Skip to main content
SpringerLink
Log in

Plant regeneration capacity of mesophyll protoplasts from Brassica napus and related species

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

Abstract

Protoplasts from a total of thirty-six genotypes of Brassica species – B. napus, B. campestris (syn. B. rapa), B. juncea, and three distant relatives, Orychophragmus violaceus, Isatis indigotica and Xinjiang wild rape – were analysed for shoot regeneration using a feeder culture system. With the exception of B. campestris and Xinjiang wild rape, some genotypes of all the species could regenerate plants with high efficiency (above 20% of isolated calli initiating shoots). Several genotypes with high regeneration ability were elite breeding lines. Culture conditions as well as genotype had a significant impact on shoot regeneration frequency. In particular, silver nitrate added to the regeneration medium at doses of 6 and 30 μM improved shoot regeneration frequency to 25.4% and 52.2% of isolated calli, respectively, compared to 7.3% percent shoot regeneration without silver nitrate in seven responsive genotypes. Addition of silver nitrate to the regeneration medium also induced shoot regeneration in non-responsive genotypes. Intact plants could be obtained within three months from protoplast isolation in the regenerative genotypes using the current culture system. Advantages of mesophyll protoplasts as compared to protoplasts isolated from hypocotyls for genetic manipulation in Brassica species are discussed.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Atanassov II, Atanassov SA, Dragoeva AI & Atanassov AI (1998) A new CMS source in Nicotiana developed via somatic cybridization between N.tabacum and N.alata.Theor. Appl. Genet. 97: 982–985

    Article  CAS  Google Scholar 

  • Chatterjee G, Sikdar SR, Das S & Sen SK (1985) Regeneration of plantlets from mesophyll protoplasts of Brassica juncea (L.) Czern. Plant Cell Rep. 4: 245–247

    Article  Google Scholar 

  • Chi GL & Pua EC (1989) Ethylene inhibitors enhanced de novo shoot regeneration from cotyledons of Brassica campestris ssp. Chinensis (Chinese cabbage) in vitro. Plant Sci. 64: 243–250

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

    Article  CAS  Google Scholar 

  • Hansen LN & Earle ED (1997) Somatic hybrids between Brassica oleracea L. and Sinapis alba L. with resistance to Alternaria brassicae (Berk.) Sacc. Theor. Appl. Genet. 94: 1078–1085

    Article  Google Scholar 

  • Heath DW & Earle ED (1997) Synthesis of high erucic acid rapeseed (Brassica napus L.) somatic hybrids with improved agronomic characters. Theor. Appl. Genet. 91: 1129–1136

    Google Scholar 

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

    Google Scholar 

  • Jourdan P & Salazar E (1993) Brassica carinata resynthesised by protoplast fusion. Theor. Appl. Genet. 86: 567–572

    Article  Google Scholar 

  • Kirti PB (1988) Somatic embryogenesis in hypocotyl protoplast culture of rapeseed (Brassica napus L.). Plant Breed. 100: 222–224

    Article  Google Scholar 

  • Kirti PB & Chopra VL (1988) Regeneration through shoot organogenesis and somatic embryogenesis in hypocotyl protoplast culture of mustard Brassica juncea (L.) Czern and Coss. Cruciferae Newsletter 13: 96

    Google Scholar 

  • Linsmaier EM & Skoog F (1965) Organic growth factor requirement 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 

  • Luo P, Lan ZQ & Li ZY (1994) Orychophragmus violaceus, a potential edible-oil crop. Plant Breed. 113: 83–85

    Article  Google Scholar 

  • Menczel L, Nagy F, Kiss ZS & Maliga P (1981) Streptomycin resistant and sensitive somatic hybrids of Nicotiana tabacum + Nicotiana knightiana 1: correlation of resistance to N.tabacum plastids. Theor. Appl. Genet. 59: 191–195

    Article  CAS  Google Scholar 

  • Müller J & Sonntag K (1998) Assessment of in vitro regeneration and differentiation for somatic hybridization of some Brassicaceae. In: Plant Biotechnology and In Vitro Biology in the 21st Century, IX International Congress on Plant Tissue and Cell Culture, Jerusalem, Israel

