Effects of Bacillus thuringiensis 0271 on individual indicators of nonspecific resistance of Origanum vulgare L. to stressful conditions

In this paper, we aim to study the effect of the Bacillus thuriengiensis strain var. darmstadiensis 0271 on the biochemical parameters of Origanum vulgare L., which determine its nonspecific resistance to unfavourable environmental conditions. The research materials were a liquid spore culture of B. Thuringiensis 0271 and the following oregano samples: 100.1 with 75.5% of carvacrol, g-4with 52.0% of carvacrol, No. 2 with 59.85% of а-terpineol, No. 1 with 21.5% of germacrene D and 19.4% of в-caryophyllene. The preservation of the strain spores on the leaf surface of Origanum vulgare was determined using A.G. Kolchevskys method. The proline content was determined by V.A. Khramov’s and E.M. Ageeva’s method, while the pigment content was measured spectrophotometrically. The total content of phenolic compounds, ascorbic acid and water-soluble carbohydrates was established according to the methods by M.N. Zaprometov, M.M. Okuntsov and M. Dubois, respectively. Ten days after treating the leaves of the 100.1 and No. 1 oregano varieties with the spore culture of B. Thuringiensis 0271, the amount of chlorophylls decreased by 27.1 and 15.2% compared to the control, respectively. At the same time, the amount of chlorophylls increased by 91.4 and 72.7% in the leaves of the g-4and No. 2 varieties, respectively. On the 10th day of the experiment, the amount of proline and phenolic compounds decreased by 4 times in the leaves of the g-4 sample. Compared to the control, the 100.1 variety demonstrated a decrease in the amount of soluble carbohydrates and phenolic compounds by 1.76 and 2.0 times, respectively. On the 10th day of the experiment, the treatment of O. vulgare plants with B. Thuringiensis 0271 promoted the accumulation of antioxidants in the leaves of the g-4sample by 14.5% compared to the control.

1. Vokou D, Kokkini S, Bessiere JM. Geographic variation of Greek oregano (Origanum vulgare ssp. hirtum) essential oils. Biochemical Systematics and Ecology. 1993;21(2):287-295.

2. Adam K, Sviropoulou A, Kokkini S, Lanaras T, Arsenakis M. Antifungal activities of Origanum vul-gare subsp. hirtum, Mentha spicata, Lavandula an-gustifolia and Salvia fructinosa essential oils against human pathogenic fungi. Journal of Agricultural and Food Chemistry. 1998;46(5):1739-1745.

3. Yoshino K, Higashi N, Koga K. Antioxidant and antiinflammatory activities of oregano extract. Journal of Health Science. 2006;52(2):169-173. https://doi.org/10.1248/jhs.52.169

4. Cosge B, Turker A, Ipek I, Gurbuz B. Chemical compositions and antibacterial activities of the essential oils from aerial parts and corollas of Origanum acutidens (Hand.-Mazz.) Ietswaart, an endemic species to Turkey. Molecules. 2009;14 (5):1702-1712. https://doi.Org/10.3390/molecules14 051702

5. Bakkali F, Averbeck S, Averbeck C, Idaomar M. Biological effects of essential oils - A review. Food and Chemical Toxicology. 2008;46(2):446-475. https://doi.org/10.1016/j.fct.2007.09.106

6. Habibi E, Shokrzadeh M, Chabra A, Naghshvar F, Keshavarz-Maleki R, Ahmadi A. Protective effects of Origanum vulgare ethanol extract against cyclophosphamide-induced liver toxicity in mice. Pharmaceutical Biology. 2015;53(1):10-15. https://doi.org/10.3109/13880209.2014.908399

7. Mohamed NA, Nassier OA. The antihypergly-caemic effect of the aqueous extract of Origanium vulgare leaves in streptozotocin-induced diabetic rats. Jordan Journal of Biological Sciences. 2013; 6(1):31-38.

8. Vujicic M, Nikolic I, Kontogianni VG, Saksida T, Charisiadis P, Orescanin-Dusic Z, et al. Methanolic extract of Origanum vulgare ameliorates type 1 diabetes through antioxidant, anti-inflammatory and anti-apoptotic activity. British Journal of Nutrition. 2015;113(5):770-782. https://doi.org/10.1017/S0007114514004048

9. Chuang L-T, Tsai T-H, Lien T-J, Huang W-C, Liu J-J, Chang H, et al. Ethanolic extract of Origanum vulgare suppresses Propionibacterium acnes-induced inflammatory responses in human monocyte and mouse ear edema models. Molecules. 2018;23(8):1987. https://doi.org/10.3390/molecules23081987

10. Ren H, Qin X, Huang B, Fernandez-Garcia V, Lv C. Responses of soil enzyme activities and plant growth in a eucalyptus seedling plantation amended with bacterial fertilizers. Archives of Microbiology. 2020;202(6):1381-1396. https://doi.org/10.1007/s00203-020-01849-4

11. Mushtaq T, Shah AA, Akram W, Yasin NA. Synergistic ameliorative effect of iron oxide nanoparticles and Bacillus subtilis S4 against arsenic toxicity in Cucurbita moschata: polyamines, antioxidants, and physiochemical studies. International Journal of Phytoremediation. 2020;22(13):1408-1419. https://doi.org/10.1080/15226514.2020.1781052

