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Callus Induction and Protoplast Isolation from Tissues of Cucumis Sativus L. and C. Melo L. Seedlings

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Biologia Plantarum

Abstract

Hypocotyls, cotyledons and true leaves of in vitro seedlings of 10 cucumber and melon genotypes resistant to downy and powdery mildew were cultured on several combinations of initiation and multiplication media to produce callus and subsequently cell suspensions as suitable sources for isolation and culture of protoplasts. Cotyledons of both species were shown to be the most responsive to variation in culture media. However, calli and cell suspensions derived from hypocotyls generally provided higher number of protoplasts by treatment with several enzymatic solutions. The protoplasts formed new cell walls after 12 h of culture in liquid culture medium and first cell division was observed 2 d later with more frequent divisions after one week of culture.

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References

  • Banks, M.S., Evans, P.K.: A comparison of the isolation and culture of mesophyll protoplasts from several Nicotianaspecies and their hybrids.-Plant Sci. Lett. 7: 409-416, 1976.

    Article  Google Scholar 

  • Bokelmann, G.S., Jarl, C.I., Kool, A.J.: Plant regeneration of protoplasts from different cultivars of melon (Cucumis melo).-Cucurbits Genet. Coop. 14: 78-80, 1991.

    Google Scholar 

  • Burza, W., Haczykowski, J., Malepszy, S.: In vitroculture of Cucumis sativusL. XVII. Direct somatic embryogenesis from protoplasts.-In: Doruchowski, R.W., Kozik, E., Niemirowicz-Szezytt, K. (ed.): Proceedings of Fifth Eucarpia CucurbitaceaeSymposium. Pp. 108-111, Research Institute for Vegetable Crops, Skierniewice 1992.

    Google Scholar 

  • Chee, P.P.: High frequency of somatic embryogenesis and recovery of fertile cucumber plants.-HortScience 25: 792-793, 1990.

    Google Scholar 

  • Cohen, Y., Eyal, H.: Downy mildew-, powdery mildew-and Fusariumwilt-resistant muskmelon breeding line PI 124111F.-Phytoparasitica 15: 187-195, 1987.

    Google Scholar 

  • Colijn-Hooymans, C.M., Bouwer, R., Dons, J.J.M.: Plant regeneration from cucumber protoplasts.-Plant Cell Tissue Organ Cult. 12: 147-150, 1988a.

    Article  Google Scholar 

  • Colijn-Hooymans, C.M., Bouwer, R., Orezyk, W., Dons, J.J.M.: Plant regeneration from cucumber (Cucumis sativus) protoplasts.-Plant Sci. 57: 63-71, 1988b.

    Article  Google Scholar 

  • Colijn-Hooymans, C.M., Hakkert, J.C., Jansen, J., Custers, J.B.M.: Competence for regeneration of cucumber cotyledons is restricted to specific developmental stages.-Plant Cell Tissue Organ Culture 39: 211-217, 1994.

    Article  Google Scholar 

  • Coutts, R.H.A., Wood, K.R.: Improved isolation and culture methods for cucumber mesophyll protoplasts.-Plant Sci. Lett. 9: 45-51, 1977.

    Article  CAS  Google Scholar 

  • Custers, J.B.M., Zijlstra, S., Jansen, J.: Somaclonal variation in cucumber (Cucumis sativusL.) plants regenerated via embryogenesis.-Acta bot. neerl. 39: 153-161, 1990.

    Google Scholar 

  • Debeaujon, I., Branchard, M.: Induction of somatic embryogenesis and caulogenesis from cotyledon and leaf protoplast-derived colonies of melon (Cucumis meloL.).-Plant Cell Rep. 12: 37-40, 1992.

    Article  Google Scholar 

  • Debeaujon, I., Branchard, M.: Somatic embryogenesis in Cucurbitaceae.-Plant Cell Tissue Organ Cult. 34: 91-100, 1993.

    Article  Google Scholar 

  • Dirks, R., Van Buggenum, M.: Regeneration of plants from mesophyll protoplasts of haploid and diploid Cucumis sativusL.-Physiol. Plant. 82: A18, 1991.

    Google Scholar 

  • Dunstan, D.I., Short, K.C.: Improved growth of tissue cultures of the onion, Allium cepa.-Physiol. Plant. 41: 70-72, 1977.

    Article  Google Scholar 

  • Ezura, H., Oosawa, K.: Ploidy of somatic embryos and the ability to regenerate plantlets in melon (Cucumis meloL.).-Plant Cell Rep. 14: 107-111, 1994.

