Cloning and functional analysis of AbCYP80F1 gene in Atropa belladonna
10.7501/j.issn.0253-2670.2020.16.025
- Author:
Wei FU
1
Author Information
1. Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Collaborative Innovation Center for Mountain Ecology & Agro-Bioengineering (CICMEAB), College of Life Sciences/Institute of Agro-bioengineering, Guizhou University
- Publication Type:Journal Article
- Keywords:
AbCYP80F1;
Atropa belladonna L.;
Promoter analysis;
RACE;
Scopolamine
- From:
Chinese Traditional and Herbal Drugs
2020;51(16):4291-4299
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone AbCYP80F1 gene and its promoter, analyze the Cis-acting elements related to control signals and transcription factors in AbCYP80F1 gene promoter and study the effect of AbCYP80F1 overexpression on scopolamine accumulation in Atropa belladonna. Methods: CYP80F1cDNA from Hyoscyamus niger was used as query sequence to do blastn search in A. belladonna transcriptome database to retrieve homologous unigenes. Full-length of AbCYP80F1 cDNA and gene promoter were acquired by RACE and hiTAIL-PCR, respectively. Agrobacterium rhizogenes-dipping method was adopted to induce AbCYP80F1- overexpressed A. belladonna hairy roots, and scopolamine content in hairy roots was detected by HPLC. Results: A full-length of 1 675 bp of AbCYP80F1 gene was obtained and the deduced protein had a closed homology with CYP80F1 from Anisodus luridus. A length of 2 000 bp promoter contained Cis-acting elements related to light, anaerobic induction, auxin, MeJA, gibberellins and low temperature, as well as recognition sites of transcription factor of MYB, AP2/ERF, WRKY and bHLH. Overexpression of AbCYP80F1 gene increased the scopolamine content in hairy roots of A. belladonna by an average of 1.8 times. Conclusion: AbCYP80F1 is a secondary root-specific gene in scopolamine biosynthetic pathway and its overexpression can enhance scopolamine accumulation. Expression of AbCYP80F1 may be regulated by WRKY and bHLH transcription factors.
