title: Determination of the precise sequences of computationally predicted miRNAs in Citrus reticulata by miR-RACE and characterization of the related target genes using RLM-RACE.
abstract: MicroRNAs play vital roles in various biological and metabolic processes by regulating the expression of their target genes in model plants. Since there are limited reports on miRNAs in Citrus reticulata (Crt-miRNAs), the determination of precise sequences of miRNAs is essential to further analyze the functions of miRNAs in Citrus reticulata. Here, miR-RACE, a recently developed technique for determination of the potential miRNAs computationally, was employed to identify the precise sequences of Crt-miRNAs. Tissue- and development-specific expression of nine miRNAs were identified by quantitative RT-PCR in the leaves, stems, flowers and fruits Subsequently, 10 potential target genes were predicated for the eight Crt-miRNAs, most of which were transcription factors and disease resistance proteins. Four target genes were experimentally validated by Poly (A) polymerase-mediated 3′ rapid amplification of cDNA ends and RNA ligase-mediated 5′ rapid amplification of cDNA ends (PPM-RACE and RLM-RACE). Our findings showed that regulatory miRNAs in C. reticulata may play a key role in regulating growth, development, and response to disease. Future work is required to study the functions of miRNAs and their targets of C. reticulata.
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Xiangpeng
authors.initials: X
authors.lastname: Leng
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Changnian
authors.initials: C
authors.lastname: Song
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Han
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Lingfei
authors.initials: L
authors.lastname: Shangguan
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Jinggui
authors.initials: J
authors.lastname: Fang
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: wangchen@njau.edu.cn.
authors.forename: Chen
authors.initials: C
authors.lastname: Wang
mesh_terms: D002957:Citrus / Q000235:genetics*; D019295:Computational Biology / Q000379:methods; D018506:Gene Expression Regulation, Plant; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D018749:RNA, Plant / Q000235:genetics* / Q000378:metabolism; D017423:Sequence Analysis, RNA / Q000379:methods*
chemical_list: D035683:MicroRNAs; D018749:RNA, Plant
keywords: Citrus reticulata; Expression; MiR-RACE; MicroRNAs; PPM-RACE; RLM-RACE