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CsHEC2 regulates fruit wart formation through interaction with CsGL3 and CsTu and directly promoting cytokinin biosynthesis in cucumber

ABSTRACT: Cucumber fruit wart composed of spine and tubercule is an important appearance quality trait, which affects product classification and market value of cucumber fruit. Although several key genes for initiation and development of spine and tubercule have been cloned, their underlying mechanisms and relationships have not been well studied. Here, we identified a cucumber basic Helix-Loop-Helix (bHLH) gene CsHEC2 that was strongly expressed in spines and tubercules of cucumber peel. Knockout lines obtained using CRISPR/Cas9 technology were used to explore the biological function of CsHEC2. Compared with the wild type, the Cshec2 mutants resulted in reduced density of wart, and decreased cytokinin accumulation in fruit peel compared to wild type. To comprehensively analyze the regulatory network, RNA sequencing (RNA-seq) experiments were conducted on female buds at 7 days before anthesis (DBA). Transcriptomic data analysis showed that 293 and 1295 genes were up- and down-regulated in Cshec2 mutants relative to WT, respectively. Several sets of genes for cytokinin biosynthesis and metabolism were expressed differently, which explained the decrease of cytokinin in Cshec2 mutants. Our results suggested that CsHEC2 is very likely to regulate the initiation of fruit wart by affecting cytokinin pathway.

ORGANISM(S): Cucumis sativus

PROVIDER: GSE166286 | GEO | 2021/11/10

REPOSITORIES: GEO

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Project description:Purpose: Cucumber (Cucumis sativus L.) is an economically important vegetable crop worldwide, and cucumber fruit spine density has an important impact on the commercial value. However, little is known about the regulatory mechanism for the fruit spine formation.In this study, the transcriptome analyses of ovaries and pericarps from numerous-spine parent and few-spine parent were conducted to identify the gene regulatory networks involved in the formation and development of numerous fruit spines in cucumber. Methods: Cucumber mRNA profiles of ovaries and pericarps from numerous-spine parent and few-spine parent were generated by deep sequencing, in triplicate, using Illumina HiSeq 4000. Then, clean data (clean reads) were obtained by removing reads containing adapters, reads containing poly-N sequences and low-quality reads from the raw data. Simultaneously, the Q20, Q30 and GC contents of the clean data were calculated. All of the downstream analyses were based on the high-quality clean data. Clean paired-end reads were mapped to the reference genome using TopHat v2.0.12 (Trapnell et al. 2012). Then, the FPKM (fragments per kilobase of transcript sequence per million base pairs sequenced) value of each gene was calculated to estimate gene expression levels (Trapnell et al. 2010). Genes with an adjusted P-value < 0.05 identified by DESeq were assigned as differentially expressed genes(DEGs). Results: We generated 42.96-57.53 million raw reads from each library, and 39.85-54.02 million clean reads were obtained after the removal of low-quality reads and adapter sequences. Among the clean reads, 79.03-80.94% were mapped to the gene database . Based on the KEGG database, pathway enrichment analysis was performed to identify significantly enriched metabolic pathways or signal transduction pathways in DEGs. Plant hormone signal transduction was significantly enriched in up-regulated genes in both F_6DBF compared with M_6DBF and F_0DAA compared with M_0DAA. Conclusions: Based on the transcriptome analysis, we excavated possible biological regulatory networks involved in the formation and development of numerous fruit spines in cucumber. This work will promote the exploration of molecular mechanisms that regulate cucumber fruit spine density.

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CsHEC2 regulates fruit wart formation through interaction with CsGL3 and CsTu and directly promoting cytokinin biosynthesis in cucumber

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