摘要
以‘小白凤’水蜜桃的果实为试材,利用miR-RACE技术验证了桃基因组中预测的ppe-miR167a的精确序列,克隆了预测的其3个ARFs靶基因PpARF6、PpARF8和PpARF6-like的ORF序列,并对预测靶基因进行了系统进化和保守结构域分析;同时降解组测序鉴定了ppe-miR167a的靶基因,并对二者间的作用模式及作用频度进行了分析。结果显示:(1)ppe-miR167a的精确序列与预测序列在5′和3′端均存在1个碱基的差异,预测的3个靶基因ARFs均含有高度保守的B3和生长素响应DNA结合域;系统进化树分析显示,桃中PpARF6、PpARF8及PpARF6-like均与巴旦杏的亲缘关系较近,其次为梅和甜樱桃。(2)降解组结果表明,预测的3个靶基因中,仅PpARF8能够被ppe-miR167a裂解,而未在PpARF6和PpARF6-like上检测到裂解位点。(3)外源IAA处理后,ppe-miR167a的表达量在处理组中低于对照组,而靶基因PpARF8及3′端裂解产物的表达均高于对照组。研究表明,桃果实中ppe-miR167a通过介导PpARF8的裂解影响生长素信号途径参与果实发育的调控。
We determined the precise sequence of peach miR167a by using miR-RACE PCR reactions of the peach(Prunus persica)cultivar‘Xiaobaifeng’,cloned three predicted target genes PpARF6,PpARF8 and PpARF6-like of ppe-miR167a and analyzed the phylogenetic and conserved domains of the three genes.The target genes of ppe-miR167a were further identified by degradome sequencing technology,and the mode and frequency of interaction between ppe-miR167a and the target genes were analyzed.The results showed:(1)the accurate sequence of ppe-miR167a was different from the predicted sequence in the 1 nucleotide at the 5′end and 3′end.All the three target genes ARFs contained highly conserved B3 and auxin response DNA binding domains.Phylogenetic tree analysis indicated that PpARF6,PpARF8 and PpARF6-like in peach were closely related to Apricot,followed by plum and sweet cherry.(2)The results of the degradome sequencing displayed that only PpARF8 could be cleavaged by ppe-miR167a among the three predicted target gene and no cleavage sites were detected on PpARF6 and PpARF6-like.(3)The analysis of gene expression of exogenous IAA treated peach showed that the expression of miR167a after treatment was lower than that in the CK,while the expression of its target gene PpARF8 and the 3′end cleavage production were higher than those in the CK.Taken together,these findings showed that ppe-miR167a in peach fruit involved in auxin signaling pathway and participated in the regulation of fruit development by mediating the cleavage of its target gene PpARF8.
作者
张彦苹
朱旭东
王晨
刘照坤
李庆魁
ZHANG Yanping;ZHU Xudong;WANG Chen;LIU Zhaokun;LI Qingkui(Suzhou Polytechnic Institute of Agriculture, Suzhou, Jiangsu 215008, China;2College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;3Suzhou Academy of Agricultural Sciences, Suzhou, Jiangsu 215000, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2021年第1期70-77,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
苏州市农业科技创新项目(SNG2020046)
江苏省科协青年科技人才托举工程(2018年度)
江苏高校“青蓝工程”
国家自然科学基金青年基金(31601727)
苏州农业职业技术学院博士提升计划科研启动基金。
