陆地棉GhSPL3基因的克隆、亚细胞定位及表达分析

Cloning,Subcellular Localization and Expression Analysis of GhSPL3 Gene in Gossypium hirsutum L.

利用生物信息学结合RT-PCR技术从陆地棉中克隆出SPL转录因子,命名为GhSPL3,在GenBank的登录号为JN795132。该基因包含一个426 bp的ORF(开放阅读框),推测编码141个氨基酸的多肽。生物信息学分析表明GhSPL3包含一个典型的SBP结构域和一个核定位信号;进化树分析发现,GhSPL3与AtSPL3聚为一组,推测棉花GhSPL3和拟南芥AtSPL3在结构和功能上可能有着一定的相似性。亚细胞定位表明,GhSPL3定位于细胞核中,荧光定量RT-PCR结果表明,GhSPL3基因在棉花各组织中都有表达,但表达量不同。在花中表达量最高,其次是在顶芽和茎中的表达量,在根和叶中表达量较低。通过分析GhSPL3在顶芽的不同发育时期的表达量发现,GhSPL3在三片真叶展平时的顶芽中表达量最高,推测GhSPL3可能在花芽的分化、生长阶段的转变和花器官的形成上起着重要作用。

GhSPL3, an SPL transcription factor, was cloned from upland cotton by bioinformation and RT-PCR technology. The sequence accession number is JN795132 in GenBank. The ORF length of GhSPL3 is 426 bp which encodes 141 amino acid residues. Bioinformatics analysis showed that GhSPL3 contains a typical SBP structure domain and a nuclear localization signal; Evolutionary tree analysis revealed that GhSPL3 and AtSPL3 were clustered in same group, thus GhSPL3 and AtSPL3 might have similar structure and function. Subcellular localization results showed that the signal of GhSPL3 was localized in the nucleus. Quantitative RT-PCR results showed that GhSPL3 expressed in all tissues, but the expression levels were different. The expression of GhSPL3 in the flowers was the highest, and the lower was that in shoot apexes and in stem, followed by that in the root and in the leaves. Among different development stages of shoot apex, the expression of GhSPL3 was the highest at the third true leaf expansion stage in shoot apex. The result of Quantitative RT-PCR indicated that GhSPL3 might play an important role in bud differentiation, the transition of the growth phase and flower formation.