The rolB gene-induced overproduction of resveratrol in Vitis amurensis transformed cells.

Area of Science:

• Plant biotechnology
• Molecular biology
• Biochemistry

Background:

• Resveratrol, a stilbene, exhibits anti-carcinogenic properties.
• Grape (Vitis amurensis Rupr.) cell cultures naturally produce low levels of resveratrol.
• Conventional methods for enhancing secondary metabolite production in plant cell cultures are often inefficient.

Purpose of the Study:

• To develop a method for significantly increasing resveratrol production in Vitis amurensis cell cultures.
• To investigate the molecular mechanisms underlying enhanced resveratrol biosynthesis.

Main Methods:

• Establishment and characterization of Vitis amurensis cell cultures.
• Genetic transformation of callus cultures using the Agrobacterium rhizogenes rolB gene.
• Quantification of resveratrol production and rolB mRNA transcript levels.
• Assessment of tyrosine phosphatase inhibitors' effects on resveratrol production.

Main Results:

• Wild-growing grape cell cultures produced low basal levels of resveratrol (up to 0.026% dry wt.).
• Transformation with the rolB gene resulted in a >100-fold increase in resveratrol production (up to 3.15% dry wt.).
• Resveratrol biosynthesis capability strongly correlated with rolB mRNA abundance.
• Tyrosine phosphatase inhibitors blocked the rolB-mediated increase in resveratrol production.

Conclusions:

• The rolB gene from Agrobacterium rhizogenes is a potent tool for enhancing resveratrol production in plant cell cultures.
• Tyrosine phosphorylation is a key signaling pathway involved in regulating resveratrol biosynthesis in plants.
• This study provides a novel approach for the biotechnological production of resveratrol.