为6.7。通过结构域分析,发现GbWOX9中含有一个由65个氨基酸组成的保守同源异型结构域。系统进化树分析结果表明GbWOX9与亚洲棉Ga WOX9亲缘关系较近。亚细胞定位预测该基因可能定位于细胞核。qRT-PCR表明,GbWOX9基因的表达量在‘新海16’胚胎发育阶段的球形胚>鱼雷胚>心形胚>胚性愈伤。[结论]根据相对表达量数值估算,GbWOX9在球形胚的表达量是鱼雷胚的1.3倍,是心形胚的3.1倍,是胚性愈伤的5.2倍。故该基因在‘新海16’体细胞胚阶段的球形胚中表达量最高。
[Objective] The gene of WOX9 plays a role in the embryonic development of plants. [Method] A pair of primers was designed based on the published CDS of WOX9 gene sequence of cotton germplasm, and a homologous gene GbWOX9 was cloned from island cotton ’Xinhai16’. Through real-time fluorescence quantitative analysis of its expression patterns in different stages of cotton somatic embryos. [Result] The open reading frame (ORF) of GbWOX9 gene of 1 038 bp was obtained and encoded 345 amino acids. The predicted molecular weight of GbWOX9 gene was 38 264.54 kDa and the isoelectric point was 6.7. By domain analysis, GbWOX9 was found to contain a conserved homeodomain consisting of 65 amino acids. Phylogenetic tree analysis showed that GbWOX9 was closely related to Ga WOX9 in Asian cotton. Subcellular localization predicts that the gene may localize to the nucleus. qRT-PCR showed that the expression level of GbWOX9 gene was in the embryonic callus> globular embryo> torpedo> cardiac embryo> embryogenic callus. [Conclusion] The expression of GbWOX9 in spherical embryos was 1.3 times that of torpedo embryos, 3.1 times of that of heart embryos and 5.2 times that of embryogenic callus according to the relative expression value. Therefore, the gene was expressed highest in the globular embryos at the stage of Xinhai 16 ’somatic embryogenesis.