Identification and validation of conserved microRNAs along with their differential expression in roots of Vigna unguiculata grown under salt stress

被引:58
作者
Paul, Sujay [1 ]
Kundu, Anirban [1 ]
Pal, Amita [1 ]
机构
[1] Bose Inst, Div Plant Biol, Kolkata 700054, India
关键词
MicroRNA; Salt stress; Vigna unguiculata (cowpea); miRNA targets; qRT-PCR; 5 ' modified RLM-RACE; ARABIDOPSIS-THALIANA; RESPONSIVE MICRORNAS; REGULATORY ROLES; SMALL RNAS; BIOGENESIS; GENES; MIRNAS;
D O I
10.1007/s11240-010-9857-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
MicroRNAs (miRNAs) are 20-24 nucleotide long non-coding RNAs known to play important regulatory roles during plant development, organ morphogenesis, and stress responses by controlling gene expression. Although Vigna unguiculata (cowpea) is an economically important salt sensitive member of legumes, very little is known about the conserved miRNAs and their expression profile during salinity stress in this plant. In the present study using comparative genomic approach and following a set of strict filtering criteria we have identified 18 conserved V. unguiculata miRNAs belonging to 16 distinct miRNA families. Using these potential miRNA sequences 15 potential target genes were predicted and all of them were identified as transcription factors. Seven of these predicted V. unguiculata miRNAs were experimentally validated in the root tissues and found to be up-regulated during salt stress as revealed by quantitative real time PCR (qRT-PCR). Perfectly cleaved Auxin response factor (ARF), the target transcript of V. unguiculata miR160 was detected successfully by modified 5' RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) method.
引用
收藏
页码:233 / 242
页数:10
相关论文
共 51 条
[1]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[2]   Conserved and novel miRNAs in the legume Phaseolus vulgaris in response to stress [J].
Arenas-Huertero, Catalina ;
Perez, Beatriz ;
Rabanal, Fernando ;
Blanco-Melo, Daniel ;
De la Rosa, Carlos ;
Estrada-Navarrete, Georgina ;
Sanchez, Federico ;
Alicia Covarrubias, Alejandra ;
Luis Reyes, Jose .
PLANT MOLECULAR BIOLOGY, 2009, 70 (04) :385-401
[3]   MicroRNAs: Genomics, biogenesis, mechanism, and function (Reprinted from Cell, vol 116, pg 281-297, 2004) [J].
Bartel, David P. .
CELL, 2007, 131 (04) :11-29
[4]   Detection of 91 potential in plant conserved plant microRNAs in Arabidopsis thaliana and Oryza sativa identifies important target genes [J].
Bonnet, E ;
Wuyts, J ;
Rouzé, P ;
Van de Peer, Y .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (31) :11511-11516
[5]  
BRESSANI R, 1985, COWPEA RES PRODUCTIO, P359
[6]   Genetic analysis of salt stress responses in asparagus bean (Vigna unguiculata (L.) ssp sesquipedalis Verdc.) [J].
Chen, Chanyou ;
Tao, Chengxue ;
Peng, Hai ;
Ding, Yi .
JOURNAL OF HEREDITY, 2007, 98 (07) :655-665
[7]   Identification of MicroRNAs in Wild Soybean (Glycine soja) [J].
Chen, Rui ;
Hu, Zheng ;
Zhang, Hui .
JOURNAL OF INTEGRATIVE PLANT BIOLOGY, 2009, 51 (12) :1071-1079
[8]   microRNA biogenesis and function in plants [J].
Chen, XM .
FEBS LETTERS, 2005, 579 (26) :5923-5931
[9]   A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development [J].
Chen, XM .
SCIENCE, 2004, 303 (5666) :2022-2025
[10]   Differential expression of miRNAs in response to salt stress in maize roots [J].
Ding, Dong ;
Zhang, Lifang ;
Wang, Hang ;
Liu, Zhijie ;
Zhang, Zuxin ;
Zheng, Yonglian .
ANNALS OF BOTANY, 2009, 103 (01) :29-38