microRNA regulates cytokinin induced parthenocarpy in Cucumber (Cucumis sativus L.)

Parthenocarpy is one of the most important agronomic traits for fruit yield in cucumbers. However, the precise gene regulation and the posttranscriptional mechanism are elusive. In the presented study, one parthenocarpic line DDX and non-parthenocarpic line ZK were applied to identify the microRNAs (miRNAs) involved in parthenocarpic fruit formation. The differential expressed miRNAs among parthenocarpic fruit of forchlorfenuron (CPPU) treated ZK (ZK-CPPU), pollinated ZK (ZK-P), non-pollinated DDX (DDX-NP) were compared with the non-parthenocarpic fruits of non-pollinated ZK (ZK-NP). It indicated 98 miRNAs exhibited differential expression were identified. Notably, a significant proportion of these miRNAs were enriched in the signal transduction pathway of plant hormones, as identified by the KEGG pathway analysis. qRT-PCR validation indicated that CsmiR156 family was upregulated in the ZK-NP while downregulated in ZK-CPPU, ZK-P, and DDX-NP at 1 day after anthesis. Meanwhile, the opposite trend was observed for CsmiR164a. In ZK-CPPU, ZK-P, and DDX-NP, CsmiRNA156 genes (CsSPL16 and CsARR9-like) were upregulated while CsmiRNA164a genes (CsNAC6, CsCUC1, and CsNAC100) were downregulated. The GUS and dual luciferase assay validated that CsmiR156a inhibited while CsmiR164a induced their target genes' transcription. This study presents novel insights into the involvement of CsmiR156a and CsmiR164a in the CK-mediated posttranscriptional regulation of cucumber parthenocarpy, which will aid future breeding programs.