芒果可可毛色二孢咪鲜胺抗药性菌株的转录组分析  

Transcriptome Analysis of Prochloraz Resistant Strains of Lasiodiplodia theobromae from Mango

作　　者：刘锦霖 李鹏声 杨叶[1,2] LIU Jinlin;LI Pengsheng;YANG Ye(Sanya Institute of Breeding and Multiplication,Hainan University,Sanya,Hainan 572022,China;School of Tropical Agriculture and Forestry,Hainan University,Haikou,Hainan 570228,China)

机构地区：[1]海南大学三亚南繁研究院,海南三亚572022 [2]海南大学热带农林学院,海南海口570228

出　　处：《热带作物学报》2024年第9期1811-1822,共12页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.32160653)。

摘　　要：为探析芒果可可毛色二孢对咪鲜胺的解毒代谢分子机制,挖掘相关功能基因。本研究使用RNA-seq技术对咪鲜胺处理和未处理的可可毛色二孢菌株进行转录组测序。结果表明:咪鲜胺对敏感菌株具有显著的抑制活性,半数有效浓度EC_(50)值为0.12 mg/L,2个抗性菌株的EC_(50)值分别为敏感菌株的7.67倍和32.67倍。对咪鲜胺敏感菌株(DF04)和抗性菌株(HL02、M108)间差异表达基因(DEG)进行GO功能富集和KEGG信号通路富集分析显示:与未用咪鲜胺处理(CK)相比,10 mg/L咪鲜胺处理后的DF04、HL02、M108菌株分别有5342、1148、2568个DEGs;其中,3个菌株之间共有566个DEGs重叠。与敏感菌株相比,抗性菌株分别有2078和3096个DEGs上调表达,1527和1793个DEGs下调表达。通过GO富集结果显示,咪鲜胺处理后的抗性和敏感菌株之间富集到细胞过程和代谢过程的DEGs最多,在HL02和M108中分别占比18%和16%、17%和15%。KEGG富集结果显示,咪鲜胺处理后抗性和敏感菌株之间DEGs显著富集在代谢通路中,HL02和M108中分别占比64.98%和63.39%。针对解毒代谢相关的DEGs的分析发现,抗性菌株中具有显著差异的细胞色素P450(Cytochrome P450)、谷胱甘肽转移酶(GST)和ABC转运蛋白(ABC transporter)基因分别为27、17、59个。随机筛选8个基因进行实时荧光定量验证,结果证实这些基因的相对表达量的变化趋势与转录组数据一致。根据抗敏菌株之间差异表达基因明显上调表达,共筛选出7个P450s基因、8个GSTs基因、11个ABC转运蛋白基因,可能与可可毛色二孢对咪鲜胺的代谢抗性机制有关。本研究结果为杀菌剂抗性机制提供新见解,也为深入探析咪鲜胺代谢抗性的分子机制奠定基础。The study was aimed to explore the molecular mechanism of detoxification of prochloraz by Lasiodiplodia theobromae from mango and to screen related functional genes.In this study,RNA-seq technology was used to analyze the transcriptome sequencing and functional annotation of prochloraz-treated and untreated strains of L.theobromae.Prochloraz had significant inhibitory activity against the sensitive strain with an EC_(50)value of 0.12 mg/L.The EC_(50)value of the two resistant strains was 7.67 and 32.67 times higher than that of the sensitive strain,respectively.GO functional enrichment and KEGG signaling pathway enrichment analyses were performed for differentially expressed genes(DEGs)between prochloraz-sensitive strains(DF04)and-resistant strains(HL02 and M108).Compared with the untreated(CK)with prochloraz,there were 5342,1148,and 2568 DEGs in DF04,HL02,and M108,respectively,after 10 mg/L prochloraz treatment.Among them,a total of 566 DEGs were overlapped among the three strains.Compared with the sensitive strains,the resistant strains had 2078 and 3096 DEGs up-regulated and 1527 and 1793 down-regulated 1812热带作物学报第45卷expression,respectively.GO enrichment showed that the highest number of genes were involved in cellular process and metabolic process between resistant and sensitive strains after prochloraz treatment,accounting for 18%and 16%,17%and 15%in HL02 and M108,respectively.KEGG enrichment showed that DEGs were significantly enriched in metabolic pathways between resistant and sensitive strains after prochloraz treatment,with 64.98%and 63.39%in HL02 and M108,respectively.The analysis for DEGs related to detoxification metabolism identified 27,17 and 59 cytochrome P450(Cytochrome P450),glutathione transferase(GST)and ABC transporter protein(ABC transporter)genes,respectively.Eight genes were randomly screened for quantitative real-time PCR validation,and the results confirmed that the trends in the relative expression of these genes were consistent with the transcriptome data.Based on the s

关 键 词：芒果 可可毛色二孢 咪鲜胺 转录组 解毒代谢酶 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]