Biocontrol Efficacy of Two Antagonistic Yeasts Against Postharvest Diseases in Peach Fruits During Storage Periods

Two antagonistic yeasts, Thichosporon pullulans and Cryptococcus laurentii, were investigated for their biocontrol potential to blue mold rot and rhizopus rot on harvested peach fruits(Prunus persica L. Batsch, cv. Okubao), alone or in combination with a low dose of iprodione(50 μg ml-1). The results indicated that T. pullulans and C. laurentii were effective at reducing disease incidence and severity of blue mold rot and rhizopus rot in peach fruits. Biocontrol efficacy of C. laurentii and T. pullulans were significantly enhanced by combination with a low dose of iprodione(50 μg ml-1)against blue mold and rhizopus rot in peach fruits. T. pullulans and C. laurentii combined with a low dose of iprodione(50 μg ml-1)resulted in better disease control than either iprodione or the yeasts used alone. Dipping fruits in suspensions of antagonist cells showed the similar control effect as the treatment with iprodione(500μg ml-1).

Sensitivity of Four Yeasts to Fungicides and CO2 Concentrations and Their Antagonistic Ability in Combination with Fungicide to Pathogenic Fungi in vitro

The yeasts Cryptococcus laurentii, Trichosporon pullulans, Rhodotorula glutinis andTrichosporon sp.were used to investigate their sensitivity to four fungicides anddifferent concentrations of CO2, as well as their antagonistic ability to Penicilliumexpansumand Alternariaalternata in vitro when applied with fungicide. There were significantdifferences in sensitivity to the fungicides among the different yeasts(P=0.05). R.glutinis was more sensitive to Deccocil, Iprodione and Stroby as compared to otheryeasts. Combination antagonistic yeasts with fungicide could more significantly enhancebiocontrol ability of the yeasts against the pathogenic fungi in vitro(P = 0.05). C.laurentii was the most effective antagonist among the four yeasts and could completelycontrol spore germination of P.expansum and A.alternata when combined with Stroby at theconcentration of 100 L L-1. The yeasts, except R.glutinis, could grow well on nutrientyeast dextrose agar (NYDA) after 8 d incubation even at 20% CO2 concentration at 25℃.Particularly Trichosporon sp. showeda better adaptability to low temperature as comparedto other antagonists.

Optimization of Media for Production of an Effective Yeast Biocontrol Agent Pichia membranefaciens

The growth of Pichia membranefaciens was studied using different nitrogen and carbonsources as substrates. Among nitrogen sources tested, soya peptone, yeast extract power,beef extract and polypeptone were relatively favorable to the growth of yeast. Thedensity of the yeast showed to be directly proportional to carbon sources supplementation.Glucose and fructose were good carbon sources for the yeast growth. However, lactoseshowed poor performance for the cell growth of the yeast. In this study, beef extractpresented a good synergic effect on the yeast growth with different carbonhydrates. Themedium for P.membranefaciens used glucose and beef extract as substrates. The higherconcentration of glucose and beef extract, the better growth of P.membranefaciens. Theaddition of chlorella growth factor (CGF) stimulated markedly the growth of P.membranefaci-ens.The increased concentration of CGF from 0.5 to 1% did not enhance the numbers ofP.membranefaciens. This result will help design a better strategy for scale-up produc-tion of P.membranefaciens.

G418-Resistance as a Dominant Selectable Marker for Heterogenous Gene Expression in Antagonist Pichia membranefaciens

Pichia membranefaciens, which was isolated from the surface of peach fruits, showed effective biocontrol capabilityagainst rhizopus rot of peach fruits. Aminoglycoside antibiotic G418 can inhibit the growth of P. membranefaciens. Theminimal inhibitory concentration of G418 to P. membranefaciens in YPD medium was 100g mL-1. We constructed aphosphoglycerate kinase (PGK) promoter-driven neoR expression cassette, which was called pFL61-neo. The biocontrolyeast P. membranefaciens was transformed with pFL61-neo by lithium acetate method. Expression vector pFL61-neoconferred P. membranefaciens drug resistance to 100g mL-1 G418. The transformant could keep a high percentage ofplasmid-containing of transformant with 67.87% after 50 generations in non-selective medium. The result showed that P.membranefaciens could recognize the promoter and terminator of PGK and direct the expression of heterologous neoRgene. Expression vector pFL61-neo could exist stably in P. membranefaciens. Therefore, it is feasible to utilize G418-resistance as a dominant selectable marker for heterogenous gene expression in antagonist P. membranefaciens.