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Int J Biol Sci 2012; 8(3):418-429. doi:10.7150/ijbs.3676

Research Paper

Isolation and Identification of miRNAs in Jatropha curcas

Chun Ming Wang1, Peng Liu1, Fei Sun1, Lei Li1, Peng Liu1, Jian Ye2, Gen Hua Yue1✉

1. Molecular Population Genetics Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 117604 Singapore;
2. Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 117604 Singapore.

Abstract

MicroRNAs (miRNAs) are small noncoding RNAs that play crucial regulatory roles by targeting mRNAs for silencing. To identify miRNAs in Jatropha curcas L, a bioenergy crop, cDNA clones from two small RNA libraries of leaves and seeds were sequenced and analyzed using bioinformatic tools. Fifty-two putative miRNAs were found from the two libraries, among them six were identical to known miRNAs and 46 were novel. Differential expression patterns of 15 miRNAs in root, stem, leave, fruit and seed were detected using quantitative real-time PCR. Ten miRNAs were highly expressed in fruit or seed, implying that they may be involved in seed development or fatty acids synthesis in seed. Moreover, 28 targets of the isolated miRNAs were predicted from a jatropha cDNA library database. The miRNA target genes were predicted to encode a broad range of proteins. Sixteen targets had clear BLASTX hits to the Uniprot database and were associated with genes belonging to the three major gene ontology categories of biological process, cellular component, and molecular function. Four targets were identified for JcumiR004. By silencing JcumiR004 primary miRNA, expressions of the four target genes were up-regulated and oil composition were modulated significantly, indicating diverse functions of JcumiR004.

Keywords: Jatropha, Biofuel, miRNA, fatty acid synthesis.

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How to cite this article:
Wang CM, Liu P, Sun F, Li L, Liu P, Ye J, Yue GH. Isolation and Identification of miRNAs in Jatropha curcas. Int J Biol Sci 2012; 8(3):418-429. doi:10.7150/ijbs.3676. Available from http://www.ijbs.com/v08p0418.htm