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Bulletin of Botanical Research ›› 2021, Vol. 41 ›› Issue (4): 522-530.doi: 10.7525/j.issn.1673-5102.2021.04.007

• Research report • Previous Articles    

Mechanism of Sm-miR858 Negatively Regulateda R2R3-MYB Transcription Factor SmPAP1 in Salvia miltiorrhiza

Fang CHEN1, Ting-Ting SHE1, Lin ZHANG1, Hao-Tian GAO2, Guo-Liang LI2, Jian WANG1,2()   

  1. 1.School Modern Agriculture and Biotechnology,Ankang University,Ankang 725000
    2.School of Food and Biological Engineering,Shaanxi University of Science and Technology,Xi’an 710021
    3.Key Laboratory of Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry,National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China,College of Life Sciences,Shaanxi Normal University,Xi’an 710062
  • Received:2020-05-07 Online:2021-07-20 Published:2021-03-24
  • Contact: Jian WANG E-mail:wangjianswj@163.com
  • About author:CHEN Fang(1979—),female,Ph.D,associate professor,engaged in gene function and molecular regulation.
  • Supported by:
    National Natural Science Foundation of China(31170281);Science and technology department of Shaanxi province(2019JQ-560);Natural science foundation of Education department of Shaanxi province(16JK1014);Ankang University Research Foundatio(2017AYQDZR05)

Abstract:

MicroRNAs(miRNAs) are a class of non-coding small RNAs that negatively regulate gene expression. Through high-throughput sequencing miRNAsof Salvia miltiorrhiza, a mature sequence miR858 named Sm-miR858 was obtained. The sequence alignment showed that the Sm-miR858 was highly conserved with the identified miR858 sequences in other plants, and the result of Small RNA northern blotting showed that the Sm-miR858 was expressed in the root, stem and leaf tissues of S. miltiorrhiza, especially relatively high in leaves. To explore the function of Sm-miR858 in S. miltiorrhiza, the Sm-miR858 was predicted first online using biological software. The result of psRNATarget analysis showed that there were 13 potential target genes of Sm-miR858. One was SmPAP1, an important transcription factor, participated in the metabolic regulation on phenolic acid active substances of S. miltiorrhiza. To further verify the targeting effect of Sm-miR858 on SmPAP1, the correlation of co-expression between Sm-miR858 and SmPAP1 was analyzed and verified by Real-time qPCR in the tobacco transient expression system and S. miltiorrhiza tissues and organs. The Real-time qPCR results showed that there was a significant negative correlation co-expression between SmPAP1 and Sm-miR858 in S. miltiorrhiza. Then the Sm-miR858 and SmPAP1 over-expressing plant vectors were constructed, and the transient co-expression of the Sm-miR858 and SmPAP1 was subsequently assayed in tobacco. The results showed that the sm-mir858 overexpression resulted in a significant decrease of the SmPAP1 mRNA expression level, compared with the control, and this indicated that sm-mir858 did actually target and negatively regulate the expression of SmPAP1 in S. miltiorrhiza. Above results laid a solid foundation for elucidating the role of sm-mir858 in the regulation phenolic acids biosynthesis in S. miltiorrhiza.

Key words: Sm-miR858, SmPAP1, Salvia miltiorrhiza, Target gene

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