Abstract

Transition of grapevine buds from paradormancy to endodormancy is coordinated by changes in gene expression, phytohormones, transcription factors, and other molecular regulators, but the mechanisms involved in transcriptional and post-transcriptional regulation of dormancy stages are not well delineated. To identify potential regulatory targets, an integrative analysis of differential gene expression profiles and their inverse relationships with miRNA abundance was performed in paradormant (long day (LD) 15 h) or endodormant (short day (SD), 13 h) Vitis riparia buds. There were 400 up- and 936 downregulated differentially expressed genes in SD relative to LD buds. Gene set and gene ontology enrichment analysis indicated that hormone signaling and cell cycling genes were downregulated in SD relative to LD buds. miRNA abundance and inverse expression analyses of miRNA target genes indicated increased abundance of miRNAs that negatively regulate genes involved with cell cycle and meristem development in endodormant buds and miRNAs targeting starch metabolism related genes in paradormant buds. Analysis of interactions between abundant miRNAs and transcription factors identified a network with coinciding regulation of cell cycle and epigenetic regulation related genes in SD buds. This network provides evidence for cross regulation occurring between miRNA and transcription factors both upstream and downstream of MYB3R1.

Highlights

  • Grapevine is a seasonally indeterminate temperate fruit crop that grows in different climates across the world

  • In the present study, RNA-seq was used to analyze differential gene expression at this pivotal stage and miRNA abundance and inverse gene expression analysis and predicted motif gene set enrichment of downregulated genes were used to identify potential regulatory networks associated with bud endodormancy

  • Hierarchal clustering indicated the transcriptome profiles of the paradormant (LD)

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Summary

Introduction

Grapevine is a seasonally indeterminate temperate fruit crop that grows in different climates across the world. C-repeat binding factor (CBF), dormancy-associated MADS-box (DAM) and flowering locus T (FT2) genes play important roles in regulating endodormancy in pear and cherry flower buds and other trees [10,17,20,22]. Both miRNA156 and miRNA172 were found to regulate seed dormancy and flowering time in lettuce (Lactuca sativa) and Arabidopsis thaliana in a delay of germination 1 (DOG1) dependent manner [21]. In the present study, RNA-seq was used to analyze differential gene expression at this pivotal stage and miRNA abundance and inverse gene expression analysis and predicted motif gene set enrichment of downregulated genes were used to identify potential regulatory networks associated with bud endodormancy

Results
Denotes in bold arethe reported with known vvi miRNA
Denotes the miR family HMM logo for the sequences
Inverse Expression Association of Predicted Target Genes and Abundant miRNAs
Predition offont
Discussion
Plant Materials and Photoperiod Treatments
RNA Sequencing and Differential Gene Expression Analysis
Transcriptome Functional Enrichment Analyses
Target Prediction for Abundant miRNAs
Transcription Factor Motif GSEA and Network
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