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本研究旨在对棉花来源的果胶甲酯酶基因进行克隆和功能分析。通过RACE技术克隆了2个棉花果胶甲酯酶基因的cDNA,命名为GhPME1和GhPME2。序列分析发现:GhPME1和GhPME2的最大开放阅读框分别为1 704和1 752bp,分别编码含有567和582个氨基酸的蛋白质。蛋白质结构预测显示这2个蛋白结构相似,包含PMEI和Pectinesterase结构域。RT-PCR和实时荧光定量PCR结果显示在陆地棉的纤维发育过程中,GhPME1和GhPME2的表达峰值分别在9和4DPA(开花后天数),然后依次降低。而在海岛棉纤维中,这2个基因的表达均呈现逐渐上升的趋势,表达峰值分别出现在开花后24和29DPA。GhPME1和GhPME2在海岛棉和陆地棉棉纤维中的表达差异初步证明该基因家族的功能可能与棉纤维品质相关。
The aim of this study was to clone and analyze the pectin methylesterase gene from cotton. The cDNA of two cotton pectin methylesterase genes was cloned by RACE technology and named as GhPME1 and GhPME2. Sequence analysis revealed that the largest open reading frames of GhPME1 and GhPME2 were 1 704 and 1 752 bp, encoding proteins containing 567 and 582 amino acids, respectively. Protein structure prediction showed that the two proteins were similar in structure and contained the PMEI and Pectinesterase domains. The results of RT-PCR and real-time fluorescence quantitative PCR showed that the expression peak of GhPME1 and GhPME2 were up to 9 and 4 DPA respectively (days after flowering) and then decreased in turn. In the island cotton fiber, the expression of these two genes showed a gradual upward trend, the peak expression appeared after flowering 24 and 29DPA. The differences in the expression of GhPME1 and GhPME2 between cotton island and upland cotton showed that the function of this gene family may be related to the quality of cotton fiber.