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Synthesis of male sterile, triazine-resistant Brassica napus by somatic hybridization between cytoplasmic male sterile B. oleracea and atrazine-resistant B. campestris

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Summary

Fusion of leaf protoplasts from an inbred line of Brassica oleracea ssp. botrytis (cauliflower, n=9) carrying the Ogura (R1) male sterile cytoplasm with hypocotyl protoplasts of B. campestris ssp. oleifera (cv “Candle”, n=10) carrying an atrazine-resistant (ATR) cytoplasm resulted in the production of synthetic B. napus (n=19). Thirty-four somatic hybrids were produced; they were characterized for morphology, phosphoglucose isomerase isoenzymes, ribosomal DNA hybridization patterns, chromosome numbers, and organelle composition. All somatic hybrids carried atrazine-resistant chloroplasts derived from B. campestris. The mitochondrial genomes in 19 hybrids were examined by restriction endonuclease and Southern blot analyses. Twelve of the 19 hybrids contained mitochondria showing novel DNA restriction patterns; of these 12 hybrids, 5 were male sterile and 7 were male fertile. The remaining hybrids contained mitochondrial DNA that was identical to that of the ATR parent and all were male fertile.

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References

  • Arús P, Orton TJ (1983) Inheritance and linkage relationships of isozyme loci in Brassica oleracea. J Hered 74:405–412

    Google Scholar 

  • Aviv D, Galun E (1986) Does pretreatment by rhodamine 6-G affect the mitochondrial composition of fusion-derived Nicotiana cybrids? Plant Cell Rep 3:227–230

    Google Scholar 

  • Bannerot H, Boilidard, Chupeau Y (1977) Unexpected difficulties met with the radish cytoplasm in Brassica oleracea. Eucarpia Cruciferae Newslett 2:16

    Google Scholar 

  • Barsby TL, Chuong PV, Yarrow SA, Wu S, Coumans M, Kemble RJ, Powell AD, Beversdorf WD, Pauls KP (1987 a) The combination of Polima cms and cytoplasmic triazine resistance in Brassica napus. Theor Appl Gen 73:809–814

    Google Scholar 

  • Barsby TL, Yarrow SA, Kemble RJ, Grant I (1987 b) The transfer of cytoplasmic male sterility to winter-type oilseed rape (Brassica napus L.) protoplast fusion. Plant Sci 53:243–248

    Google Scholar 

  • Bartkowiak-Broda I, Rousselle P, Renard M (1979) Investigations of two kinds of cytoplasmic male sterility in rapeseed (Brassica napus L.). Genet Pol 20:487–497

    Google Scholar 

  • Boeshore ML, Lifshitz I, Hanson MR, Izhar S (1985) Novel composition of mitochondrial genomes in Petunia somatic hybrids derived from cytoplasmic male sterile and fertile plants. Mol Gen Genet 190:459–467

    Google Scholar 

  • Chetrit P, Mathieu C, Vedel F, Pelletier G, Primard C (1985) Mitochondrial DNA polymorphism induced by protoplast fusion in Cruciferae. Theor Appl Genet 69:361–366

    Google Scholar 

  • Chiang MS, Créte R (1985) Male fertile and male sterile cabbage, broccoli, and cauliflower clubroot resistant breeding lines. Hort Science 20:457–458

    Google Scholar 

  • Chourey PS, Lloyd RE, Sharpe DZ, Isola NR (1986) Molecular analysis of hypervariability in the mitochondrial genome of tissue cultured cells of maize and sorghum. In: Mantell SH, Chapman GP, Street PFS (eds) The Chondriome-chloroplast and mitochondrial genomes. Longman Scientific, Harlow, pp 177–191

    Google Scholar 

  • Clark EM, Izhar S, Hanson MR (1985) Independent segregation of the plastid genome and cytoplasmic male sterility in Petunia somatic hybrids. Mol Gen Genet 199:440–445

    Google Scholar 

  • 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

    Google Scholar 

  • Cséplö A, Medgyesy P, Hideg E, Demeter S, Márton L, Maliga P (1985) Triazine-resistant Nicotiana mutants from photomixotrophic cell cultures. Mol Gen Genet 200:508–510

    Google Scholar 

  • Dickson MH (1985) Male sterile persistent white curd cauliflower NY7642A and its maintainer NY7642B. HortScience 20:957

