Potential Role of Exoglucanase Genes (WaEXG1 and WaEXG2) in the Biocontrol Activity of Wickerhamomyces anomalus.

The use of yeasts, including Wickerhamomyces anomalus, as biocontrol agents of fungi responsible for postharvest diseases of fruits and vegetables has been investigated for the past two decades. Among a variety of mechanisms, the production of glucanases coded by the "killer genes" WaEXG1 and WaEXG2 have been reported to play a role in the ability of yeast to inhibit other fungi. The objective of the present study was to determine the expression of these genes by RT-qPCR, utilizing gene-specific primers, when W. anomalus was grown on grape berries and oranges that were either non-inoculated or inoculated with Botrytis cinerea or Penicillium digitatum, or in minimal media supplemented with cell walls of various plant pathogens and different amounts of glucose. Results indicated that WaEXG2 was more responsive than WaEXG1 to the nutritional environment (including the addition of glucose to cell wall-amended media) in vitro and appeared to play a greater role in the cellular metabolism of W. anomalus. WaEXG2 expression also appeared to be more responsive to the presence of cell walls of P. digitatum and B. cinerea than other fungal species, whereas the same level of induction was not seen in vivo when the yeast was grown in wounded/pathogen-inoculated fruits.

Enhancing in vitro degradation of alfalfa hay and corn silage using feed enzymes.

A series of in vitro fermentation experiments was performed to assess the effects of 4 feed enzyme products (FE) that varied in enzymatic activities on the degradation of alfalfa hay and corn silage. The FE contained a range of endoglucanase, exoglucanase, xylanase, and protease activities, and a range of dose rates (DR) was used. The objective of the study was to identify effective formulations and optimum DR, and to establish if combining FE would further improve fiber degradation. For alfalfa hay, quadratic increases in gas production and degradation of dry matter (DM) and fiber were observed for all FE, with maximum responses at low to medium DR. For corn silage, none of the FE increased gas production or DM degradation, but all FE increased NDF degradation, with optimum DR in the low to medium range. The proteolytic enzyme papain improved fiber degradation of alfalfa hay and corn silage in a manner similar to that observed for polysaccharidase FE. Among the polysaccharidase FE, added activities of endoglucanase and exoglucanase were positively correlated with improvement in neutral detergent fiber (NDF) degradability of corn silage, whereas only added endoglucanase activity tended to be correlated with improvement in NDF degradability of alfalfa hay. Combining effective polysaccharidase FE further improved fiber degradation of both forages, with greater improvements for corn silage. Combining polysaccharidase and proteolytic FE further improved NDF degradation of corn silage, but not alfalfa hay. Combination treatments generally resulted in additive effects with increases in fiber degradation equal to the sum of the improvements for the individual enzyme components. Improved fiber degradation of corn silage was associated with decreased acetate to propionate ratios. Enzyme products that improve in vitro degradation of forages may have the potential to improve lactational performance of dairy cows.