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Global identification of Ginkgo biloba microRNAs and insight into their role in metabolism regulatory network of terpene trilactones by high-throughput sequencing and degradome analysis
Industrial Crops and Products ( IF 5.9 ) Pub Date : 2020-03-03 , DOI: 10.1016/j.indcrop.2020.112289
Jiabao Ye , Xian Zhang , Junping Tan , Feng Xu , Shuiyuan Cheng , Zexiong Chen , Weiwei Zhang , Yongling Liao

Terpene trilactones (TTLs) are important secondary metabolites and healthy components of Ginkgo biloba. They are essential for the quality of G. biloba and have been widely used in food, medicine and natural health products. However, there are no reports of miRNA associated with the biosynthesis of TTLs. To screen and identify the potential miRNA that participates in TTL metabolism in G. biloba, we performed high-throughput sequencing and bioinformatics methods of microRNAs and the degradome in 10 libraries of G. biloba. In total, 3,314 miRNAs (1,197 conserved miRNAs and 2,117 novel miRNAs) were identified among five organs, which were used to test those miRNAs by northern blot, quantitative real-time PCR (qRT-PCR), RLM-RACE, and degradome sequencing. In addition, we evaluated the correlation among the miRNA and mRNA expression level and TTL content in different organs of G. biloba. The results showed that four conserved miRNAs and five novel miRNAs might participate in TTL biosynthesis pathways by predicting the targets of those miRNAs to 12 TTL biosynthesis structure genes. Furthermore, 25 miRNAs were identified to potentially play vital role in TTL accumulation by targeting transcript factor genes, bHLH, WRKY, and AP2, respectively, which are candidate regulatory genes in the pathway of TTL biosynthesis. qRT-PCR analysis verified the expression patterns of 31 miRNAs and corresponding 27 target genes. This study is the first to authenticate miRNAs as well as their potential target genes involved in TTL biosynthesis, thereby providing molecular evidence for revealing the comprehensive miRNA-mRNA regulatory network participated in TTL metabolism of G. biloba.



中文翻译:

通过高通量测序和降解组学分析全球鉴定银杏microRNA及其对萜三内酯代谢调控网络的作用的了解

萜内酯(TTLs)是银杏的重要次生代谢产物和健康成分。它们对于银杏的质量至关重要,已被广泛用于食品,药品和天然保健产品中。但是,没有报道与TTLs的生物合成有关的miRNA。为了筛选和鉴定参与G. biloba的TTL代谢的潜在miRNA ,我们在10个G. biloba文库中进行了microRNA和降解组的高通量测序和生物信息学方法。总共在五个器官中鉴定出3,314个miRNA(1,197个保守miRNA和2,117个新miRNA),用于通过Northern blot,定量实时PCR(qRT-PCR),RLM-RACE和降解组测序来测试这些miRNA。此外,我们评估了银杏不同器官中miRNA和mRNA表达水平以及TTL含量之间的相关性。结果表明,通过预测那些miRNA靶向12个TTL生物合成结构基因的目标,四个保守的miRNA和五个新颖的miRNA可能参与TTL生物合成途径。此外,通过靶向转录因子基因bHLHWRKYAP2,已鉴定出25种可能在TTL积累中起重要作用的miRNA。分别是TTL生物合成途径中的候选调控基因。qRT-PCR分析验证了31个miRNA和相应的27个靶基因的表达模式。这项研究是第一个鉴定miRNA及其可能参与TTL生物合成的潜在靶基因的鉴定,从而为揭示参与G. biloba TTL代谢的全面miRNA-mRNA调控网络提供分子证据。

更新日期:2020-03-03
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