Discovery of the REN11 Locus From Vitis aestivalis for Stable Resistance to Grapevine Powdery Mildew in a Family Segregating for Several Unstable and Tissue-Specific Quantitative Resistance Loci.

Race-specific resistance loci, whether having qualitative or quantitative effects, present plant-breeding challenges for phenotypic selection and deciding which loci to select or stack with other resistance loci for improved durability. Previously, resistance to grapevine powdery mildew (GPM, caused by Erysiphe necator) was predicted to be conferred by at least three race-specific loci in the mapping family B37-28 × C56-11 segregating for GPM resistance from Vitis aestivalis. In this study, 9 years of vineyard GPM disease severity ratings plus a greenhouse and laboratory assays were genetically mapped, using a rhAmpSeq core genome marker platform with 2,000 local haplotype markers. A new qualitative resistance locus, named REN11, on the chromosome (Chr) 15 was found to be effective in nearly all (11 of 12) vineyard environments on leaves, rachis, berries, and most of the time (7 of 12) stems. REN11 was independently validated in a pseudo-testcross with the grandparent source of resistance, "Tamiami." Five other loci significantly predicted GPM severity on leaves in only one or two environments, which could indicate race-specific resistance or their roles in different timepoints in epidemic progress. Loci on Chr 8 and 9 reproducibly predicted disease severity on stems but not on other tissues and had additive effects with REN11 on the stems. The rhAmpSeq local haplotype sequences published in this study for REN11 and Chr 8 and 9 stem quantitative trait locus (QTL) can be used directly for marker-assisted selection or converted to SNP assays. In screening for REN11 in a diversity panel of 20,651 vines representing the diversity of Vitis, this rhAmpSeq haplotype had a false positive rate of 0.034% or less. The effects of the other foliar resistance loci detected in this study seem too unstable for genetic improvement regardless of quantitative effect size, whether due to race specificity or other environmental variables.

Survival of foodborne pathogens on commercially packed table grapes under simulated refrigerated transit conditions.

We examined the survival of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica Thompson inoculated on commercially packed table grapes under simulated refrigerated transit conditions (1.1 ± 0.5 °C; 90% RH). Grapes were placed in perforated polyethylene cluster bags, within a commercial expanded polystyrene box equipped with either a SO2-generating pad; a perforated polyethylene box liner; a SO2-generating pad and a box liner; or none of them. L. monocytogenes was most sensitive to SO2-generating pad. SO2-generating pad or SO2-generating pad with box liner inactivated this pathogen completely on day 12 following the inoculation. S. enterica Thompson displayed a similar cold sensitivity as L. monocytogenes, but was more resistant to SO2-generating pad than L. monocytogenes. While SO2-generating pad eliminated S. enterica Thompson on day 20, a combination of box liner with SO2-generating pad inactivated this pathogen completely on day 13. E. coli O157:H7 had the highest tolerance to transit temperature and to SO2-generating pad; SO2-generating pad inactivated this pathogen completely on Day 20. Our data suggest that use of SO2-generating pad combined with box liner is effective in reducing foodborne pathogens L. monocytogenes and S. enterica Thompson, while the use of SO2-generating pad alone was more effective on E. coli O157:H7.

Effect of sulfur dioxide fumigation on survival of foodborne pathogens on table grapes under standard storage temperature.

We examined the fate of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica Thompson inoculated on freshly-harvested table grapes under standard cold storage with initial and weekly sulfur dioxide (SO2) fumigation. L. monocytogenes and S. enterica Thompson were much more sensitive to cold temperature than E. coli O157:H7. Furthermore, L. monocytogenes was highly susceptible to SO2. Initial fumigation with 100 or 200 ppm-hr was sufficient to eliminate this pathogen on grapes with low (10(4) cells/grape) and high (10(6) cells/grape) inocula, respectively. Initial fumigation with 300 ppm-hr reduced S. enterica Thompson population about 300- and 10-fold on grapes with low and high inocula, respectively. Initial fumigation with 300 ppm-hr reduced E. coli O157:H7 population to less than 10-fold, regardless of inoculum density. When grapes were inoculated with the high inoculum and fumigated on days 0 and 7 with 200 or 300 ppm-hr SO2, S. enterica Thompson and E. coli O157:H7 were completely inactivated between days 8 and 14 of cold storage. Standard cold storage combined with SO2 fumigation was effective in reducing and eliminating all three pathogens on table grapes, however, depending on the dose, two or three fumigations were needed for elimination of S. enterica Thompson and E. coli O157:H7.

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.

A single dominant locus, ren4, confers rapid non-race-specific resistance to grapevine powdery mildew.

