Abstract
Somatic hybrids were produced by protoplast fusion between Arabidopsis thaliana ecotype Columbia and a male-sterile radish line MS-Gensuke (Raphanus sativus) with the Ogura cytoplasm. Forty-one shoots were differentiated from the regenerated calli and established as shoot cultures in vitro. About 20 of these shoots were judged to be hybrids based on growth characteristics and morphology. Molecular analyses of 11 shoots were performed, confirming the hybrid features. Of these 11 shoots, eight were established as rooted plants in the greenhouse. Polymerase chain reaction and randomly amplified polymorphic DNA analyses of the nuclear genomes of all analyzed shoots and plants confirmed that they contained hybrid DNA patterns. Their chromosome numbers also supported the hybrid nature of the plants. Investigations of the organelles in the hybrids revealed that the chloroplast (cp) genome was exclusively represented by radish cpDNA, while the mitochondrial DNA configuration showed a combination of both parental genomes as well as fragments unique to the hybrids. Hybrid plants that flowered were male-sterile independent of the presence of the Ogura CMS-gene orf138.
Similar content being viewed by others
Abbreviations
- CMS :
-
Cytoplasmic male sterility
References
Balder A, Gaikwad K, Kirti PB, Mohapatra T, Prakash S, Chopra VL (1998) Recombination between chloroplast genomes of Trachystoma ballii and Brassica juncea following protoplast fusion. Mol Gen Genet 260:357–361
Banks MS, Evans PK (1976) A comparison of the isolation and culture of mesophyll protoplasts from several Nicotiana species and their hybrids. Plant Sci Lett 7:409–416
Bauer-Weston B, Keller W, Webb J, Gleddie S (1993) Production and characterization of asymmetric somatic hybrids between Arabidopsis thaliana and Brassica napus. Theor Appl Genet 86:150–158
Forsberg J, Landgren M, Glimelius K (1994) Fertile somatic hybrids between Brassica napus and Arabidopsis thaliana. Plant Sci 95:213–223
Forsberg J, Dixelius C, Lagercrantz U, Glimelius K (1998) UV-dose dependent DNA elimination in asymmetric somatic hybrids between Brassica napus and Arabidopsis thaliana. Plant Sci 137:65–76
Gleba YY, Hoffmann F (1980) "Arabidobrassica": a novel plant obtained by protoplast fusion. Planta 149:112–117
Glimelius K (1984) High growth rate and regeneration capacity of hypocotyl protoplasts in some Brassicaceae. Physiol Plant 61:38–44
Hagimori M, Nagaoka M, Kato N, Yoshikawa H (1992) Production and characterization of somatic hybrids between the Japanese radish and cauliflower. Theor Appl Genet 82:819–824
Kaneko Y, Matsuzawa, Y (1993) Radish. In: Kalloo G, Bergh BO (eds) Genetic improvement of vegetable crops. Pergamon, London, pp 487–510
Lelivelt C, Krens FA (1992) Transfer of resistance to the beet cyst nematode (Heterodera schachtii Schm.) into the Brassica napus L. gene pool through intergeneric somatic hybridization with Raphanus sativus L. Theor Appl Genet 83:887–894
Quiros CF (1999) Genome structure and mapping. In: Gómez-Campo C (ed) Biology of brassica coenospecies. Elsevier, Amsterdam, pp 217–245
Sakai T, Imamura J (1990) Intergeneric transfer of cytoplasmic male sterility between Raphanus sativus (CMS line) and Brassica napus through cytoplast-protoplast fusion. Theor Appl Genet 80:421–427
Sakai T, Liu HJ, Iwabuchi M, Kohno-Murase J, Imamura J (1996) 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 iodoacetoamide-treated protoplasts from a cytoplasmic male sterile cybrid of B. napus. Theor Appl Genet 93:373–379
Sato S, Nakamura Y, Kaneko T, Asamizu E, Tabata S (1999) Complete structure of the chloroplast genome of Arabidopsis thaliana. DNA Res 6:283–290
Shih MC (1994) Cloning and sequencing of a cDNA clone encoding the cytosolic triose-phosphate isomerase from Arabidopsis thaliana. Plant Physiol 104:1103–1104
Siemens J, Sacristán MD (1995) Production and characterization of somatic hybrids between Arabidopsis thaliana and Brassica napus. Plant Sci 111:95–106
Sundberg E, Glimelius K (1991) Effects of parental ploidy level and genetic divergence on chromosome elimination and chloroplast segregation in somatic hybrids within Brassicaceae. Theor Appl Genet 83:81–88
Terachi T (1996) Sequence analysis of cDNA encoding cytosolic phosphoglucose isomerase from radish. Bull Inst Nat Land Util Dev Kyoto Sangyo Univ 17:108–118
The Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408:796–815
Thomas BR, Ford US, Pichersky E, Gottlieb LD (1993) Molecular characterization of duplicate cytosolic phosphoglucose isomerase genes in Clarkia and comparison to the single gene in Arabidopsis. Genetics 135:895–905
Unseld M, Marienfeld JR, Brandt P, Brennicke A (1997) The mitochondrial genome of Arabidopsis thaliana contains 57 genes in 366,924 nucleotides. Nat Genet 15:57–61
Yamagishi H, Landgren M, Forsberg J, Glimelius K (2002) Production of asymmetric hybrids between Arabidopsis thaliana and Brassica napus utilizing an efficient protoplast culture system. Theor Appl Genet 104:959–964
Acknowledgements
The authors thank J. Sasaki for skilful assistance. This work was supported in part by the Foundation of Bio-Venture Research Center of Ministry of Education, Culture, Sports and Technology, Japan and by the Swedish Council for Forestry and Agricultural Research, SJFR.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by M.R. Davey
Rights and permissions
About this article
Cite this article
Yamagishi, H., Glimelius, K. Somatic hybrids between Arabidopsis thaliana and cytoplasmic male-sterile radish (Raphanus sativus). Plant Cell Rep 22, 52–58 (2003). https://doi.org/10.1007/s00299-003-0655-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-003-0655-0