Genetic Manipulation in Brassica

  • F. Trail
  • C. Richards
  • F.-S. Wu
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 9)


Members of the family Cruciferae are sources of condiments, fodder, spices, and vegetables. Their contributions to human food supply range from the edible roots of the radish plant (Raphanus sativa) to the seeds of Brassica napus, which provide 10% of the world’s vegetable oil (Austin 1986). The six species of Brassica commonly cultivated worldwide are: B. campestris L. (turnip, turnip rape, chinese cabbage), B. carinata Braun (Abyssinian mustard), B. juncea Coss. (leaf mustard), B. napus (oil rape, canola, rutabaga), B. nigra Koch (black mustard), and B.oleracea L. (cabbage, cauliflower, broccoli, brussel sprouts). The family Cruciferae also includes the genera Sinapis (mustard), Eruca (salad greens) and Crambe (source for industrial oils). The less familiar Moricandia has been the object of recent studies as a physiological intermediate to C3 and C4 plants (Hunt et al. 1987). Arabidopsis is a common weed, but an important model plant for genetic and developmental studies because of its small genome size and rapid life-cycle (Meyerowitz and Pruitt 1985).


Cytoplasmic Male Sterility Brassica Napus Oilseed Rape Somatic Hybrid Protoplast Fusion 
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  1. Austin RB (1986) Oilseed rape. In: Austin RB, Flavell RB, Henson IE, Lowe HJB (eds): Molecular biology and crop improvement: a case study of wheat, oilseed rape and faba beans. Univ Press, Cambridge, pp 70–87CrossRefGoogle Scholar
  2. Balazs E, Bonneville JM (1987) Chloramphenicol acetyl transferase activity in Brassica spp. Plant Sci 50:65 — 68Google Scholar
  3. Bannerot H, Boulidard L, Cauderon Y, Tempé J (1974) Transfer of cytoplasmic male sterility from Raphanus sativa to Brassica oleracea. Proc Eucarpia Meet Cruciferae Scottish Hortic Res Inst, Invergowrie, GB, pp 52–54Google Scholar
  4. Bannerot H, Boulidard L, Chupeau Y (1977) Unexpected difficulties met with the radish cytoplasm. Eucarpia Cruciferae Newslett 2: 2–16Google Scholar
  5. Barsby TL, Yarrow SA, Shepard JF (1984) Heterokaryon identification through simultaneous fluorescence of fluorescein isothiocyanate and tetramethylrhodamine isothiocyanate labelled protoplasts. Stain Technol 59: 217–220PubMedGoogle Scholar
  6. Barsby TL, Yarrow SA, Shepard JF (1986) A rapid and efficient alternative procedure for regeneration of plants from hypocotyl protoplasts of Brassica napus. Plant Cell Rep 5: 101–103CrossRefGoogle Scholar
  7. Barsby TL, Chuong PV, Yarrow SA, Wu SC, 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: 809–814CrossRefGoogle Scholar
  8. Beversdorf WD, Weiss-Lerman J, Erikson LR, Souza-Machado V (1980) Transfer of cytoplasmically inherited triazine resistance from birds-rape to cultivated oilseed rape (Brassica campestris and B. napus). Can J Genet Cytol 22: 167–172Google Scholar
  9. Brisson N, Paszkowski J, Penswick JR, Groennborn B, Potrykus I, Hohn T (1984) Expression of a bacterial gene in plants by using a viral vector. Nature (Lond) 310: 511–514CrossRefGoogle Scholar
  10. Chetrit P, Mathieu C, Vedel F, Pelletier G, Primard C (1985) Mitochondrial DNA polymorphism induced by protoplast fusion in Cruciferae. Theor Appl Genet 69: 361–366CrossRefGoogle Scholar
  11. Chiang MS, Crete R (1983) Transfer of resistance to race 2 of Plasmodiophera Brassicae from Brassica napus to cabbage (B. oleracea spp. Capitata). V. The inheritance of resistance. Euphytica 32:479— 483Google Scholar
