农业生物技术学报2018，Vol.26Issue(2)：194-204,11.DOI:10.3969/j.issn.1674-7968.2018.02.002
甜瓜雄性不育两用系转录组分析与抗氧化酶活性研究
Transcriptome Analysis and Study of Antioxidant Enzyme Activities on Male Sterile AB Lines in Melon (Cucumis melo)
摘要
Abstract
Melon (Cucumis melo) was mainly cultivated in the temperate zone to the tropics of the world.In order to explor in the relationship between the occurrence of male sterility in melon and the metabolism of superoxide and the related oxidase activities from the transcriptional level,this study used the male sterile AB line of stamens of the fertile and sterile plants of 5 mm in melon to do RNA sequencing (RNA-seq),and screened differentially expressed genes which related to the oxidase.In addition,the study carried out the real time fluorescence quantitative analysis in 2 stages of microspore development to identify the expression quantities of differential genes,and determined enzymatic activities of the buds of male sterile and fertile plants.The results showed that there were total of 1 364 differentially expressed genes in transcriptome sequencing,including 834 up-regulated genes and 530 down regulated genes.Then,it did cluster analysis to the screened differentially expressed genes according to the same or similar expression patterns,it showed that these differentially expressed genes divided into 28 categories,and eighth of them have 18 differentially expressed genes,these genes had extremely significant correlation (P＞0.001) with pathway of superoxide metabolism.Gene Ontology (GO) function analysis of differentially expressed genes showed that a total of 1 118 genes were involved in the 3 branches including the biosynthesis,molecular function and cell component,576 differentially expressed genes were involved in the catalytic process of catalase,of which 17 genes were differentially expressed significantly.Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation results showed that there were 10 genes related to the oxidoreductase among them including 9 peroxidase homologous genes and a catalase homologous gene.In the biosynthetic pathway of phenylpropanoidand,compared with the male fertile plants,the gene expression of peroxidase (POD) related gene MELO3C012183 was significantly up regulated in male sterile plants,and the expression of gene MELO3C014656 was down regulated.Furthermore,in the pathway of tryptophan metabolism,compared with the male fertile plants,the gene expression of catalase (CAT) related gene MELO3C017024 was significantly up regulated in male sterile plants.Quantitative real time polymerase chain reaction (qRT-PCR) validation showed that fertile plants' expression quantities of peroxidase 2-like precursor gene and peroxidase 11 gene which related to POD were lower than sterile plants,on the contrary,fertile plants' expression quantities of catalase isozyme l-like gene which related to CAT were higher than sterile plants,the enzymatic activities of POD and CAT in male sterile plants were higher than that of fertile plants.The results supposed that the differential performances of enzymatic activities of POD and CAT in the male sterile line were caused by the self protection and inducing of plants,the accumulations of oxygen free radicals caused the abnormal increase in the level of membrane lipid peroxidation in male sterile lines,this phenomenon was likely to cause damage to microspore development and pollen abortion.This study would provid a scientific basis for the internal cause and mechanism of the pollen abortion in melon.关键词
甜瓜/雄性不育/转录组测序/抗氧化酶活性/差异表达基因Key words
Melon/Male sterility/RNA sequencing (RNA-seq)/Antioxidant enzyme activities/Differentially expressed genes分类
农业科技引用本文复制引用
王昱丹,何兴佳,康艺楠,周姜睿航,王喜庆,盛云燕,靳亚忠,廉华..甜瓜雄性不育两用系转录组分析与抗氧化酶活性研究[J].农业生物技术学报,2018,26(2):194-204,11.基金项目
国家自然基金项目(No.31401892和No.31772330)、教育部回国留学人员科研启动项目、黑龙江省自然基金(No.C2017054)、黑龙江省教育厅科研项目(No.12541575)和黑龙江八一农垦大学校级培育重点课题(No.XA201701) （No.31401892和No.31772330）