    Article  CAS  Google Scholar 

  • Fellner, M., Binarová, P., Lebeda, A.: Isolation and fusion of Cucumis sativusand Cucumis meloprotoplasts.-In: Gómez-Guillamón, M.L., Soria, C., Cuartero, J., Torés, J.A., Fernandez-Muñoz, R. (ed.): Cucurbits Towards 2000. Proceedings of the VIthEucarpia Meeting on Cucurbit Genetics and Breeding Pp. 202-209. Estación Experimental "La Mayora", Málaga 1996.

    Google Scholar 

  • Fellner, M., Havránek, P.: Culture of protoplasts isolated from leaves and callus cultures of Allium sativumand Allium longicuspis: A preliminary report.-Biol. Zentralbl. 113: 317-328, 1994.

    Google Scholar 

  • Hejtmánek, M., Doležel, J., Holubová, I.: Staining of fungal cell walls with fluorescent brighteners: flow-cytometric analysis.-Folia microbiol. 35: 437-442, 1990.

    Google Scholar 

  • Jarl, C.I., Bokelmann, G.S., De Haas, J.M.: Protoplast regeneration and fusion in Cucumis: melon × cucumber.-Plant Cell Tissue Organ Cult. 43: 259-265, 1995.

    Google Scholar 

  • Kantharajah, A.S., Dood, W.A.: Factors that influence the yield and viability of cucumber (Cucumis sativusL.) cotyledon protoplasts.-Aust. J. Bot. 38: 169-175, 1990.

    Article  Google Scholar 

  • Kim, S.G., Chang, J.R., Cha, H.C., Lee, K.W.: Callus growth and plant regeneration in diverse cultivars of cucumber (Cucumis sativusL.).-Plant Cell Tissue Organ Cult. 12: 67-74, 1988.

    Article  Google Scholar 

  • Knerr, L.D., Staub, J.E., Holder, D.J., May, B.P.: Genetic diversity in Cucumis sativusL. assessed by variation at 18 allozyme coding loci.-Theor. Appl. Genet. 78: 119-128, 1989.

    Article  Google Scholar 

  • Kubaláková, M., Doležel, J., Lebeda, A.: Ploidy instability of embryogenic cucumber (Cucumis sativusL.) callus culture.-Biol. Plant. 38: 475-480, 1996.

    Google Scholar 

  • Láska, P., Lebeda, A.: Resistance in wild Cucumisspecies to the glasshouse whitefly (Trialeurodes vaporariorum). Arch. Züchtungsforsch. 19: 89-93, 1989.

    Google Scholar 

  • Lebeda, A.: Screening of wild Cucumisspecies for resistance to cucumber powdery mildew (Erysiphe cichoracearumand Sphaerotheca fuliginea).-Scientia Hort. 24: 241-249, 1984.

    Article  Google Scholar 

  • Lebeda, A.: Resistance in muskmelons to Czechoslovak isolates of Pseudoperonospora cubensisfrom cucumbers.-Scientia Hort. 45: 255-260, 1991.

    Article  Google Scholar 

  • Lebeda, A.: Search for resistance in wild Cucumisspecies to twospotted spider mite (Tetranychus urticae).-In: Doruchowski, R.W., Kozik, E., Niemirowicz-Szezytt, K. (ed.): Proceedings of Fifth Eucarpia CucurbitaceaeSymposium. Pp. 171-174. Research Institute for Vegetable Crops, Skierniewice 1992a.

    Google Scholar 

  • Lebeda, A.: Susceptibility of accessions of Cucumis sativusto Pseudoperonospora cubensis.-Ann. appl. Biol. 120(Suppl.: Tests Agrochem. Cult. No. 13): 102-103, 1992b.

    Google Scholar 

  • Lebeda, A., Křístková, E., Kubaláková, M.: Interspecific hybridization of Cucumis sativus× Cucumis meloas potential way to transfer resistance to Pseudoperonospora cubensis.-In: Gómez-Guillamón, M.L., Soria, C., Cuartero, J., Torés, J.A., Fernandez-Muñoz, R. (ed.): Cucurbits Towards 2000. Proceedings of the VIthEucarpia Meeting on Cucurbit Genetics and Breeding. Pp. 31-37. Estación Experimental "La Mayora", Málaga 1996.

    Google Scholar 

  • Lebeda, A., Křístková, E., Kubaláková, M., Havlický, T., Vagera, J., Binarová, P.: [Biotechnological methods in breeding of cucurbit vegetables].-Rost. Výroba (Praha) 39: 171-179, 1993. [In Czech.]

    Google Scholar 

  • Lebeda, A., Prášil, J.: Susceptibility of Cucumis sativuscultivars to Pseudoperonospora cubensis.-Acta phytopathol. entomol. hung. 29: 89-94, 1994.

    Google Scholar 

  • Leppik, E.E.: Searching gene centers of the genus Cucumisthrough host-parasite relationship.-Euphytica 15: 323-328, 1966.

