Summary
The effect of the type of gelling agent and of several antibiotics on the adventitious bud regeneration from in vitro leaves was tested on eight pear genotypes. The use of gellan gum (Phytagel™) in the medium instead of agar had a very strong positive effect on the rate of adventitious bud regeneration for all pear genotypes tested in this study. This gelling agent induced faster cell divisions than agar, thus more callus was produced on wound sites and subsequently more buds regenerated. Incubation on gellan gum medium during the first 20 d of bud induction was sufficient to induce a stimulatory effect on regeneration and limited the production of hyperhydric buds. In the prospect of Agrobacterium transformation, the effect of several antibiotics was tested. Cefotaxime (200 mg/l) plus ticarcillin/clavulanic acid (100 mg/l) could be used in the culture medium without affecting the frequency of bud regeneration. The inhibition of bud regeneration was obtained with different kanamycin concentrations according to the gelling agent in the medium. On gellan gum medium, a concentration of 100 mg/l of kanamycin was suitable. These conditions can be recommended for experiments on Agrobacterium-mediated transformation of pear, where bacterial inoculation and presence of antibiotics generally reduce and delay bud regeneration.
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Abu-Quaoud, H.; Skirvin, R. M.; Below, F. E. Influence of nitrogen form and NH4-N/NO3-N ratios on adventitious shoot formation from pear (Pyrus communis) leaf explants in vitro. Plant Cell Tissue Organ Cult. 27:315–319; 1991.
Chevreau, E.; Leblay, C. The effect of mother plant pretreatment and explant choice on regeneration from in vitro pear leaves. Acta Hortic. 336:263–268; 1993.
Chevreau, E.; Skirvin, R. M. Pear. In: Hammerschlag, F. A.; Litz, R. E., eds. Biotechnology of perennial fruit crops. Biotechnology in Agriculture No. 8. C.A.B. International 263–276; 1992.
De Bondt, A.; Eggermont, K.; Penninckx, I., et al. Agrobacterium-mediated transformation of apple (Malus × domestica Borkh.): an assessment of factors affecting regeneration of transgenic plants. Plant Cell Rep. 15:549–554; 1996.
Hadrami, E.; Housti, F.; Michaux-Ferriere, N., et al. Effects of gelling agents and liquid medium on embryogenic potential, polyamines and enzymatic factors in browning in Hevea brasiliensis calli. J. Plant Physiol. 141:230–233; 1993.
Holford, P.; Newbury, H. J. The effects of antibiotics and their breakdown products on the in vitro growth of Antirrhinum majus. Plant Cell Rep. 11:93–96; 1992.
Joersbo, M.; Okkels, F. T. Calcium reduces toxicity of aminoglycoside antibiotics in sugar beet explants in vitro. Physiol. Plant. 97:245–250; 1996.
Leblay, C.; Chevreau, E.; Raboin, L. M. Adventitious shoot regeneration from in vitro leaves of several pear cultivars (Pyrus communis L.). Plant Cell Tissue Organ Cult. 25:99–105; 1991.
Luft, J. H. Improvements in epoxy resin embedding methods. J. Biophys. Biochem. Cytol. 9:409–414; 1961.
Maheshwaran, G.; Welander, M.; Hutchinson, J., et al. Transformation of apple rootstock M26 with Agrobacterium tumefaciens. J. Plant Physiol. 139:560–568; 1992.
Mathews, H.; Wagoner, W.; Cohen, C., et al. Efficient genetic transformation of red raspberry, Rubus ideaus L. Plant Cell Rep. 14:471–476; 1995.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Paques, M. Vitrification and micropropagation: causes, remedies and prospects. Acta Hortic. 289:283–290; 1991.
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. 111(6):976–980; 1986.
Predieri, S.; Fasolo Fabbri Malavasi, F.; Passey, A. J., et al. Regeneration from in-vitro leaves of Conference and other pear cultivars (Pyrus communis L.). J. Hortic. Sci. 64(5):553–559; 1989.
Reed, B. M. Screening Pyrus germplasm for in vitro rooting response. HortScience 30(6):1292–1294; 1995.
Sabatini, D. D.; Bebsch, K.; Barnett, R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol. 17:19–58; 1963.
Scherer, P. A. Standardization of plant micropropagation by usage of a liquid medium with polyurethane foam plugs or a solidified medium with the gellan gum gelrite instead of agar. Acta Hortic. 226:107–114; 1988.
Scherer, P. A.; Muller, E.; Lippert, H., et al. 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 the gellan gum gelrite. Acta Hortic. 226:655–658; 1988.
Seingre, D.; O’Rourke, J.; Gavillet, S., et al. Influence of gelling agent and carbon source on the in vitro proliferation rate of apple rootstock EM IX. Acta Hortic. 289:151–155; 1991.
Turner, S. R.; Singha, S. Vitrification of crabapple, pear and Geum on gellan gum-solidified culture medium. HortScience 25:1648–1650; 1990.
Weising, K.; Schell, J.; Kahl, G. Foreign genes in plants: transfer, structure, expression and applications. Annu. Rev. Genet. 22:421–477; 1988.
Ye, X.; Brown, S.; Scorza, R., et al. Genetic transformation of peach tissues by particle bombardment. J. Am. Soc. Hortic. Sci. 119(2):367–373; 1994.
Yeo, D. Y.; Reed, B. M. Micropropagation of three Pyrus rootstocks. HortScience 30(3):620–623; 1995.
Yepes, L. M.; Aldwinckle, H. S. Factors that affect leaf regeneration efficiency in apple, and effects of antibiotics in morphogenesis. Plant Cell Tissue Organ Cult. 37:257–269; 1994.
Zimmerman, R.; Bhardwaj, S. V.; Fordham, I. M. Use of starch-gelled medium for tissue culture of some fruit crops. Plant Cell Tissue Organ Cult. 43:207–213; 1995.
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Chevreau, E., Mourgues, F., Neveu, M. et al. Effect of gelling agents and antibiotics on adventitious bud regeneration from In vitro leaves of pear. In Vitro Cell.Dev.Biol.-Plant 33, 173–179 (1997). https://doi.org/10.1007/s11627-997-0017-7
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DOI: https://doi.org/10.1007/s11627-997-0017-7