Skip to main content
Log in

Genetic analysis of protoplast regeneration ability in Brassica oleracea

  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

In Brassica oleracea L., plant regeneration from protoplasts is genotype-dependent and colony formation can be obtained routinely. In order to identify genes for regenerability, we performed a genetic analysis of the characteristic in the F2 generation of a cross between two accessions selected for high and low regenerability. Regeneration frequencies were obtained from protoplast culture of 248 individual F2 plants after 5, 10, and 15 weeks of culture on regeneration medium. Broad-sense heritability estimate was 0.492 at the early stage and lower (0.046–0.149) at advanced stages. The frequency distribution observed during short-term culture can be explained by two independent loci with duplicate dominant genes controlling regeneration. In long-term culture, one additional dominant gene can confer regeneration; i.e., three independent loci are responsible for regenerability. The results suggest that selection for high regeneration response may be efficient at early stages because of the lower environmental influence on the characteristic, and because fewer genes are involved at this stage than at advanced stages. The control of regenerability by a few dominant genes facilitates incorporation of the trait into breeding material. Molecular markers linked to the genes may assist in the selection of genotypes with high regeneration percentage in the future.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Armstrong CL, Romero-Severson J & Hodges TK (1992) Improved tissue culture response of an elite maize inbred through backcross breeding, and identification of chromosomal regions important for regeneration by RFLP analysis. Theor. Appl. Genet. 84: 755–762

    Article  Google Scholar 

  • Ben Amer IM, Korzum V, Worland AJ & Börner A (1997) Genetic mapping of QTL controlling tissue-culture response on chromosome 2B of wheat (Triticum aestivum L.) in relation to major genes and RFLP markers. Theor. AppI. Genet. 94: 1047–1052

    Article  CAS  Google Scholar 

  • Birhman RK, Laublin G & Cappadocia M (1994) Genetic control of in vitro shoot regeneration from leaf explants in Solanum chacoense Bitt. Theor. Appl. Genet. 88: 535–540

    Article  Google Scholar 

  • Bonfils AC, Sproule A, Webb JA & Keller WA (1992) Plant regeneration from stem cortex explant and protoplast cultures of Brassica juncea (mustard). Plant Cell Rep. 11: 614–617

    Article  Google Scholar 

  • Buiatti M, Baroncelli S, Bennici A, Pagliai M & Tesi R (1974) Genetics of growth and differentiation in vitro of Brassica oleracea var. botrytis. II. An in vitro and in vivo analysis of a diallel cross. Z. Pflanzenzüchtg. 72: 269–274

    Google Scholar 

  • Cheng J & Veilleux RE (1991) Genetic analysis of protoplast culturability in Solanum phureja. Plant Science 75: 257–265

    Article  Google Scholar 

  • Coleman M, Waugh R & Powell W (1990) Genetical analysis of in vitro cell and tissue culture response in potato. Plant Cell Tiss. Org. Cult. 23: 181–186

    Google Scholar 

  • Crea F, Bellucci M, Damiani F & Arcioni S (1995) Genetic control of somatic embryogenesis in alfalfa (Medicago sativa L. cv. Adriana). Euphytica 81: 151–155

    Article  Google Scholar 

  • Deglene L, Lesignes P, Alibert G & Sarrafi A (1997) Genetic control of organogenesis in cotyledons of sunflower (Helianthus annuus). Plant Cell Tiss. Org. Cult. 48: 127–130

    Article  Google Scholar 

  • Glimelius K (1984) High growth rate and regeneration capacity of hypocotyl protoplasts in some Brassicaceae. Physiol. Plant. 61: 38–44

    Article  CAS  Google Scholar 

  • Hansen LH (1998) Intertribal somatic hybridization between rapid cycling Brassica oleracea L. and Camelina sativa (L.) Crantz. Euphytica 104: 173–179

    Article  Google Scholar 

  • Hansen LN & Earle ED (1994) Regeneration of plants from protoplasts of rapid cycling Brassica oleracea L. Plant Cell Rep. 13: 335–339

    Article  CAS  Google Scholar 

  • Izhar S & Power JB (1977) Genetical studies with petunia leaf protoplasts. I. Genetic variation to specific growth hormones and possible genetic control on stages of protoplast development in culture. Plant Sci. Let. 8: 375–383

    Article  CAS  Google Scholar 

  • Jaiswal SK, Hammatt N, Bhojwani SS, Cocking EC & Davey MR (1990) Plant regeneration from cotyledon protoplasts of Brassica carinata.Plant Cell Tiss. Org. Cult. 22: 159–165

    Article  Google Scholar 

  • Jourdan PS & Earle ED (1989) Genotypic variability in the frequency of plant regeneration from leaf protoplasts of four Brassica spp. and of Raphanus sativus. J. Amer. Soc. Hort. Sci. 114: 343–349

    Google Scholar 

  • Jourdan PS, Earle ED & Mutschler MA (1990) Improved protoplast culture and stability of cytoplasmic traits in plants regenerated from leaf protoplasts of cauliflower (Brassica oleracea ssp. botrytis). Plant Cell Tiss. Org. Cult. 21: 227–236

    Article  Google Scholar 

  • Kik C & Zaal Macm(1993) Protoplast culture and regeneration from Brassica oleracea ‘rapid cycling’ and other varieties. Plant Cell Tiss. Org. Cult. 35: 107–114

    Article  Google Scholar 

  • Kirti PB & Chopra VL (1990) Rapid plant regeneration through organogenesis and somatic embryogenesis from cultured protoplasts of Brassica juncea.Plant Cell Tiss. Org. Cult. 20: 65–67

    Article  CAS  Google Scholar 

  • Komatsuda T, Enomoto S & Nakajima K (1989) Genetics of callus proliferation and shoot differentiation in barley. Journal of Heredity 80: 345–350