In the present study we screened the progeny of Vitis vinifera × V. romanetii populations segregating for resistance to powdery mildew and determined the presence of a single, dominant locus, Ren4, conferring rapid and extreme resistance to the grapevine powdery mildew fungus Erysiphe necator. In each of nine Ren4 pseudo-backcross 2 (pBC(2)) and pBC(3) populations (1,030 progeny), resistance fit a 1:1 segregation ratio and overall segregated as 543 resistant progeny to 487 susceptible. In full-sib progeny, microscopic observations revealed the reduction of penetration success rate (as indicated by the emergence of secondary hyphae) from 86% in susceptible progeny to below 10% in resistant progeny. Similarly, extreme differences were seen macroscopically. Ratings for Ren4 pBC(2) population 03-3004 screened using natural infection in a California vineyard and greenhouse and using artificial inoculation of an aggressive New York isolate were fully consistent among all three pathogen sources and environments. From 2006 to 2010, Ren4 pBC(2) and pBC(3) vines were continuously screened in California and New York (in the center of diversity for E. necator), and no sporulating colonies were observed. For population 03-3004, severity ratings on leaves, shoots, berries, and rachises were highly correlated (R(2) = 0.875 to 0.996) in the vineyard. Together, these data document a powdery mildew resistance mechanism not previously described in the Vitaceae or elsewhere, in which a dominantly inherited resistance prevents hyphal emergence and is non-race-specific and tissue-independent. In addition to its role in breeding for durable resistance, Ren4 may provide mechanistic insights into the early events that enable powdery mildew infection.

Effect of chitosan dissolved in different acids on its ability to control postharvest gray mold of table grape.

Chitosan is a natural biopolymer that must be dissolved in an acid solution to activate its antimicrobial and eliciting properties. Among 15 acids tested, chitosan dissolved in 1% solutions of acetic, L-ascorbic, formic, L-glutamic, hydrochloric, lactic, maleic, malic, phosphorous, and succinic acid. To control gray mold, table grape berries were immersed for 10 s in these chitosan solutions that had been adjusted to pH 5.6. The reduction in decay among single berries of several cultivars (Thompson Seedless, Autumn Seedless, and grape selection B36-55) inoculated with Botrytis cinerea at 1 x 10(5) conidia/ml before or after immersion in chitosan acetate or formate, followed by storage at 15 degrees C for 10 days, was approximately 70%. The acids alone at pH 5.6 did not control gray mold. Decay among clusters of two cultivars (Thompson Seedless and Crimson Seedless) inoculated before treatment was reduced approximately 60% after immersion in chitosan lactate or chitosan acetate followed by storage for 60 days at 0.5 degrees C. The viscosity of solutions was 1.9 centipoises (cp) (ascorbate) to 306.4 cp (maleicate) and the thickness of chitosan coating on berries was 4.4 microm (acetate) to 15.4 microm (ascorbate), neither of which was correlated with solution effectiveness. Chitosan acetate was the most effective treatment which effectively reduced gray mold at cold and ambient storage temperatures, decreased CO2 and O2 exchange, and did not injure the grape berries.

Near-Harvest Applications of Metschnikowia fructicola, Ethanol, and Sodium Bicarbonate to Control Postharvest Diseases of Grape in Central California.

The yeast Metschnikowia fructicola, ethanol, and sodium bicarbonate (SBC), alone or in combinations, were applied to table grapes on vines 24 h before harvest to control the incidence of postharvest diseases. In four experiments, all significantly reduced the total number of decayed berries caused by Botrytis cinerea, Alternaria spp., or Aspergillus niger after storage for 30 days at 1°C followed by 2 days at 20°C. In three experiments, a mean gray mold incidence (caused by B. cinerea) of 34.2 infected berries per kilogram among untreated grape was reduced by Metschnikowia fructicola at 2 × 107 CFU/ml, ethanol at 50% (vol/vol), or SBC at 2% (wt/vol) to 12.9, 8.1, or 10.6 infected berries per kilogram, respectively. Ethanol, SBC, and SO2 generator pads were similarly effective. M. fructicola effectiveness was not improved when combined with ethanol or SBC treatments. Ethanol and yeast treatments did not harm the appearance of the grapes. M. fructicola and SBC left noticeable residues, and SBC caused some visible phytotoxicity to the rachis and berries. Ethanol applied at 50% (vol/vol) reduced epiphytic fungal and bacterial populations by about 50% compared with controls. M. fructicola populations persisted on berries during storage when applied alone or after ethanol treatments, whereas SBC reduced its population significantly.

Correlations of Morphological, Anatomical, and Chemical Features of Grape Berries with Resistance to Botrytis cinerea.

ABSTRACT Resistance of mature berries of grapevine cultivars and selections to postharvest infection by Botrytis cinerea was assessed. Little or no resistance existed in most popular table grape Vitis vinifera cultivars, except in moderately resistant 'Emperor' and 'Autumn Black'. Highly resistant grapes were V. rotundifolia, V. labrusca, or other complex hybrids. Morphological, anatomical, and chemical characteristics of 42 genetically diverse cultivars and selections with various levels of resistance to B. cinerea were examined to determine which features were associated with resistance. We quantified the (i) density of berries within a cluster; (ii) number of pores and lenticels on the berry surface; (iii) thickness and number of cell layers in the epidermis and external hypodermis; (iv) amount of cuticle and wax; (v) berry skin protein content; (vi) total phenolic content of the skin before and after B. cinerea inoculation; and (vii) catechin and trans- and cis-resveratrol contents of the skin before and after inoculation. The number of pores was negatively correlated with resistance. Highly resistant cultivars had few or no pores in the berry surface. The number and thickness of epidermal and hypodermal cell layers and cuticle and wax contents were positively correlated with resistance. Other characteristics evaluated were not associated with resistance. trans-Resveratrol and cis-resveratrol were induced by B. cinerea inoculation only in sensitive and moderately resistant cultivars and selections.