Identification of race-specific resistance in North American Vitis spp. limiting Erysiphe necator hyphal growth.

Race-specific resistance against powdery mildews is well documented in small grains but, in other crops such as grapevine, controlled analysis of host-pathogen interactions on resistant plants is uncommon. In the current study, we attempted to confirm powdery mildew resistance phenotypes through vineyard, greenhouse, and in vitro inoculations for test cross-mapping populations for two resistance sources: (i) a complex hybrid breeding line, 'Bloodworth 81-107-11', of at least Vitis rotundifolia, V. vinifera, V. berlandieri, V. rupestris, V. labrusca, and V. aestivalis background; and (ii) Vitis hybrid 'Tamiami' of V. aestivalis and V. vinifera origin. Statistical analysis of vineyard resistance data suggested the segregation of two and three race-specific resistance genes from the two sources, respectively. However, in each population, some resistant progeny were susceptible in greenhouse or in vitro screens, which suggested the presence of Erysiphe necator isolates virulent on progeny segregating for one or more resistance genes. Controlled inoculation of resistant and susceptible progeny with a diverse set of E. necator isolates clearly demonstrated the presence of fungal races differentially interacting with race-specific resistance genes, providing proof of race specificity in the grape powdery mildew pathosystem. Consistent with known race-specific resistance mechanisms, both resistance sources were characterized by programmed cell death of host epidermal cells under appressoria, which arrested or slowed hyphal growth; this response was also accompanied by collapse of conidia, germ tubes, appressoria, and secondary hyphae. The observation of prevalent isolates virulent on progeny with multiple race-specific resistance genes before resistance gene deployment has implications for grape breeding strategies. We suggest that grape breeders should characterize the mechanisms of resistance and pyramid multiple resistance genes with different mechanisms for improved durability.

Mutation at ß-Tubulin Codon 200 Indicated Thiabendazole Resistance in Penicillium digitatum Collected from California Citrus Packinghouses.

Thiabendazole (TBZ) is commonly applied to harvested citrus fruit in packinghouses to control citrus green mold, caused by Penicillium digitatum. Although TBZ is not used before harvest, another benzimidazole, thiophanate methyl, is commonly used in Florida and may be introduced soon in California to control postharvest decay of citrus fruit. Isolates from infected lemons and oranges were collected from many geographically diverse locations in California. Thirty-five isolates collected from commercial groves and residential trees were sensitive to TBZ, while 19 of 74 isolates collected from 10 packinghouses were resistant to TBZ. Random amplified polymorphic DNA analysis indicated that the isolates were genetically distinct and differed from each other. Nineteen TBZ-resistant isolates and a known TBZ-resistant isolate displayed a point mutation in the ß-tubulin gene sequence corresponding to amino acid codon position 200. Thymine was replaced by adenine (TTC → TAC), which changed the phenylalanine (F) to tyrosine (Y). In contrast, for 49 TBZ-sensitive isolates that were sequenced, no mutations at this or any other codon positions were found. All of the isolates of P. digitatum resistant to TBZ collected from a geographically diverse sample of California packinghouses appeared to have the same point mutation conferring thiabendazole resistance.

Control of Citrus Green Mold by Carbonate and Bicarbonate Salts and the Influence of Commercial Postharvest Practices on Their Efficacy.

The toxicity to Penicillium digitatum and practical use of carbonate and bicarbonate salts to control green mold were determined. The effective dose (ED50) concentrations to inhibit the germination of P. digitatum spores of sodium carbonate (SC), potassium carbonate, sodium bicarbonate (SBC), ammonium bicarbonate, and potassium bicarbonate were 5.0, 6.2, 14.1, 16.4, and 33.4 mM, respectively. All were fungistatic because spores removed from the solutions germinated in potato dextrose broth. SC and SBC were equal and superior to the other salts for control of green mold on lemons and oranges inoculated 24 h before treatment. When sodium content and high pH must be minimized, SBC could replace SC. Furthermore, because a higher proportion of NaOCl would be present in the active hypochlorous acid at the lower pH of SBC compared to SC, sanitation of the SBC solution should be easier to maintain. NaOCl (200 µg/ml) added to SBC at pH 7.5 improved green mold control. Rinse water as high as 50 ml per fruit applied after SC did not reduce its effectiveness; however, high-pressure water cleaning after SC did. Conversely, high-pressure water cleaning of fruit before SC improved control of green mold. The risk of injury to fruit posed by SC treatment was determined by immersing oranges for 1 min in 3% (wt/vol) SC at 28, 33, 44, 50, 56, or 61°C (±1°C) and followed by storage for 3 weeks at 10°C. Rind injuries occurred only after treatment at 56 and 61°C. The risk of injury is low because these temperatures exceed that needed for control of green mold. SC was compatible with subsequent imazalil and biological control treatments.

Combination of Hot Water and Ethanol to Control Postharvest Decay of Peaches and Nectarines.

Spores of Monilinia fructicola or Rhizopus stolonifer were immersed in water or 10% ethanol (EtOH) for 1, 2, 4, or 8 min at temperatures of 46 or 50°C to determine exposure times that would produce 95% lethality (LT95). EtOH reduced the LT95 by about 90%. Peaches and nectarines infected with M. fructicola were immersed in hot water alone or with EtOH to control decay. EtOH significantly increased the control of brown rot compared to water alone. Immersion of fruit in water at 46 or 50°C for 2.5 min reduced the incidence of decayed fruit from 82.8% to 59.3 and 38.8%, respectively. Immersion of fruit in 10% ethanol at 46 or 50°C for 2.5 min further reduced decay to 33.8 and 24.5%, respectively. Decay after triforine (1,000 µg ml-1) treatment was 32.8%. Two treatments, 10% EtOH at 50°C for 2.5 min and 20% EtOH at 46°C for 1.25 min, were selected for extensive evaluation. The flesh of EtOH-treated fruit was significantly firmer, approximately 4.4 N force, than that of control fruit among seven of nine cultivars evaluated. No other factor evaluated was significantly influenced by heated EtOH treatments. The EtOH content of fruit treated with 10 or 20% EtOH was approximately 520 and 100 µg g-1 1 day and 14 days after treatment, respectively.

Reduction of Microbial Populations on Prunes by Vapor-Phase Hydrogen Peroxide †.

Vapor-phase hydrogen peroxide (VPHP) was used to disinfect prunes. Concentrated hydrogen peroxide solution (35%, wt/wt) was volatilized into a stream of dried air to approximately 3.1 mg/l (wt/vol) of hydrogen peroxide. Dried prunes obtained from commercial dehydrators were treated with VPHP and compared to untreated prunes. Microbial populations were determined for treatment comparisons. Untreated dried prune microbial populations were 155, 107, and 111 CFU/g of prunes on aerobic plate count agar, potato dextrose agar, and dichloran rose bengal agar, respectively. In contrast, VPHP-treated prune microbial populations were reduced to near zero on all media after 10 minutes of VPHP exposure. The color of prunes exposed for 20 min or longer, however, showed oxidation damage. No hydrogen peroxide residues were detected 90 days after treatment.