Discrimination between nongenetically modified (Non-GM) and GM plant tissue expressing cysteine-rich polypeptide using FT-raman spectroscopy.
Fourier transform (FT)-Raman spectroscopy was applied to the analysis of genetically modified (GM) plant tissue. Transgenic carrot callus and tobacco plants possessing a novel StSn1 gene coding for a cysteine-rich snakin-1 polypeptide were obtained after Agrobacterium-mediated transformation. The presence of the StSn1 gene and its expression were confirmed by polymerase chain reactions using plant DNA and cDNA as templates for the amplification of the transgenes. Raman measurements were taken from lyophilized GM carrot callus tissue, fresh GM tobacco leaves, and from seeds produced by GM tobacco plants as well as from the nontransformed controls. Cluster analysis applied to the obtained spectra allowed clear separation of the GM samples expressing the StSn1 gene and the nontransformed control to distinct groups. Such discrimination was achieved only when wavenumber ranges around 500 cm (-1) were analyzed. The results indicate that discrimination between the GM and non-GM materials was related to S-S stretching vibrations in snakin-1, as it contained six sulfur bridges. Other introduced genes, neomycine phosphotransferase ( nptII) and Chitinase ( chit36), did not cause any detectable changes by Raman spectroscopy in plant tissue. This is the first report on the use of Raman spectroscopy for a nondestructive analysis of GM plant material expressing the gene coding for a cysteine-rich polypeptide.
Baranski
Rafal
R
Department of Genetics, Plant Breeding, and Seed Science, Agricultural University of Krakow, Al. 29 Listopada 54, 31-425 Krakow, Poland. baranski@ogr.ar.krakow.pl
Baranska
Malgorzata
M
D003545:Cysteine / Q000032:analysis*; D018744:DNA, Plant / Q000032:analysis; D018552:Daucus carota / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012231:Rhizobium / Q000235:genetics; D012639:Seeds / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D014026:Nicotiana / Q000737:chemistry / Q000235:genetics
D018744:DNA, Plant; D010940:Plant Proteins; D003545:Cysteine