黑莓RuEGs基因克隆及其不同采后处理下的表达特性  

Cloning of RuEG Genes from Blackberry and Its Expression Characteristics Under Different Postharvest Treatments

作　　者：何嘉琪 吴雅琼 吴文龙[2] 闾连飞[2] 刘洪霞 李维林 He Jiaqi;Wu Yaqiong;Wu Wenlong;Liu Lianfei;Liu Hongxia;Li Weilin(Co-Innovation Center for the Sustainable Forestry in Southern China,College of Forestry,Nanjing Forestry University,Nanjing,210037;Institute of Botany,Jiangsu Province and Chinese Academy of Sciences,Nanjing,210014)

机构地区：[1]南京林业大学林学院,南方现代林业协同创新中心,南京210037 [2]江苏省中国科学院植物研究所,南京210014

出　　处：《分子植物育种》2024年第9期2829-2836,共8页Molecular Plant Breeding

基　　金：江苏省自然科学青年基金项目(BK20190273);江苏省重点研发计划(现代农业)项目(BE2020344)共同资助。

摘　　要：黑莓果实柔软多汁,采后不耐贮藏,因此针对这一性状调控的基因进行深入研究具有实际意义。内切葡聚糖酶(endo-1,4-β-glucanase,EG)是纤维素酶中的重要酶类,能够分解细胞壁,导致果实的成熟软化,从而影响果实的耐贮性。基于阿魏酸和那他霉素采后处理下黑莓‘Hull’品种果实转录组测序的结果,克隆了差异表达的2个EGs基因,将其命名为RuEG1和RuEG2,并对这2个基因在阿魏酸和那他霉素处理下的表达情况进行分析。结果表明,RuEG1和RuEG2分别含1578和1491 bp开放阅读框,分别编码了525个和496个氨基酸。RuEG1和RuEG2蛋白均属于糖基水解酶第9家族,即含有该家族特有的4个半胱氨酸(Cys)残基、2个氨基酸保守区和1个启动位点。在阿魏酸和那他霉素采后处理下,黑莓成熟果实中RuEG1和RuEG2的表达量均呈现显著下降,说明其表达被抑制。本研究结果将为后续研究EGs基因的结构和功能提供理论基础,同时对探明EG酶在果实成熟软化中的作用具有重要意义。Blackberry fruit is soft and juicy with poor storability during postharvest period.Therefore,it is of practical significance to investigate the genes regulating this trait.Endoglucanase(endo-1,4-β-glucanase,EG)is an important enzyme in cellulase,which can decompose cell wall and affect fruit ripening and softening.Based on the results of transcriptome sequencing of blackberry'Hull'treated with ferulic acid and natamycin after harvest,two differentially expressed EG genes were cloned and named RuEG1 and RuEG2.In addition,the expression of these two genes in treated samples was analyzed.The results showed that RuEG1 and RuEG2 contained 1578 bp and 1491 bp open reading frame,which encoded 525 and 496 amino acids,respectively.RuEG1 and RuEG2 proteins belong to glycosylhydrolase family 9,containing four cysteine(Cys)residues,two amino acid conserved regions and one promoter site.The significant decrease in the expression of RuEG1 and RuEG2 genes in mature blackberry indicated that the expression of these genes was inhibited under postharvest treatment with ferulic acid and natamycin.This study will be a foundation for further study on the structure and function of EG genes as well as being of great significance to explore the role of EG enzyme in fruit ripening and softening.

关 键 词：黑莓 内切葡聚糖酶 基因克隆 表达量 

分 类 号：S663.2[农业科学—果树学]