Antimicrobial and anti-cancer activities of extracts obtained after applying elisitor to Salvia hispanica L. callus produced in vitro conditions

Salvia hispanica L., which is in the Lamiaceae family, is a plant widely used in complementary medicine. In this study, the root, leaf and petiole parts of the S. hispanica L. plant grown in vitro were used as explant source, and after UV-C application to the developing callus, methanol extracts were extracted and the antimicrobial and anticancer activities of these extracts were investigated. Firstly, the seeds of S. hispanica L. were soaked in commercial bleach for 6 minutes to ensure surface sterilization, and the seeds were germinated in gelrite solidified MS nutrient medium. Root, leaf and petiole parts of germinating plants were used as explant source and explants were cultured by applying two different protocols. Callus development from explants was encouraged by using different doses of 2,4-D (0,25, 0,5, 0,75, 1, 1,25 mg/l) in the first protocol, and BAP (0,2 mg/l) together with 2,4-D in the second protocol. The most effective callus growth rate and callus weight were obtained in MS nutrient medium containing 0,25 mg/l 2,4-D + 0,2 mg/l BAP at 100% and 27,19 ± 1,50 g weight in leaf explants in the second protocol. After obtaining methanol extracts of calli exposed to UV-C treatment, 3 mg/ml concentrations of the extracts were used. 7 different gram positive (Bacillus subtilis ATCC 6337, Brevibacillus brevis, Bacillus megaterium DSM 32, Bacillus subtilis IM 622, Bacillus cereus EMC 19, Staphylococcus aureus 6538 P, Listeria monocytogenes NCTC 5348), 9 different gram-negative (Salmonella typhimurium NRRLE 4413, Pseudomonas fluorescens, Enterobacter aerogenes CCM 2531, Klebsiella pneumoniae EMCS, Escherichia coli ATCC 25922, Proteus vulgaris FMC II, Pseudomonas aeruginosa DSM 50070, Proteus vulgaris, Salmonella enterica ATCC 13311) bacteria antimicrobial activity was investigated using disk diffusion method. While the most effective antimicrobial activity on Grampositive bacteria was observed with a zone diameter of 12,5±2,04 mm against Listeria monocytogenes NCTC 5348 bacteria, in callus extracts obtained using petiole explants of the second protocol, the least effect was observed with a zone diameter of 7±0 mm for Staphylococcus aureus 6538 P, Bacillus subtilis ATCC 6337 and Bacillus cereus EMC 19 bacteria were detected in the calli of the leaf explant in the first protocol and in the extracts obtained from the calli of the leaf explant in the second protocol. The most effective antimicrobial activity on Gram-negative bacteria, with a zone diameter of 11±0,82 mm, was detected in callus extracts obtained from the petiole explant in the second protocol against Klebsiella pneumoniae EMCS bacteria, while the least effect was observed in Salmonella typhimurium NRRLE 4413 with 5,5±0,41 mm. Anticancer activities on SH-SY5Y human neuroblastoma cells were investigated by using 1000, 500, 250, 125, 62,5, 31,25, 15,625, 7,8125 µg/mL doses of the extracts, and the most effective cytotoxic activity was determined at the 1000 µg/mL dose of the extracts obtained from both protocols. Keywords: Salvia hispanica L., callus, UV-C, antimicrobial activity, anticancer activity