12. Mahmood F, Shahid M, Hussain S, Haider MZ, Shahzad T, Ahmed T, et al. Bacillus firmus strain FSS2C ameliorated oxidative stress in wheat plants induced by azo dye (reactive black-5). 3 Biotech. 2020;10(2):40. https://doi.org/10.1007/s13205-019-2031-y

13. Khan MA, Asaf S, Khan AL, Jan R, Kang S-M, Kim K-M, et al. Extending thermotolerance to tomato seedlings by inoculation with SA1 isolate of Bacillus cereus and comparison with exogenous humic acid application. PLoS ONE. 2020;15(4). https://doi.org/10.1371/journal.pone.0232228

14. Yoo S-J, Weon H-Y, Song J, Sang MK. Induced tolerance to salinity stress by halotolerant bacteria Bacillus aryabhattai H19-1 and B. mesonae H20-5 in tomato plants. Journal of Microbiology and Biotechnology. 2019;29(7):1124-1136. https://doi.org/10.4014/jmb.1904.04026

15. Saad MMG, Kandil M, Mohammed YMM. Isolation and Identification of Plant GrowthPromoting Bacteria Highly Effective in Suppressing Root Rot in Fava Beans. Current Microbiology. 2020;77(9):2155-2165. https://doi.org/10.1007/s00284-020-02015-1

16. Shreya D, Jinal HN, Kartik VP, Amaresan N. Amelioration effect of chromium-tolerant bacteria on growth, physiological properties and chromium mobilization in chickpea (Cicer arietinum) under chromium stress. Archives of Microbiology. 2020;202(4):887-894. https://doi.org/10.1007/s00203-019-01801-1

17. Raddadi N, Cherif A, Ouzari HI, Marzorati M, Brusetti L, Boudabous A, et al. Bacillus thuringiensis beyond insect biocontrol: plant growth promotion and biosafety of polyvalent strains. Annals of Microbiology. 2007;57(4):481-494. https://doi.org/10.1007/bf03175344

18. Makonde HM, Lenga FK, Masiga D, Mugo S, Boga HI. Effects of Bacillus thuringiensis CRY1A(c) d-endotoxin on growth, nodulation and productivity of beans [Phaseolus vulgaris (L.) and siratro (Macroptilium atropurpureum DC.)]. African Journal of Biotechnology. 2010;9(1):017-024.

19. Azizoglu U. Bacillus thuringiensis as a biofertilizer and biostimulator: a mini-review of the little-known plant growth-promoting properties of Bt. Current Microbiology. 2019;76(11):1379-1385. https://doi.org/10.1007/s00284-019-01705-9

20. Belousova ME, Grishechkina SD, Ermolova VP, Antonets KS, Mardanov AV, Rakitin AL, et al. Whole genome sequencing of bacillus thuringiensis var. darmstadiensis 56 strain and the study of insecticidal activity of the biological preparation on its basis. Sel'skohozyaistvennaya biologiya = Agricultural Biology. 2020;55(1):87-96. https://doi.org/10.15389/agrobiology.2020.1.87rus (In Russian)

21. Bejaoui A, Chaabane H, Jemli M, Boulila A, Boussaid M. Essential oil composition and antibacterial activity of Origanum vulgare subsp. glandu-losum Desf. at different phenological stages. Journal of Medicinal Food. 2013;16(12):1115-1120. https://doi.org/10.1089/jmf.2013.0079

22. Rodriguez-Garcia I, Silva-Espinoza BA, Ortega-Ramirez LA, Leyva JM, Siddiqui MW, Cruz-Valenzuela MR., et al. Oregano essential oil as an antimicrobial and antioxidant additive in food products. Critical Reviews in Food Science and Nutrition. 2016;56(10):1717-1727. https://doi.org/10.1080/10408398.2013.800832

23. Putievsky E, Ravid U, Husain SZ. Differences in the yield of plant material, essential oils and their main components during the life cycle of Origanum vulgare L. In: Essential oils and aromatic plants: Proseedings of the International Symposium on Essential Oils, 1984, Noordwijkerhout, Netherlands. Noordwijkerhout; 1984. p. 185-189.

24. Khramov VA, Ageeva EM. Colorimetric methods for determining the content of free proline and amino nitrogen in dormant wheat seeds and their analytical activity. Sel'skokhozyaistvennaya biologiya.1986;10:122-124.(In Russian)

25. Khalafyan AA. Modern statistical methods in medical research. Moscow: Lenard, 2014, 320 p. (In Russian)

26. Taylor W, Camilleri E, Craft DL, Korza G, Granados MR, Peterson J, et al. DNA Damage Kills Bacterial Spores and Cells Exposed to 222-Nanometer UV Radiation. Applied and Environmental Microbiology. 2020;86(8):e03039-19 (14 p.)

27. Hasanuzzaman M, Bhuyan MHMB, Zulfiqar F, Raza A, Mohsin SM, Mahmud JA, et al. Reactive oxygen species and antioxidant defense in plants under abiotic stress: revisiting the crucial role of a universal defense regulator. Antioxidants. 2020; 9(8):681. https://doi.org/10.3390/antiox9080681