Chinese Bulletin of Botany ›› 2022, Vol. 57 ›› Issue (5): 635-648.DOI: 10.11983/CBB22004

• EXPERIMENTAL COMMUNICATIONS • Previous Articles     Next Articles

Hrh-miRn458 Regulates Oil Biosynthesis of Sea Buckthorn via Targeting Transcription Factor WRI1

Yu Miao1, Ruan Chengjiang1,*(), Ding Jian1, Li Jingbin1, Lu Shunguang2, Wen Xiufeng2   

  1. 1Institute of Plant Resources, Dalian Minzu University, Dalian 116600, China
    2Seabuckthorn Development Management Center, Ministry of Water Resources, Beijing 100038, China
  • Received:2022-01-06 Accepted:2022-03-18 Online:2022-09-01 Published:2022-09-09
  • Contact: Ruan Chengjiang
  • About author:*E-mail:

Abstract: Transcription factor WRINKLED 1 (WRI1) plays important regulatory role in oil biosynthesis. Most studies focused on the regulation mechanism of its transcription, translation, and downstream target genes. Very few reports on its post transcriptional regulation. To explore the relationship between hrh-miRn458 and transcription factor WRI1 of sea buckthorn, and analyze their expression dynamics in oil biosynthesis of pulp and seeds, the bioinformatic method was used to predict the binding sites in the CDS region of the candidate target gene WRI1 by hrh-miRn458 mature sequence. Double luciferase reporter system and RNA pull down assay were used to verify whether hrh-miRn458 could target the CDS region of WRI1. The expression of hrh-miRn458 and WRI1 were analyzed using real-time fluorescent PCR technology in developing sea buckthorn pulp and seeds. The results showed that the positions between 309-327 of the CDS region of WRI1 gene were complementary to 15 bases of the mature hrh-miRn458 sequence. The vector of pmirGLO-WRI1-WT+hrh-miRn458 mimics significantly inhibited the luciferase activity (P<0.001), and RNA pull down assay further verified that hrh-miRn458 could pull down WRI1 in the system. In pulp and seeds of different developing stages, the expression level of hrh-miRn458 decreased at first and then increased, but the expression level of its target gene WRI1 first increased and then decreased. At the same developmental stage, the hrh-miRn458 expression level in pulp was less than that of seeds, but the expression level of its target gene WRI1 in pulp was more than that of seeds. Thus, sea buckthorn hrh-miRn458 may target the WRI1, and there is negatively regulative relationship between them. These data provide scientific references for further understanding the seed oil biosynthesis mechanism of sea buckthorn and breeding for high oil varieties.

Key words: sea buckthorn, oil biosynthesis, hrh-miRn458, transcription factor WRI1, qRT-PCR