Screening and Functional Analysis of CsCOL5,the Upstream Regulator of CsCER1,in Cucumber Pericarp Wax Synthesis
A 70 bp promoter sequence of wax synthesis gene CsCER1 was used as bait to screen the upstream regulators in Cucumis sativus L.by yeast one hybrid assay.Combined with transcriptome data analysis,HSPA2,ZFP,COL5,GLO1 and PGK were considered as possible candidate genes,and CsCOL5 selected as a candidate gene for further study.qRT-PCR showed that CsCOL5 was expressed in all tissues of cucumber,with the highest expression in male flowers.Then,CsCOL5 was cloned and col5 mutants were generated by CRISPR/Cas9 technology.qRT-PCR showed that knockout of CsCOL5 reduced CsCER1 expression,and the total wax content of col5 knockout lines decreased by 84％-86％compared with the control.Based on the findings above,it is concluded that CsCOL5 might act as an upstream regulator of CsCER1 to regulate the wax content in cucumber.
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