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  • Agrobacterium tumefaciens  (1)
  • ELS (embryo-like structures)  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 78 (1989), S. 831-835 
    ISSN: 1432-2242
    Keywords: Haploid ; Transformation ; Canola ; Octopine ; Agrobacterium tumefaciens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Microspore-derived embryos of Brassica napus were transformed using the disarmed octopine-producing LBA4404 strain of Agrobacterium tumefaciens containing the binary vector pBin19. Octopine-producing strains have previously been reported to be ineffective in transforming Brassica. Four actively growing yellow/ green sectors were selected from the embryos on 50 mg/l kanamycin and plants regenerated. Analysis for NPT-II activity in these young plants initially indicated no expression of the bacterial NPT-II gene. The plants were nevertheless grown to maturity, selfed and S1 seed was collected. Three of the S1 plants produced microspores which were from 4 to 20 times more tolerant to kanamycin than the original parent. Southern analysis revealed that one plant (EC-1) had a single site of insertion and the other two plants (EC-2 and EC-6) had two sites of insertion with sequence homology to the bacterial NPT-II gene. Microspores from the EC-2 and EC-6 transgenics produced embryos on approximately five times the level of kanamycin tolerated by microspores from untransformed plants, while the EC-1 transgenic produced microspores with more than 20 times the tolerance to kanamycin. Analysis of S1 progeny of the EC-1 transgenic indicated that 100% of the progeny exhibited the trait through both Southern analysis and by expressing tolerance to kanamycin in microspore-derived embryos.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5044
    Keywords: ELS (embryo-like structures) ; greenhouse ; growth chamber ; Seneca-60
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microspore cultures were initiated from the North American sweet corn hybrid, ‘Seneca 60’. Donor plants were grown under two environments. One treatment comprised plants that matured completely in the greenhouse (GH) (28°C/23°C: day/night), while in a second treatment donor plants were isolated and divided into two treatment sets: cultured directly at 25°C, or given a heat treatment of 32°C for 10 days. Greenhouse-grown plants produced fewer embryo-like structures (ELS) than growth chambertreated plants regardless of the culture temperature treatment. If the microspores isolated from GC plants were subsequently provided with the initial high culture temperature, the number of ELS that could be recovered was more than doubled compared to the cultures incubated at 25°C continuously. The high culture temperature treatment also resulted in a higher quality of ELS (more compact), which led to a higher frequency of ELS that survived and were subsequently transferred to regeneration medium. However, while plant regeneration and subsequent selfed seeds were obtained, the frequency was very low and further research is required in this area.
    Type of Medium: Electronic Resource
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