摘要
MicroRNAs(miRNAs)是一类小的非编码RNA,在植物逆境及生物胁迫中通过调节靶基因来发挥重要的调控作用。半夏内源miR167是否参与了大豆花叶病毒(soybean mosaic virus,SMV)的胁迫及相互作用关系尚不清楚。本研究利用茎环法扩增半夏内源miR167基因,命名为pt-miR167,采用生物信息学软件对其进行保守性分析,系统进化性分析及靶基因预测。在此基础上,采用qRT-PCR技术检测病毒积累量、miR167及其潜在的靶基因的表达水平,分析miR167及靶基因响应病毒侵染的表达模式。结果表明,病毒积累量在5~15 d时迅速增加,其中10 d时增加最快,随后呈现缓慢上升趋势。病毒侵染后,pt-miR167相比对照组,表达量呈下调,并且在10 d时表达量最低。进化树分析表明,pt-miR167与番茄(Solanum lycopersicum)中的sly-miR167a聚为一族,高度同源。预测的潜在主要靶基因ARF6表达量与pt-miR167表达量呈现正好相反的趋势,其在10 d时表达量达到最高。本研究表明,miRNA167参与了寄主半夏与病毒SMV的相互作用关系,研究结果有助于揭示SMV与寄主半夏之间的相互作用机制,为今后深入研究病毒与植物提供了参考,为半夏生产提供了研究的基础。
MicroRNAs(miRNAs)are small non-coding RNAs that play an important role in regulating target genes in stress response and biotic stress in plants.It is unclear whether endogenous miR167 of Pinellia ternate is involved in virus stress.In this study,the endogenous miR167 of Pinellia ternate was firstly amplified and named pt-miR167.The conservative analysis,phylogenetic analysis and target gene prediction were carried out using bioinformatics software.On this basis,qRT-PCR was used to detect the accumulation of soybean mosaic virus(SMV),the expression level of miR167 and its potential target genes,analysis of miR167 and target gene expression patterns in response to viral infection.The results showed that the virus accumulation increased rapidly at 5~15 d,with the fastest increase at 10 d,and then showed a slow upward trend.After virus infection,compared with the control group,the expression of pt-miR167 was down-regulated,and the expression was the lowest at 10 d.Evolutionary tree analysis shows that,pt-miR167 is highly homologous to sly-miR167a in tomato(Solanum lycopersicum).The predicted expression of ARF6 target genes is in the opposite trend to that of pt-miR167,and its expression reached the highest level at 10 d.This study shows that miRNA participates in the interaction between the host and the virus.The results are helpful to reveal the interaction mechanism between SMV and the host Pinellia ternate.It will provide a reference for further studies of viruses and plants in the future,and a research basis for the production of Pinellia ternate.
作者
王晨燕
杜江
马振男
张丽
王德富
牛颜冰
WANG Chen-Yan;DU Jiang;MA Zhen-Nan;ZHANG Li;WANG De-Fu;NIU Yan-Bing(College of Life Sciences,Shanxi Agricultural University,Taigu 030801,Shanxi,China;College of Grassland Science,Shanxi Agricultural University,Taigu 030801,Shanxi,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2023年第5期724-733,共10页
Chinese Journal of Biochemistry and Molecular Biology
基金
山西农业大学科技创新基金项目(No.2020BQ51)
国家现代农业产业技术体系(No.CARS-21)资助。
关键词
半夏
大豆花叶病毒
miR167
靶基因
表达模式
Pinellia ternate(Thunb)Briet
soybean mosaic virus(SMV)
microRNAs 167(miR167)
target gene
expression
