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Grapevine fanleaf virus (GFLV) is the causal agent of a widespread disease that affects vineyards. Since it is difficult to culture viruses, the availability of an easy and efficient method of virus maintenance in the laboratory would be of interest to virologists. The objective of this research was to determine an adequate culture medium that promotes callus growth and permits the preservation of GFLV on Vitis vinifera tissue. Fragments of in vitro cultured leaves (25 mm 2 ), originated from Cabernet Sauvignon positive for GFLV, were cultivated on a Murashige and Skoog (1962) medium, and the callus was monitored for the presence of GFLV every two weeks using ELISA. Higher 2,4-D concentration induced a higher growth, particularly when combined with a low BA concentration. The medium enriched with 1.0 ppm of 2,4-D combined with 0.5 ppm of BA showed the best result, with the callus area reaching more than 250 mm 2 after 8 weeks in culture. ELISA absorbance observed on callus tissues during the whole period was, at least, three times higher than that observed on leaves positive for GFLV kept either in vivo or in vitro and more than 18 times higher than that of the negative control. Any remarkable difference in absorbance was recorded during the period of callus cultivation. It was concluded that the viral load on the callus was not affected during this time, suggesting that this kind of in vitro culture is an efficient method to preserve GFLV.
365 372 Brown, P. T. H., Gobel, E., Lorz, H. (1991) RFLP analysis of Zea mays callus cultures and their regenerated plants. Theor. Appl. Genet. 81 , 227
263 265 Bajji, M., Lutts, S., Kinet, J. M. (2000): Physiological changes after exposure to and recovery from polyethylene glycol-induced water deficit in callus cultures issued
A simple and efficient protocol has been developed for high frequency plant regeneration through callus cultures derived from leaf bases of abiotic stress sensitive Asian indica rice variety IR 64. Leaf base segments (4–5 mm diameter) were obtained from 6-day-old dark grown seedlings germinated on halfstrength Murashige and Skoog medium and cultured on MS medium supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2.2–18 μM) and Kinetin (0.2–1.7 μM). Among the various combinations, 13.5 μM 2,4-D and 1.3 μM Kn resulted in high callus induction frequency (87.5%) with a maximum fresh weight of 0.22 g per segment. The regeneration frequency was 75.5% with multiple shoots within 3 weeks of transfer on MS medium supplemented with 13.3 μM 6-benzylamino purine and 8 μM Naphthaleneacetic acid. The shoots readily rooted on half-strength MS medium without any hormonal supplements. In vitro regenerated plantlets with multiple shoots and roots were transferred to sterile soil and vermiculite mix and maintained in shade house for 30 days. Complete plantlets were then transferred to nursery and acclimatized to the external environment until seed set. RAPD profile reveals monomorphism and thus confirming the genetic stability of the regenerated plants. This method has the potential for both direct as well as indirect method of transformation for the production of genetically modified plants.
54 Dahleen LS (1995) Improved plant regeneration from barley callus cultures by improved copper levels. Plant Cell Tiss Organ Cult 43
229 236 György, Z., Tolonen, A., Neubauer P. & Hohtola, A. (2005): Enhanced biotransformation capacity of Rhodiola rosea callus cultures for glycosid production
Endogenous carbohydrate (fructose, glucose and sucrose) fractions were measured in calli of potato genotypes with different field tolerance to drought. Under in vitro stress conditions induced by 0.8 M mannitol, sucrose level of calli increased extremely in medium-tolerant (by 424.5%) and sensitive (by 302.7%) genotypes whilst the rate of increase was 12–18-times lower in the drought tolerant variety. Results suggest the applicability of sucrose as biochemical marker for distinguish drought tolerant genotypes from great population in callus culture.
.) callus cultures. Food Res. Int. 38 , 937–942. Giovinazzo G. Characterization of in vitro anthocyanin-producing sour cherry (Prunus cerasus L.) callus cultures
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
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
