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 1-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.
[1]陈贤丰，梁承邺.HGDNR不育花药能量代谢、H2O2的积累与雄性不育关系 [J].植物生理学通讯, 1991. , 27(1):21-24[2]陈全求，蓝家样，韩光明，等. 核不育系杂交棉不同发育时期抗氧化酶活性变化及其杂种优势分析[J].湖北农业科学, 2015., 54(24):6173-6177[3]曹墨菊, 程江, 汪静, 等.太空诱变玉米核雄性不育与植物激素的关系[J].核农学报, 2010., 24(3):447-452[4]高营营.结球甘蓝细胞质雄性不育系细胞学及生理生化分析[J].东北农业大学, 2014, :-[5]何天明.梨雄性不育系‘新梨 号’花器官内氨基酸含量和过氧化氢酶活性分析[J].果树学报, 2012, 29(3):338-342[6]侯磊, 肖月华, 李先碧, 等.棉花洞雄性不育系花药发育的差别显示[J].遗传学报, 2002, 29(4):359-363[7]刘永明，张玲，邱涛，等.高通量转录组测序技术在植物雄性不育研究中的应用[J].遗传, 2016, 38(8):677-687[8]刘淑娟，朱 祺，幸学俊，等.植物雄性不育影响因素研究进展[J].中国农学通报, 2014. , 30(34):46-50[9]刘红艳，吴坤，杨敏敏，等. 芝麻显性细胞核雄性不育系内源激素、可溶性糖和淀粉含量变化[J].中国油料作物学报, 2014., 36(2):175-180[10]刘齐元，程元强，朱肖文，等. 雄性不育烟草花蕾中SOD、POD和CAT活性研究[J].中国烟草学报, 2011., 17(5):34-39[11]那海燕，邓祖湖，张木清，等.植物丝氨酸/精氨酸丰富(SR)蛋白的结构和功能及其在植物发育中的作用[J].植物生理学通讯, 2008. , 44(6):1209-1215[12]沈亮余，李荣冲，王瑞雪，等.油菜雄性不育系160S育性转换中保护酶活性变化[J].重庆师范大学学报：自然科学版, 2012. , 29(1):82-86[13]盛云燕, 常薇, 矫士琦, 等. 甜瓜雄性不育植株雄蕊发育结构及生理生化特征[J].植物生理学报, 2016., 52(7):1028-1034[14]吴则东, 王华忠, 韩英, 2009.甜菜细胞质雄性不育系和保持系花期几种酶活性的比较[J].中国糖科, 2009, (1):27-31[15]王永琦，杨小振，莫言玲，等. 西瓜雄性不育系‘Se18’抗氧化酶活性和内源激素含量变化分析[J].园艺学报, 2016., 43(11):2161-2172[16]张文静，刘 亮，黄正来，等.低温胁迫对稻茬小麦根系抗氧化酶活性及内源激素含量的影响[J].麦类作物学报, 2016. , 36(4):501-506[17]Ali G S, Palusa S G, Golovkin M, et al. Regulatian of plant developmental processes by a novel splicing factor[J].PLoS ONE, 2007., 2(5):e471-[18]An H, Yang Z H, Yi B, et al.Comparative transcript profiling of the fertile and sterile flower buds of pol CMS in B. napus[J].BMC Genomics, 2014. , 15(1):258-[19]Arora A, Singh V P.Cysteine protease gene expression and proteolytic activity during floral development andsenescence in ethylen-insenstive Gladiolus grandiflora[J].Plant Biochern Biotechnol, 2004. , 13(2):23-126[20]Chen C M, Chen G J, Cao B H, et al.Transcriptional profiling analysis of genic male sterile-fertile Capsicum annuum reveal candidate genes for pollen development and maturation by RNA-Seq technology[J].Plant Cell Tiss Organ Cult (PCTOC), 2015. , 122(2):465-476[21]Elowitz M B, Levine A J, Siggia E D, et al. Stochastic gene expression in a single cell[J].Science, 2002., (297):1183-1186[22]Hansen K D, Wu Z J, Irizarry R A, et al.Sequencing technology does not eliminate biological variability[J].Nature Biotechnology, 2011. , 29(7):572-573[23]Li J J, Han S H, Ding X L, et al.Comparative transcriptome analysis between the cytoplasmic male sterile line NJCMS1A and its maintainer NJCMS1B in soybean (Glycine max (L) Merr.)[J].PLoS One, 2015, 10(5):e126771-[24]Qu C M, Fu F Y, Liu M, et al.Comparative transcriptomeanalysis of recessive male sterility (RGMS) in sterile andfertile Brassica napus lines[J].PLoS One, 2015, 10(12):e0144118-[25]Shemesh-Mayer E, Ben-Michael T, Rotem N, et al. Garlic (Allium sativum L.)fertility: transcriptome and proteome analyses provide insightinto flower and pollen development[J].Front Plant Science, 2015., (6):271-[26]Wei M M, Wei H L, Wu M, et al.Comparative expression profiling ofmiRNA during anther development in genetic male sterileand wild type cotton[J].BMC Plant Biology, 2013. , 13(1):66-
