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Thiols are known to play crucial roles in plant defense responses, e.g. against pathogens. In the present study, the efficiency of these molecules was evaluated in regenerated Cucurbita pepo L. subsp. pepo var. styriaca Greb. plants in response to compatible Zucchini yellow mosaic virus infection. Thiol contents were modulated by applying the cysteine precursor L-2-oxothiazolidine-4-carboxylic acid (OTC) to callus cells. Cytological and biochemical methods were used for determination of glutathione within callus cells and leaves of plants regenerated via somatic embryogenesis. Reduced glutathione levels (GSH) significantly increased in callus cells one week after 1.0 mM OTC treatment by about 56% and resulted in enhanced GSH levels throughout the investigation period of 13 weeks. Three weeks after ZYMV infection untreated plants regenerated from callus culture exhibited stunted growth forms, yellowing, mosaic and blistering on the leaves whereas 68% of 1.0 mM OTC pretreated regenerated plants were characterized by a complete absence of symptoms. Treatment with OTC stimulated the synthesis, conversion and translocation of thiols during advanced ZYMV infection. Compared to untreated infected leaves, a graduate increase in cysteine (cys) and γ-glutamyl-cysteine (γ-glucys) contents was measured in 0.5 mM and 1.0 mM OTC treated older leaves, whereas total glutathione (tGSH) decreased significantly after infection. In OTC treated younger leaves cys and tGSH concentrations remained at control levels and were unaffected by ZYMV. In contrast, OTC treatment caused a dramatic induction of γ-glucys, up to 191% in younger infected leaves. The results show that altered levels of thiols by OTC were accompanied by enhanced tolerance against ZYMV, suppression of ZYMV induced symptoms as well as delayed symptom development.

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., 1986. Substitution analysis of plant regeneration from callus culture in wheat. Plant Breeding 97: 261–263 Sutka J. Substitution analysis of plant regeneration from callus culture in

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163 169 Galiba G, Kovacs G and Sutka J (1986) Substitution analysis of plant regeneration from callus culture in wheat. Plant Breed. 97: 261

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.) callus cultures. Food Res. Int. 38 , 937–942. Giovinazzo G. Characterization of in vitro anthocyanin-producing sour cherry (Prunus cerasus L.) callus cultures

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Copper D.B., R.G. Sears, G.L. Lookhart & B.L. Jones, 1986. Heritable somaclonal variation in gliadin proteins of wheat plants derived from immature embryo callus culture. Theor Appl Genet 71: 784–790. Jones B

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salt tolerance in callus cultures. Plant Sci. , 73 , 227-232. Responses of osmotic and NaCl stress of wheat varieties differing in drought and salt tolerance in callus cultures Plant Sci

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Dolgykh, Y. I. (1994): Establishment of callus cultures and regeneration of maize plants. In: Bajaj, Y. P. S. (ed.), Biotechnology in Agriculture and Forestry, Maize, Vol. 2. Springer, Berlin, pp. 197

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Köves, E., Szabó, M. (1987) Ethylene production in habituated and auxin-requiring tobacco callus-cultures — Does ethylene play a role in the habituation. Physiol. Plant. 69 , 351–355. Szabó M

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473 477 Amitha, K., Reddy, T. P. (1996) Regeneration of plant lets from different explants and callus cultures of cowpea ( Vigna unguiculata L.). Phytomorph. 46 , 207

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Cheng, T.-Y., Smith, H. H. (1975) Organogenesis from callus culture of Hordeum vulgare . Planta 123 , 307–310. Smith H. H. Organogenesis from callus culture of Hordeum

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