    Google Scholar 

  • Fluhr R, Aviv D, Edelman M, Galun E (1983 a) Cybrids containing mixed and sorted-out chloroplasts following interspecific somatic fusions in Nicotiana. Theor Appl Genet 65:289–294

    Google Scholar 

  • Fluhr R, Aviv D, Edelman M, Galun E (1983 b) Generation of heteroplastidic Nicotiana cybrids by protoplast fusion analysis for plastid recombination. Theor Appl Genet 67:491–497

    Google Scholar 

  • Gear ARL (1974) Rhodamine 6G a potent inhibitor of mitochondrial oxidative phosphorylation. J Biol Chem 249:3628–3637

    Google Scholar 

  • Gengenbach BG, Connelly JA, Pring DR, Conde MF (1981) Mitochondrial DNA variation in maize plants regenerated during tissue culture selection. Theor Appl Genet 59:161–167

    Google Scholar 

  • Gleddie S, Keller WA, Setterfield S (1986) Production and characterization of somatic hybrids between Solanum melongena L. and S. sisymbriifolium. Lam. Theor Appl Genet 71:613–621

    Google Scholar 

  • Glimelius K (1984) High growth rates and regeneration capacity of hypocotyl protoplasts in some Brassicaceae. Physiol Plant 61:38–44

    Google Scholar 

  • Grayburn WS, Bendich A (1987) Variable abundance of a mitochondrial DNA fragment in cultured tobacco cells. Curr Genet 12:257–261

    Google Scholar 

  • Harms CT (1983) Somatic incompatibility in the development of higher plant somatic hybrids. Q Rev Biol 53:325–353

    Google Scholar 

  • Jourdan PS (1988) Manipulation of cytoplasmic traits in Brassica by somatic hybridization of male sterile B. oleracea with atrazine-resistant B. campestris and B. napus. PhD Thesis, Cornell University, Ithaca/NY

    Google Scholar 

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

    Google Scholar 

  • Kemble RJ, Shepard JF (1984) Cytoplasmic DNA variation in a potato protoclonal population. Theor Appl Genet 69:211–216

    Google Scholar 

  • Kemble RJ, Yarrow SA, Wu SC, Barsby TL (1988) Absence of mitochondrial and chloroplast DNA recombinations in Brassica napus plants regenerated from protoplasts, protoplast fusions and anther culture. Theor Appl Genet 75:875–881

    Google Scholar 

  • Kumar A, Cocking EC (1987) Protoplast fusion: a novel approach to organelle genetics in higher plants. Am J Bot 74:1289–1303

    Google Scholar 

  • Linsmaier M, Skoog F (1965) Organic growth factor requirements of tobacco tissue culture. Physiol Plant 18:100–127

    Google Scholar 

  • Makaroff CA, Palmer JD (1987) Extensive mitochondrial specific transcription of the Brassica campestris mitochondrial genome. Nucleic Acids Res 15:5141–5156

    Google Scholar 

  • McCollum (1981) Induction of an alloplasmic male sterile Brassica oleracea by substituting cytoplasm from ‘Early Scarlet Globe’ radish (Raphanus sativus). Euphytica 30:855–859

    Google Scholar 

  • Medgyesy P, Fejes E, Maliga P (1985) Interspecific chloroplast recombination in a Nicotiana somatic hybrid. Proc Natl Acad Sci USA 82:6960–6964

    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

    Google Scholar 

  • Menczel L, Morgan A, Brown S, Maliga P (1987) Fusion-mediated combination of Ogura-type cytoplasmic male sterility with Brassica napus plastids using X-irradiated cms protoplasts. Plant Cell Rep 6:98–101

    Google Scholar 

  • Morgan A, Maliga P (1987) Rapid chloroplast segregation and recombination of mitochondrial DNA in Brassica cybrids. Mol Gen Genet 209:240–246

    Google Scholar 

  • O'Connell MA, Hanson MR (1986) Regeneration of somatic hybrid plants formed between Lycopersicon esculentum and Solanum rickii. Theor Appl Genet 72:59–65

    Google Scholar 

  • Ogura H (1968) Studies on the new male-sterility in Japanese radish, with special reference to the utilization of this sterility towards practical raising of hybrid seed. Mem Fac Agric Kagoshima Univ 6:39–78

    Google Scholar 

  • Palmer JD, Shields CR (1984) Tripartite structure of Brassica campestris mitochondrial genome. Nature 307:437–440

    Google Scholar 

  • Palmer JD, Shields CR, Cohen DB, Orton TJ (1983) Chloroplast DNA evolution and the origin of amphidiploid Brassica species. Theor Appl Genet 65:181–189

