红肉苹果愈伤组织生长素信号相关基因MdARF3的克隆与表达分析

发布时间：2018-06-26 00:51

本文选题：苹果 + 生长素　；参考：《园艺学报》2017年04期

【摘要】：以红肉苹果‘紫红3号’(新疆红肉苹果与‘富士’F_1代)的红色幼嫩叶片为外植体诱导的红色愈伤组织为试材,初步探讨生长素调控苹果花青苷代谢机理。根据拟南芥AtARF3蛋白序列在苹果基因组中Blast比对得到一个生长素信号相关基因(MDP0000173151),暂命名为MdARF3。克隆测序发现该基因的开放阅读框长度为2 127 bp,编码708个氨基酸。进化树分析表明,MdARF3与AtARF3在同一进化支上,推测它们具有相似的功能。愈伤组织在含有0.3 mg·L~(-1) NAA的MS培养基培养2 h之后其MdARF3上调表达,表达量极显著高于无NAA培养基(对照),而CHS、CHI、F3H、DFR、UFGT及LDOX等花青苷合成结构基因的表达量均极显著低于对照,并且MdARF3表达量与培养基中NAA浓度呈显著正相关(相关系数为0.98),与愈伤组织花青苷含量呈显著负相关(相关系数为 0.89),推测MdARF3对培养基中生长素快速做出反应调控花青苷合成;通过原核诱导获得了MdARF3的重组蛋白,为进一步研究MdARF3蛋白在花青苷代谢途径中的功能奠定了基础。
[Abstract]:The red young leaves of red meat apple 'Zihong 3' (Xinjiang red meat apple and 'Fujiao F1 generation) were used as explants to induce red callus. The mechanism of auxin regulating anthocyanin metabolism was studied. According to Arabidopsis thaliana AtARF3 protein sequence, a auxin signaling related gene (MDP0000173151) was obtained by blast alignment in the apple genome, and was tentatively named MdARF3. Cloning and sequencing showed that the open reading frame length of the gene was 2 127 BP, encoding 708 amino acids. The phylogenetic tree analysis shows that MdARF3 and AtARF3 are in the same evolutionary branch, and it is inferred that MdARF3 and AtARF3 have similar functions. The expression of MdARF3 in callus cultured in MS medium containing 0.3 mg / L ~ (-1) NAA for 2 h was significantly higher than that in non-NAA medium (control). The expression of MdARF3 was positively correlated with NAA concentration (correlation coefficient was 0.98) and negatively correlated with anthocyanin content in callus (correlation coefficient was 0.89). It was inferred that MdARF3 could react quickly with auxin to regulate anthocyanin synthesis. The recombinant protein of MdARF3 was obtained by prokaryotic induction, which laid a foundation for further study of the function of MdARF3 protein in anthocyanin metabolism pathway.
【作者单位】：山东农业大学作物生物学国家重点实验室;
【基金】：国家重点研发计划项目(2016YFC0501505) 国家自然科学基金项目(31572091)
【分类号】：S661.1

【参考文献】