Abstract
The Venus flytrap (Dionaea muscipula J. Ellis) is a carnivorous plant able to synthesize large amounts of phenolic compounds, such as phenylpropanoids, flavonoids, phenolic acids, and 1,4-naphtoquinones. In this study, the first genetic transformation of D. muscipula tissues is presented. Two wild-type Rhizobium rhizogenes strains (LBA 9402 and ATCC 15834) were suitable vector organisms in the transformation process. Transformation led to the formation of teratoma (transformed shoot) cultures with the bacterial rolB gene incorporated into the plant genome in a single copy. Using high-pressure liquid chromatography, we demonstrated that transgenic plants were characterized by an increased quantity of phenolic compounds, including 1,4-naphtoquinone derivative, plumbagin (up to 106.63 mg × g−1 DW), and phenolic acids (including salicylic, caffeic, and ellagic acid), in comparison to non-transformed plants. Moreover, Rhizobium-mediated transformation highly increased the bactericidal properties of teratoma-derived extracts. The antibacterial properties of transformed plants were increased up to 33% against Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli and up to 7% against Pseudomonas aeruginosa. For the first time, we prove the possibility of D. muscipula transformation. Moreover, we propose that transformation may be a valuable tool for enhancing secondary metabolite production in D. muscipula tissue and to increase bactericidal properties against human antibiotic-resistant bacteria.Key points• Rhizobium-mediated transformation created Dionaea muscipula teratomas.• Transformed plants had highly increased synthesis of phenolic compounds.• The MBC value was connected with plumbagin and phenolic acid concentrations.
Highlights
Rhizobium rhizogenes is bacteria from the family Rhizobiaceae and is a “natural genetic engineer” because of the ability to transfer T-DNA and incorporate bacteria-derived genes into the plant genome (Georgiev et al 2007)
D. muscipula tissue was inoculated with three different strains of wild-type R. rhizogenes: LBA 9402, ATCC 15834, and A4
Based on preliminary results from growth observations and screening for phenolic compound accumulation in teratomas tissue compared to non-transformed plants (NT plants), four transformed clones of the Venus flytrap were taken for further analysis: clones P, K, L, and E
Summary
Rhizobium rhizogenes (former: Agrobacterium rhizogenes) is bacteria from the family Rhizobiaceae and is a “natural genetic engineer” because of the ability to transfer T-DNA (transfer DNA) and incorporate bacteria-derived genes into the plant genome (Georgiev et al 2007). During T-DNA transfer to plant cells, R. rhizogenes pass on the set of rol genes occurring in Ri plasmids (root-inducing plasmids). These genes encode specific proteins responsible for control over auxin and cytokinin synthesis in plant cells (Guillon et al 2006). The consequence of R. rhizogenes infection is the change in plant hormone balance and rise of the hairy root phenotype. Depending on the plant genotype, transformed shoots (teratomas) may occur instead of hairy roots (Królicka et al 2010). Despite the infection mechanism of plants by R. rhizogenes being known for a few decades, the physiological consequences of such events related to plant species need to be studied
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