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Influence of agar on in vitro cultures: II. Biological performance of ranunculus on media solidified with three different agar brands

  • Physiology
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Summary

The type of gelling agent can influence to a large extent clonal propagation of Ranunculus asiaticus L. through axillary bud stimulation. In preliminary experiments we identified three agar brands (Oxoid=OX, Merck=MK, and Roth=RT) which affect the availability of water and minerals to tissues in different ways. In the present study we investigate the influence of these agars on the in vitro performance of Ranunculus. On OX and MK gels, growth was satisfactory, although the former had a more promotive effect on fresh and dry weight production and on multiplication rate. Growth and development of shoots were poor on RT; shoot clumps showed symptoms of hyperhydricity, with shoots having large dark-green malformed leaves and very elongated petioles. Epidermal strips of leaves from shoots grown on the different gels and collected at the end of the culture period revealed differences according to the agar brand on which the plantlets were cultured. Severe structural deformations of stomata could be detected on RT-grown shoots. The analyses of the sugar content of the gel at the end of the culture period demonstrated that the explants grown on RT gels are strictly dependent on the carbohydrates in the medium. On OX and MK gels the heterotrophic metabolism was lowered compared to RT-grown explants. The agar brand on which plantlets were grown also influenced water retention capacity and water content of the shoots. Experiments with tritiated water were undertaken to better understand the water fluxes inside the vessel and to investigate the difference in “pump function” exerted by shoots cultured on the three gels. Shoots grown on OX media showed the best “pump function,” which would account for the better results obtained on this gel. On the basis of the relationship between gel properties and the growth of Ranunculus shoots, we conclude that the different physiological responses on the three gels are a reflection of different water and nutrient availability in the different media.

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References

  • Beruto, M. Agar and gel characteristics with special reference to micropropagation systems of Ranunculus asiaticus L. Ph.D. thesis, Gent, Faculteit van de Landbouwkundige en Toegepaste Biologische Wetenschappen, University Gent, Belgium; 1997: 251 P.

  • Beruto, D.; Beruto, M.; Ciccarelli, C.; Debergh, P. Matric potential evaluations and measurements for gelled substrates. Physiol. Plant. 84:151–157; 1995.

    Article  Google Scholar 

  • Beruto, M.; Beruto, D.; Debergh, P. Influence of agar on in vitro cultures: I. Physico-chemical properties of agar and agar gelled media. In Vitro Cell. Dev. Biol. Plant:35:1; 1998.

    Google Scholar 

  • Beruto, M.; Cane, G.; Debergh, P. Field performance of tissue-cultured plants of Ranunculus asiaticus L. Sci. Hortic. 66:229–239; 1996a.

    Article  Google Scholar 

  • Beruto, M.; Curir, P.; Debergh, P. Callus growth and somatic embryogenesis in thalamus tissue of Ranunculus asiaticus L. cultivated in vitro: cytokinin effect and phenol metabolism. In Vitro Cell. Dev. Biol. Plant 32:154–160; 1996b.

    CAS  Google Scholar 

  • Beruto, M.; Debergh, P. Differences in availability of water to in vitro cultures using different brands of agars. Acta Hortic. 289:331–333; 1991.

    Google Scholar 

  • Beruto, M.; Debergh, P. Somatic embryogenesis in Ranunculus asiaticus L. hybr. thalamus cultivated in vitro. Plant Cell Tissue Organ Cult. 29:161–165; 1992.

    Article  CAS  Google Scholar 

  • Beruto, M.; Portogallo, C. Factors affecting the growth of Ranunculus asiaticus L. in vitro. Proc. ISHS Congress on Science and Horticulture Interfaces and Interactions, Brussels, Belgium, 2–7 August; 1998. Acta Hortic., in press.

  • Beruto, M.; Siccardi, G.; Lercari, G. Risultati preliminari nella propagazione in vitro del ranuncolo. In: Proceeding of Symposium on Colture in vitro e micropropagazione in ortoflorofrutticoltura, Abstracts, Cesena, Italy. G. Gaggioli, Edt. Bologna: Lo Scarabeo Press; 1989: p. 33.

    Google Scholar 

  • Bornman, C. H.; Vogelmann, T. C. Effect of rigidity of gel medium on benzyladenine-induced adventitious bud formation and vitrification in vitro in Picea abies. Physiol. Plant 61:505–512; 1984.

    Article  CAS  Google Scholar 

  • Capellades, M. Histological and ecophysiological study of the changes occurring during the acclimatization of in vitro cultured roses. Ph.D. Thesis, Gent, Faculteit van de Landbouwwetenschappen; 1989: 98 P.

  • Debergh, P. Effects of agar brand and concentration on the tissue culture medium. Physiol. Plant. 59:270–276; 1983.

