An E. coli ada transgenic clone of Nicotiana tabacum var. Xanthi has increased sensitivity to the mutagenic action of alkylating agents, maleic hydrazide and gamma-rays.
Two transgenic clones X3 and X15 of Nicotiana tabacum var. Xanthi, heterozygous in two genes (a1 and a2) for chloroplast differentiation and transformed with the E. coli DNA repair gene ada cloned downstream from the 1' direction of the dual mas promoter, differed in the expression of the ada gene, in the number of copies of integrated T-DNA and in the response to the mutagenic action of alkylating and non-alkylating agents. The X3 genome contained four copies and the X15 genome one copy of T-DNA, nevertheless the expression of the ada gene, measured by the activity of O6-alkylguanine DNA alkyltransferase (ATase), was about six times higher in X15 than in X3. ATase activity in both clones was highest in extracts from callus whereas very low (X15) or no (X3) activity was detected in leaf extracts. This may explain the lack of difference between X15 and non-transformed tobacco (NTX) in the frequency of N-methyl-N-nitrosourea (MNU)-induced somatic mutations in leaves. In contrast, the frequency of somatic mutations in X3 was about 2-5 times higher than in NTX and X15 after the same doses of MNU, methyl methanesulfonate, maleic hydrazide and gamma-rays. Alteration of plant gene(s) essential in mutation pathway(s) by insertion of T-DNA or by somaclonal variation may explain the higher sensitivity of the X3 clone.
Velemínský
J
J
Institute of Experimental Botany, Academy of Sciences, Prague, Czech Republic.
Angelis
K
K
Babůrek
I
I
Gichner
T
T
Satava
J
J
Bríza
J
J
Margison
G P
GP
D000477:Alkylating Agents / Q000378:metabolism / Q000633:toxicity*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005720:Gamma Rays; D005798:Genes, Bacterial; D008300:Maleic Hydrazide / Q000378:metabolism / Q000633:toxicity*; D008780:Methyltransferases / Q000378:metabolism; D008969:Molecular Sequence Data; D009153:Mutagens / Q000378:metabolism / Q000633:toxicity*; D019853:O(6)-Methylguanine-DNA Methyltransferase; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D014157:Transcription Factors; D014170:Transformation, Genetic
D000477:Alkylating Agents; D001426:Bacterial Proteins; D004274:DNA, Recombinant; D029968:Escherichia coli Proteins; D009153:Mutagens; D014157:Transcription Factors; D008780:Methyltransferases; C045675:Ada protein, E coli; D019853:O(6)-Methylguanine-DNA Methyltransferase; D008300:Maleic Hydrazide