  12. Chuong PV, Pauls KP, Beversdorf WD (1985) A simple culture method for Brassica hypocotyl protoplasts. Plant Cell Rep 4: 4–6CrossRefGoogle Scholar
  13. Cornai L, Facciotti D, Miatt MR, Thompson G, Rose RE, Stalker DM (1985) Expression in plants of a mutant aroA gene from Salmonella typhimurium confers tolerance to glyphosate. Nature (Lond) 317:741 — 744Google Scholar
  14. Evans DA, Sharp WR (1986) Applications of somaclonal variation. Biotechnology 4: 528–532CrossRefGoogle Scholar
  15. Fan Z, Stefansson BR (1986) Influence of temperature on sterility of two cytoplasmic male-sterile systems in rape (Brassica napus L.). Can J Plant Sci 66: 221–227CrossRefGoogle Scholar
  16. Fraley RT, Rogers SG, Horsh RB (1986) Genetic transformation in higher plants. CRC Crit Rev Plant Sci 4: 1–46CrossRefGoogle Scholar
  17. Fromm M, Taylor LP, Walbot V (1985) Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci USA 82: 5824–5828PubMedCrossRefGoogle Scholar
  18. Fu TD (1981) Production and research of rapeseed in the People’s Republic of China. Eucarpia Cruciferae Newslett 6: 6–7Google Scholar
  19. Gleba YY, Hoffmann F (1978) Hybrid cell lines Arabidopsis thaliana+Brassica campestris: no evidence for specific chromosome elimination. Mol Gen Genet 165: 257–264CrossRefGoogle Scholar
  20. Gleba YY, Hoffmann F (1980) “Arabidobrassica”: A novel plant obtained by protoplast fusion. Planta 149: 112–117Google Scholar
  21. Glimelius K (1984) High growth rate and regeneration capacity of hypocotyl protoplasts in some Brassicaceae. Physiol Plant 61: 38–44CrossRefGoogle Scholar
  22. Glimelius K, Djupsjobacka M, Fellner-Feldegg H (1986) Selection and enrichment of plant protoplast heterokaryons of Brassicaceae by flow sorting. Plant Sci 45: 133–141CrossRefGoogle Scholar
  23. Griesbach RJ, Sink KC (1983) Evacuolation of mesophyll protoplasts. Plant Sci Lett 30: 297–301CrossRefGoogle Scholar
  24. Grimsley N, Hohn B, Hohn T, Walden R (1986) “Agroinfection”, an alternative route for viral infection of plants by using the Ti plasmid. Proc Natl Acad Sci USA 83: 3282–3286Google Scholar
  25. Guerche P, Charbonnier M, Jouanin L, Pelletier G (1986) Direct gene transfer in Brassica napus. Eucarpia Cruciferae Newslett 11: 98–99Google Scholar
  26. Guerche P, Jouanin L, Tepfer D, Pelletier G (1987) Genetic transformation of oilseed rape (Brassica napus) by the Ri T-DNA of Agrobacterium rhizogenes and analysis of inheritance of the transformed phenotype. Mol Gen Genet 206: 382–386CrossRefGoogle Scholar
  27. Harms CT (1983) Somatic incompatibility in the development of higher plant somatic hybrids. Q Rev Biol 58: 325–353CrossRefGoogle Scholar
  28. Hoffmann F, Adachi T (1981) “Arabidobrassica”: Chromosomal recombination and morphogenesis in asymmetric intergeneric hybrid cells. Planta 153: 586–593Google Scholar
  29. Holbrook LA, Miki BL (1985) Brassica grown gall tumourigenesis and in vitro of transformed tissue. Plant Cell Rep 4: 329–332Google Scholar
  30. Howell SH, Walker LL, Dudley RK (1980) Cloned Cauliflower Mosaic Virus DNA infects turnips (Brassica rapa). Science 208:1265 —1267Google Scholar
  31. Hunt S, Smith AM, Woolhouse HW (1987) Evidence for a light-dependent system for reassimilation of photo-respiratory CO2, which does not include a C4 cycle, in the C3–C4 intermediate species Moricandia arvensis. Planta 171: 227–234CrossRefGoogle Scholar
