Mapping of leaf water content using near-infrared hyperspectral imaging.

In this study, near-infrared hyperspectral imaging was applied to predict the water content of golden pothos (Epipremnum aureum) leaves, after which partial least squares regression (PLSR) analysis was performed to predict their averaged water content. The resulting predictive model was then applied to each single-pixel spectra in order to construct a water content image that could be used to evaluate the model's applicability to the single-pixel spectra through partial least squares score comparisons between the averaged spectra used for calibration and the single-pixel spectra. In the next phase, it was determined that a rebuilt PLSR predictive model based on the averaged spectra of an applicable pixel showed higher prediction accuracy than that of the original model. This study provides effective information about the limitations of prediction mapping and the optimization of pixel selections for better calibrations.

Characterization of Afp1, an antifreeze protein from the psychrophilic yeast Glaciozyma antarctica PI12.

The psychrophilic yeast Glaciozyma antarctica demonstrated high antifreeze activity in its culture filtrate. The culture filtrate exhibited both thermal hysteresis (TH) and ice recrystallization inhibition (RI) properties. The TH of 0.1 °C was comparable to that previously reported for bacteria and fungi. A genome sequence survey of the G. antarctica genome identified a novel antifreeze protein gene. The cDNA encoded a 177 amino acid protein with 30 % similarity to a fungal antifreeze protein from Typhula ishikariensis. The expression levels of AFP1 were quantified via real time-quantitative polymerase chain reaction (RT-qPCR), and the highest expression levels were detected within 6 h of growth at -12 °C. The cDNA of the antifreeze protein was cloned into an Escherichia coli expression system. Expression of recombinant Afp1 in E. coli resulted in the formation of inclusion bodies that were subsequently denatured by treatment with urea and allowed to refold in vitro. Activity assays of the recombinant Afp1 confirmed the antifreeze protein properties with a high TH value of 0.08 °C.

Cryoprotection and cryosterilization effects of type I antifreeze protein on E. coli cells.

To establish the effects of type I antifreeze protein (AFP) on E. coli cells, we have focused on the survival rate of the E. coli cells using type I AFP at various concentrations under rapid cooling conditions using liquid N2 at atmospheric or low pressure. The survival rate of E. coli was enhanced by the addition of type I AFP at a concentration of 10 microg/ml, and its value shifted from 0.73% to 2.96%. When the concentration of type I AFP was 100 microg/ml, the cell survival rate markedly decreased to 0.090%. This low survival rate was further decreased (0.022%) by the application of the same freeze-thaw treatment for four times. Also, the effect of type I AFP as a bactericidal agent did not vary according to the varying initial cell densities from 10(4) to 10(8) cells / ml. Furthermore, the effects of using type I AFP at 1.0 MPa with N2 gas under conditions of low pressure and low oxygen tension using a simple device were examined. When the actions of type I AFP as a cryoprotectant were stimulated, the survival rate of the E. coil cells increased to 57.8%. In addition, the bactericidal effect of type I AFP at 100 micro g/ml of protein concentration could also be enhanced. The survival rate using 100 g/ml of type I AFP under low pressure was 0.35% of that using 10 microg/ml under the same conditions. This is the first report on the cryoprotectant and cryosterilization effects of type I AFP of E. coli cells under various conditions.

A novel, intracellular antifreeze protein in an antarctic bacterium, Flavobacterium xanthum.

One strain of Antarctic bacteria, Flavobacterium xanthum IAM12026, has a highly active antifreeze protein (AFP) in the intracellular space. The cell-free extract from strain IAM12026 after culturing at 4 degree C for 7 days in TSB medium, had activity of 0.04 degree C at a concentration of 0.7 mg/ml. The ice crystals formed do not have distinct facets without typically rounded shape and the changes of their morphology during the course of the thermal hysteresis (TH) measurement. The ice crystal 'burst' occurring at the end-point of the TH is dendritic with hexagonal symmetry. Also, this activity was not affected by the treatment of dialysis and the addition of EDTA. Furthermore, this cell-free extract had high levels of ice recrystallization-inhibiting (RI) activity like those of Fish AFPs. The AFP (FlAFP) was homogeneity purified using chromatography. A relative molecular mass of approximately 59,000 was calculated from gel filtration and SDS-PAGE data. The thermal stability of FlAFP was below 50 degree C, and TH value was absent above 60 degree C. The TH value of FlAFP was activated at 5.2 degree C by the addition of 0.5 M malate. This activation was decreased with increasing protein concentration. To our knowledge this is the first report on the high level of TH and RI activities of bacterial intracellular AFP.