    Article  Google Scholar 

  • Lhotský, B., Lebeda, A., Zvára, J.: Resistance of wild Cucumisspecies to gummy stem blight (Didymella bryoniae).-Acta phytopathol. entomol. hung. 26: 303-306, 1991.

    Google Scholar 

  • Molina, R.V., Nuez, F.: Correlated response of in vitroregeneration capacity from different source of explants in Cucumis melo.-Plant Cell Rep. 15: 129-132, 1995a.

    Article  CAS  Google Scholar 

  • Molina, R.V., Nuez, F.: Characterization and classification of different genotypes in a population of Cucumis melobased on their ability to regenerate shoots from leaf explants.-Plant Cell Tissue Organ Cult. 43: 249-257, 1995b.

    Google Scholar 

  • Moreno, V., Zubeldia, L., Roig, L.A.: A method for obtaining callus cultures from mesophyll protoplasts of melon (Cucumis meloL.).-Plant Sci. Lett. 34: 195-201, 1984.

    Article  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 

  • Nijs Den, A.P.N., Custers, J.B.M.: Introducing resistances into cucumbers by interspecific hybridization.-In: Bates, D.M., Robinson, R.W., Jeffrey, C. (ed.): Biology and Utilization of the Cucurbitaceae. Pp. 382-396. Comstock Publ. Assoc., Cornell Univ. Press, Ithaca-London 1990.

    Google Scholar 

  • Orezyk, W., Malepszy, S.: In vitroculture of Cucumis sativusL. V. Stabilizing effect of glycine on leaf protoplasts.-Plant Cell Rep. 4: 269-273, 1985.

    Article  Google Scholar 

  • Oridate, T., Oosawa, K.: Somatic embryogenesis and plant regeneration from suspension callus culture in melon (Cucumis meloL.). Jap. J. Breed. 36: 424-428, 1986.

    Google Scholar 

  • Pitrat, M.: Gene list for Cucumis meloL.-Cucurbits Genet. Coop. 3: 58-68, 1990.

    Google Scholar 

  • Punja, Z.K., Tang, F.A., Sarmento, G.G.: Isolation, culture and plantlet regeneration from cotyledon and mesophyll protoplasts of two pickling cucumber (Cucumis sativusL.) genotypes.-Plant Cell Rep. 9: 61-64, 1990.

    Article  Google Scholar 

  • Raamsdonk Van, L.W.D., Nijs Den, A.P.M., Jongerius, M.C.: Meiotic analyses of Cucumishybrids and an evolutionary evaluation of the genus Cucumis(Cucurbitaceae).-Plant System. Evol. 163: 133-146, 1989.

    Article  Google Scholar 

  • Roig, L.A., Zubeldia, L., Orts, M.C., Roche, M.V., Moreno, V.: Plant regeneration from cotyledon protoplasts of Cucumis melocv. Cantaloup Charentais.-Cucurbits Genet. Coop. 9: 74-77, 1986.

    Google Scholar 

  • Staub, J.E., Frederick, L., Marty, T.L.: Electrophoretic variation in cross-compatible wild diploid species of Cucumis.-Can. J. Bot. 65: 792-798, 1987.

    Article  CAS  Google Scholar 

  • Tabei, Y., Kanno, T., Nishio, T.: Regulation of organogenesis and somatic embryogenesis by auxin in melon, Cucumis meloL.-Plant Cell Rep. 10: 225-229, 1991.

    Article  CAS  Google Scholar 

  • Tabei, Y., Nishio, T., Kanno, T.: Shoot regeneration from cotyledonary protoplasts of melon (Cucumis melocv. Charentais).-J. jap. Soc. hort. Sci. 61: 317-322, 1992.

    Article  Google Scholar 

  • Tatlioglu, T.: Cucumber Cucumis sativusL.-In: Kalloo, G., Bergh, B.O. (ed.): Genetic Improvement of Vegetable Crops. Pp. 197-234. Pergamon Press, Oxford 1993.

    Google Scholar 

  • Thomas, C.E.: Downy and powdery mildew resistant muskmelon breeding line MR-1.-HortScience 21: 329, 1986.

    Google Scholar 

  • Widholm, J.M.: The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells.-Stain Technol. 47: 189-194, 1972.

    PubMed  CAS  Google Scholar 

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Authors
  1. M. Fellner
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  2. A. Lebeda
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Fellner, M., Lebeda, A. Callus Induction and Protoplast Isolation from Tissues of Cucumis Sativus L. and C. Melo L. Seedlings. Biologia Plantarum 41, 11–24 (1998). https://doi.org/10.1023/A:1001791812746

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  • DOI: https://doi.org/10.1023/A:1001791812746

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