Multiple microRNAs Regulate the Floral Development and Sex Differentiation in the Dioecious Cucurbit Coccinia grandis (L.) Voigt
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Despite having a small genome and heteromorphic sex chromosomes, the molecular basis of sex differentiation in the dioecious cucurbit, Coccinia grandis, largely remains unclear. Increasing evidences indicate a vital regulatory role of microRNAs in the reproductive development of plants. In this study, we used a combination of high-throughput small RNA sequencing and computational analysis to identify sex-specific miRNAomes from male and female buds of C. grandis. Ninety-eight conserved miRNAs from 22 families and 44 novel miRNAs specific to C. grandis were detected. Comparative profiling together with Northern blot and qRT-PCR analysis revealed 41 significantly differentially expressed (DE) miRNAs, of which 16 could be fundamental to the regulation of sexual dimorphism. One hundred six target genes were predicted for 35 DE miRNAs that were significantly involved in flower organogenesis, phytohormone signaling, metabolism, transcription regulation, and DNA methylation. Temporal analysis of a set of 16 target genes at three stages of the bud development revealed a reciprocal alteration in their expression pattern with the complementary miRNAs. Further, the miR167a-1, miR393a, miR398b, and miRn9 mediated down-regulation of four predicted targets associated with reproductive organ development in C. grandis was confirmed through transient co-expression in Nicotiana benthaminana. Taken together, the present study represents the first report suggesting that multiple miRNA-mediated gene silencing cascade could be involved in regulating the molecular basis of sex differentiation in C. grandis.
KeywordsCoccinia grandis Dioecy Sex differentiation sRNA-seq miRNA Transcription factors Methyltransferase
We thank DST-FIST, Govt. of India, for the research infrastructure facilities provided to Centre of Biotechnology, Siksha O Anusandhan University.
RKJ conceived, designed, and supervised the research work. JNM performed the experiments and SKC analyzed the data. JNM and SKC wrote the manuscript. RKJ provided inputs on data presentation and critically reviewed the manuscript. All authors read and approved the final manuscript.
This research is supported by grants from Dept. of Biotechnology (DBT), Govt. of India (grant no. BT/PR3919/PBD/16/959/2011). JNM is supported by research fellowships from Dept. of Biotechnology (DBT), Govt. of India.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- Chen J, Zheng Y, Li Q, Wang Y, Chen L, He Y, Fei Z, Lu G (2016) Identification of miRNAs and their targets through high-throughput sequencing and degradome analysis in male and female Asparagus officinalis. BMC Plant Biol:16–80Google Scholar
- Conesa A, Götz S (2008) Blast2GO: a comprehensive suite for functional analysis in plant genomics. Int J Plant Genomics 619832Google Scholar
- Li WT, He M, Wang J, Wang Y (2013) Zinc finger protein (ZFP) in plants—a review. Plant Omics J 6:474–480Google Scholar
- Lin PC, Lu CW, Shen BN, Lee GZ, Bowmen JL, Artega-Vazquez MA, Liu LY, Hong SF, Lo CF, Su GM, Kohchi T, Ishizaki K, Zachgo S, Althoff F, Takenaka M, Yamato KT, Lin SS (2016) Identification of miRNAs and their targets in the liverwort Marchantia polymorpha by integrating RNA-Seq and degradome analyses. Plant Cell Physiol 57:339–358CrossRefGoogle Scholar
- Perry LM, Metzger J (1980) Medicinal plants of east and Southeast Asia 29: 218Google Scholar
- Song C, Zhang D, Zheng L, Zhang J, Zhang B, Luo W, Li Y, Li G, Ma J, Han M (2017) miRNA and degradome sequencing reveal miRNA and their target genes that may mediate shoot growth in spur type mutant “Yanfu 6”. Front Plant Sci 8:441Google Scholar
- Vyskot B, Araya A, Veuskens J, Negrutu I, Mouras A (1993) DNA methylation of sex chromosomes in a dioecious plant, Melandrium album. Mol Gen Genet 239:219–224Google Scholar