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Plant Molecular Biology Reporter

, Volume 37, Issue 1–2, pp 111–128 | Cite as

Multiple microRNAs Regulate the Floral Development and Sex Differentiation in the Dioecious Cucurbit Coccinia grandis (L.) Voigt

  • Jatindra Nath Mohanty
  • Subodh Kumar Chand
  • Raj Kumar JoshiEmail author
Original Paper
  • 95 Downloads

Abstract

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.

Keywords

Coccinia grandis Dioecy Sex differentiation sRNA-seq miRNA Transcription factors Methyltransferase 

Notes

Acknowledgements

We thank DST-FIST, Govt. of India, for the research infrastructure facilities provided to Centre of Biotechnology, Siksha O Anusandhan University.

Author Contributions

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.

Funding Information

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.

Supplementary material

11105_2019_1143_MOESM1_ESM.jpg (649 kb)
Figure S1 Abundance of known (A) and novel (B) miRNA families identified from Coccinia grandis. (JPG 648 kb)
11105_2019_1143_MOESM2_ESM.jpg (216 kb)
Figure S2 Gene ontology (GO) classification of the target genes for differentially expressed microRNAs. GO terms were assigned based on significant hits and classified into three main categories: Biological process, cellular component and molecular function. (JPG 215 kb)
11105_2019_1143_MOESM3_ESM.docx (22 kb)
Table S1 Primers and probes used in the present study. (DOCX 21 kb)
11105_2019_1143_MOESM4_ESM.xlsx (37 kb)
Table S2 Detailed information of conserved miRNAs isolated from Coccinia grandis. (XLSX 36 kb)
11105_2019_1143_MOESM5_ESM.xls (41 kb)
Table S3 Novel miRNAs identified from Coccinia grandis. (XLS 41 kb)
11105_2019_1143_MOESM6_ESM.xlsx (15 kb)
Table S4 Differentially expressed members of C. grandis miRNA families. (XLSX 15 kb)
11105_2019_1143_MOESM7_ESM.xlsx (20 kb)
Table S5 List of predicted target genes for the differentially expressed miRNAs. (XLSX 20 kb)
11105_2019_1143_MOESM8_ESM.xlsx (12 kb)
Table S6 Gene ontology (GO) based functional classification of conserved and novel miRNA targets with miRNA. (XLSX 12 kb)
11105_2019_1143_MOESM9_ESM.xlsx (10 kb)
Table S7 Functional classification of conserved and novel miRNA targets using Kyoto Enclycopedia of Genes and Genomes (KEGG) analysis. (XLSX 10 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Functional Genomics Laboratory, Centre of BiotechnologySiksha O Anusandhan UniversityBhubaneswarIndia
  2. 2.Department of BiotechnologyRama Devi Women’s UniversityBhubaneswarIndia

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