关键词: 苹果, 乙烯响应因子, 酵母单杂, 荧光素酶报告, 花青苷

Abstract: The expression levels of genes involved in anthocyanin and ethylene biosythesis were analyzed via qRT-PCR using uncolored and colored fruits of‘Taishan Zaoxia’apple at different developmental stages. An ethylene response factor was isolated and tentatively named as MdERF1B-like （MDP0000167207），with an open reading frame of 690 bp encoding a protein of 229 amino acid residues. The expression level of MdERF1B-like was consistent with the expression trends of anthocyanin synthesis related genes MdDFR，MdANS and the regulatory gene MdMYB9 in the colored and uncolored‘Taishan Zaoxia’fruit. Phylogenic analysis indicated that MdERF1B-like was located in group IX and was closely related to PyERF1B-like. Overexpression of MdERF1B-like in‘Orin’calli led to the significantly higher anthocyanin accumulation than in the control. The MdMYB9 promoter sequence of 746 bp was analyzed，which contained an ERF binding RAA motif in its promoter，and yeast one-hybrid assay showed that MdERF1B-like bound with MdMYB9 promoter. Furthermore，luciferase reporter assay was used to verify that MdERF1B-like can promote the transcriptional activity of MdMYB9 promoter. Therefore，MdERF1B-like may promote the accumulation of anthocyanin through regulating the expression of MdMYB9.

Key words: apple, ethylene response factor, yeast one-hybrid assay, luciferase reporter assay, anthocyanin