    Article  CAS  Google Scholar 

  • Debergh, P.; De Riek, J; Matthys, D. Nutrient supply and growth of plants in culture. In: Lumsden, P. J.; Nicholas, J. R.; Davies, W. J., ed. Physiology, growth and development of plants in culture. Dordrecht, The Netherlands: Kluwer Academic Publishers; 1994:58–68.

    Google Scholar 

  • Debergh, P.; Harbaoui, Y.; Lemeur, R. Mass propagation of globe artichoke (Cynara scolymus): evaluation of different hypotheses to overcome vitrification with special reference to water potential. Physiol. Plant. 53:181–187; 1981.

    Article  Google Scholar 

  • De Gryze, C. Waterhuishouding door in vitro geteelde Rosa multiflora MSc thesis, Gent, Faculteit van de Landbouwwetenschappen; 1993: 100 P.

  • De Proft, M. P.; Maene, L.; Debergh, P. Carbon dioxide and ethylene evolution in the culture atmosphere of Magnolia cultured in vitro. Physiol. Plant. 65:375–379; 1985.

    Article  Google Scholar 

  • De Riek, J. Interaction between sucrose uptake and photosynthesis in micropropagated Rosa multiflora L. Ph.D. thesis, Gent, Faculteit van de Landbouwwetenschappen; 1995: 150 P.

  • George, E. F.; Sherrington, P. D. Plant propagation by tissue culture. Exegetics Ltd., Eversley, Basingstoke, Hants, England; 1984: 709 P.

  • Ghashghaie, J.; Brenckmann, F.; Saugier, B. Effects of agar concentration on water status and growth of rose plants cultured in vitro. Physiol. Plant. 82:73–78; 1991.

    Article  Google Scholar 

  • Kevers, C.; Coumans, M.; Coumans-Gilles, F.; Gaspar, T. Physiological and biochemical events leading to vitrification of plants cultured in vitro. Physiol. Plant. 61:69–74; 1984.

    Article  CAS  Google Scholar 

  • Kozai, T.; Smith, M. A. L. Environmental control in plant tissue culture—general introduction and overview. In: Aitken-Christie, J.; Kozai, T.; Smith, M. A. L., ed. Automation and environmental control in plant tissue culture. Dordrecht, The Netherlands: Kluwer Academic Press; 1995:301–318.

    Google Scholar 

  • Ladyman, J. A. R.; Girard, B. Cucumber somatic embryo development on various gelling agents and carbohydrate sources. HortScience 27:164–165; 1992.

    Google Scholar 

  • Lee, N.; Wetzstein, H. Y.; Sommer, H. E. The effect of agar vs liquid medium on rooting in tissue-cultured sweetgum. HortScience 21:317–318; 1986.

    Google Scholar 

  • Lercari, G. B.; Littardi, M.; Garibaldi, A. E. In vitro propagation of Ranunculus: experimental results. Riv. Ortoflorofrutt. It. 69:321–326; 1985.

    Google Scholar 

  • Mackay, W. A.; Kitto, S. L. Factors affecting in vitro shoot proliferation of French tarragon. J. Am. Soc. Hortic. Sci. 113:282–287; 1988.

    CAS  Google Scholar 

  • Maia, E.; Bettachini, B.; Beck, D.; Marais, A. Régéneration de renoncules par culture d’apex in vitro. Ann. Phytopathol. 5:125–129; 1973.

    Google Scholar 

  • Meynet, J.; Duclos, A. Culture in vitro de la renoncule de fleuristes (R. asiaticus L.) I. Néoformation et multiplication végétative in vitro de plantes à partir de tronçon de thalamus. Agronomie 2:157–162; 1990a.

    Google Scholar 

  • Meynet, J.; Duclos, A. Culture in vitro de la renoncule de fleuristes (R. asiaticus L.) II. Production de plantes par culture d’anthères in vitro. Agronomie 10:213–218; 1990b.

    Google Scholar 

  • Meynet, J.; Duclos, A. Culture in vitro de la renoncule de fleuristes (R. asiaticus L.) III. Etude des plantes produites par embryogenèse somatique à partir des tissus superficiels de l’anthère. Agronomie 10:285–290; 1990c.

    Google Scholar 

  • Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.

    Article  CAS  Google Scholar 

  • Nachi, N.; Le Guen, J. Dry matter accumulation and seed yield in faba bean (Vicia faba L.) genotypes. Agronomie 16:47–59; 1996.

    Google Scholar 

  • Owens, L. D.; Wozniak, C. A. Measurement and effects of gel matric potential and expressibility on production of morphogenic callus by cultured sugarbeet leaf discs. Plant Cell Tissue Organ Cult. 26:127–133; 1991.

