FaRALF1基因调控‘红颜’草莓果实成熟的功能分析

Function Analysis of FaRALF1 in Regulating Fruit Ripening of’Benihoppe’Strawberry

多肽信号广泛存在于高等植物体内,并对植物的生长发育和果实成熟起重要的调控作用,为探究多肽信号调控植物果实成熟机制,本研究以八倍体‘红颜’草莓(Fragaria×ananassa’Benihoppe’)为试验材料,通过生物信息学技术在草莓全基因组中鉴定出一类植物多肽信号家族RALF(Rapid alkalinization factors),并根据染色体上的定位命名将其为FaRALF1-FaRALF9,通过分析FaRALFs基因家族成员在草莓果实不同发育时期的表达差异,以及其对脱落酸(ABA)、生长素(IAA)、油菜素内酯(BR)、蔗糖处理(Suc)的响应特征分析,初步推测家族成员FaRALF1可能参与调控草莓果实成熟。通过构建FaRALF1基因的超表达FaRALF1-OE(Over Expression)和RNA干扰FaRALF1-RNAi(RNA interference)载体瞬时侵染大绿时期的草莓果实,通过体视荧光显微镜观察侵染5 d后草莓果实的荧光标签和成熟表型,并通过RT-qPCR(Quantitative real-time PCR)分析基因表达量,其中FaRALF1-OE促进草莓果实成熟;相反,FaRALF1-RNAi抑制果实成熟。本研究初步表明植物多肽信号家族成员FaRALF1基因参与调控‘红颜’草莓果实成熟,且表达量与果实成熟程度呈正相关,为研究植物多肽信号调控果实成熟机理提供了基础。

Plant hormones and polypeptides signal widely exist in higher plants,and play an important role in regulating plant growth and fruit ripening.In order to explore the mechanism of peptide signaling regulating fruit ripening in plants,we used octoploid strawberry(Fragaria×ananassa’Benihoppe’)as experimental material to iden tify a class of plant polypeptide signal family RALF(rapid alkalinization)in the whole genome of strawberry by bioinformatics technology.By analyzing the expression differences of members of the FaRALFs family at different developmental stages of strawberry fruit and their response characteristics to abscisic acid(ABA),auxin(IAA),brassinolide(BR)and sucrose(Suc)treatment,it is preliminarily speculated that FaRALF1 may be involved in regulating strawberry fruit ripening.In this study,we constructed the over expression and RNA interference(RNAi)vectors of FaRALF1 to infect strawberry fruits in the big green stage.The fluorescent label and mature phenotype of strawberry fruits were observed by stereoscopic fluorescence microscope after 5 days of infection,and the quan titative real-time PCR(RT-q PCR)was used to detect the fluorescent label and mature phenotype of strawberry fruits PCR analysis showed that FaRALF1-OE promoted strawberry fruit ripening.On the contrary,FaRALF1-RNAi inhibited fruit ripening.This study preliminarily showed that FaRALF1,a member of plant polypeptide signaling family was involved in regulating fruit ripening of strawberry(Fragaria×ananassa’Benihoppe’),and the expression level was positively correlated with fruit ripening degree,which provided a basis for studying the mechanism of plant polypeptide signaling regulating fruit ripening.