摘要
【目的】葡萄白腐病是危害最严重的病害之一,不同的葡萄种质对白腐病的抗性存在差异。在前期的研究中筛选到了1份抗病种质刺葡萄0941,该种质为中国野生葡萄,具有耐湿热、抗病性强的特点。旨在通过转录组测序和WGCNA筛选,挖掘刺葡萄抗白腐病关键基因。【方法】以抗病的刺葡萄0941(Vd)和感病的欧亚种美人指(Vv)为试材,通过室内接菌,设置0 h、24 h、48 h三个时间点,研究病原菌诱导下感病和抗病种质的基因表达差异,筛选出抗病基因,并利用qRT-PCR进行验证。【结果】根据|Log2Fold Change|≥2和p≤0.05的筛选条件,在0 h、24 h、48 h抗病品种刺葡萄和感病品种美人指之间筛选到差异表达基因分别有5266、4725、4653个。其中上调基因数分别为1737、1788、1727个,下调基因数为3529、2937、2926个;进一步对这些差异基因进行GO和KEGG富集分析,发现了具有防御反应、跨膜运输和一些植物激素信号转导等功能的基因或者通路。为了明确不同品种葡萄果实抗病之间的调控网络,进行了WGCNA分析,得到7个基因模块,筛选出2个与抗病高度相关的关键的模块并且通过hub基因分析明确模块内基因的互作网络关系,鉴定出2个类黄酮代谢相关基因、2个植物激素信号转导相关基因以及2个hub基因,并且通过荧光定量验证其表达情况与转录组数据一致。【结论】获得了刺葡萄与美人指响应白腐病侵染的差异表达基因,并显著富集在防御反应、跨膜运输和一些植物激素信号转导等通路,筛选出6个抗病候选基因,包括F3’5’H、EIN和MYB等。
【Objective】White-rot[Coniella diplodiella(Speg.)Petrk&Sydow.]of grapes is one of the most dangerous diseases,and different grapevine germplasm differs in their resistance to white-rot.In the previous study,the disease-resistant germplasm of Vitis davidii 0941 was screened,which was a wild type of grape in China with the characteristics of heat and humidity tolerance and good disease resistance.In this study,we screened the key genes of resistance to white rot in V.davidii by transcriptome sequencing and WGCNA.【Methods】The disease-resistant V.davidii 0941(Vd)and the diseasesusceptible V.vinifera‘Manicure Finger’(Vv)were used as test materials to study the differences in gene expression between disease-susceptible and disease-resistant germplasm resources under pathogen induction conditions by inoculation indoors at three time points of 0 h,24 h and 48 h.The disease resistance genes were screened and validated using qRT-PCR.【Results】According to the p≤0.05 and|Log2Fold Change|≥2,the 5266,4725 and 4653 differentially expressed genes were screened 0 h,24 h,and 48 h with the resistant cultivar(Vd)and susceptible cultivar(Vv).Among them,the numbers of upregulated genes were 1737,1788 and 1727,and the numbers of down-regulated genes were 3529,2937 and 2926,respectively.Further,these differential genes were enriched by GO(Gene Ontology)and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis. Some defense responses wereobserved, such as transmembrane transport, some plant hormone signal transduction and other functionsor pathways. In cellular components, the membranes, cells, cell parts, etc. had a high degree of enrichment,and in biological processes, the defense reactions, transmembrane transport, metabolic processes,cellular processes, etc. had a high degree of enrichment. In molecular functions, there was a high degreeof enrichment in transcriptional regulatory activity and catalytic activity, and the results indicated thatthese functions played an important role in the defense response
作者
谭西北
李鹏
孙磊
樊秀彩
刘崇怀
姜建福
张颖
TAN Xibei;LI Peng;SUN Lei;FAN Xiucai;LIU Chonghuai;JIANG Jianfu;ZHANG Ying(Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou 450009,Henan,China)
出处
《果树学报》
CAS
CSCD
北大核心
2023年第4期653-668,共16页
Journal of Fruit Science
基金
国家重点研发计划(2021YFD1200202)
国家自然科学基金项目(31872057)
国家重点研发专项(2019YFD1001405)
中国农业科学院科技创新工程专项经费项目(CAAS-ASTIP-2016-ZFRI-02)
现代农业产业技术体系建设专项资金(CARS-29-yz-1)。
关键词
葡萄
抗病
转录组
基因挖掘
WGCNA分析
Grape
Disease resistance
Transcriptome
Gene mining
WGCNA analysis
