Makowski Wojciech, Królicka Aleksandra, Hinc Krzysztof, Szopa Agnieszka, Kubica Paweł, Sroka Julia, Tokarz Barbara, Tokarz Krzysztof Michał
Department of Botany, Physiology and Plant Protection, Faculty of Biotechnology and Horticulture, University of Agriculture in Kraków, 29 Listopada 54, 31-425 Kraków, Poland.
Laboratory of Biologically Active Compounds, Intercollegiate Faculty of Biotechnology UG and MUG, University of Gdańsk, Abrahama 58, 80-307 Gdańsk, Poland.
Int J Mol Sci. 2025 Jan 3;26(1):362. doi: 10.3390/ijms26010362.
Houtt. is the source of various phenolic compounds: phenolic acids, flawan-3-ols, and stilbenes, with a broad range of biological activity. The rhizome (underground organ of these plants) is abundant in secondary metabolites but, in natural conditions, may accumulate various toxic substances (such as heavy metals) from the soil. The principal objective of this research was to produce transformed cultures of hairy roots that would serve as a valuable source of phenolic compounds, independent of environmental resources. The transformation was performed using a variety of wild strains of bacteria, of which only strain A4 (ATCC 31798) proved effective. The molecular characterization of transformed clones was performed using PCR. The biometric parameters (growth index and dry weight content), phenolic compounds accumulation (DAD-HPLC), antioxidant capacity (DPPH, CUPRAC), and bactericidal properties against with various sensitivity to antibiotics were evaluated. Two obtained transformed clones (RJ 9 and 30) exhibited the incorporation of the entire bacterial T-DNA into genomic DNA, while clones RJ 10 and 11 demonstrated only the presence of the LT-DNA sequence. The results demonstrated an increase in flawan-3-ols (catechins) accumulation in hairy root tissue relative to non-transformed (NT) plants. Moreover, hairy roots exhibited enhanced antioxidant activity and bactericidal properties compared with NT roots and NT shoots, respectively.
酚酸、黄烷-3-醇和芪类化合物，具有广泛的生物活性。根茎（这些植物的地下器官）富含次生代谢产物，但在自然条件下，可能会从土壤中积累各种有毒物质（如重金属）。本研究的主要目的是培育毛状根转化培养物，使其成为独立于环境资源的酚类化合物宝贵来源。转化过程使用了多种野生菌株，其中只有A4菌株（ATCC 31798）被证明是有效的。使用聚合酶链反应（PCR）对转化克隆进行分子表征。评估了生物统计学参数（生长指数和干重含量）、酚类化合物积累（二极管阵列-高效液相色谱法）、抗氧化能力（二苯基苦味酰基自由基法、铜离子还原抗氧化能力法）以及对各种抗生素敏感性不同的杀菌特性。获得的两个转化克隆（RJ 9和30）显示整个细菌转移DNA整合到基因组DNA中，而克隆RJ 10和11仅显示存在LT-DNA序列。结果表明，相对于未转化（NT）植株，毛状根组织中黄烷-3-醇（儿茶素）的积累有所增加。此外，与NT根和NT茎相比，毛状根分别表现出增强的抗氧化活性和杀菌特性。
