Antibacterial, Antihemolytic, Cytotoxic, Anticancer, and Antileishmanial Effects of Ajuga bracteosa Transgenic Plants

Herbal and traditional medicines can play a pivotal role in combating cancer and neglected tropical diseases. Ajuga bracteosa, family Lamiaceae, is an important medicinal plant. The genetic transformation of A. bracteosa with rol genes of Agrobacterium rhizogenes further enhances its metabolic content. This study aimed at undertaking the molecular, phytochemical, and in vitro biological analysis of A. bracteosa extracts. We transformed the A. bracteosa plant with rol genes and raised the regenerants from the hairy roots. Transgenic integration and expression of rolB were confirmed by conventional polymerase chain reaction (PCR) and qPCR analysis. The methanol: chloroform crude extracts of wild-type plants and transgenic regenerants were screened for in vitro antibacterial, antihemolytic, cytotoxic, anticancer, and leishmanial activity. Among all plants, transgenic line 3 (ABRL3) showed the highest expression of the rolB gene. Fourier transform infra-red (FTIR) analysis confirmed the enhanced number of functional groups of active compounds in all transgenic lines. Moreover, ABRL3 exhibited the highest antibacterial activity, minimum hemolytic activity (CC50 = 7293.05 ± 7 μg/mL) and maximum antileishmanial activity (IC50 of 56.16 ± 2 μg/mL). ABRL1 demonstrated the most prominent brine shrimp cytotoxicity (LD5039.6 ± 4 μg/mL). ABRL3 was most effective against various human cancer cell lines with an IC50 of 57.1 ± 2.2 μg/mL, 46.2 ± 1.1 μg/mL, 72.4 ± 1.3 μg/mL, 73.3 ± 2.1 μg/mL, 98.7 ± 1.6 μg/mL, and 97.1 ± 2.5 μg/mL against HepG2, LM3, A549, HT29, MCF-7, and MDA-MB-231, respectively. Overall, these transgenic extracts may offer a cheaper therapeutic source than the more expensive synthetic drugs.

Hairy root induction in Catharanthus roseus by various strains of Agrobacterium rhizogenes isolated in Vietnam

Catharanthus roseus is a well known medicinal plant. It produces several phytocompounds and many of which show anticancerous properties. However the yields of these compounds are very low. Recently, induction of C. roseus hairy roots by Agrobacterium rhizogenes, is interested as a promising tool for the enhanced production of these metabolites. In this research, wounded leaves from four strains of C. roseus were infected with various strains of A. rhizogenes isolated in Vietnam to provide more information about the induction efficiency of hairy roots. In this experiment, after 3 weeks of infection, the presence of rolB gene in hairy roots were analysed by polymerase chain reaction. All of 13 A. rhizogenes strains could induce the formation of hairy root in C. roseus. The A. rhizogenes C18 strain had the highest induction percentage in C. roseus VIN002, VIN005, and VIN072 with 59.4 %, 50.3 %, and 40.0 % respectively. And the same result was ontained at 26.7% by A. Rhizogenes C26 for C. roseus VIN077 rhizogenes strains. This result identified two A. rhizogenes strains C18 and C26 as potential transformation tools for hairy root production from C. roseus.