摘要
【目的】研究NAC转录因子FaNAC56在草莓果实成熟中的作用。【方法】根据前期果实成熟过程中的蛋白组数据,以八倍体红颜草莓为试材,克隆FaNAC56基因及其启动子。在蛋白水平,利用MEGA构建FaNAC56系统进化树,并通过生物信息学预测其编码蛋白的二级结构,以及利用烟草进行亚细胞定位;在转录调控水平,利用预测网站分析FaNAC56启动子区顺式作用元件以及下游靶基因,并通过RT-qPCR分析FaNAC56的时空表达模式。最后,利用果实圆片温育方法分析FaNAC56基因受外源激素诱导情况。【结果】FaNAC56基因全长1035 bp,编码344个氨基酸,具有NAC转录因子NAM保守结构域。系统发育分析表明FaNAC56与月季NAC56同源性最高。亚细胞定位显示FaNAC56定位于细胞核内。FaNAC56在果实中高度表达并随果实成熟表达量急剧增加。FaNAC56启动子区含有ABA、赤霉素、生长素、乙烯等激素和胁迫相关响应元件,其表达水平受这些激素诱导。靶基因分析发现FaNAC56可能响应多种激素、花色苷和蔗糖调控元件。【结论】FaNAC56可能通过多种激素调控草莓果实的发育和成熟。
【Objective】Strawberry is a very valuable horticultural crop, and its fruit ripening is regulated by a complex process. To find the important regulatory factor involved in strawberry fruit ripening, the proteome of Fragaria × ananassa‘Benihoppe’was analyzed around the onset of fruit ripening. The data have showed that a NAC transcription factor has high expression level during fruit ripening, which is named as FaNAC56. NAC is one of the largest families of plant-specific transcription factors, which play an important role in plant growth and development. In order to determine the function of FaNAC56 in strawberry fruit ripening, we first cloned the FaNAC56 gene and then investigated its tissue expression pattern and subcellular localization.【Methods】Firstly, we screened differentially expressed proteins around the onset of fruit ripening on the basis of our proteome, and found a NAC transcription factor that increased rapidly during strawberry ripening. According to phylogenetic analysis, this transcription factor was named as FaNAC56. The full-length ORF sequence of the FaNAC56 gene was obtained by RT-PCR. After sequencing the amino acid sequence of the FaNAC56 gene based on NCBI database, these genes with high homology were screened and downloaded. MEGA software was used to construct the phylogenetic tree. The molecular weight, isoelectric point, liposoluble index of encoding protein were analyzed by the Expasy website. The secondary structure of its protein, cis-acting elements in the promoter region and downstream target genes were predicted through bioinformatics. Secondly,we used real-time PCR to detect the expression level of FaNAC56 in strawberry including various organs and developmental fruits, including the root, stem, leaf, flower, seed, small green fruit, large green fruit, de-greening fruit, white fruit, initial red fruit, partial red fruit, and full red fruit. Meanwhile, the white fruit was made into cylinder-shaped cut with 8-mm diameter. We treated the fruits with the solution contain
作者
许田
吴敏
陈雪雪
黄芸
沈元月
XU Tian;WU Min;CHEN Xuexue;HUANG Yun;SHEN Yuanyue(College of Plant Science and Technology,Beijing University of Agriculture/Beijing Key Laboratory of New Technology in Agricultural Application,Beijing 102206,China)
出处
《果树学报》
CAS
CSCD
北大核心
2021年第12期2072-2081,共10页
Journal of Fruit Science
基金
国家自然科学基金(32030100,32102362)。
关键词
草莓
FaNAC56
表达分析
亚细胞定位
激素诱导
Strawberry
FaNAC56
Expression analysis
Subcellular localization
Hormone induction
