EFFECT OF PLANT GROWTH REGULATORS ON GROWTH PATTERNS AND ENZYMATIC ANTIOXIDANT ACTIVITIES IN HYPERICUM CALYCINUM SHOOT CULTURES
Keywords:
Hypericum calycinum, shoot culture, enzymatic antioxidant defense, polyphenolics in vitroAbstract
Hypericum calycinum L. (section Ascyreia) is a species of the Hypericum genus, relative to the medicinal plant H. perforatum (St. Jonn’s worth). Unlike the widely studied H. perforatum, it is characterized by the lack of hypericins production. However, research has revealed its commensurable antidepressant activity, as compared with the latter species, and shown the presence of polyphenolics with marked radical scavenging activity. In order to develop an in vitro system with the potential of a novel source of bioactive constituent’s characteristic for the Hypericum genus, we initiated in vitro cultures of H. calycinum. In the present work we studiy the effect of exogenous BA and IBA treatments on the developmental patterns, enzymatic and non-enzymatic antioxidant defence of hypericin non-producing Hypericum calycinum shoot cultures. It was shown that supplementation of PGR led to stimulation of phenylalanine ammonia-lyase and superoxide dismuthase, as well as non-enzymatic antioxidant glutathione. However, they inhibited glutathione reductase, as well as lowered the levels of flavonoids, ascorbate and dehydroascorbate in comparison with plant growth regulators-free control. Further on, it was established that elevation of IBA concentration slightly stimulated axillary shoot formation
and shoot length, but inhibited polyphenolic levels in vitro. These results are in agreement with our previous results of interrelations between biomass formation and polyphenolics production in other Hypericum species in vitro. An in vitro culture system optimization is in progress in order to increase biomass production and retain biosynthetic capacity of the species.
References
Aebi, H., 1984. Catalase in vitro. Methods of Enzymology, 105: 121–126.
Azizi, M., A. Ghani, T. Ebadi and S. Crockett, 2011. The ex situ comparison of two improved St. John’s Wort (Hypericum perforatum) cultivars with an Iranian wild population. Acta Horticulturae, 925: 163–170.
Bäcker, W., H. Bart, F. Bischoff, S. Grabley, R. Goedecke, W. Johannisbauer, V. Jordan, R. Stockfl eth, J. Strube and V. Wiesmet, 2006. Phytoextrakte – Produkte und Prozesse. Available online: http://www.dechema.de/dechema_media/Downloads/Extraktion/Phytoextrakte.pdf
Benson, E. E., 2000. Special symposium: in vitro plant recalcitrance do free radicals have a role in plant tissue culture recalcitrance? In Vitro Cell Dev. Biol. Plant., 36: 163–170.
Butterweck, V. and M. Schmidt, 2007. St. John’s wort: role of active compounds for its mechanism of action and efficacy. Wien Med. Wochenschr., 157: 356–361.
Cui, X. H., H. N. Murthy, C. H. Wu and K. Y. Paek, 2010. Sucrose-induced osmotic stress affects biomass, metabolite, and antioxidant levels in root suspension cultures of Hypericum perforatum L. Plant Cell Tiss. Organ Cult., 103: 7–14.
Danova, K., 2010. Production of polyphenolic compounds in shoot cultures of Hypericum species characteristic for The Balkan Flora. Botanica Serbica, 34: 29–36.
Danova, K., E. Čellárová and V. Kapchina-Toteva, 2010. Impact of growth regulators on in vitro regeneration of Hypericum rumeliacum Boiss. Journal of Environmental Prot. and Ecol., 11: 1285–1292.
Danova, K., E. Čellárová, A. Macková, Z. Daxnerová and V. Kapchina-Toteva, 2010. In vitro culture of Hypericum rumeliacum Boiss. and production of phenolics and flavonoids.
In Vitro Cellular and Developmental Biology – Plant, 46: 422–429.
Danova, K., B. Nikolova-Damianova, R. Denev and D. Dimitrov, 2012a. Influence of vitamins on polyphenolic content, morphological development, and stress response in shoot cultures of Hypericum spp. Plant Cell Tiss. Organ Cult., 110: 383–393.