    Article  Google Scholar 

  • Pasqualetto, P. L.; Zimmerman, R. H.; Fordham, I. Gelling agent and growth regulator effects on shoot vitrification of “Gala” apple in vitro. J. Am. Soc. Hortic. Sci. 11:976–980; 1986.

    Google Scholar 

  • Pasqualetto, P. L.; Zimmerman, R. H.; Fordham, I. The influence of cation and gelling agent concentrations on vitrification of apple cultivars in vitro. Plant Cell Tissue Organ Cult. 14:31–40; 1988a.

    Article  CAS  Google Scholar 

  • Pasqualetto, P. L.; Wergin, W. P.; Zimmerman, R. H. Changes in structure and elemental composition of vitrified leaves of ‘Gala’ apples in vitro. Acta Hortic. 227:352–357; 1988b.

    Google Scholar 

  • Pochet, B.; Scoman, V.; Mestdagh, M. M.; Moreau, B.; Andre, P. Influence of agar gel properties on the in vitro micropropagation of different clones of Thuja plicata. Plant Cell Rep. 10:406–409; 1991.

    Article  CAS  Google Scholar 

  • Pugliesi, C.; Rabaglio, M.; Cecconi, F.; Baroncelli, S. Plant regeneration from tissue cultures of Persian buttercup (Ranunculus asiaticus L.). Plant Cell Tissue Organ Cult. 28:125–128; 1992.

    Article  CAS  Google Scholar 

  • Romberger, J.; Tabor, C. A. The Picea abies shoot apical meristem in culture: I. Agar and autoclaving effects. Am. J. Bot. 58:131–140; 1971.

    Article  Google Scholar 

  • Singha, S. Influence of two commercial agars on in vitro shoot proliferation of “Almey” crabapple and “Seckel” pear. HortScience 19:227–228; 1984.

    CAS  Google Scholar 

  • Singha, S.; Townsend, C.; Oberly, G. H. Mineral nutrient status of crabapple and pear shoots cultured in vitro on varying concentrations of three commercial agars. J. Am. Soc. Hortic. Sci. 110:407–411; 1985.

    CAS  Google Scholar 

  • Scherer, P. A.; Muller, E.; Lipper, H.; Wolff, G. Multielement analysis of agar and gelrite impurities investigated by inductively coupled plasma emission spectrometry as well as physical properties of tissue culture media prepared with agar or gellan gum gelrite. Acta Hortic. 226:655–658; 1988.

    Google Scholar 

  • Sholten, H. J.; Pierik, R. L. M. Effect of agar quality on in vitro culture of plants. Acta Bot. Neerl. 40:230; 1991.

    Google Scholar 

  • Scholten, H. J.; Pierik, R. L. M. Quality parameters of agars for regeneration and micropropagation. In: Scaramuzzi, O., ed. Abstracts VIIIth International Congress of Plant Tissue and Cell Culture, June 12–17, Firenze, Italy; 1994: p. 11.

  • Stolz, L. P. Agar restriction of the growth of excised mature Iris embryos. J. Am. Soc. Hortic. Sci. 96:681–684; 1971.

    Google Scholar 

  • Williams, R. R.; Taji, A. M. Auxin type, gel concentration, rooting and survival of Cheiranthera volubilis in vitro. HortScience 24:305–307; 1989.

    CAS  Google Scholar 

  • Ziv, M. Quality of micropropagated plants—vitrification. In Vitro Cell. Dev. Biol. 27P:329–339; 1991.

    Google Scholar 

  • Ziv, M.; Ariel, T. Vitrification in relation to stomatal deformation and malfunction in carnation leaves in vitro. In: Lumsden, P. J.; Nicholas, J. R.; Davies, W. J., ed. Physiology, growth and development of plants in culture. Dordrecht, The Netherlands: Kluwer Academic Publishers; 1994:143–154.

    Google Scholar 

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Authors and Affiliations

  1. Regional Institute for Floriculture, Via Carducci 12, 18038, Sanremo (IM), Italy

    Margherita Beruto

  2. Istituto Chimica Agraria, University of Turin, Via P. Giuria 15, 10100, Torino, Italy

    Paolo Curir

  3. Laboratory of Horticulture, Faculty of Agronomical and Applied Biological Sciences, University of Gent, Coupure links 653, B-9000, Gent, Belgium

    Pierre Debergh

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  1. Margherita Beruto
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  2. Paolo Curir
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  3. Pierre Debergh
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Beruto, M., Curir, P. & Debergh, P. Influence of agar on in vitro cultures: II. Biological performance of ranunculus on media solidified with three different agar brands. In Vitro Cell.Dev.Biol.-Plant 35, 94–101 (1999). https://doi.org/10.1007/s11627-999-0017-x

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