Abstract
Intertribal Brassica napus (+) Lesquerella fendleri hybrids have been produced by polyethylene glycol-induced fusions of B. napus hypocotyl and L. fendleri mesophyll protoplasts. Two series of experiments were performed. In the first, symmetric fusion experiments, protoplasts from the two materials were fused without any pretreatments. In the second, asymmetric fusion experiments, X-ray irradiation at doses of 180 and 200 Gy were used to limit the transfer of the L. fendleri genome to the hybrids. X-ray irradiation of L. fendleri mesophyll protoplasts did not suppress the proliferation rate and callus formation of the fusion products but did significantly decrease growth and differentiation of non-fused L. fendleri protoplasts. In total, 128 regenerated plants were identified as intertribal somatic hybrids on the basis of morphological criteria. Nuclear DNA analysis performed on 80 plants, using species specific sequences, demonstrated that 33 plants from the symmetric fusions and 43 plants from the asymmetric fusions were hybrids. Chloroplast and mitochondrial DNA analysis revealed a biased segregation that favoured B. napus organelles in the hybrids from the symmetric fusion experiments. The bias was even stronger in the hybrids from the asymmetric fusion experiments where no hybrids with L. fendleri organelles were found. X-ray irradiation of L. fendleri protoplasts increased the possibility of obtaining mature somatic hybrid plants with improved fertility. Five plants from the symmetric and 24 plants from the asymmetric fusion experiments were established in the greenhouse. From the symmetric fusions 2 plants could be fertilised and set seeds after cross-pollination with B. napus. From the asymmetric fusions 9 plants could be selfed as well as fertilised when backcrossed with B. napus. Chromosome analysis was performed on all of the plants but 1 that were transferred to the greenhouse. Three plants from the symmetric fusions contained 50 chromosomes, which corresponded to the sum of the parental genomes. From the asymmetric fusions, 11 hybrids contained 38 chromosomes. Among the other asymmetric hybrids, plants with 50 chromosomes and with chromosome numbers higher than the sum of the parental chromosomes were found. When different root squashes of the same plant were analysed, a total of 6 plants were found that had different chromosome numbers.
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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
Bennett MD, Finch RA, Barclay IR (1976) The time rate and mechanism of chromosome elimination in Hordeum hybrids. Chromosoma 54:175–200
Bonnema AB, Melzer JM, Murray LW, O'Connell MA (1992) Nonrandom inheritance of organellar genomes in symmetric and asymmetric somatic hybrids between Lycopersicon esculentum and L. pennellii. Theor Appl Genet 84:435–442
Bonnett HT, Glimelius K (1983) Somatic hybridization in Nicotiana:behavior of organelles after fusion of protoplasts from male-fertile and male-sterile cultivars. Theor Appl Genet 65:213–217
Butterfass T (1988) Nuclear control of cell division. In:Boffey SA, Lloyd D (eds) Division and segregation of organelles. (Society for experimental biology seminar series, vol 35). Cambridge University Press, Cambridge, pp 21–38
Clark E, Schnabelrauch L, Hanson MR, Sink KC (1986) Differential fate of plastid and mitochondrial genomes in Petunia somatic hybrids. Theor Appl Genet 72:748–755
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21
Dudits D, Maroy E, Praznovsky T, Olah Z, Gyorgyey J, Cella R (1987) Transfer of the resistance traits from carrot into tobacco by asymmetric somatic hybridization regeneration of fertile plants. Proc Natl Acad Sci USA 84:8434–8438
Fahleson J, Eriksson I, Landgren M, Stymne S, Glimelius K (1994) Intertribal somatic hybrids between Brassica napus and Thlaspi perfoliatum with high content of the T. perfoliatum-specific nervonic acid. Theor Appl Genet 87:795–804
Feher A, Preiszner J, Litkey Z, Csanádi Gy, Dudits D (1992) Characterization of chromosome instability in interspecific somatic hybrids obtained by X-ray fusion between potato (Solanum tuberosum L.) and S. brevidens Phil. Theor Appl Genet 84:880–890
Forsberg J, Landgren M, Glimelius K (1994) Fertile somatic hybrids between Brassica napus and Arabidopsis thaliana. Plant Sci 95:213–223
Gentry HS, Barclay AS (1962) The search for new industrial crops. III. Prospects of Lesquerella fendleri. Econ Bot 16:206–211
Gleba YY, Sytnik KM(1984) Protoplast fusion. Genetic engineering in higher plants. Springer. Berlin Heidelberg New York, 203 pp
Gleba YY, Parokonny A, Kotov V, Negrutiu I, Momot V (1987) Spatial separation of parental genomes in hybrids of somatic plant cells. Proc Natl Acad Sci USA 84:3709–3713
Gleba YY, Hinnisdaels S, Sidorov VA, Kaleda VA, Parokonny AS, Boryshuk NV, Cherep NN, Negrutiu I, Jacobs M (1988) Intergenetic asymmetric hybrids between Nicotiana plumbaginifolia and Atropa belladonna obtained by “gamma-fusion”. Theor Appl Genet 76:760–766
