南方农业学报2025，Vol.56Issue(8)：2363-2374,12.DOI:10.3969/j.issn.2095-1191.2025.08.001
海棠花青苷调控基因MsMYB44-likes的克隆与鉴定
Cloning and identification of the anthocyanin-regulated MsMYB44-likes genes in crabapple
摘要
Abstract
[Objective]This study aimed to clone and identify the transcription factors(MYB44-likes)that related to the anthocyanin biosynthesis in crabapple,with the goal of providing a reference for elucidating the regulatory mecha-nism MsMYB44-likes genes in the leaf coloring process of crabapple and improving the ornamental traits of crabapple.[Method]Using the crabapple cultivar Malus spectabilis as the research object,tissue-cultured seedlings of Malus specta-bilis were sprayed with 25 μmol/L abscisic acid(ABA)treatment,while the seedlings sprayed with water as the control(CK).Three MsMYB44-likes genes in the tender leaves of Malus spectabilis were cloned by PCR(MsMYB44-like1,MsMYB44-like2,and MsMYB44-like3),respectively.Bioinformatics analysis,encoded protein subcellular localization and phylogenetic analysis were performed on these three MsMYB44-likes genes.Transient overexpression analysis of the MsMYB44-like2 gene was conducted in apple epidermis,and the binding of MsMYB44-like2 protein to the promoter of the structural gene UFGT in the anthocyanin biosynthesis pathway was studied using electrophoretic mobility shift as-say(EMSA).[Result]Three MsMYB44-likes genes(MsMYB44-like1,MsMYB44-like2,and MsMYB44-like3)were cloned,whose full-length cDNA sequences of were 954,960 and 942 bp,and they encoded 317,319,and 313 amino acid residues,respectively.In the three MsMYB44-likes protein structures,α-helix and random coil accounted for rela-tively high proportions.The protein sequences of MsMYB44-like1,MsMYB44-like2,MsMYB44-like3 and the three pro-teins shared typical MYB-DNA binding domains with protein sequences of Arabidopsis thaliana,wheat,rice,and pear.MsMYB44-like1 and MsMYB44-like2 clustered with the homologous proteins of pear into one clade.Three MsMYB44-likes proteins were localized in the nucleus.The transient transformation experiment showed that after injecting the Agro-bacterium suspension contained pCAMBIA2300-MsMYB44-like2,the area around the injection point on the apple epider-mis turned yellowish-green,indicating that the gene inhibited the anthocyanins biosynthesis of the apple epidermis.Com-pared with the leaves of CK,the relative expression of the MsMYB44-like2 gene in leaves after ABA treatment decreased significantly(P<0.05).The results of EMSA showed that the binding of MsMYB44-like2 protein to the promoter frag-ment of the MsUFGT gene led to increased mobility.[Conclusion]The three MsMYB44-likes proteins contain typical MYB-DNA binding domains and are localized in the nucleus.The expression of the MsMYB44-like2 gene is negatively regulated by ABA,and the protein of this gene suppresses anthocyanin biosynthesis in crabapple by directly binding to the promoter of the gene MsUFGT.关键词
海棠/花青苷/MsMYB44-likes转录因子/脱落酸Key words
crabapple/anthocyanin/MsMYB44-likes transcription factors/abscisic acid分类
农业科技引用本文复制引用
陈雅晴,薛星月,符嘉庆,马宇航,李增林,段瑛..海棠花青苷调控基因MsMYB44-likes的克隆与鉴定[J].南方农业学报,2025,56(8):2363-2374,12.基金项目
国家自然科学基金项目(32300306) （32300306）
西北农林科技大学博士启动经费项目(2452024011)National Natural Science Foundation of China(32300306) （2452024011）
Doctoral Startup Fund of Northwest A&F University(2452024011) （2452024011）
