甜瓜CmERFIV-3基因cDNA的克隆及功能初步分析

he CmERFIV-3(MELO3C021306) gene, one of the ERF subfamily members, was cloned by RT-PCR method using cDNA as template from the fruit of Cucumis melon to analyze the function of ERF subfamily genes during melon fruit ripening. The ORF of this CmERFIV-3 gene was 702 bp, which encoded 233 amino acids. The full-length of the CmERFIV-3 gene was combined with overexpression vector pPZP221 and two interfering sequences against CmERFIV-3 gene were constructed into RNAi vector pART27 respectively. These vectors were then transformed into Agrobacterium AGL1 and injected into mature green fruits of melon separately using the method of transient expression inducted with Agrobacterium. The phenotypes of melon pericarp showed that the tissue transformed by overexpression vecors turned yellow ahead of the control group which took the proportion of 23.3% and the tissue transformed by RNAi vectors maintained in green in comparison to the control group which took the proportion of 20%. The expression level of the CmERFIV-3 gene was analyzed by RT-qPCR. The results revealed the mRNA level of the CmERFIV-3 gene in the tissue transformed by overexpression vector was 2.37 times in comparison to the control group. In addition, the expression level of the CmERFIV-3 gene in the tissue transformed by RNAi vectors was reduced about to 0.24 times compared with the control group. These results indicated that CmERFIV-3 gene may have a function of acceleration during fruit ripening in melon.