黄瓜蜡质基因CsCER1调控因子CsCOL5的筛选及其功能分析  

Screening and Functional Analysis of CsCOL5,the Upstream Regulator of CsCER1,in Cucumber Pericarp Wax Synthesis

作　　者：王文娇[1] 邢军杰 申成丞 李斌 WANG Wenjiao;XING Junjie;SHEN Chengcheng;LI Bin(College of Horticulture,Shanxi Agricultural University,Taigu,Shanxi 030800,China)

机构地区：[1]山西农业大学园艺学院,山西太谷030800

出　　处：《园艺学报》2024年第5期1005-1016,共12页Acta Horticulturae Sinica

基　　金：山西省自然科学基金面上项目(202303021221089,20210302123401);山西省重点研发计划项目(202102140601013);国家自然科学基金项目(31902029)。

摘　　要：以黄瓜(Cucumis sativus L.)蜡质合成基因CsCER1启动子序列中的70 bp片段作为诱饵进行酵母单杂文库筛选,结合转录组数据分析筛选得到HSPA2、ZFP、COL5、GLO1、PGK共5个基因,并将Cs COL5作为候选基因进一步研究。荧光定量结果显示,CsCOL5在黄瓜的各个组织部位均有表达,其中在雄花中表达最高。克隆CsCOL5并通过CRISPR/Cas9技术创制敲除CsCOL5的株系col5,荧光定量结果表明敲除CsCOL5使得CsCER1的表达降低,并且col5敲除株系的蜡质总含量比对照下降了84%~86%。据此推测CsCOL5可能通过与CsCER1启动子结合来调控黄瓜果实蜡质的合成。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.

关 键 词：黄瓜 蜡质调控 CsCOL5 基因编辑 

分 类 号：S642.2[农业科学—蔬菜学]