Abstract
Protoplast fusions were performed between hypocotyl protoplasts of Brassica napus and mesophyll protoplasts of Thlaspi perfoliatum. The two species are members of the Lepidieae and Brassiceae tribes, respectively, in the family of Brassicaceae. Seeds of T. perfoliatum are rich in the fatty acid C24∶1 (nervonic acid), an oil valuable for technical purposes. In the search for renewable oils to replace the mineral oils, plant breeders have been trying to develop oil crops with a high content of long-chain fatty acids. After fusion of B. napus protoplasts with non-irradiated as well as irradiated protoplasts of T. perfoliatum selection was carried out by flow cytometry and cell sorting. Of the shoots regenerated from different calli 27 were verified as hybrids or partial hybrids using the isoenzyme phosphoglucose isomerase (PGI) as a marker. Another 6 plants were identified as partial hybrids using a T. perfoliatum-specific repetitive DNA sequence. Slot blot experiments were performed to estimate the copy number of the repetitive DNA sequence in the parental species and in the hybrids. In T. perfoliatum there were approximately 105 copies per haploid genome, and the range in the hybrids was 1–37% of the value in T. perfoliatum. When the nuclear DNA content of the regenerated shoots was analysed we found partial as well as symmetric hybrids. Even though the rooting and establishment of hybrid shoots in the greenhouse were difficult, resulting in the death of many plants, 19 plants were cultured to full maturity. Seeds obtained from 15 plants were analysed to determine whether they contained nervonic acid, and 5 of the hybrids were found to contain significantly greater amounts of nervonic acid than B. napus.
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References
Appelquist LÅ (1976) Lipids in the Cruciferae. In: Vaughan JG, MacLeod AJ, Jones BMG (eds) The biology and chemistry of the Cruciferae. Academic Press, London New York San Francisco, pp 221–277
Babiychuk E, Kushnir S, Gleba YY (1992) Spontaneous extensive chromosome elimination in somatic hybrids between somatically congruent species Nicotiana tabacum L. and Atropa belladonna L. Theor Appl Genet 84:87–91
Bernatzky R, Tanksley SD (1986) Genetics of actin-related sequences in tomato. Theor Appl Genet 72:314–321
Chatterjee G, Sikdar SR, Das S, Sen SK (1988) Intergeneric somatic hybrid production through protoplast fusion between Brassica juncea and Diplotaxis muralis. Theor Appl Genet 76:915–922
Dewey RE, Levings CS III, Timothy DH (1986) Novel recombinations in the maize mitochondrial genome produce a unique transcriptional unit in the Texas male-sterile cytoplasm. Cell 44:439–449
Fahleson J, Rahlén L, Glimelius K (1988a) Analysis of plant regenerated from protoplast fusion between Brassica napus and Eruca sativa. Theor Appl Genet 76:507–512
Fahleson J, Dixelius J, Sundberg E, Glimelius K (1988b) Correlation between flow cytometric determination of nuclear DNA content and chromosome number in somatic hybrids within Brassicaceae. Plant Cell Rep 7:74–77
Fahleson J, Eriksson I, Glimelius K (1994) Intertribal somatic hybrids between Brassica napus and Barbarea vulgaris. Plant Cell Rep (in press)
Forsberg J, Landgren M, Glimelius K (1994) Fertile somatic hybrids between Brassica napus and Arabidopsis thaliana. Plant Sci (in press)
Gleba YY, Hoffmann F (1980) “Arabidobrassica”: A novel plant obtained by protoplast fusion. Planta 149:112–117
Glimelius K (1984) High growth and regeneration capacity of hypocotyl protoplasts in some Brassicacaceae. 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 Apl Genet 84:819–824
Hinnisdaels S, Negrutiu I, Jacobs M, Sidorov V (1988) Plant somatic cell hybridizations: evaluations and prospectives. IAPTC Newsl 55:2–10
Hoffman F, Adachi T (1981) “Arabidobrassica”: Chromosomal recombination and morphogenesis in asymmetric intergeneric hybrid cells. Planta 153:586–593
Håkansson G, Glimelius K (1991) Extensive nuclear influence on mitochondrial transcription and genome structure in malefertile and male-sterile alloplasmic Nicotiana materials. Mol Gen Genet 229:380–388
Iwabuchi M, Itoh K, Shimamoto K (1991) In situ hybridization with species-specific DNA probes gives evidence for asymmetric nature of Brassica hybrids obtained by X-ray fusion. Theor Appl Genet 81:356–362
Jansen RJ, Palmer JD (1987) Chloroplast DNA from lettuce and Barnadesia (Asteraceae): structure, gene localis ation and characterization of a large inversion. Curr Genet 11:553–564
Jones JDG, Flavell RB (1982) The structure, amount and chromosomal localisation of defined repeated DNA sequences in species of the genus Secale. Chromosoma 86:613–641
Jourdan PS, Earle ED, Mutschler MA (1989a) Atrazine-resistant cauliflower obtained by somatic hybridization between Brassica oleracea and ATR-B. napus. Theor Appl Genet 78:271–279
