大白菜叶片响应菌核病侵染的转录组分析  

Transcriptome Analysis of Chinese Cabbage Responding to Infections of Sclerotinia sclerotiorum

作　　者：冯冬林[1] 丁玲[1] 刘美琴[1] 钟开勤 FENG Donglin;DING Ling;LIU Meiqin;ZHONG Kaiqin(Fujian Vocational College of Agriculture/Fuzhou Institute of Vegetable Science,Fuzhou,Fujian 350001,China)

机构地区：[1]福建农业职业技术学院/福州市蔬菜科学研究所,福建福州350001

出　　处：《热带作物学报》2023年第6期1106-1113,共8页Chinese Journal of Tropical Crops

基　　金：福建省科技厅引导性项目(No.2020N0022);福建农业职业技术学院科技项目(No.2019JS008)。

摘　　要：由核盘菌引起的大白菜菌核病是严重危害十字花科蔬菜品质和产量的病害之一。为深入研究菌核病抗病机制提供候选基因,从分子水平探索核盘菌侵染后大白菜的转录水平变化情况,以抗菌核病自交系H72和易感自交系Y26为材料,利用转录组测序技术分析抗感材料在接种核盘菌后不同时间的差异基因。结果表明,转录组测序共获得121.77 Gb的clean data,对原始测序数据进行过滤,各样品clean data均达6 Gb以上,Q_(20)介于97.21%~97.99%之间,平均值为97.61%,Q_(30)介于92.59%~93.98%之间,平均值为93.20%,表明测序质量较好,可以用于后续分析。接种的抗感材料在36、48 h的共同差异表达基因分别为13、11个,去除各接种点共同差异表达的基因,共获得18个差异表达基因。依据基因功能注释,筛选到11个防御反应相关基因,其中转录因子MYB34上调表达,乙烯响应因子ERF003下调表达,ERF003可能负向调控大白菜菌核病的侵染;生长素早期响应基因GH3.3、病程相关蛋白TLP1均上调表达;此外,F-box、氨基酸转运酶ANT1、咖啡酰辅酶A氧甲基转移酶Cco AOMT、天冬氨酸蛋白酶AED3、热激蛋白DnaJ 11均为上调表达,可能参与了大白菜对核盘菌的防御反应;上皮硫特异蛋白基因ESP和类钙调磷酸酶B蛋白CBL1为下调表达,其在大白菜抗菌核病中的具体功能需进一步分析验证。选择8个差异表达基因,利用qPCR验证了转录组分析的可靠性,其中ERF003、CBL1、GH3.3、MYB34、ANT1、TLP1基因的表达趋势与转录组测序结果基本一致,研究结果有助于进一步分析大白菜应答菌核病的抗病分子机制。Chinese cabbage Sclerotinia rot is caused by Sclerotinia sclerotiorum,which seriously endangers the yield and quality of Chinese cabbage.In this study,the transcription level of resistance genes to the infection of S.scle-rotiorum were explored at the molecular level and provided candidate genes for further study of disease resistance mechanism.The leaves of resistant inbred line H72 and susceptible strain Y26 were inoculated for transcriptome se-quencing analysis.The results show that a total of 121.77 Gb clean reads were obtained by transcriptome sequencing analysis.Filtering the raw sequencing data,the clean reads of each sample were over 6 Gb,Q_(20) ranged from 97.21%to 97.99%,and the average percentage was 97.61%,Q_(30) ranged from 92.59%to 93.98%%,and the average percentage was 93.20%,indicating that the sequencing quality could be used for subsequent analysis.The number of common differen-tially expressed genes were 13 and 11 after inoculation for 36 h and 48 h,respectively.After removing the common differentially expressed genes at each time point,we obtained 18 differentially expressed genes.Combined with gene function annotation,11 genes involved in defense response.MYB34 transcription factor was up-regulated,while ERF003 was down-regulated,which might negatively regulate the infection of S.sclerotiorum.Gretchen hagen 3.3(GH3.3),PR proteins(TLP1),F-box,amino acid transporter(ANT1),caffeoyl-CoA O-methyltransferase(Cco AOMT),aspartyl protease(AED3),chaperone protein(DnaJ 11)were up-regulated,which were proposed to response to Chinese cabbage pathogen.The expression of epithiospecifier protein(ESP)and calcineurin B-like protein gene(CBL1)were down-regulated,and the disease resistance function in Chinese cabbage needs further analysis and verification.A further qPCR analysis was used to verify the transcriptome,The expression trends of differentially expressed genes ERF003,CBL1,GH3.3,MYB34,ANT1,TLP1 were consistent with that of transcriptome sequencing.These results will be helpful for further research the

关 键 词：大白菜 菌核病 转录组 差异表达基因 

分 类 号：S436.341.1[农业科学—农业昆虫与害虫防治]