Laboratory of Adaptational Biotechnology, Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv, Ukraine.
Institute of Microbiology and Virology, NAS of Ukraine, Kyiv, Ukraine.
Curr Microbiol. 2021 Dec 14;79(1):14. doi: 10.1007/s00284-021-02709-0.
The aim of the study was to evaluate the long-term effect of Agrobacterium rhizogenes genes transfer on plant antioxidant system by the study of superoxide dismutase (SOD) activity in "hairy" roots of Artemisia and Althaea spp plants. PCR analyses revealed stability of the transformation and presence of bacterial rol B and rol C genes in the "hairy" roots after 4-6 years from the transformation event. SOD activity in the roots of untransformed in vitro cultivated plants used for the initiation of "hairy" roots growth was in the range of 45.8 ± 8.7 U/μg (Althaea officinalis) and 275 ± 97.1 U/μg (Artemisia ludoviciana). After a long-term in vitro cultivation more than half of tested "hairy" root lines (54%) showed a significant increase in SOD activity values compared to untransformed roots. The highest SOD activity values of "hairy" root lines (24-fold increase) were founded in A. officinalis (1105 ± 174 U/μg) and A. dracunculus (1356 ± 402 U/μg). The increase of the activity was found also in "hairy" roots of A. vulgaris (up to 375 ± 28.2 U/μg, sevenfold increase), A. ludoviciana (1001 ± 191 U/μg, 3.6-fold increase), and A. tilesii (438 ± 104 U/μg, 1.6-fold increase). The results of our study indicate that transformation by wild-type A. rhizogenes not harboring any foreign genes implementing in SOD activity regulation can often stably activate plant antioxidant enzyme system. This effect, observed in the "hairy" roots of five plant species in 4-6 years of the transformation event, obviously, should be taken into account in works aimed at creating transgenic plants by Agrobacterium-mediated transformation.
本研究的目的是通过研究青蒿和蜀葵植物“毛状”根中超氧化物歧化酶(SOD)活性,评估根癌农杆菌基因转移对植物抗氧化系统的长期影响。PCR 分析显示,在转化事件发生 4-6 年后,“毛状”根中仍稳定存在细菌 rolB 和 rolC 基因。未转化的体外培养植物用于“毛状”根生长起始的根中 SOD 活性范围为 45.8±8.7 U/μg(蜀葵)和 275±97.1 U/μg(青蒿)。经过长期的体外培养,超过一半的测试“毛状”根系(54%)与未转化的根相比,SOD 活性值显著增加。在 A. officinalis(24 倍增加)和 A. dracunculus(1356±402 U/μg)中发现“毛状”根系的 SOD 活性最高值(1105±174 U/μg)。还发现 A. vulgaris(高达 375±28.2 U/μg,增加 7 倍)、A. ludoviciana(1001±191 U/μg,增加 3.6 倍)和 A. tilesii(438±104 U/μg,增加 1.6 倍)的“毛状”根中的活性增加。我们的研究结果表明,野生型根癌农杆菌的转化不含有任何参与 SOD 活性调节的外源基因,通常可以稳定激活植物抗氧化酶系统。这种效应在 4-6 年的转化事件中在五种植物的“毛状”根中观察到,显然在通过根癌农杆菌介导的转化创建转基因植物的工作中应予以考虑。