Makowski Wojciech, Królicka Aleksandra, Nowicka Anna, Zwyrtková Jana, Tokarz Barbara, Pecinka Ales, Banasiuk Rafał, Tokarz Krzysztof Michał
Department of Botany, Physiology and Plant Protection, Faculty of Biotechnology and Horticulture, University of Agriculture in Krakow, Krakow, Poland.
Intercollegiate Faculty of Biotechnology UG and MUG, Laboratory of Biologically Active Compounds, University of Gdansk, Gdansk, Poland.
Appl Microbiol Biotechnol. 2021 Feb;105(3):1215-1226. doi: 10.1007/s00253-021-11101-8. Epub 2021 Jan 15.
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 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.
捕蝇草（Dionaea muscipula J. Ellis）是一种能够合成大量酚类化合物的食虫植物，这些酚类化合物包括苯丙烷类、黄酮类、酚酸类和1,4-萘醌类。在本研究中，展示了捕蝇草组织的首次遗传转化。两种野生型发根农杆菌菌株（LBA 9402和ATCC 15834）是转化过程中合适的载体生物。转化导致形成了畸胎瘤（转化芽）培养物，细菌rolB基因以单拷贝形式整合到植物基因组中。使用高压液相色谱法，我们证明与未转化的植物相比，转基因植物的特征在于酚类化合物的含量增加，包括1,4-萘醌衍生物、白花丹素（高达106.63 mg×g干重）和酚酸（包括水杨酸、咖啡酸和鞣花酸）。此外，发根农杆菌介导的转化极大地提高了畸胎瘤衍生提取物的杀菌性能。转化植物对金黄色葡萄球菌、粪肠球菌和大肠杆菌的抗菌性能提高了33%，对铜绿假单胞菌的抗菌性能提高了7%。我们首次证明了捕蝇草转化的可能性。此外，我们提出转化可能是增强捕蝇草组织中次生代谢产物产量以及提高对人类抗生素耐药菌的杀菌性能的有价值工具。要点：• 发根农杆菌介导的转化产生了捕蝇草畸胎瘤。• 转化植物的酚类化合物合成大幅增加。• 最低杀菌浓度值与白花丹素和酚酸浓度相关。
