Abstract
To establish a cytoplasmic male-sterile/restored fertility (cms-Rf) system for F1 seed production in Brassica napus, we transferred a gene from fertillity restored radish to B. napus by protoplast fusion. X-irradiated protoplasts, isolated from shoots of Raphanus sativus cv Kosena (Rf line), were fused with iodoacetamide-treated protoplasts of a B. napus cms cybrid. Among 300 regenerated plants, six were male-fertile. The fertile plants were characterized for petal color, chromosome number and the percentage of viable pollen grains. Three fertile plants had aneuploid chromosome numbers and white or cream petals, which is a dominant marker in radish. Of these three plants, one which had 2n = 47 chromosomes and white petals was used for further backcrosses. After two backcrosses, chromosome number and petal color became identical to that of B. napus. No female sterility was observed in the BC3 generations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bates GW, Hansekampf CA, Contolini CL, Piastuch WC (1987) Asymmetric hybridization in Nicotiana by fusion of irradiated protoplasts. Theor Appl Genet 74:718–726
Barsby TL, Chong PV, Yarrow SA, Sau-Ching Wu, Coumans M, Kemble RJ, Powell AD, Beversdorf WD, Pauls KP (1987) The combination of Polima cms and cytoplasmic triazine resistance in Brassica napus. Theor Appl Genet 73:809–814
Bauer-Weston B, Keller W, Webbe J, Gleddie S (1993) Production and characterization of asymmetric somatic hybrids between Arabidopsis thaliana and Brassica napus. Theor Appl Genet 86:150–158
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Bio Rep 4:19–21
Delourme R, Bouchereau A, Hubert N, Renard M, Landry BS (1994) Identification of RAPD markers linked to a fertility restorer gene for the Ogura radish cytoplasmic male sterility of rapeseed (Brassica napus L.). Theor Appl Genet 88:741–748
Dolstra O (1982) Synthesis and fertility of x Brassicoraphanus and ways of transferring Raphanus characters to Brassica. In: Foundation for Agricultural Plant Breeding (SVP), Wageningen, pp 1–86
Dudits D, Maroy E, Parznovszky T, Olah Z, Gyorgyey J, Cella R (1987) Transfer of resistance traits from carrot into tobacca by asymmetric somatic hybridization: regeneration of fertile plants. Proc Natl Acad Sci USA 84:8434–8438
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soyabean roots cells. Exp Cell Res 50:151–158
Heyn FW (1976) Transfer of restorer gene from Raphanus to cytoplasmic male-sterile B. Napus. Cruciferae Newslett 1:15–16
Ichikawa H, Tanno-Suenaga L, Imamura J (1989) Transfer of mitochondria through protoplast fusion: In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 9. Plant protoplasts and genetic engineering II. Springer, Berlin Heidelberg New York, pp 360–375
Jacobsen E, Reinhout P, Bergervoet JEM, de Looff J, Abidin PE, Huigen DJ, Ramanna MS (1992) Isolation and characterization of potato-tomato somatic hybrids using an amylose-free potato mutant as parental genotype. Theor Appl Genet 85:159–164
Leivelt CLC, Krens FA (1992) Transfer of resistance to the beet cyst nametode (Heterodera schachtii Schm.) into the Brassica napus L. gene pool through intergeneric somatic hybridization with Raphanus sativus L. Theor Appl Genet 83:887–894
Masuelli RW, Tanimoto EY, Brown CR, Cornai L (1995) Irregular meiosis in a somatic hybrid between S. Bulbocastanum and S. Tuberosum by species-species PCR markers and cytological analysis. Theor Appl Genet 91:401–408
Melzer JM, O'Connell MA (1992) Effect of radiation does on the production and the extent of asymmetry in tomato asymmetric somatic hybrids. Theor Appl Genet 83:337–344
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Paulmann W, Robbelen G (1988) Effective transfer of cytoplasmic male sterility from radish (Raphanus sativus L.) to rape (Brassica napus L.). Plant Breed 100:299–309
Pelletier G, Primard C, Vedel F, Chetrit P, Remy R, Rousselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191:244–250
Pellen-Delourme R, Renard M (1988) Cytoplasmic male sterility in rapeseed (Brassica napus L.): female fertility of restored rapeseed with “Ogura” and cybrids cytoplasm. Genome 30:234–238
Ratushynyak YI, Cherep NN, Zavgorodnyaya AV, Latypov SA, Borozenko IV, Rachkovskaya RI, Gleba YY (1993) Fertile asymmetric somatic hybrids between Lycopersicon esculentum Mill, and Lycopersicon peruvianum var. dentatum. Dun. Mol Gen Genet 236:427–432
Ringdahl EA, McVetty PBE, Serynk JL (1987) Intergeneric hybridization of Diplotaxis spp. with Brassica napus: a source of new CMS systems? Can J Plant Sci 67:239–247
Sakai T, Imamura J (1991) Intergeneric transfer of cytoplasmic male stertility between Raphanus sativus (cms line) and Brassica napus through cytoplast-protoplast fusion. Theor Appl Genet 80:421–427
Sakai T, Imamura J (1992) Alteration of mitrochondrial genome containing atpA genes in the sexual progeny of cybrids between Raphanus sativus cms line and Brassica napus cv Westar. Theor Appl Genet 84:923–929
Shiga T, Baba S (1973) Cytoplasmic male stertility in oil seed rape, Brassica napus L. and its utilization for breeding. Jpn J Breed 23:187–197
Somers DA, Narayanan KR, Kleihofs A, Cooper-Blaud S, Cocking EC (1986) Immunological evidence for transfer of the barley nitrate reductase structural gene to Nicotiana tabacum by protoplast fusion. Mol Gen Genet 204:296–301
Trick H, Zelcer A, Bates GW (1994) Chromosome elimination in asymmetric somatic hybrids: effect of gamma dose and time in culture. Theor Appl Genet 88:965–972
Yarrow SA, Wu SC, Barsby TL, Kemble RJ, Shepard JF (1986) The introduction of CMS mitochondria of triazine-tolerant Brassica napus L., “Regent”, by micromanipulation of individual heterokaryons. Plant Cell Rep 5:415–418
Yamashita Y, Terada R, Nishibayashi S, Shimamoto K (1989) Asymmetric somatic hybrids of Brassica: partial transfer of B. campestris genome into B. oleracea by cell fusion. Theor Appl Genet 77:189–194
Xu YS, Murto M, Dunckley R, Jones MGK, Pehu E (1993) Production of asymmetric hybrids between Solanum tuberosum and irradiated S.Brevidens. Theor Appl Genet 85:729–734
Author information
Authors and Affiliations
Additional information
Communicated by Y. Gleba
Rights and permissions
About this article
Cite this article
Sakai, T., Liu, H.J., Iwabuchi, M. et al. Introduction of a gene from fertility restored radish (Raphanus sativus) into Brassica napus by fusion of X-irradiated protoplasts from a radish restorer line and iodacetoamide-treated protoplasts from a cytoplasmic male-sterile cybrid of B. napus . Theoret. Appl. Genetics 93, 373–379 (1996). https://doi.org/10.1007/BF00223179
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00223179