  32. Jourdan PS, Earle ED (1986) Influence of genotype on the regeneration of plants from seedling mesophyll protoplasts of three Brassica species (Abstr). Cruciferae genetics Worksh I II, Univ Guelph, Ontario, CanGoogle Scholar
  33. Kameya T (1983) Studies on plant cell fusion by dextran: Effect of pH, inorganic salts and electric stimulus. Cytologia 48: 873–878CrossRefGoogle Scholar
  34. Karp A, Bright SWJ (1985) On the causes and origins of somaclonal variation. Oxford Sury Plant Mol Cell Biol 2: 199–234Google Scholar
  35. Kartha KK, Michayluk MR, Kao KN, Gamborg OL, Constabel F (1974) Callus formation and plant regeneration from mesophyll protoplasts of rape plants (Brassica napus L. cv. Zephyr ). Plant Sci Lett 3: 265–271Google Scholar
  36. Kemble RJ (1987) A rapid, single leaf, nucleic acid assay for determining the cytoplasmic organelle complement of rapeseed and related Brassica species. Theor Appl Genet 73: 364–370CrossRefGoogle Scholar
  37. Klimaszewska K, Keller WA (1985) High frequency plant regeneration from thin layer explants of Brassica napus. Plant Cell, Tissue Org Cult 4: 184–197Google Scholar
  38. Klimaszweska K, Keller WA (1987) Plant regeneration from stem cortex protoplasts of Brassica napus. Plant Cell, Tissue Org Cult 8: 225–233Google Scholar
  39. Kolenbach HW, Wenzel G, Hoffmann F (1982) Regeneration of Brassica napus plantlets in cultures from isolated protoplasts of haploid stem embryos as compared with lead protoplasts. Z Pflanzenphysiol 105: 131–142Google Scholar
  40. Lammerick J (1970) Inter-specific transfer of clubroot resistance from Brassica campestris L. to B. napus L. N Z J Agric Res 13:105 —110Google Scholar
  41. Mackay GR (1977) The introgression of S-alleles into forage rape, Brassica napus L. from turnip, Brassica campestris L. spp. rapifera. Euphytica 26: 511–519CrossRefGoogle Scholar
  42. Mathews H, Bhatia CR, Mitra R, Krisha TG, Rao PS (1985) Regeneration of shoots from Brassica juncea (Linn) Czern and Coss cells transformed by Agrobacterium tumefaciens and expression of nopaline dehydrogenase genes. Plant Sci 39: 49–54CrossRefGoogle Scholar
  43. Mathews H, Rao PS, Bhatia CR (1986) Transformation of Brassica juncea by Agrobacterium tumefaciens harbouring plasmid pTiT37 and its `rooty’ mutant pTiT37.14a/a. J Genet 65: 37–44CrossRefGoogle Scholar
  44. Matsuzawa Y, Sarashima M (1986) Intergeneric hybridization between Raphanus sativa L. and Brassica nigra Koch and alloplasmic radish derivative. Jpn J Breed 36: 122–130Google Scholar
  45. Menczel L, Wolfe K (1984) High frequency of fusion induced in freely suspended protoplast mixtures by polyethylene glycol and dimethylsulfoxide at high pH. Plant Cell Rep 3: 196–198CrossRefGoogle Scholar
  46. 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: 98–101Google Scholar
  47. Meyerowitz EM, Pruitt RE (1985) Arabidopsis thaliana and plant molecular genetics. Science 229: 1214–1218Google Scholar
  48. Morgan A, Maliga P (1987) Rapid chloroplast segregation and recombination of mitochondrial DNA in Brassica cybrids. Mol Gen Genet 209: 240–246PubMedCrossRefGoogle Scholar
  49. Morinaga T (1934) Interspecific hybridization in Brassica. VI. The cytology of Fl hybrids of B. juncea and B. nigra. Cytologia 6: 62–67CrossRefGoogle Scholar
  50. Nasrallah JB, Kao T-H, Goldberg ML, Nasrallah ME (1985) A cDNA clone encoding an S-locusspecific glycoprotein from Brassica oleracea. Science 318: 263–267Google Scholar
  51. Negrutiu I, Jacobs M, Cattoir-Reynaerts A (1984) Progress in cellular engineering of plants: biochemical and genetic assessment of selectable markers from cultured cells. Plant Mol Biol 3: 289–302CrossRefGoogle Scholar
