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目的:克隆甜瓜Cm-EILs基因cDNA,并分析甜瓜果实发育过程中的表达特性,为研究其功能奠定基础。方法:根据已报道的甜瓜(Cucumis melo L.cv.Andes)Cm-EIL1和Cm-EIL2基因cDNA序列设计合成特异性引物,应用RT-PCR技术从甜瓜品种河套蜜瓜(Cucumis melo L.cv.Hetao)成熟果实中克隆得到Cm-EIL1和Cm-EIL2 cDNA全长编码区序列,并通过定量PCR法检测果实不同发育时期该基因的表达特性。结果:Cm-EIL1和Cm-EIL2 cDNA编码区长度分别为2 082bp和1 848bp。核苷酸序列比对分析表明,得到的cDNA序列与已报道的Andes甜瓜相应基因的cDNA序列高度同源。不同发育时期果实样品定量PCR检测结果表明,Cm-EIL1、Cm-EIL2基因在授粉后15~30 d,表达水平较低,变化幅度较小,Cm-EIL1基因在35 d表达量最高,Cm-EIL2基因在授粉后40 d表达量最高,2个基因的表达量分别为15 d的2倍和2.5倍。结论:Cm-EIL1和CmEIL2表达模式基本相同,并且与甜瓜果实成熟过程及乙烯生成量显著正相关,表明该基因在果实成熟过程中可能起着重要作用。
OBJECTIVE: To clone the cDNA of Cm-EILs from Muskmelon and analyze the expression characteristics of Muskmelon during fruit development to lay a foundation for the study of its function. Methods: The specific primers were designed according to the reported cDNA sequences of Cm-EIL1 and Cm-EIL2 in melon (Cucumis melo L. cv. Ann des). The melon variety Cucumis melo L.cv. Hetao) were used to clone the full-length coding region of Cm-EIL1 and Cm-EIL2 cDNA. The expression of this gene was detected by quantitative PCR at different developmental stages. Results: The cDNA coding regions of Cm-EIL1 and Cm-EIL2 were 2 082 bp and 1 848 bp, respectively. Nucleotide sequence alignment analysis showed that the obtained cDNA sequence was highly homologous with the reported cDNA sequence of the corresponding gene of Andes melon. The quantitative PCR results of the fruit samples at different developmental stages showed that the expressions of Cm-EIL1 and Cm-EIL2 genes were lower at 15-30 days after pollination, The highest expression level of EIL2 gene 40 days after pollination was observed. The expression level of EIL2 gene was 2-fold and 2.5-fold higher than that of 15-day. CONCLUSION: The expression patterns of Cm-EIL1 and CmEIL2 are basically the same, and there is a significant positive correlation between the expression of Cm-EIL1 and ethylene production, indicating that the gene may play an important role in the fruit ripening process.