摘要
甘薯黑斑病是由长喙壳菌(Ceratocystis fimbriata Ellis et Halsted)侵染引起的,该病害是甘薯重要病害之一,在田间生产和贮藏期均可发生,严重影响甘薯的产量和品质。为了明确长喙壳菌对甘薯黑斑病的致病机理,本研究以甘薯黑斑病抗病品种南京92和感病品种徐薯18为试验材料,采用针刺接种孢子悬浮液的方法人工接种甘薯长喙壳菌,分别于接种当天与接种后第1 d、第3 d、第7 d取样,利用转录组测序技术进行测序分析。转录组测序结果显示,与接种当天相比,长喙壳菌接种至南京92薯块后第1 d、第3 d和第7 d上调差异表达基因(DEGs)分别为1124个、1713个和2032个,下调DEGs分别为869个、1042个和1005个;长喙壳菌接种至徐薯18薯块后第1 d、第3 d和第7 d上调DEGs分别为901个、1625个和1957个,下调DEGs分别为1017个、976个和1032个。GO功能富集分析结果表明,差异表达基因主要富集于生物过程类别的生物进程、代谢进程、单一生物进程以及细胞组分类别的细胞、细胞部分、细胞器等。KEGG通路富集的主要通路为代谢途径。维恩图分析结果表明,长喙壳菌接种至南京92薯块后不同时间点共有DEGs为1545个,接种至徐薯18薯块后不同时间点共有DEGs为1116个,对这些DEGs进行交互分析,发现接种至2个甘薯品种薯块后在各时间点共有的DEGs有925个。对925个共有DEGs进行KEGG聚类分析,结果表明,这些DEGs主要被富集到代谢通路中,如碳水化合物代谢、脂质代谢、氨基酸代谢、核苷酸代谢、辅因子和维生素代谢以及能量代谢等途径中,进一步对这些DEGs进行筛选,得到相对表达量高于1.00、持续上调表达并且差异表达倍数达到40倍以上的关键致病基因共20个。本研究结果为深入研究长喙壳菌致病分子机理奠定了重要基础。
Sweet potato black rot is caused by the infection of Ceratocystis fimbriata Ellis et Halsted.As one of the important diseases of sweet potato,it can occur during field production and storage,seriously affecting the yield and quality of sweet potato.To clarify the pathogenic mechanism of Ceratocystis fimbriata on sweet potato,this study used Nanjing 92(a resistant variety to sweet potato black rot)and Xushu 18(a susceptible variety)as test materials.The method of needle prick inoculation with spore suspension was adopted to artificially inoculate Ceratocystis fimbriata on sweet potato.Samples were collected at 0 d,1 d,3 d and 7 d after inoculation,and transcriptome sequencing technology was used for sequencing analysis.The results of transcriptome sequencing showed that compared with the day of inoculation,the numbers of up-regulated differentially expressed genes(DEGs)at 1 d,3 d and 7 d after inoculation of Ceratocystis fimbriata on Nanjing 92 tubers were 1124,1713 and 2032,respectively,and the numbers of down-regulated DEGs were 869,1042 and 1005,respectively.For Xushu 18 tubers,the numbers of up-regulated DEGs at 1 d,3 d and 7 d after inoculation were 901,1625 and 1957 respectively,and the numbers of down-regulated DEGs were 1017,976 and 1032 respectively.The results of GO functional enrichment analysis revealed that the differentially expressed genes were predominantly enriched in biological process categories such as biological process,metabolic process and single-organism process,as well as in cellular component categories including cell,cell part and organelle.The main pathways enriched by KEGG pathway were metabolic pathways.Venn diagram analysis showed that the common DEGs at different time points after inoculation of Ceratocystis fimbriata on Nanjing 92 tubers were 1545,and those on Xushu 18 tubers were 1116.Through interactive analysis of these DEGs,it was found that there were 925 common DEGs in the two sweet potato varieties at each time period.KEGG cluster analysis of the 925 common DEGs showed that these DEGs were mainly enriched in metabolic pathways,such as carbohydrate metabolism,lipid metabolism,amino acid metabolism,nucleotide metabolism,cofactor and vitamin metabolism,and energy metabolism.Further screening of these DEGs identified 20 key pathogenic genes with relative expression levels higher than 1.00,continuous up-regulation and a fold change of more than 40 times.The results of this study lay an important foundation for the in-depth study of the molecular pathogenic mechanism of Ceratocystis fimbriata.
作者
杨冬静
高方园
陈晶伟
马居奎
唐伟
梁昭
张成玲
孙厚俊
谢逸萍
YANG Dongjing;GAO Fangyuan;CHEN Jingwei;MA Jukui;TANG Wei;LIANG Zhao;ZHANG Chengling;SUN Houjun;XIE Yiping(Xuzhou Institute of Agricultural Sciences of the Xuhuai District of Jiangsu Province/Key Laboratory of Sweetpotato Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs,Xuzhou 221131,China)
出处
《江苏农业学报》
北大核心
2025年第7期1320-1331,共12页
Jiangsu Journal of Agricultural Sciences
基金
国家现代农业产业技术体系资助项目(CARS-10)
国家自然科学基金青年基金项目(32001599)。
关键词
甘薯
长喙壳菌
转录组测序
差异表达基因
致病机理
sweet potato
Ceratocystis fimbriata
transcriptome sequencing
differentially expressed genes(DEGs)
pathogenic mechanism
