Characterization of grapevine microR164 and its target genes

Abstract

MicroRNAs (miRNAs) are an extensive class of newly identified small RNAs that regulate gene expression at post-transcription level by mRNA cleavage or translation. In our study, we used qRT-PCR and found that Vv-miR164 is expression in grapevine leaves, stems, tendrils, inflorescences, flowers and fruits. In addition, two potential target genes for Vv-miR164 were also found and verified by PPM-RACE and RLM-RACE. The results not only maps the cleavage site of the target mRNA but allowed for detection the expression pattern of cleaved fragments that can indicate the regulatory function of this miRNA on its target genes. These target genes were explored by qRT-PCR where some exhibited different expression patterns from their corresponding miRNA, indicating the cleavage mode of the miRNA on its target genes. The efficient and powerful approach used in this study can help in further understanding of how miRNAs cleaved their target mRNAs. Results from this study prove the importance of Vv-miR164 in regulating development and growth of grapes, and adds to the existing knowledge of small RNAmediated regulation in grapes.

Integrated Network Analysis Identifies Fight-Club Nodes as a Class of Hubs Encompassing Key Putative Switch Genes That Induce Major Transcriptome Reprogramming during Grapevine Development

“…Interestingly, among the putative miRNA:mRNA pairs, we identified Vv-miR164:NAC33, whose interaction has already been experimentally validated in grapevine (Sun et al, 2012).…”

“…The NAC33 expression profile is generally inversely correlated with that of Vv-miR164, which would be expected for a miRNA target (Sun et al, 2012). The recent transcriptome-wide analysis of grapevine miRNAs in six different organs during development revealed the spatiotemporal expression profile of the miRNAs involved in the predicted miRNA/switch gene pairs (Wang et al, 2014).…”

“…ZFWD proteins contain a C3H-type zinc finger and seven WD40 repeats, and a role in development has been suggested based on their structural similarity to the Arabidopsis thaliana CONSTITUTIVE PHOTOMORPHOGENESIS1 and PLEIOTROPIC REGULATORY LOCUS1 proteins, which regulate photomorphogenesis (Németh et al, 1998;Torii et al, 1998), whereas NAC transcription factors have been shown to regulate vegetative and reproductive development in Arabidopsis (Raman et al, 2008), tomato (Hendelman et al, 2013), papaya (Carica papaya) (Fabi et al, 2012), and grapevine (Sun et al, 2012;Wang et al, 2013), suggesting a broad role as master regulators of fruit ripening. The parameters z g and K p represent a normalized measure of intramodule communication and the mode of communication between nodes in different modules, respectively.…”

“…The interaction between miR164 and NAC-domain mRNAs has been experimentally validated in grapevine (Sun et al, 2012), revealing that Vv-miR164 is expressed strongly in leaves (vegetative/green tissues) but only weakly in fruits (mature tissues). The NAC33 expression profile is generally inversely correlated with that of Vv-miR164, which would be expected for a miRNA target (Sun et al, 2012).…”

Effect of Constitutive miR164 Expression on Plant Morphology and Fruit Development in Arabidopsis and Tomato

“…Furthermore, it has been observed that miR164 is also involved in senescence of leaves in Arabidopsis [13,41]. In addition, interestingly, miR164 is one of the most abundant miRNAs in some fruits such as orange [42], grape [43,44], tomato [45], and prickly pear fruit (tuna) [46]. In orange, it was observed that miR164 has remarkably higher expression in fruits in comparison with other organs [42].…”