转录因子FabHLH148参与草莓果实的颜色发育  

Transcription factor FabHLH148 is involved in the color development of strawberry fruit

作　　者：郑珍珍 陈雪雪 沈元月[1] 黄芸 ZHENG Zhenzhen;CHEN Xuexue;SHEN Yuanyue;HUANG Yun(College of Plant Science and Technology,Beijing University of Agriculture/Beijing Key Laboratory of New Technology in Agricultural Application,Beijing 102206,China)

机构地区：[1]北京农学院植物科学技术学院·农业应用新技术北京市重点实验室,北京102206

出　　处：《果树学报》2022年第8期1358-1367,共10页Journal of Fruit Science

基　　金：国家自然科学基金项目(32102362,32030100)。

摘　　要：【目的】探究碱性螺旋-环-螺旋(basic helix-loop-helix,bHLH)转录因子在草莓果实成熟过程中的功能。【方法】基于转录组测序和基因组数据库,克隆FabHLH148基因;分析其理化性质、保守结构域、预测编码的蛋白质结构、系统进化树、亚细胞定位等;采用RT-qPCR检测其在草莓中的时空表达水平,并构建过表达(over expression,OE)和RNA干扰(RNAinterference,RNAi)载体,利用农杆菌介导瞬时侵染草莓果实,观察记录表型,检测FabHLH148表达水平,并测定花色素苷含量。【结果】FabHLH148基因CDS区全长687 bp,编码228个氨基酸,预测蛋白分子质量24.89 ku,理论等电点(pI)为11.65;该基因属于bHLH超家族,因其与二倍体草莓FvbHLH148序列相似度最高,所以将其命名为FabHLH148;亚细胞定位显示,FabHLH148主要定位在细胞核。RT-qPCR结果表明,FabHLH148在草莓不同组织部位均有表达,在果实中有较高表达并随果实成熟表达量显著升高。过表达FabHLH148能够促进果实着色以及花色素苷的积累,通过RNAi调低Fab-HLH148的表达则作用结果相反,且FabHLH148在OE组果实中表达水平显著高于对照,RNAi组果实中表达水平显著低于对照(p<0.01)。【结论】转录因子FabHLH148属于bHLH超家族,可促进草莓果实着色。【Objective】Strawberry is the most widely cultivated small berry fruit in the world due to its unique flavor as well as high nutritional and economic value.Meanwhile,it is a prominent problem to improve the quality of strawberry in cultivation.Coloring is an important evaluation index of fruit quality.Anthocyanin is the main component affecting the color of strawberry fruit.The basic helix-loop-helix(bHLH)transcription factor family as the second largest transcription factor families in plants has been proved to be involved in many processes of plant growth,development,morphogenesis and stress response.Some members of bHLH transcription factors play crucial regulatory roles in plant anthocyanin synthesis.To analyze the function of bHLH transcription factors during strawberry fruit coloring,the FabHLH148 gene was cloned and we used physiological and molecular biology methods to reveal the function of FabHLH148.【Methods】Firstly,according to the reported transcriptome data of strawberry fruits in five development periods(SG,LG,Wt,IR and PR),a gene increasing rapidly with fruit ripening was screened.As it contained a bHLH superfamily conserved domain,it had the highest sequence similarity with diploid strawberry FvbHLH148(GenBank accession:XM_004295010.2),and the gene was named FabHLH148.The total RNA from strawberry cultivar Bebihoppe fruit was extracted using the plant total RNA extraction kit(Huayueyang,China)according to the manufacturer’s protocol.Firststrand cDNA was synthesized and the reverse transcription was carried out using Hifair®Ⅲ1st Strand cDNA Synthesis SuperMix(YEASEN,China)and then,the full-length CDS sequence of the Fab-HLH148 gene was obtained by PCR.Secondly,some bioinformatics techniques were used in this research.ExPASy website was referred to analyze the molecular weight,isoelectric point and liposoluble index of encoding protein.The conserved domains of FabHLH148 were predicted using NCBI Conserved Domains.The secondary and tertiary structure of FabHLH148 was analyzed by DNA star

关 键 词：草莓 FabHLH148 转录因子 表达模式 花色素苷 

分 类 号：S668.4[农业科学—果树学]