Abstract
The aim of this investigation was to develop a protocol to be used in asymmetric protoplast fusions with breeding material of melon and cucumber. Efficient methods for plant regeneration from unfused protoplasts of commercial melon lines were established and are reported. Ploidy levels of explant material and plants, regenerated from protoplasts were analyzed. Electrofusion was carried out between melon protoplasts and irradiated and non-irradiated donor protoplasts of cucumber. Although initial plating efficiencies of heterokaryons were high, development stopped after a few divisions. In control experiments, shoots were regenerated at high frequencies. In only two fusion experiments, development continued to the callus stage, but further development was not observed. When analyzed with PCR using arbitrary primers, the majority of these calli DNA were identical to the melon DNA. However, a few of the examined calli, although being mainly homologous to melon, were observed to have new bands corresponding to bands specific for cucumber. Due to sexual incompatibility, successful hybridization between cucumber and melon was never obtained by sexual crosses. We suggest that the failure to regenerate plants in our fused material could be explained partially by an analogous somatic incompatibility reaction.
Similar content being viewed by others
Abbreviations
- BA:
-
benzyl adenine
- 2,4-d :
-
2,4-dichlorophenoxyacetic acid
- FDA:
-
fluorescein diacetate
- FITC:
-
fluorescein isothiocyanate
- IAA:
-
indoleacetic acid
- MS:
-
Murashige & Skoog
- NAA:
-
naphtalene acetic acid
- PEG:
-
polyethylene glycol
- PE:
-
plating efficiency
- RAPD:
-
random amplified polymorphic DNA
References
Branchard M & Debeaujon I (1989) Somatic embryogenesis in Cucurbitaceae with specific reference to muskmelon (Cucumis melo L.). In: Proc. Cucurbitaceae 89: Evaluation and Enhancement of Cucurbit Germplasm Nov 29–Dec 2, 1989, (pp 9–20) Charleston, SC, USA.
Bokelmann GS, Jarl CI & Kool AJ (1990) An efficient method for plant regeneration of protoplasts of commercial varieties of melon. In: Nijkamp HJJ et al., (eds) Abstracts of VIIth International Congress on Plant Tissue and Organ Culture (p 8) Kluwer Academic Publishers Dordrecht
Bokelmann GS, Jarl CI & Kool AJ (1991) Plant regeneration of protoplasts of different cultivars of melon (Cucumis melo L.). Cuc. Gen. Coop. Rep. 14: 78–80
Colijn-Hooymans CM, Bouwer R, Orczyk W & Dons JJM (1988) Plant regeneration from cucumber (Cucumis sativus L.) protoplasts. Plant Sci. 57: 63–71
Debeaujon I & Branchard M (1992) Induction of somatic embryogenesis and caulogenesis from cotyledon and leaf protoplast-derived colonies of melon (Cucumis melo L.). Plant Cell Rep. 12: 37–40
Dellaporte SL, Wood J & Hicks JB (1983) A plant molecular DNA minipreparation. Version II. Plant Mol. Biol. Rep. 1: 19–21
Dudits D, Fejer O, Hadlaczky Gy, Koncz Cs, Lazar GB & Horvath G (1980) Intergeneric gene transfer mediated by protoplast fusion. Mol. Gen. Genet. 179: 283–288
Gupta PP, Schieder OvGupta M (1982) Correction of nitrase reductase defect in auxotrophic plant cells through protoplast-mediated intergeneric gene transfer. Mol. Gen. Genet. 188: 378–383
Gupta PP, Schieder O & Gupta M (1984) Intergeneric nuclear gene transfer between somatically and sexually incompatible plants through asymmetric protoplast fusion. Mol. Gen. Genet. 197: 30–35
Harms CT (1983) Somatic incompatibility in the development of higher plant somatic hybrids. Q. Rev. Biol. 58: 325–353
Kao KN & Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at very low population density in liquid media. Planta 126: 105–110
Li R, Sun Y, Zhang L & Li X (1990) Plant regeneration from cotyledon protoplasts of Xinjiang muskmelon. Plant Cell Rep. 9: 199–203
Murashige T & Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Plant Physiol. 15: 473–497
den Nijs APM & Custers JBM (1990) Introducing resistances into cucumbers by interspecific hybridization. In: Bates DM, Robinson RW, Jeffrey C (eds) Biology and Utilization of the Cucurbitaceae, Cornell University Press (pp 382–396). New York
Provvidenti R (1987) Inheritance of resistance to a strain of Zucchini Yellow Mosaic virus in cucumber. HortSci. 22(1): 102–103
Shahin EA (1985) Totipotency of tomato protoplasts. Theor. Appl. Genet. 69: 235–240
Sree Ramulu K, Dijkhuis P, Roest S, Bokelmann GS & de Groot B (1984) Early occurrence of genetic stability in protoplast cultures of potato. Plant Sci. L. 36: 79–86
Tabei Y, Nishio T & Kanno T (1992) Shoot regeneration from cotyledonary protoplasts of melon (Cucumis melo L. cv. Charentais). J. Japan. Soc. Hort. Sci. 61 (2): 317–322
Visser DL, van Leeuwen L & Kho YO (1980) The IVT Cucumis species collection, some aspects of taxonomical and crossability studies. In: Eucarpia meeting on “Breeding of Cucumber and Melons” (pp 44–48) Wageningen
Williams JGK, Kubelik AR, Livak KJ, Rafalski JA & Tingey SV (1991) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18 (22): 6531–6535
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jarl, C.I., Bokelmann, G.S. & De Haas, J.M. Protoplast regeneration and fusion in Cucumis: melon × cucumber. Plant Cell Tiss Organ Cult 43, 259–265 (1995). https://doi.org/10.1007/BF00039953
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00039953