High frequency Agrobacterium tumefaciens-mediated plant transformation induced by ammonium nitrate.
Success in plant genetic transformation depends on the efficiency of explant regeneration and transgene integration. Whereas the former one depends on explant totipotency, the latter depends on the activity of host DNA repair and chromatin organisation factors. We analyzed whether factors that result in an increase in recombination frequency can also increase transformation efficiency. Here, we report that a threefold increase in the concentration of NH(4)NO(3) in the growth medium results in more than a threefold increase in the Agrobacterium tumefaciens-mediated transformation frequency of Nicotiana tabacum plants. Regeneration of calli without selection showed that the increase in transformation frequency was primarily due to the increase in transgene integration efficiency rather than in tissue regeneration efficiency. PCR analysis of insertion sites showed a decrease in the frequency of truncations of the T-DNA right border and an increase on the left border. We hypothesize that exposure to ammonium nitrate modifies the activity of host factors leading to higher frequency of transgene integrations and possibly to the shift in the mechanism of transgene integrations.
Boyko
Alex
A
Department of Biological Sciences, University of Lethbridge, Lethbridge, Canada.
Matsuoka
Aki
A
Kovalchuk
Igor
I
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics*; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D009566:Nitrates / Q000494:pharmacology*; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics; D012038:Regeneration; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics*; D014170:Transformation, Genetic; D019076:Transgenes
D003470:Culture Media; D004269:DNA, Bacterial; D009566:Nitrates; C036483:T-DNA; C006568:ammonium nitrate