    Google Scholar 

  • Koornneef M, Hanhart C, Jongsma M, Toma I, Weide R, Zabel P & Hille J (1986)Breeding of a tomato genotype readily accessible to genetic manipulation. Plant Sci. 45: 201–208

    Article  Google Scholar 

  • Koornneef M, Hanhart CJ & Martinelli L (1987)A genetic analysis of cell culture traits in tomato. Theor. Appl. Genet. 74: 633–641

    Article  Google Scholar 

  • Lange CE, Federizzi LC, Carvalho FIF, Tavares MJCMS, Dornelles ALC & Handel CL (1995) Genetic analysis of somatic embryogenesis and plant regeneration of wheat (Triticum aestivum L.). J. Genet. Breed. 49: 195–200

    Google Scholar 

  • Linsmaier EM & Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol. Plant. 18: 100–127

    Article  CAS  Google Scholar 

  • Loudon PT, Nelson RS & Ingram DS (1989) Studies of protoplast culture and plant regeneration from commercial and rapid-cycling Brassica species. Plant Cell Tiss. Org. Cult. 19: 213–224

    Article  Google Scholar 

  • Molina RV & Nuez F (1996) The inheritance of organogenic response in melon. Plant Cell Tiss. Org. Cult. 46: 251–256

    Article  Google Scholar 

  • Morocz S, Donn G, Nemeth J & Dudits D (1990) An improved system to obtain fertile regenerants via maize protoplasts isolated from a highly embryogenic maize suspension culture. Theor. Appl. Genet. 80: 721–726

    Google Scholar 

  • Nadolska-Orczyk A & Malepszy S (1989) In vitro culture of Cucumis sativus L. 7. Genes controlling plant regeneration. Theor. Appl. Genet. 78: 836–840

    Article  Google Scholar 

  • Narasimhulu SB, Kirti PB, Prakash S & Chopra VL (1993) Rapid and high frequency shoot regeneration from hypocotyl protoplasts of Brassica nigra. Plant Cell Tiss Org. Cult. 32: 35–39

    Article  CAS  Google Scholar 

  • Ockendon DJ & Sutherland RA (1987) Genetic and non-genetic factors affecting anther culture of Brussels sprouts (Brassica oleracea var. gemmifera). Theor. Appl. Genet. 74: 566–570

    Article  Google Scholar 

  • Pelletier G, Primard C, Vedel F, Chetrit P, Remy R, Rouselle P & Renard M (1983) Intergeneric cytoplasm hybridization in Cruciferae by protoplast fusion. Mol. Gen. Genet. 191: 244–250

    Article  CAS  Google Scholar 

  • Ray IM & Bingham ET (1989)Breeding diploid alfalfa for regeneration from tissue culture. Crop Sci. 29: 1545–1548

    Article  Google Scholar 

  • Rosati C, Landi P & Tuberosa R (1994) Recurrent selection for regeneration capacity from immature embryo-derived calli in maize. Crop Sci. 34: 343–347

    Article  Google Scholar 

  • Tan Mmc, Colijn-Hooymans CM, Lindhout WH & Kool AJ (1987) A comparison of shoot regeneration from protoplasts and leaf discs of different genotypes of the cultivated tomato. Theor. Appl. Genet. 75: 105–108

    Article  Google Scholar 

  • Taylor TE & Veilleux RE (1992) Inheritance of competencies for leaf disc regeneration, anther culture, and protoplast culture in Solanum phureja and correlations among them. Plant Cell Tiss. Org. Cult. 31: 95–103

    Article  Google Scholar 

  • Van Sint Jan V, Laublin G, Birhman RK & Cappadocia M (1996) Genetic analysis of leaf explant regenerability in Solanum chacoense. Plant Cell Tiss. Org. Cult. 47: 9–13

    Article  Google Scholar 

  • Walters TW & Earle ED (1990) A simple versatile feeder layer system for Brassica oleracea protoplast culture. Plant Cell Rep. 9: 316–319

    Article  Google Scholar 

  • Wan Y, Sorensen EL & Liang GH (1988) Genetic control of in vitro regeneration in alfalfa (Medicago sativa L.). Euphytica 39: 3–9

    Article  Google Scholar 

  • Willman MR, Schroll SM & Hodges TK (1989) Inheritance of somatic embryogenesis and plantlet regeneration from primary (type 1) callus in maize. In Vitro Cell. Develop. Biol. 25: 95–100

    Google Scholar 

  • Yamashigi M, Otani M, Higashi M, Fukuta Y, Fukui K, Yano M & Shimada T (1998) Chromosomal regions controlling anther culturability in rice (Oryza sativa L.). Euphytica 103: 227–234

    Article  Google Scholar 

  • Zhang L & Hattori K (1996) Genetic analysis of regeneration ability in rice seed-callus. Genes Genet. Syst. 71: 313–3l7

    Article  Google Scholar 

  • Zhao K-N, Bittisnich DJ, Halloran GM & Whitecross MI (1995) Studies of cotyledon protoplast cultures from B.napus, B. campestris and B.oleracea. II: Callus formation and plant regeneration. Plant Cell Tiss. Org. Cult. 40: 73–84

    Article  CAS  Google Scholar 

  • Zhao K-N, Whitecross MI & Bittisnich DJ (1994) Studies on plant regeneration from cotyledonary protoplasts of B.campestris. Plant Cell Rep. 13: 164–170

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hansen, L.N., Ortiz, R. & Andersen, S.B. Genetic analysis of protoplast regeneration ability in Brassica oleracea. Plant Cell, Tissue and Organ Culture 58, 127–132 (1999). https://doi.org/10.1023/A:1006359804328

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006359804328

Navigation