The Validation of the Dai Nippon Medi·Ca SA Method for AOAC Research Institute Performance Tested MethodsSM Certification.

A ready-made dry medium method for Staphylococcus aureus count, the Medi·Ca SA method incubated at 35 or 37°C, was compared with the Baird-Parker method (AOAC Official MethodSM 975.55) for 11 food matrices: raw beef, raw ground beef, raw lamb, cooked ham, raw salmon, frozen prawn, fresh chilled pasta, pasteurized milk, natural cheese, cream puff, and potato salad. The mean difference between the two methods at each contamination level for each matrix was <0.5 log10, and the 95% confidence intervals on the mean differences fell within the range of -0.50 to 0.50. Standard deviation of repeatability and RSDr values of the Medi·Ca SA method were generally the same level as those of the Baird-Parker method, and r2 ranged from 0.98 to 1.00. Product consistency and stability studies showed little variability between productions lots and a shelf-life of 16 months. Incubation time within the range of 22-26 h and variations to the sample volume did not adversely affect the results. These results showed that the Medi·Ca SA method is a reasonable alternative to the reference method for selected food matrices and makes it possible to simultaneously detect and enumerate S. aureus in only 24 h.

Rice DEP1, encoding a highly cysteine-rich G protein γ subunit, confers cadmium tolerance on yeast cells and plants.

A rice cDNA, OsDEP1, encoding a highly cysteine (Cys)-rich G protein γ subunit, was initially identified as it conferred cadmium (Cd) tolerance on yeast cells. Of the 426 aa constituting OsDEP1, 120 are Cys residues (28.2%), of which 88 are clustered in the C-terminal half region (aa 170-426). To evaluate the independent effects of these two regions, two truncated versions of the OsDEP1-expressing plasmids pOsDEP1(1-169) and pOsDEP1(170-426) were used to examine their effects on yeast Cd tolerance. Although OsDEP1(170-426) conferred a similar level of Cd tolerance as the intact OsDEP1, OsDEP1(1-169) provided no such tolerance, indicating that the tolerance effect is localized to the aa 170-426 C-terminal peptide region. The Cd responses of transgenic Arabidopsis plants constitutively expressing OsDEP1, OsDEP1(1-169) or OsDEP1(170-426), were similar to the observations in yeast cells, with OsDEP1 and OsDEP1(170-426) transgenic plants displaying Cd tolerance but OsDEP1(1-169) plants showing no such tolerance. In addition, a positive correlation between the transcript levels of OsDEP1 or OsDEP1(170-426) in the transgenics and the Cd content of these plants upon Cd application was observed. As several Arabidopsis loss-of-function heterotrimeric G protein ß and γ subunit gene mutants did not show differences in their Cd sensitivity compared with wild-type plants, we propose that the Cys-rich region of OsDEP1 may function directly as a trap for Cd ions.

A novel plant cysteine-rich peptide family conferring cadmium tolerance to yeast and plants.

We have identified a novel cDNA clone, termed DcCDT1, from Digitaria ciliaris, that confers cadmium (Cd)-tolerance to yeast (Saccharomyces cerevisiae). The gene encodes a predicted peptide of 55 amino acid residues of which 15 (27.3%) are cysteine residues. We found that monocotyledonous plants possess multiple DcCDT1 homologues, for example rice contains five DcCDT1 homologues (designated OsCDT1~5), whereas dicotyledonous plants, including Arabidopsis thaliana, Brassica rapa, poplar (Populus tremula x Populus alba) and Picea sitchensis, appear to possess only a single homologue. GFP fusion experiments demonstrate that DcCDT1 and OsCDT1 are targeted to both the plant cytoplasmic membranes and cell walls. Constitutive expression of DcCDT1 or OsCDT1 confers Cd-tolerance to transgenic A. thaliana plants by lowering the accumulation of Cd in the cells. The functions of the DcCDT1 family members are discussed in the light of these findings.