Effect of the rolC gene on biosynthesis of tropane alkaloids in Atropa belladonna
10.16438/j.0513-4870.2021-1431
- VernacularTitle:rolC基因对颠茄托品烷生物碱合成的影响
- Author:
Xing-xing LU
1
,
2
;
Ai-juan TAN
1
,
2
;
Hao ZHOU
1
,
2
;
Wen-wen AO
1
,
2
;
Shi-ming LÜ
3
;
Jian-min MAN
1
,
2
;
Ming-sheng ZHANG
1
,
2
;
Wei QIANG
1
,
2
Author Information
1. Ministry of Education, Collaborative Innovation Center for Mountain Ecology &
2. Agro-Bioengineering (CICMEAB), College of Life Sciences/Institute of Agro-bioengineering, Guizhou University, Guiyang 550025, China
3. College of Animal Science, Guizhou University, Guiyang 550025, China
- Publication Type:Research Article
- Keywords:
italic>Atropa belladonna L.;
italic>rolC gene;
hyoscyamine;
scopolamine;
anisodamine
- From:
Acta Pharmaceutica Sinica
2022;57(2):533-540
- CountryChina
- Language:Chinese
-
Abstract:
Hair roots induced by Agrobacterium rhizogene produce higher levels of secondary metabolites than non-induced plants, and the enhanced metabolic capacity is driven by the rol gene. We hypothesized that rol genes can be utilized to improve the biosynthesis of tropane alkaloids (TAs) in Atropa belladonna. In this study, the rolC gene from Agrobacterium rhizogene pRiA4 plasmid, driven by a CaMV35S promoter, was overexpressed in A. belladonna. The phenotypes, TAs content and transcriptional expression of key genes in TAs biosynthesis were analyzed in transgenic A. belladonna plants. Results show that transgenic A. belladonna exhibited a well-developed root system, male sterility, higher stamen column length than pistil, early flowering, internode shortening, smaller but more flowers, increased axillary buds and lateral buds, decreased apical dominance, and long and narrow leaves as compared to wild-type plants. Transgenic A. belladonna produced more TAs than wild-type plants, with the content of hyoscyamine, anisodamine and scopolamine reaching 2.58, 3.59 and 15.77-fold that of the control group, respectively. The gene expression of putrescine N-methyltransferase (PMT), tropinone reductase I (TRⅠ) and hyoscyamine 6-β-hydroxylase (H6H), key enzymes in TAs biosynthesis, were up-regulated compared with the control group. The above results indicate that the rolC gene enhances TAs biosynthesis in A. belladonna by up-regulating the expression of key enzymes in the TAs biosynthesis pathway, laying a foundation for genetic manipulation of A. belladonna to increase TAs content by increasing rolC gene expression.
