Thermally Stimulated Depolarization Currents (TSDC): A sensitive technique for analyzing protein structure

The water molecules surrounding proteins as a thin layer and those packaged in pockets and cavities shape and control their structure. Thermally Stimulated Depolarization Currents (TSDC) technique has been applied to investigate the hydration structure of six proteins with different structural motifs: pepsin,β-lactoglobulin,α-chymotrypsin, bovine serum albumin, human serum albumin and myoglobin, at very low hydration level (water vapor activity aw≈0.80) both in the native state and after treatment in trifluoroethanol/water mixture 80% (v/v). A combined approach based on the use of the TSDC technique, able to distinguish H2O dipoles belonging to the solvation shell in terms of their order degree and mobility, and of FTIR and CD spectroscopies has allowed us to reexamine the problem of conformational stability of macromolecules as a function of their hydration.

Vapor Activity and Systemic Movement of Mefenoxam Control Grapevine Downy Mildew

Metalaxyl is translocated from roots to leaves to control a number of oomycete pathogens, but systemic movement from vegetative organs into fruit and vapor activity against Plasmopara viticola, the causal agent of grapevine downy mildew, has not been examined experimentally. We inoculated fruit clusters of grapevines with P. viticola at prebloom, bloom, or 1 week postbloom. We then selectively applied mefenoxam (288 mg/liter), the active enantiomer of metalaxyl, to the leaves or stem tissue 12 to 48 h after inoculation. Little to no downy mildew developed on fruit when mefenoxam was applied to leaf tissue, stem tissue, or both. In contrast, downy mildew symptoms were severe on inoculated clusters on untreated shoots. When potential vapor activity was blocked, we observed fungicidal activity on seedling foliage in response to apparent systemic movement from treated stems and soil, but not from leaves. However, when vapor activity was permitted, mefenoxam residues on treated leaves controlled disease on other, untreated leaves. In subsequent vineyard experiments, vapor and systemic activity provided equivalent and near-complete suppression of downy mildew on clusters 48 h post inoculation. Furthermore, inoculated grape seedlings that were placed near mefenoxam-treated seedlings in open and closed systems developed nil to trace levels of downy mildew compared with controls, further indicating that the material has strong vapor activity.

Comparative Physical Modes of Action of Azoxystrobin, Mancozeb, and Metalaxyl Against Plasmopara viticola (Grapevine Downy Mildew)

The physical modes of action of azoxystrobin, mancozeb, and metalaxyl were evaluated on grapevine seedlings using Plasmopara viticola as a model pathogen. The protectant, postinfection, postsymptom, translaminar, and vapor activities of azoxystrobin were evaluated at a rate of 250 μg/ml. Azoxystrobin provided 100% disease control when applied 1 to 5 days before inoculation. Postinfection applications of azoxystrobin had little effect on the incidence of disease, but colony area and sporulation from the resultant lesions was reduced by 47 and 96%, respectively, relative to the check treatment when applied up to 5 days after inoculation. Postsymptom applications (6 days after inoculation) of azoxystrobin resulted in an 85% mean reduction of resporulation from diseased tissue relative to the check when seedlings were evaluated 1 to 14 days after treatment. Translaminar activity was greatest when the upper surface of the leaf was treated 7 days before inoculation of the lower leaf surface (94% disease control). In contrast, control was <50% when leaves were similarly inoculated 1 and 3 days after treatment. Vapor activity was not pronounced, providing maximum reductions of 5, 11, and 37%, with regard to incidence, colony area, and sporulation, relative to the check when seedlings were treated 1 to 7 days before inoculating adjacent, untreated leaves. Comparatively, mancozeb (1,790 μg/ml) provided complete control of the disease when applied 1 to 5 days before inoculation, but showed little postinfection activity in reducing disease incidence, although it exhibited moderate to high antisporulant activity when applied in postinfection and postsymptom modes (mean reductions of 38 and 89%, respectively, compared with the check treatments). Metalaxyl (260 μg/ml) also provided complete control of the disease when used in protectant mode, and also when applied 1 day after inoculation. Applications at 3 to 5 days after inoculation provided substantial reductions in disease severity and sporulation (mean reductions of 46 and 94%, respectively, compared with the check treatments), and postsymptom applications resulted in a mean 84% reduction in resporulation. Collectively, the results of this study illustrate the unique physical modes of action for azoxystrobin in comparison to that of two traditional protectant and systemic fungicides, and provide information on how azoxystrobin and other strobilurin fungicides with similar physical modes of action should be best used in disease management programs.

WATER BALANCE IN FLESH FLY PUPAE AND WATER VAPOR ABSORPTION ASSOCIATED WITH DIAPAUSE

We report the water balance characteristics for diapausing and nondiapausing pupae of the flesh fly Sarcophaga crassipalpis. The challenge of maintaining water balance is particularly acute for pupae that spend 9–10 months in diapause, without access to drinking water. While diapausing pupae can tolerate a loss of up to 24.5% of their total body water content (67.3%), they have also acquired several other physiological attributes that have enhanced their capacity for maintaining water balance. Net transpiration rates for diapausing pupae (0.008 % h−1) are far lower than rates for nondiapausing pupae (0.023 % h−1). In addition, diapausing pupae can counter water loss with their ability to absorb water vapor from lower humidities (approximate water vapor activity, av, 0.58 at 20°C) than can nondiapausing pupae (approximate av 0.74 at 20°C). The high ritical transition temperature for diapausing pupae (39 °C, compared to 30 °C for nondiapausing pupae) suggests that epicuticular lipids have been modified to restrict water loss during diapause.