  52. Negrutiu I, DeBrouwer D, Watts JW, Sidorov VI, Dirks R, Jacobs M (1986) Fusion of plant protoplasts: a study using auxotrophic mutants of Nicotiana plumbaginifolia, Viviani. Theor Appl Genet 72:279 — 286Google Scholar
  53. Newell CA, Rhoads ML, Bidney DL (1984) Cytogenetic analysis of plants regenerated from tissue ex-plants and mesophyll protoplasts of winter rape, Brassica napus L. Can J Genet Cytol 26: 752–761Google Scholar
  54. Ogura H (1968) Studies of the new male sterility in japanese radish with special reference to the utilization of this sterility towards the practical raising of hybrid seed. Mem Fac Agric Kagoshima Univ 6: 39–78Google Scholar
  55. Olsson G (1960) Species crosses within the genus Brassica. 1. Artificial Brassica juncea Coss. Hereditas 46:171— 223Google Scholar
  56. Ooms G, Bains A, Burrell M, Karp A, Tuell D, Wilcox E (1985) Genetic manipulation in cultivars of oilseed rape (Brassica napus) using Agrobacterium. Theor Appl Genet 71: 325–329Google Scholar
  57. Paszkowski J, Pisan B, Shillito RD, Hohn T, Hohn B, Potrykus I (1986) Genetic transformation of Brassica campestris var. rapa protoplasts with an engineered Cauliflower Mosaic Virus genome. Plant Mol Biol 6: 303–312CrossRefGoogle Scholar
  58. Patnaik G, Cocking EC, Hamill J, Pental D (1982) A simple procedure for the manual isolation and identification of plant heterokaryons. Plant Sci Lett 24:105 —110Google Scholar
  59. Pauls KP, Chuong PV (1986) Flow cytometric identification of Brassica napus protoplast fusion products. Can J Bot 65: 834–838CrossRefGoogle Scholar
  60. Pelletier C, Primand C, Vedel F, Chetrie P, Remy R, Rousselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191: 244–250CrossRefGoogle Scholar
  61. Pontecorvo G (1971) Induction of directional chromosome elimination in somatic cell hybrids. Nature (Lond) 230: 367–369CrossRefGoogle Scholar
  62. Pua E-C, Mehra-Palta A, Nagy F, Chua N-H (1987) Transgenic plants of Brassica napus L. Biotechnology 5:815 — 817Google Scholar
  63. Robertson D, Earle ED (1986) Plant regeneration from leaf protoplasts of Brassica oleracea var. italica cv. green comet broccoli. Plant Cell Rep 5: 61–64CrossRefGoogle Scholar
  64. Robertson D, Earle ED (1987) Nitro-blue tetrazolium: A specific stain for photosynthetic activity in protoplasts. Plant Cell Rep 6: 70–73Google Scholar
  65. 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 — 309Google Scholar
  66. Roy NN (1984) Interspecific transfer of Brassica juncea-type high blackleg resistance to Brassica napus. Euphytica 33: 295–304CrossRefGoogle Scholar
  67. Schenck HR, Röbbelen G (1982) Somatic hybrids by fusion of protoplasts from Brassica oleracea and B. campestris. Z Pflanzenz 89: 278–288Google Scholar
  68. Shah DM, Horsch RB, Klee HJ, Kishore GM, Winter JA, Tumer NE, Hironaka CM, Sanders PR, Gasser CS, Aykent S, Siegel NR, Rogers SG, Fraley RT (1986) Engineering herbicide tolerance in transgenic plants. Science 233: 478–481PubMedCrossRefGoogle Scholar
  69. Shahin EA, Spivey R (1986) A dominant gene for Fusarium wilt resistance in protoplast-derived tomato plants. Theor Appl Genet 73: 164–169CrossRefGoogle Scholar
  70. Shiga T, Baba S (1971) Cytoplasmic male sterility in rape plants (Brassica napus L.). Jpn J Breed 21: 16–17Google Scholar
  71. Shiga T, Baba S (1973) Cytoplasmic male sterility in oilseed rape, Brassica napus L., and its utilization to breeding. Jpn J Breed 23: 187–197Google Scholar
