荧光实时定量PCR研究GA负调控因子基因在棉花纤维不同发育时期的表达模式

摘要

棉花纤维发育经历起始期、延伸期、次级细胞壁加厚期、脱水期共4个发育时期.为了了解GA负调控因子基因在整个纤维发育时期的调节作用,采用敏感的实时定量PCR对GA负调控因子基因GhRGL在棉花纤维不同发育时期中的转录水平进行了定量研究,通过提取来源于不同纤维发育阶段[包括-3~0 DPA(开花后)胚珠, 3 DPA胚珠,5 DPA胚珠,10 DPA胚珠,15 DPA纤维,25 DPA纤维]的样品总RNA,探讨不同发育时期GhRGL基因的表达模式.结果显示,GhRGL基因在所有被选的样品中都有表达,其中在5 DPA胚珠和10 DPA胚珠中转录水平较高,在10 DPA胚珠中表达水平最高,在-3~0 DPA胚珠、3 DPA胚珠、15 DPA纤维、25 DPA纤维中的表达水平较低,表明GhRGL基因在纤维伸长期表达水平较高,暗示GhRGL可能对纤维伸长起着重要作用.%Cotton fibers, which are highly elongated epidermal cells that grow from the seed integument, undergo a developmental process that includes cell fate determination,initiation,elongation,specialization, and finally, programmed cell death. Transcript levels of GhRGL at different stages of cotton fiber development were quantified and analyzed by sensitive and real-time PCR technique to understand the expression pattern of GhRGL gene at all stages of cotton fiber development. Total RNA samples were extracted from different stages of cotton ovules and fibers including -3~0 DPA ovules, 3 DPA ovules, 5 DPA ovules,10 DPA ovules,15 DPA fibers,and 25 DPA fibers to analyze the expression pattern of GhRGL gene in cotton fiber development. The results showed that the GhRGL gene was expressed in all selected cotton samples. Higher levels of GhRGL mRNA were detected in 5 DPA and 10 DPA ovules, and the highest levels of GhRGL mRNA were detected in 10 DPA ovules.Lower levels of mRNA were detected in-3~0 DPA ovules,3 DPA ovules and 15 DPA fibers,and 25 DPA fibers. gene was expressed at the highest levels at the fiber elongation stage,suggesting the important role of GhRGL in cotton fiber elongation.