  • Olin-Fatih M (1996) The morphology, cytology, and C-banded karyotypes of Brassica campestris, B.oleracea, and B.napus plants regenerated from protoplasts. Theor. Appl. Genet. 93: 414–420

    Article  Google Scholar 

  • Pauk J, Fekete S, Vilkki J & Pulli S (1991) Protoplast culture and plant regeneration of different agronomically important Brassica species and varieties. J. Science Finland, 63: 371–378

    Google Scholar 

  • Pellitier 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  Google Scholar 

  • Prakash S, Kirti PB, Bhat SR, Gaikwad K, Kumar VD & Chopra VL (1998) A Moricandia arvensis-based cytoplasmic male sterility and fertility restoration system in Brassica juncea. Theor. Appl. Genet. 97: 488–492

    Article  CAS  Google Scholar 

  • Purnhauser L, Medgyesy P, Czako M, Dix PJ & Marton L (1987) Stimulatiom of shoot regeneration in Triticum aestivum and Nicotiana plumbaginifolia Viv. tissue culture using the ethylene inhibitor AgNO3. Plant Cell Rep. 6: 1–4

    Article  CAS  Google Scholar 

  • Pua EC (1990) Somatic embryogenesis and plant regeneration from hypocotyl protoplasts of Brassica juncea (L.) Czern & Coss. Plant Sci. 68: 231–238

    Article  CAS  Google Scholar 

  • Qian XZ & Guan CY (1988) Review on investigation and collection of Xinjiang and Yunnan wild rape. Oil Crops in China, Supplement: 1–3

  • Sigareva MA & Earle ED (1999) Camalexin induction in intertribal somatic hybrids between Camelina sativa and rapid-cycling Brassica oleracea. Theor. Appl. Genet. 98: 164–170

    Article  CAS  Google Scholar 

  • Sjödin C & Glimelius K (1989) Transfer of resistance against Phoma lingam to Brassica napus by asymmetric somatic hybridization combined with toxin selection. Theor. Appl. Genet. 78: 513–520

    Article  Google Scholar 

  • Songtad DD, Duncan DR & Wildholm JM (1988) Effect of 1-aminocyclopropane-1-carboxylic acid, silver nitrate, and norbornadiene on plant regeneration from maize callus cultures. Plant Cell Rep. 7: 262–265

    Article  Google Scholar 

  • Vamling K & Glimelius K (1990) IV.2 Regeneration of plants from protoplasts of oilseed Brassica crops. In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry 10, Legumes and Oilseed Crops I (pp 386–417). Springer-Verlag, Berlin

    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 

  • Wang QY & Wan XB (1988) A TMV-resistant material-Banlangen (Isatis indigotica Fort.). Acta-Agriculturae-Boreali-Sinica 3: 92–95 (in Chinese)

    Google Scholar 

  • Zhao KN, Whitecross MI & Bittisnich DJ (1994) Studies on plant regeneration from cotyledonary protoplasts in Brassica campestris. Plant Cell Rep. 13: 164–170

    Article  CAS  Google Scholar 

  • Zhao KN, Bittisnich DJ, Halloran GM & Whitecross MI (1995a) 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 HJ, Huang YJ & Wang YY (1995b) Breeding materials from intergeneric hybridization Brassica napus × Isatis indigotica. J. of Huazhong Agricultural University, Abstract Collection of 94' Sino-British Workshop on Brassica Genetics and Breeding, 17: 20

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Agricultural Sciences, The Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, DK-1871, Frederiksberg C, Denmark

    Qiong Hu, Sven B. Andersen & Lise N. Hansen

  2. Institute of Oil Crops, Chinese Academy of Agricultural Sciences, 430062, Wuhan, P.R. China

    Qiong Hu

Authors
  1. Qiong Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sven B. Andersen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lise N. Hansen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hu, Q., Andersen, S.B. & Hansen, L.N. Plant regeneration capacity of mesophyll protoplasts from Brassica napus and related species. Plant Cell, Tissue and Organ Culture 59, 189–196 (1999). https://doi.org/10.1023/A:1006417530587

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006417530587

Search

Navigation