Another study used modified winter flounder AFP genes, plus
and minus a signal peptide, to transform spring wheat. Results were similar to those reported by
Wallis et al. (1997) for transgenic potato. A synthetic gene encoding a larger form of the winter flounder AFP was used and this was suggested to be responsible for the increased thermal stability of the expressed protein relative to an earlier study by Kenward
et al. (1993). In the study reported by Khanna and Daggard (2006),
substantial amounts of the AFP accumulated in both the cytoplasm and the apoplast when plants were grown at 14–18 ◦ C. The apoplast targeted AFP represented 1.61% of the total soluble protein and 90% of the AFP was apoplastic. This is important as ice is typically initiated in the apoplast. Recrystallization inhibition was observed in whole plant extracts and in apoplastic fluid, indicating the presence and accumulation of active AFP. Electrolyte
leakage tests indicated that transgenic lines expressing AFP had improved freeze tolerance relative to wild-type plants. This was especially true in the line demonstrating the highest level of AFP (measured by Western blot), with 90% of the AFP being secreted to the apoplast. While wild-type plants exhibited >50% electrolyte leakage at −5 ◦ C, the highest AFP producing line exhibited only 27% leakage at −5 ◦ C, 40% at −6 ◦ C, and approximately 55% at −7 ◦ C.
Consequently, once again the best AFP producing lines provided
about 1 ◦ C of frost protection relative to the wild-type. The reduction in freezing injury in the transgenic lines could not be absolutely attributed to inhibition of freezing (avoidance) or to less freezing damage (tolerance).
Another study used modified winter flounder AFP genes, plusand minus a signal peptide, to transform spring wheat. Results were similar to those reported byWallis et al. (1997) for transgenic potato. A synthetic gene encoding a larger form of the winter flounder AFP was used and this was suggested to be responsible for the increased thermal stability of the expressed protein relative to an earlier study by Kenwardet al. (1993). In the study reported by Khanna and Daggard (2006),substantial amounts of the AFP accumulated in both the cytoplasm and the apoplast when plants were grown at 14–18 ◦ C. The apoplast targeted AFP represented 1.61% of the total soluble protein and 90% of the AFP was apoplastic. This is important as ice is typically initiated in the apoplast. Recrystallization inhibition was observed in whole plant extracts and in apoplastic fluid, indicating the presence and accumulation of active AFP. Electrolyteleakage tests indicated that transgenic lines expressing AFP had improved freeze tolerance relative to wild-type plants. This was especially true in the line demonstrating the highest level of AFP (measured by Western blot), with 90% of the AFP being secreted to the apoplast. While wild-type plants exhibited >50% electrolyte leakage at −5 ◦ C, the highest AFP producing line exhibited only 27% leakage at −5 ◦ C, 40% at −6 ◦ C, and approximately 55% at −7 ◦ C.Consequently, once again the best AFP producing lines providedabout 1 ◦ C of frost protection relative to the wild-type. The reduction in freezing injury in the transgenic lines could not be absolutely attributed to inhibition of freezing (avoidance) or to less freezing damage (tolerance).
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Another study used modified winter flounder AFP genes, plus
and minus a signal peptide, to transform spring wheat. Results were similar to those reported by
Wallis et al. (1997) for transgenic potato. A synthetic gene encoding a larger form of the winter flounder AFP was used and this was suggested to be responsible for the increased thermal stability of the expressed protein relative to an earlier study by Kenward
et al. (1993). In the study reported by Khanna and Daggard (2006),
substantial amounts of the AFP accumulated in both the cytoplasm and the apoplast when plants were grown at 14–18 ◦ C. The apoplast targeted AFP represented 1.61% of the total soluble protein and 90% of the AFP was apoplastic. This is important as ice is typically initiated in the apoplast. Recrystallization inhibition was observed in whole plant extracts and in apoplastic fluid, indicating the presence and accumulation of active AFP. Electrolyte
leakage tests indicated that transgenic lines expressing AFP had improved freeze tolerance relative to wild-type plants. This was especially true in the line demonstrating the highest level of AFP (measured by Western blot), with 90% of the AFP being secreted to the apoplast. While wild-type plants exhibited >50% electrolyte leakage at −5 ◦ C, the highest AFP producing line exhibited only 27% leakage at −5 ◦ C, 40% at −6 ◦ C, and approximately 55% at −7 ◦ C.
Consequently, once again the best AFP producing lines provided
about 1 ◦ C of frost protection relative to the wild-type. The reduction in freezing injury in the transgenic lines could not be absolutely attributed to inhibition of freezing (avoidance) or to less freezing damage (tolerance).
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