  72. Spangenberg G, Schweiger H-G (1986) Controlled electrofusion of different types of protoplasts and subprotoplasts including cell reconstitution in Brassica napus L. Eur J Cell Biol 41: 51–56Google Scholar
  73. Spangenberg G, Neuhaus G, Schweiger H-G (1986) Expression of foreign genes in a higher plant cell after electrofusion-mediated cell reconstitution of a microinjected karyoplast and a cytoplast. Eur J Cell Biol 42: 236–238Google Scholar
  74. 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 — 162Google Scholar
  75. Taguchi T, Kameya T (1986) Production of somatic hybrid plants between cabbage and chinese cabbage through protoplast fusion. Jpn J Breed 36: 186–189Google Scholar
  76. Tanaka N, Hayakawa M, Mano Y, Ohkawa H, Matsui C (1985) Infection of turnip and radish storage roots with Agrobacterium rhizogenes. Plant Cell Rep 4: 74–77CrossRefGoogle Scholar
  77. 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: 379–384CrossRefGoogle Scholar
  78. Thompson KF (1972) Cytoplasmic male sterility in oilseed rape. Heredity 29: 253–257CrossRefGoogle Scholar
  79. Toriyama K, Kameya T, Hinata K (1987) Selection of a universal hybridizer in Sinapis turgida Del. and regeneration of plantlets from somatic hybrids with Brassica species. Planta 170: 308–313CrossRefGoogle Scholar
  80. Toriyama K, Hinata K, Kameya T (1987) Production of somatic hybrid plants, “Brassicomoricandia”, through protoplast fusion between Moricandia arvensis and Brassica oleracea. Plant Sci 48:123 —128Google Scholar
  81. Trail F, Wu FS (1986) Transformation in Brassica spp.: Sensitivity to antibiotics in vitro and tumor formation in vivo (Abstract). VI th Int Congr Plant tissue and cell culture, Univ Minnesota, Minneapolis, p 203Google Scholar
  82. U N (1935) Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization. Jpn J Bot 7: 389–452Google Scholar
  83. Vaeck M, Reynaerts A, Hofte H, Jansens S, De Beuckeleer M, Dean C, Zabeau M, Van Montagu M, Leemans J (1987) Transgenic plants protected from insect attack. Nature (Lond) 328: 33–37CrossRefGoogle Scholar
  84. Williams P, Hill CB (1986) Rapid-cycling populations of Brassica. Science 232: 1385–1389PubMedCrossRefGoogle Scholar
  85. Wu FS (1987) Localization of mitochondria in plant cells by vital staining with rhodamine 123. Planta 171: 346–357CrossRefGoogle Scholar
  86. Wu FS, Richards C, Trail F (1987) Calli formation and regeneration of hypocotyl protoplasts fused with evacuolated and enucleated mesophyll protoplasts in Brassica oleracea (Abstract). In Vitro 23:45 AGoogle Scholar
  87. Xu ZH, Davey MR, Cocking EC (1982) Plant regeneration from root protoplasts of Brassica. Plant Sci Lett 24:117 —121Google Scholar
  88. 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: 415–418CrossRefGoogle Scholar
  89. Zachrisson A, Bornman CH (1984) Application of electric field fusion in plant tissue culture. Physiol Plant 61: 314–320CrossRefGoogle Scholar
  90. Zachrisson A, Bornman CH (1986) Electromanipulation of plant protoplasts. Physiol Plant 67:507 —516Google Scholar
  91. Zimmermann U, Scheurich P (1981) High frequency fusion of plant protoplasts by electric fields. Planta 151: 26–32CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1989

Authors and Affiliations

  • F. Trail
    • 1
  • C. Richards
    • 2
  • F.-S. Wu
    • 2
  1. 1.Dept. of Plant PathologyCornell UniversityGenevaUSA
  2. 2.Zoecon Research InstituteSandoz Crop Protection CorporationPalo AltoUSA

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