蓝标型核不育小麦花药转录物组学及花色素苷合成相关基因的表达分析  

Anther Transcriptomics and Expression Analysis of Genes Related to Anthocyanin Synthesis in Blue Labeled Genic Male Sterile Wheat

作　　者：冯小雨 张建朝 牛娜[1] 宋瑜龙[1] 马守才[1] 王鹏科[1] 张改生[1] 王军卫[1] 董剑[1] FENG Xiao-Yu;ZHANG Jian-Chao;NIU Na;SONG Yu-Long;MA Shou-Cai;WANG Peng-Ke;ZHANG Gai-Sheng;WANG Jun-Wei;DONG Jian(College of Agronomy,Northwest A&F University,National Yang ling Agricultural Biotechnology&Breeding Center/Yangling Branch of State Wheat Improvement Center/Wheat Breeding Engineering Research Center,Ministry of Education/Key Laboratory of Crop Heterosis of Shaanxi Province,Yangling 712100,Shaanxi,China;Institute of Grain Crops,Xinjiang Academy of Agricultural Sciences,Urumqi 830001,China)

机构地区：[1]西北农林科技大学农学院,国家杨凌农业生物技术育种中心/国家小麦改良中心杨凌分中心/小麦育种教育部工程研究中心/陕西省作物杂种优势研究与利用重点实验室,陕西杨凌712100 [2]新疆农科院粮食作物研究所,乌鲁木齐830001

出　　处：《中国生物化学与分子生物学报》2021年第4期504-515,共12页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家小麦产业技术体系项目(No.K3070217023);陕西省作物杂种优势研究与利用重点实验室后补助项目(No.2018SZS-22)资助。

摘　　要：为了揭示蓝标型小麦核雄性不育的分子机制,更好地利用隐性核不育小麦杂种优势,本研究以蓝标型白粒小麦WS(不育)和浅蓝粒小麦WF(育性正常)植株花药为试验材料,利用转录物组学技术对两者差异表达基因进行了分析,并对其中涉及花色素苷合成相关基因进行了验证。结果表明:WF与WS相比,共检测到2352个差异表达基因,这些基因经GO功能注释分为3大类43个小类,主要涉及生物合成、苯丙烷代谢、L-苯丙氨酸分解代谢、膜组成部分、质膜、细胞质、ATP结合和蛋白质丝氨酸/苏氨酸激酶活性等。KEGG通路分析结果显示,苯丙烷类生物合成通路富集基因最多,有159个,其次是苯丙氨酸代谢通路,包含136个显著差异表达基因,其他还涉及多种氨基酸代谢、嘌呤代谢、嘧啶代谢及糖代谢通路;与花青素代谢直接相关的通路中,多个控制关键酶结构基因存在差异表达,且大多数在WF中上调表达,只有黄烷酮3-羟化酶基因(flavanone 3-hydroxylase,F3H)和无色花青素双加氧酶基因(anthocyanin dioxygenase,ANS)下调表达;实时荧光定量分析显示,10个与花青素代谢相关基因实际表达情况和转录物组测序数据中基因表达情况具有相同的上下调趋势;差异基因序列同源性分析显示,筛选出的2个转录因子(DN48762c2g1、DN25944c0g1)与玉米、水稻及拟南芥花色素苷合成调控转录因子聚为同一簇,可能是蓝标型小麦浅蓝粒植株蓝色糊粉层性状的候选基因。并且荧光定量分析表明,DN48762c2g1和DN25944c0g1在WF中的表达量要明显高于WS。综上认为,花青素的生物合成途径相关基因不仅与籽粒蓝色性状有关,而且可能参与了蓝标型核不育系的花药败育。In order to reveal the molecular mechanism of blue labeled genic male sterility(BM-type GMS)and utilize the heterosis of BM-type GMS,we used the anthers of white-seed plants WS(sterile)and light blue seed plants WF(normal fertility)as experimental materials to analyze the differences in gene expression between them by transcriptome technology.And we also verified the genes expressed in anthocyanin synthesis in this study.Compared with WF,a total of 2352 differentially expressed genes were detected in WS.According to GO functional annotation,these genes could be divided into 3 categories and 43 subgroups.They are mainly involved in biosynthesis,phenylpropane metabolism,L-phenylalanine catabolism,membrane components,plasma membrane,cytoplasm,ATP binding,protein serine/threonine kinase activity,etc.KEGG pathway analysis showed that there were 159 genes enriched in the phenylpropanoid biosynthesis pathway,followed by the phenylalanine pathway,including 136 differentially expressed genes.Other genes are also involved a variety of amino acid metabolism,purine metabolism,pyrimidine metabolism and sugar metabolism pathway.Related to anthocyanin metabolism,several structural genes of key enzymes were differentially expressed,and most of them were up-regulated in WF,while only Flavanone 3-hydroxylase(F3 H)and colorless anthocyanin dioxygenase(ANS)were down-regulated.Quantitative real-time PCR showed that the expression of 10 genes related to anthocyanin metabolism had the same trend as that in transcriptome sequencing data.Sequence homology analysis showed that the two selected transcription factors(DN48762 c2 g1 and DN25944 c0 g1)are clustered into the same cluster as the transcription factors regulating anthocyanin biosynthesis in maize,rice and Arabidopsis thaliana,which might be candidate genes for the blue aleurone layer of light blue seed plants in wheat.And fluorescence quantitative analysis showed that the expression level of DN48762 c2 g1 and DN25944 c0 g1 in WF was significantly higher than that in WS.In conclusi

关 键 词：蓝标型小麦 隐性核不育 花色素苷 转录物组学 定量PCR 

分 类 号：S512.1[农业科学—作物学] S330