    Google Scholar 

  • Pearson OH (1972) Cytoplasmically inherited male sterility characters and flavor components from the species cross Brassica nigra (L) Koch x B. oleracea L. J Am Soc Hortic Sci 97:397–402

    Google Scholar 

  • Pelletier G (1986) Plant organelle genetics through somatic hybridization. Oxford Surv Plant Mol Cell Biol 3:96–121

    Google Scholar 

  • Pelletier G, Primard C, Vedel F, Chetrit P, Remy R, Rouselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191:244–252

    Google Scholar 

  • Reich TJ, Iyer VN, Haffner M, Holbrook LA, Miki BL (1986) The use of fluorescent dyes in the microinjection of alfalfa protoplasts. Can J Bot 64:1259–1267

    Google Scholar 

  • Robertson D, Earle ED (1987) Nitro-blue tetrazolium: a stain for photosynthesis in protoplasts. Plant Cell Rep 6:70–73

    Google Scholar 

  • Robertson D, Palmer JD, Earle ED, Mutschler MA (1987) Analysis of organelle genomes in a somatic hybrid derived from cytoplasmic male-sterile Brassica oleracea and atrazine-resistant B. campestris. Theor Appl Genet 74:303–309

    Google Scholar 

  • Rode A, Hartmann C, Falconet D, Lejeune B, Quétier F, Benslimane A, Henry Y, Buyser J de (1987) Extensive mitochondrial DNA variation in somatic tissue cultures initiated from wheat immature embryos. Curr Genet 12:369–376

    Google Scholar 

  • Rothenberg M, Boeshore ML, Hanson MR, Izhar S (1985) Intergenomic recombination of mitochondrial genomes in a somatic hybrid plant. Curr Genet 9:615–618

    Google Scholar 

  • Schenck HR, Röbbelen G (1982) Somatic hybrids by fusion of protoplasts from Brassica oleracea and B. campestris. Z Pflanzenzuecht 89:278–288

    Google Scholar 

  • Shirzagedan M, Christey M, Earle ED, Palmer JD (1989) Rearrangement, amplification and assortment of mitochondrial DNA molecules in cultured cells of Brassica campestris. Theor Appl Genet 77:17–25

    Google Scholar 

  • 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

    Google Scholar 

  • Sundberg E, Landgren M, Glimelius K (1987) Fertility and chromosome stability in Brassica napus resynthesized by protoplast fusion. Theor Appl Genet 75:96–104

    Google Scholar 

  • Taguchi T, Kameya T (1986) Production of somatic hybrid plants between cabbage and Chinese cabbage through protoplast fusion. Jpn J Breed 36:185–189

    Google Scholar 

  • Tanksley SD (1979) An efficient and economical design for starch gel electrophoresis. Rep Tomato Genet Coorp 29:37–38

    Google Scholar 

  • Tanksley SD (1980) Pgi-1, a single gene in tomato responsible for variable number of isozymes. Can J Genet Cytol 22:271–278

    Google Scholar 

  • Terada R, Yamashita Y, Nishibayashi S, Shimamoto K (1987) Somatic hybrids between Brassica oleracea and B. campestris: selection by the use of iodoacetamide inactivation and regeneration ability. Theor Appl Genet 73:379–384

    Google Scholar 

  • Vedel F, Chetrit P, Mathieu C, Pelletier G, Primard C (1986) Several different mitochondrial DNA regions are involved in intergenomic recombination in Brassica napus cybrid plant. Curr Genet 11:17–24

    Google Scholar 

  • Yarrow SA, Wu SC, Barsby TL, Kemble RJ, Shepard JF (1986) The introduction of CMS mitochondria to triazine-tolerant Brassica napus L., var. “Regent”, by micromanipulation of individual heterokaryons. Plant Cell Rep 5:415–418

    Google Scholar 

  • Young E, Hanson MR (1987) A fused mitochondrial gene associated with cytoplasmic male sterility is developmentally regulated. Cell 50:41–49

    Google Scholar 

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Communicated by P. Maliga

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Jourdan, P.S., Earle, E.D. & Mutschler, M.A. Synthesis of male sterile, triazine-resistant Brassica napus by somatic hybridization between cytoplasmic male sterile B. oleracea and atrazine-resistant B. campestris . Theoret. Appl. Genetics 78, 445–455 (1989). https://doi.org/10.1007/BF00265310

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