miR164c and miR168a regulate seed vigor in rice.

MicroRNAs (miRNAs) are key regulators of gene expression in many important biological processes of plants. However, few miRNAs have been shown to regulate seed vigor. Here, we conducted microarray assays to analyze miRNA expression levels in seeds of the rice (Oryza sativa L.) cultivar ZR02. Results showed significant differences in the expression of 11 miRNAs between artificially aged and untreated control seeds. Among these, osa-miR164c was transcriptionally upregulated, while osa-miR168a was downregulated in artificially aged seeds; this was verified by quantitative real-time PCR analysis. Under the same aging condition, osa-miR164c overexpression in OE164c transgenic seeds and osa-miR168a silencing in MIM168a transgenic seeds of the rice cultivar Kasalath led to lower germination rates, whereas osa-miR164c silencing in MIM164c and osa-miR168a overexpression in OE168a resulted in higher seed germination rates compared with wild-type seeds. Meanwhile, changes in cytomembrane permeability of seeds and in the expression level of osa-miR164c target genes (OsPM27 and OsPSK5) and osa-miR168a target genes (OsAGO1 and OsPTR2) under aging conditions coincided with changes in seed vigor induced by osa-miR164c and osa-miR168a. Thus, genetic manipulation of miRNAs has important implications in the development of crop cultivars with high vigor and extended life span of seeds.