Abstract
Black rot caused by the bacterium Xanthomonas campestris pv campestris is one of the most serious diseases of Brassica oleracea. Since sources of resistance to the disease within B. oleracea are insufficient and control means are limited, the development of resistant breeding lines is extremely desirable. Certain lines of B. napus contain very high resistance controlled by a dominant gene, but crossing the two species sexually is very difficult. Therefore, somatic hybrids were produced by protoplast fusion between rapid cycling B. oleracea and a B. napus line highly resistant to X. campestris pv campestris. Hybrid identity was confirmed by morphological studies, flow cytometric estimation of nuclear DNA content, and analysis of random amplified polymorphic DNA (RAPD). Inoculations with the pathogen identified four somatic hybrids with high resistance. The resistant hybrid plants were fertile and set seed when selfed or crossed reciprocally to the bridge line ‘15’ (Quazi 1988). Direct crosses to B. oleracea were unsuccessful, but embryo rescue facilitated the production of a first-backcross generation. The BC1 plants were resistant to the pathogen. Progeny from the crosses to ‘line 15’ were all susceptible. Embryo rescue techniques were not obligatory for the development of a second-backcross generation, and several resistant BC2 plants were obtained.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arumuganathan K, Earle ED (1991) Estimation of nuclear DNA content of plants by flow cytometry. Plant Mol Biol Rep 9:221–231
Ayotte R, Harney PM, Machado VS (1987) The transfer of triazine resistance from Brassica napus L. to B. oleracea L. I. Production of F1 hybrids through embryo rescue. Euphytica 36:615–624
Ayotte R, Harne, PM, Machad, VS (1988a) The transfer of triazine resistance from Brassica napus L. to B. oleracea L. II. Morphology, fertility and cytology of the F1 hybrid. Euphytica 37:189–197
Ayotte R, Harney PM, Machado VS (1988b) The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First back-cross to parental species. Euphytica 38:137–142
Ayotte R, Harney PM, Machado VS. (1989) The transfer of triazine resistance from Brassica napus L. to B. oleracea L. IV. second and third backcrosses to B. oleracea and recovery of an 18-chromosome, triazine resistant BC3. Euphytica 40:15–19
Bain DC (1952) Reaction of Brassica seedlings to black rot. Phytopathology 42:497–500
Chiang MS, Chiang BY, Grant WF (1977) Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea var. capitata). I. Interspecific hybridization between B. napus and B. oleracea var. capitata. Euphytica 26:319–336
Chiang MS, Chiang BY, Grant WF (1978) Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea var. capitata). II. Meiosis in the interspecific hybrids between B. napus and 2x and 4x cabbage. Euphytica 27:81–93
Dickson MD, Hunter JE (1987) Inheritance of resistance in cabbage seedlings to black rot. HortSci 22:108–109
Ferreira ME, Dias JS, Mengistu A, Williams PH (1993) Screening of Portuguese cole landraces (Brassica oleracea L.) with Leptosphaeria maculans and Xanthomonas campestris pv campestris. Euphytica 65:219–227
Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220:1049–1051
Grabiec B (1971) Evaluation of semisynthetic values of forms of the Brassica napus L. species from the standpoint of plant breeding. Genet Polon 12:233–237
Guo H, Dickson MH, Hunter JE (1991) Brassica napus sources of resistance to black rot in crucifers and inheritance of resistance. HortSci 26:1545–1547
Hansen LN, Earle ED (1994a) Regeneration of plants from protoplasts of rapid cycling Brassica oleracea L. Plant Cell Rep 13:335–339
Hansen LN, Earle ED (1994b) Novel flowering and fatty acid characters of rapid cycling Brassica napus L. resynthesized by protoplast fusion. Plant Cell Rep 14:151–156
Honma S, Summers WL (1976) Interspecific hybridization between Brassica napus L. (Napobrassica group) and B. oleracea L. (Botrytis group). J Am Soc Hort Sci 101:299–302
Hu J, Quiros CF (1991) Identification of broccoli and cauliflower cultivars with RAPD markers. Plant Cell Rep 10:505–511
Hunter JE, Dickson MH, Ludwig JW (1987) Source of resistance to black rot of cabbage expressed in seedlings and adult plants. Plant Dis 71:263–266
Jamwal RS, Sharma PP (1986) Inheritance of resistance to black rot (Xanthomonas campestris pv campestris) in cauliflower (Brassica oleracea var. botrytis). Euphytica 35:941–943
Jourdan PS, Earle ED, Mutschler MA (1989) Atrazine-resistant cauliflower obtained by somatic hybridization between Brassica oleracea and ATR-B. napus. Theor Appl Genet 78:271–279
Kao HM, Keller WA, Gleddie S, Brown GG (1992) Synthesis of Brassica oleracea/Brassica napus somatic hybrid plants with novel organelle DNA compositions. Theor Appl Genet 83:313–320
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
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497
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 191:244–250
Quazi MH (1988) Interspecific hybrids between Brassica napus L. and Brassica oleracea L. developed by embryo culture. Theor Appl Genet 75:309–318
Shelton AM, Hunter JE (1985) Evaluation of the potential of the flea beetle Phyllotreta cruciferae to transmit Xanthomonas campestris pv campestris, causal agent of black rot of crucifers. Can J Plant Pathol 7:308–310
Staub T, Williams PH (1972) Factors influencing black rot lesion development in resistant and susceptible cabbage. Phytopathology 62:722–728
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
U N (1935) Genome analysis in Brassica with special reference to the experimental formation of Brassica napus and peculiar mode of fertilization. Jpn J Bot 7:389–452
Walters TW, Earle ED (1990) A simple, versatile feeder layer system for Brassica oleracea protoplast culture. Plant Cell Rep 9:316–319
Williams PH (1980) Black rot: a continuing threat to world crucifers. Plant Dis 64:736–742
Williams PH, Staub T, Sutton JC (1972) Inheritance of resistance in cabbage to black rot. Phytopathology 62:247–252
Yarrow SA, Burnett LA, Wildeman RP, Kemble RJ (1990) The transfer of ‘Polima’ cytoplasmic male sterility from oilseed (Brassica napus) to broccoli (B. oleracea) by protoplast fusion. Plant Cell Rep 9:185–188
Author information
Authors and Affiliations
Additional information
Communicated by P. Maliga
Rights and permissions
About this article
Cite this article
Hansen, L.N., Earle, E.D. Transfer of resistance to Xanthomonas campestris pv campestris into Brassica oleracea L. by protoplast fusion. Theoret. Appl. Genetics 91, 1293–1300 (1995). https://doi.org/10.1007/BF00220944
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00220944