Jourdan PS, Earle ED, Mutschler MA (1988b) Synthesis of male-sterile, triazine-resistant Brassica napus by somatic hybridization between cytoplasmic male-sterile B. oleracea and atrazine-resistant B. campestris. Theor Appl Genet 78:445–455
Kirti PB, Prakash S, Chopra VL (1991) Interspecific hybridization between Brassica juncea and B. spinescens through protoplast fusion. Plant Cell Rep 9:639–642
Klimaszewska K, Keller WA (1988) Regeneration and characterization of somatic hybrids between Brassica napus and Diplotaxis harra. Plant Sci 58:211–222
Köhler RH, Horn R, Lössl A, Zetsche K (1991) Cytoplasmatic male sterility in sunflower is correlated with the co-transcription of a new open reading frame with the atpA gene. Mol Gen Genet 227:369–376
Landgren M, Glimelius K (1990) Analysis of chloroplast and mitochondrial segregation in three different combinations of somatic hybrids produced within Brassicaceae. Theor Appl Genet 80:776–784
Landgren M, Glimelius K (1994) A high frequency of intergenomic recombination and an overall biased segregation of B. campestris or recombined B. campestris mitochondria were found in somatic hybrids made within Brassicaceae. Theor Appl Genet (in press)
Lapitan NLV (1991) Organization and evolution of higher plant genomes. Genome 35:171–181
Lonsdale DM (1987) Cytoplasmic male sterility: a molecular perspective. Plant Physiol Biochem 25:265–271
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Sprin Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Mathias R, Espinosa S, Röbbelen G (1990) A new embryo resuce procedure for interspecific hybridization. Plant Breed 104:258–261
Menczel L, Nagy F, Kiss ZR, Maliga P (1981) Streptomycin resistant and sensitive somatic hybrids of Nicotiana tabacum + Nacotiana knightiana: correlation of resistance to N. tabacum plastids. Theor Appl Genet 59:191–195
Miller RW, Earle FR, Wolff IA (1965) Search for new industrial oils. XIII. Oils from 102 species of Cruciferae. J Am Oil Chem Soc 42:817–821
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissues. Physiol Plant 15:473–497
Murphy DJ (1992) Modifying oilseed crops for non-edible products. Trends Biotechnol 10:84–87
Primard C, Vedel F, Mathieu C, Pelletier G, Chevre AM (1988) Interspecific somatic hybridization between Brassica napus and Brassica hirta (Sinapis alba L.). Theor Appl Genet 75:546–552
Rosén B, Haldén C, Heneen WK (1988) Diploid Brassica napus somatic hybrids: Characterization of nuclear and organellar DNA. Theor Appl Genet 76:197–203
Schenck HR, Röbbelen G (1982) Somatic hybrids by fusion of protoplasts from Brassica oleracea and B. campestris. Z Pflanzenzuecht 89:278–288
Sikdar SR, Chatterjee G, Das S, Sen SK (1990) “Erussica”, the intergeneric fertile somatic hybrid developed through protoplast fusion between Eruca sativa Lam. and Brassica juncea (L.) Czern. Theor Appl Genet 79:561–567
Sjödin C, Glimelius K (1989a) Brassica naponigra, a somatic hybrid resistant to Phoma lingam. Theor Appl Genet 77:651–656
Sjödin C, Glimelius K (1989b) Transfer of resistance against Phoma lingam to Brassica napus by asymmetric somatic hybridization combined with toxin selection. Theor Appl Genet 78:513–520
Sundberg E, Glimelius K (1986) A method for production of interspecific hybrids within Brassiceae via somatic hybridization, using resynthesis of Brassica napus as a model. Plant Sci 43:155–162
Sundberg E, Landgren M, Glimelius K (1987) Fertility and chromosome stability in Brassica napus resynthesised by protoplast fusion. Theor Appl Genet 75:96–104
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
Sundberg E, Lagercrantz U, Glimelius K (1991) Effects of cell type used for fusion on chromosome elimination and chloroplast segregation in Brassica oleracea (+) Brassica napus hybrids. Plant Sci 78:89–98
Toriyama K, Hinata K, Kameya T (1987a) Production oef somatic hybrid plants, ‘Brassicomoricandia’ through protoplast fusion between Moricandia arvensis and Brassica oleracea. Plant Sci 48:123–128
Toriyama K, Kameya T, Hinata K (1987b) Selection of a universal hybridizer in Sinapis turgida Del. and regeneration of plantlets from somatic hybrids with Brassica species. Planta 170:308–313
Wiberg E, Rahlén L, Hellman M, Tillberg E, Glimelius K, Stymne S (1991) The microspore-derived embryo of Brassica napus L. as a tool for studying embryo-specific lipid biogenesis and regulation of oil quality. Theor Appl Genet 82:515–520
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Communicated by I. Potrykus
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Fahleson, J., Eriksson, I., Landgren, M. et al. Intertribal somatic hybrids between Brassica napus and Thlaspi perfoliatum with high content of the T. perfoliatum-specific nervonic acid. Theoret. Appl. Genetics 87, 795–804 (1994). https://doi.org/10.1007/BF00221131
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DOI: https://doi.org/10.1007/BF00221131