Danova, K., 2014. Biotechnological Utilization of the Indigenous Biosynthetic Capacity of Medicinal and Aromatic Plants: Experience in the Genera Hypericum Pulsatilla and Essential Oil Bearing Artemisia alba Characteristic for the Balkan Region. Chapter 14, In: J.N. Govil (Editor), Recent Progress in Medicinal Plants Biotechnology and Genetic Engineering II & Vol.39, Studium Press LLC, USA, pp. 355–392.
Danova, K., B. Nikolova-Damianova, R. Denev and Y. Markovska, 2012b. Impact of pre-culture on short- and long-term in vitro recovery of the biosynthetic potential and enzymatic and non-enzymatic antioxidant defense of Hypericum rumeliacum Boiss. after cryostorage. Plant Growth Regulation, 68: 447–457.
Decosterd, L. A., E. Hoffmann, R. Kyburz, D. Bray and K. Hostettmann, 1991. A new phloroglucinol derivative from Hypericum calycinum with antifungal and in vitro antimalarial activity. Planta Medica, 57: 548–551.
Fadzilla, N. M., R. P. Finch and R. H. Burdon, 1997. Salinity, oxidative stress and antioxidant responses in root culture of rice. J. Exp. Bot., 48: 325–331.
Foyer, C., J. Rowell and D. Walker, 1983. Measurement of the ascorbate content of spinach leaf protoplasts and chloroplasts during illumination. Planta, 157: 239–244.
Griffi th, O. W., 1980. Determination of glutathione disulphide using glutathione reductase and 2-vinylpyridine. Anal. Biochem., 106: 207–212.
Kartning, T., I. Göbel and B. Heydel, 1996. Production of hypericin, pseudohypericin and flavonoids in cell cultures of various Hypericum species and their chemotypes. Planta Med., 62: 51–53.
Kirmizibekmez, H., C. Bassarello, S. Piacente, E. Celep, I. Atay, G. Mercanoğlu and E. Yeşilada, 2009. Phenolic compounds from Hypericum calycinum and their antioxidant activity. Natural Product Communications, 4: 531–534.
Kirakosyan, A., T. M. Sirvent, D. M. Gibson and P. B. Kaufman, 2004. The production of hypericins and hyperforin by in vitro cultures of St. John’s wort (Hypericum perforatum). Biotechnol. Appl. Biochem., 39: 71–81.
Kitanov, G. M. and K. F. Blinova, 1987. Modern state of the chemical study of species of the genus Hypericum. Chemistry of Natural Compounds, 23: 151–166.
Marklund, S. and G. Marklund, 1974. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase, Eur. J. Biochem., 47: 469-474.
Nakano, Y. and K. Asada, 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol., 22: 867–880.
Savio, L. E. B., L. V. Astarita and E. R. Santarém, 2012. Secondary metabolism in micropropagated Hypericum perforatum L. grown in nonaerated liquid medium. Plant Cell Tiss. Organ Cult., 108: 465–472.
Sherwin, H. W. and J. M. Farrant, 1998. Protection mechanisms against excess light in the resurrection plants Craterostigma wilmsii and Xerophyta viscosa. Plant Growth Reg., 24: 203–210.
Singleton, V. L., R. Orthofer and R. M. Lamuela-Raventós, 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Method Enzymol., 299: 152–178.
Öztürk, Y., S. Aydin, R. Beis, K. H. C. Başer and H. Berberoğlu, 1996. Effects of Hypericum perforatum L and Hypericum calycinum L extracts on the central nervous system in mice. Phytomedicine, 3: 139–146.
Wolfram, E., L. Hostettler, S. Peter, M. Todorova, A. Trendofilova and K. Danova, 2013. Phytochemical assessment of the effect of stimulated in vitro multiplication on the metabolic profile of in vitro cultured Hypericum richeri and Artemisia alba. Planta Med., 79: PN119.
Yuan, Y. J., C. Li, Z. D. Hu, J. C. Wu and A. P. Zeng, 2002. Fungal elicitor induced cell apoptosis in suspension cultures of Taxus chinensis var. mairei for taxol production. Process Biochem.,38:193–198.
Zhishen, J., T. Mengcheng and W. Jianming, 1999. The determination of flavonoid content in mulberry and their scavenging effects on superoxide radicals. Food Chem., 64: 555–559.
Zobayed, S. M. A., F. Afreen and T. Kozai, 2005. Temperature stress can alter the photosynthetic efficiency and secondary metabolite concentrations in St. John’s wort. Plant Physiology and Biochemistry, 43: 977–984.
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