title: Mining, identification and function analysis of microRNAs and target genes in peanut (Arachis hypogaea L.).
abstract: In the present investigation, a total of 60 conserved peanut (Arachis hypogaea L.) microRNA (miRNA) sequences, belonging to 16 families, were identified using bioinformatics methods. There were 392 target gene sequences, identified from 58 miRNAs with Target-align software and BLASTx analyses. Gene Ontology (GO) functional analysis suggested that these target genes were involved in mediating peanut growth and development, signal transduction and stress resistance. There were 55 miRNA sequences, verified employing a poly (A) tailing test, with a success rate of up to 91.67%. Twenty peanut target gene sequences were randomly selected, and the 5' rapid amplification of the cDNA ends (5'-RACE) method were used to validate the cleavage sites of these target genes. Of these, 14 (70%) peanut miRNA targets were verified by means of gel electrophoresis, cloning and sequencing. Furthermore, functional analysis and homologous sequence retrieval were conducted for target gene sequences, and 26 target genes were chosen as the objects for stress resistance experimental study. Real-time fluorescence quantitative PCR (qRT-PCR) technology was applied to measure the expression level of resistance-associated miRNAs and their target genes in peanut exposed to Aspergillus flavus (A. flavus) infection and drought stress, respectively. In consequence, 5 groups of miRNAs & targets were found accorded with the mode of miRNA negatively controlling the expression of target genes. This study, preliminarily determined the biological functions of some resistance-associated miRNAs and their target genes in peanut.
authors.affiliation: Shandong Peanut Research Institute, Qingdao, 266100, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Shandong Peanut Research Institute, Qingdao, 266100, China; Shandong University, Weihai, 264200, China.
authors.forename: Shuhao
authors.initials: S
authors.lastname: Hu
authors.affiliation: Shandong Peanut Research Institute, Qingdao, 266100, China.
authors.forename: Caixia
authors.initials: C
authors.lastname: Yan
authors.affiliation: Shandong Peanut Research Institute, Qingdao, 266100, China.
authors.forename: Chunjuan
authors.initials: C
authors.lastname: Li
authors.affiliation: Shandong Peanut Research Institute, Qingdao, 266100, China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Shandong Academy of Agricultural Sciences, Jinan, 250100, China. Electronic address: wansb@saas.ac.cn.
authors.forename: Shubo
authors.initials: S
authors.lastname: Wan
authors.affiliation: Shandong Peanut Research Institute, Qingdao, 266100, China. Electronic address: shhshan@sina.com.
authors.forename: Shihua
authors.initials: S
authors.lastname: Shan
mesh_terms: D010367:Arachis / Q000235:genetics* / Q000382:microbiology; D001231:Aspergillus flavus / Q000502:physiology; D001483:Base Sequence; D057225:Data Mining; D055864:Droughts; D004587:Electrophoresis, Agar Gel; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics* / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D035683:MicroRNAs; D012322:RNA Precursors; D018749:RNA, Plant
keywords: Bioinformatics; Expression; MicroRNA; Resistance; qRT-PCR