杨树响应胶孢炭疽菌侵染的转录组学研究  

Transcriptome analysis of poplar leaves infected with Colletotrichum gloeosporioides

作　　者：李娴 陈思思 谢剑波[1] Li Xian;Chen Sisi;Xie Jianbo(School of Biological Sciences and Biotechnology,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学生物科学与技术学院,北京100083

出　　处：《北京林业大学学报》2024年第4期91-102,共12页Journal of Beijing Forestry University

基　　金：国家自然科学基因项目(31872671、31972954、32022057);中国科协项目(2018QNRC001);国家林业草原科技创新青年拔尖人才项目(2020132607)。

摘　　要：【目的】通过生理指标和高通量转录组数据解析杨树响应病原菌侵染的分子机制,为探究杨树叶片在胶孢炭疽菌侵染下生理及分子响应模式奠定重要的理论基础。【方法】以毛白杨无性系LM50为试验材料,对胶孢炭疽菌侵染后3种抗氧化酶活性(多酚氧化酶、超氧化物歧化酶和过氧化氢酶)变化模式进行分析;基于RNA高通量测序技术分析关键基因及其表达模式。【结果】病原菌侵染叶片6 d后,丙二醛含量和3种抗氧化酶的活性显著提高。共检测到4547个杨树差异表达基因,其中2262个基因上调,2285个基因下调,差异基因主要富集到糖类、脂类、次生代谢产物、苯丙烷、谷胱甘肽、多种氨基酸、不饱和脂肪酸合成及代谢等生物学通路。WRKY、ERF等转录因子家族表达量明显改变,可能在杨树响应病原菌胁迫过程中发挥了重要作用,其中27个WRKY和23个ERF转录因子表达明显受到激活。MapMan分析结果表明病原菌侵染导致氧化胁迫的产生,植物激素含量也增加,这些激素作为信号激活防御反应,最终诱导大量抗病通路相关基因表达。【结论】27个WRKY和23个ERF转录因子可能在杨树响应病原菌胁迫的过程中起到重要的调控作用,谷胱甘肽、苯丙烷和类黄酮次生代谢通路可能参与杨树响应胶孢炭疽菌侵染过程。[Objective]Our objective was to explore the key genes and the molecular mechanism during the infection of Colletotrichum gloeosporioides,which could provide new insights into the genetic basis on poplar defense pathways.[Method]The healthy poplar(Populus tomentosa LM50)leaves were inoculated with C.gloeosporioides.The dynamic changes of antioxidase activities of poplar leaves and key genes in the plant-interaction pathway were investigated by physiological and biochemical assays and high-throughput transcriptome sequencing.[Result]Malondialdehyde content,polyphenol oxidase activity,superoxide dismutase activity and catalase activity in poplar leaves were increased at 6 d after inoculation.A total of 4547 differential genes were screened between healthy and infected leaves,among which 2262 were upregulated and 2285 were down-regulated.Differential expression genes were distributed in many functional categories,including sugar,lipid,secondary metabolite,glutathione,phenylpropanoid,amino acid,unsaturated fatty acids metabolites and so on.A large number of transcription factors were detected to be activated during the infection,such as 27 WRKY,23 ERF,suggesting that transcription factors play important roles in response to pathogen stress.During the infection,the reactive oxygen species may act as signals that modulate the activating of plant stress responses and disease resistance pathways.The results were further confirmed by real-time quantitative PCR of six differential expressed genes detected by RNAseq,despite differences in magnitude.[Conclusion]The above results uncover several essential genes that may play crucial roles in response to biotic stress.Pathways of glutathione,phenylpropanoid and flavonoid biosynthesis may also be activated during the infection.

关 键 词：杨树 生物胁迫 转录组 胶孢炭疽菌 

分 类 号：S792.11[农业科学—林木遗传育种] S718.85[农业科学—林学]