Glimelius K (1984) High growth and regeneration capacity of hypocotyl protoplasts in some Brassicaceae. Physiol Plant 61:38–44
Glimelius K, Djupsjöbacka M, Fellner-Feldegg H (1986) Selection and enrichment of plant protoplast heterokaryons of Brassicaceae by flow sorting. Plant Sci 45:133–141
Glimelius K, Fahleson J, Landgren M, Sjödin C, Sundberg E (1991) Gene transfer via somatic hybridization in plants. Trend Biotechnol 9:24–30
Hinnisdaels S, Negrutiu I, Jacobs M, Sidorov VA (1988) Plant somatic cell hybridizations:evaluations and perspectives. IAPTC Newsl 55:2–10
Imamura J, Saul MW, Potrykus I (1987) X-ray irradiation promoted asymmetric somatic hybridization and molecular analysis of the products. Theor Appl Genet 74:445–450
Jansen RJ, Palmer JD (1987) Chloroplast DNA from lettuce and Barnadesia (Asteraceae):structure, gene localization and characterization of a large inversion. Curr Genet 11:553–564
Kao KN (1977) Chromosomal behaviour in somatic hybrids of soybean-Nicotiana glauca. Mol Gen Genet 150:225–230
Kovtun YV, Korostash MA, Butsko YV, Gleba YY (1993) Amplification of repetitive DNA from Nicotiana plumbaginifolia in asymmetric somatic hybrids between Nicotiana sylvestris and Nicotiana plumbaginifolia. Theor Appl Genet 86:221–228
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 mitochondrial 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 87:854–862
Landgren M, Sundberg E, Glimelius K (1994) Biased mitochondrial segregation, independent of cell type used for fusion and of hybrid nuclear DNA content, was found in Brassica napus (+)B. oleracea somatic hybrids. Plant Sci 103:51–57
Lelivelt CLC, Leunissen EHM, Frederiks HJ, Helsper JPFG, Krens FA (1993) Transfer of resistance to the beet cyst nematode (Heterodera schachtii Schm.) from Sinapis alba L. (white mustard) to the Brassica napus L. gene pool by means of sexual and somatic hybridization. Theor Appl Genet 85:688–696
Levi A, Ridley BL, Sink KC (1988) Biased organelle transmission in somatic hybrids of Lycopersicon esculentum and Solanum lycopersicoides. Curr Genet 14:177–182
Liu J-H, Dixelius C, Eriksson I, Glimelius K (1995) Brassica napus (+)B. tournefortii, a somatic hybrid containing traits of agronomic importance for rapeseed breeding. Plant Sci 109:75–86
Menczel L, Nagy F, Kiss ZR, Maliga P (1981) Streptomycin-resistant and sensitive somatic hybrids of Nicotiana tabacum (+)N. knightiana:correlation resistance to N. tabacum plastids. Theor Appl Genet 59:191–195
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–479
Muuse BG, Cuperus FP, Derksen JTP (1992) Composition and physical properties of oils from new oilseed crops. Ind Crops Prod 1:57–65
Parokonny AS, Kenton A, Gleba YY, Bennett MD (1992) Genome reorganization in Nicotiana asymmetric somatic hybrids analyzed by in-situ hybridization. Plant J 2:695–704
Pehu E, Karp A, Moore K, Steele S, Dunckley R, Jones MGK (1989) Molecular, cytogenetic and morphological characterization of somatic hybrids of dihaploid Solanum tuberosum and diploid S. brevidens. Theor Appl Genet 78:696–704
Perl A, Aviv D, Galun E (1991) Nuclear-organelle interaction in Solanum:interspecific cybridizations and their correlation with a plastome dendrogram. Mol Gen Genet 228:193–200
Piastuch WC, Bates GW (1990) Chromosomal analysis of Nicotiana asymmetric somatic hybrids by dot blotting and in situ hybridization. Mol Gen Genet 222:97–103
Potrykus I (1991) Gene transfer to plants. Assessement of published approaches and results. Annu Rev Plant Physiol Plant Mol Biol 42:205–225
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, Landgren M, Glimelius K (1987) Fertility and chromosome stability in Brassica napus resynthesised by protoplast fusion. Theor Appl Genet 75:96–104
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
White J, Rees H (1985) The chromosome cytology of a somatic hybrid petunia. Heredity 55:53–59
Wolters AMA, Schoenmakers HCH, van der Meulen-Muisers JJM, van der Knaap E, Derks FHM, Koornneef M, Zelcer A (1991) Limited DNA elimination from the irradiated potato parent in fusion products of albino Lycopersicon esculentum and Solanum tuberosum. Theor Appl Genet 83:225–232
Wolters AMA, Vergunst AC, van de Werff F, Koornneef M (1993) Analysis of nuclear and organellar DNA of somatic hybrid calli and plants between Lycopersicon spp. and Nicotiana spp. Mol Gen Genet 241:707–718
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Communicated by Yu. Gleba
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Skarzhinskaya, M., Landgren, M. & Glimelius, K. Production of intertribal somatic hybrids between Brassica napus L. and Lesquerella fendleri (Gray) Wats. Theoret. Appl. Genetics 93, 1242–1250 (1996). https://doi.org/10.1007/BF00223456
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DOI: https://doi.org/10.1007/BF00223456