中文摘要：菜豆金色花叶病毒是最庞大且最重要的一类植物病毒，仅由介体烟粉虱以循持久、循环的方式传播。在此过程中，菜豆金色花叶病毒和烟粉虱介体间产生了密切、复杂的互作关系。然而，互作关系下潜在的分子机制仍然未知，且尚未有带毒和未带毒烟粉虱microRNA性能方面的研究。本研究从烟粉虱中鉴定出了Ago1和Dcr1的同源基因，这表明烟粉虱中存在miRNA介导的基因沉默。通过对比分析携带中国黄化曲叶病毒和未带毒烟粉虱的miRNAs，揭示了烟粉虱失调的miRNAs与转录后的基因调控之间的关系。本研究对所有表达miRNAs的潜在靶标进行了预测。本研究结果将有助于更好地理解菜豆金色花叶病毒与烟粉虱复杂的互作关系下潜在的分子机制。
外文摘要：Background: The begomoviruses are the largest and most economically important group of plant viruses exclusively vectored by whitefly (Bemisia tabaci) in a circulative, persistent manner. During this process, begomoviruses and whitefly vectors have developed close relationships and complex interactions. However, the molecular mechanisms underlying these interactions remain largely unknown, and the microRNA profiles for viruliferous and nonviruliferous whiteflies have not been studied.
Methods: Sequences of Argonaute 1(Ago1) and Dicer 1 (Dcr1) genes were cloned from B. tabaci MEAM1 cDNAs. Subsequently, deep sequencing of small RNA libraries from uninfected and Tomato yellow leaf curl China virus (TYLCCNV)-infected whiteflies was performed. The conserved and novel miRNAs were identified using the release of miRBase Version 19.0 and the prediction software miRDeep2, respectively. The sequencing results of selected deregulated and novel miRNAs were further confirmed using quantitative reverse transcription-PCR. Moreover, the previously published B. tabaci MEAM1 transcriptome database and the miRNA target prediction algorithm miRanda 3.1 were utilized to predict potential targets for miRNAs. Gene Ontology (GO) analysis was also used to classify the potential enriched functional groups of their putative targets.
Results: Ago1 and Dcr1orthologs with conserved domains were identified from B. tabaci MEAM1. BLASTn searches and sequence analysis identified 112 and 136 conserved miRNAs from nonviruliferous and viruliferous whitefly libraries respectively, and a comparison of the conserved miRNAs of viruliferous and nonviruliferous whiteflies revealed 15 up-and 9 down-regulated conserved miRNAs. 7 novel miRNA candidates with secondary pre-miRNA hairpin structures were also identified. Potential targets of conserved and novel miRNAs were predicted using GO analysis, for the targets of up-and down-regulated miRNAs, eight and nine GO terms were significantly enriched.
Conclusions: We identified Ago1 and Dcr1 orthologs from whiteflies, which indicated that miRNA-mediated silencing is present in whiteflies. Our comparative analysis of miRNAs from TYLCCNV viruliferous and nonviruliferous whiteflies revealed the relevance of deregulated miRNAs for the post-transcriptional gene regulation in these whiteflies. The potential targets of all expressed miRNAs were also predicted. These results will help to acquire a better understanding of the molecular mechanism underlying the complex interactions between begomoviruses and whiteflies.
 
外文关键词：Tomato yellow leaf curl China virus；Whitefly Bemisia tabaci；Gene silencing machinery；Differentially regulated miRNA profiling
作者：Wang, Bi；Wang, Lanlan；Chen, Fangyuan；等
作者单位：浙江大学
期刊名称：VIROLOGY JOURNAL
期刊影响因子：2.362
出版年份：2016
出版刊次：2
点击下载：携带中国番茄黄化曲叶病毒的B型（MEAM1）烟粉虱MicroRNA性能分析