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There is a demonstrated and paramount need for rapid, reliable infectious disease diagnostics, particularly those for invasive fungal infections. Current clinical determinations for an appropriate antifungal therapy can take up to 3 days using current antifungal susceptibility testing methods, a time-to-readout that can prove detrimental for immunocompromised patients and promote the spread of antifungal resistant pathogens. Herein, we demonstrate the application of intensity-based reflectometric interference spectroscopic measurements (termed iPRISM) on microstructured silicon sensors for use as a rapid, phenotypic antifungal susceptibility test. This diagnostic platform optically tracks morphological changes of fungi corresponding to conidia growth and hyphal colonization at a solid-liquid interface in real time. Using Aspergillus niger as a model fungal pathogen, we can determine the minimal inhibitory concentration of clinically relevant antifungals within 12 h. This assay allows for expedited detection of fungal growth and provides a label-free alternative to broth microdilution and agar diffusion methods, with the potential to be used for point-of-care diagnostics.

There is a demonstrated and paramount need for rapid, reliable infectious disease diagnostics, particularly those for invasive fungal infections. Current clinical determinations for an appropriate antifungal therapy can take up to 3 days using current antifungal susceptibility testing methods, a time-to-readout that can prove detrimental for immunocompromised patients and promote the spread of antifungal resistant pathogens. Herein, we demonstrate the application of intensity-based reflectometric interference spectroscopic measurements (termed iPRISM) on microstructured silicon sensors for use as a rapid, phenotypic antifungal susceptibility test. This diagnostic platform optically tracks morphological changes of fungi corresponding to conidia growth and hyphal colonization at a solid–liquid interface in real time. Using Aspergillus niger as a model fungal pathogen, we can determine the minimal inhibitory concentration of clinically relevant antifungals within 12 h. This assay allows for expedited detection of fungal growth and provides a label-free alternative to broth microdilution and agar diffusion methods, with the potential to be used for point-of-care diagnostics.

Asteriscus graveolens is an aromatic desert shrub which holds medicinal potential. This species belongs to the Asteraceae family and is endemic to the Mediterranean region. In the present study, wild plants were sampled from eleven locations throughout southern Israel and the volatile profiles from leaves and flowers were analyzed using GC/MS. Three methods for volatile sampling were tested for a representative population: solvent extraction (methyl tert-butyl ether), hydrodistillation of the essential oil and headspace solid-phase microextraction. In all methods, the majority of volatiles were characterized as oxygenated mono- and sesquiterpenes. Only solvent extraction was able to detect asteriscunolides that were previously reported as anticancer molecules. Hence, that method was chosen for further analyses. The leaves were dominated by three asteriscunolide isomers, cis-chrysanthenyl acetate and intermedeol. The flowers were dominated by bisabolone, 6-hydroxybisabol-2-en-1-one, cis-chrysanthenyl acetate, epi-α-cadinol, and germacrene-D. k-Means clustering analysis of these data divided the population into four clusters that significantly differ in their volatile composition as was further demonstrated by MANOVA analysis. Geographically, A. graveolens populations growing in Israel were found to be chemically diverse with unique varieties in the Dead Sea basin and the Arava region. This work demonstrates that chemo-geographic variation of volatile composition exists within A. graveolens population growing in Israel, so future research evaluating the medicinal potential of that plant should take this into consideration.

Soil pollution in Israel, due to diesel contamination, is a major concern, with gas stations, factories and refineries being the main polluters (>60%). Vetiver grass (Vetiveria zizanioides L.) is a perennial grass belonging to the Poaceae family, and is recognized world-wide for its potential as a plant with phytoremediation traits to contaminated soils. It is demonstrated here to decrease diesel contamination in field and court-yard trials. Chemical soil analysis indicated up to a 79% decrease (P < .05) in diesel pollution of contaminated soil planted with Vetiver; and at high soil contamination levels of 10 L/m2, a significant (P < .05) reduction of 96, 96 and 87% was recorded at soil depths of 0-20, 20-40 and 40-60 cm, respectively. Furthermore, in field plots contaminated with diesel and planted with Vetiver, weeds' biomass recovered to non-polluted levels following 8 to 9 months of Vetiver treatment. An economic evaluation conducted based on the cost-benefit analysis (CBA) principles, utilizing the Net Present Value (NPV) compared phytoremediation to other currently used decontamination procedures. The economic comparison showed that phytoremediation cleanup costs are lower and more beneficial to society at large, primarily from an ecosystem services perspective. Combining the results of the agronomic examination with the economic valuation, this research pointed out that phytoremediation with Vetiver has a non-negligible potential, making it a good solution for cleansing diesel from soils on a state-wide scale in Israel and worthy of further research and development.

Basil, better known as “the king of herbs,” is widespread today in almost every home garden and is a leading spice worldwide, used for culinary purposes both dried and fresh. It is a source of essential oils and other valuable antioxidant and phenolic compounds and a medicinal herb with a history of tales and lore. In India, basil is considered sacred; in Italy, it is a symbol of love; in ancient Greece, it is a symbol of hatred; and in Judaism, it is one of the scented spices used during the “Havdalah” service, the ceremony performed to mark the end of the holy Sabbath. In the following chapter, we will discuss the genus Ocimum and concentrate on the breeding efforts and approaches that are undertaken by many scientists and researchers worldwide that made the “king of herbs” into a global economy and profitable industry.

The increasing number of invasive fungal infections among immunocompromised patients and the emergence of antifungal resistant pathogens has resulted in the need for rapid and reliable antifungal susceptibility testing (AFST). Accelerating antifungal susceptibility testing allows for advanced treatment decisions and the reduction in future instances of antifungal resistance. In this work, we demonstrate the application of a silicon phase grating as sensor for the detection of growth ofAspergillus niger(A. niger) by intensity-based reflectometric interference spectroscopy and its use as an antifungal susceptibility test. The silicon gratings provide a solid-liquid interface to capture micron-sizedAspergillusconidia within microwell arrays. Fungal growth is optically tracked and detected by the reduction in the intensity of reflected light from the silicon grating. The growth ofA. nigerin the presence of various concentrations of the antifungal agents voriconazole and amphotericin B is investigated by intensity-based reflectometric interference spectroscopy and used for the determination of the minimal inhibitory concentrations (MIC), which are compared to standard broth microdilution testing. This assay allows for expedited detection of fungal growth and provides a label-free alternative to standard antifungal susceptibility testing methods, such as broth microdilution and agar diffusion methods.

Roughly, one-third of the food produced globally for human consumption is lost or wasted. These losses occur at all stages of the food value chain and across all types of food. Active packaging already plays a vital role in preventing wastage and further innovation is imperative to streamlining the food supply chain. Herein, we present an antimicrobial packaging based on polyamide (Nylon 6), containing a model essential oil (carvacrol). The volatile carvacrol molecules are encapsulated with Halloysite nanotubes (HNTs), which are naturally occurring aluminosilicate. The resulting polyamide films had an outstanding in vitro antifungal properties, with a broad spectrum of inhibitory activity against a wide range of fungal molds: Alternaria alternata, Botrytis cinerea, Penicillium digitatum, Penicillium expansum and Aspergillus niger. Furthermore, the active polyamide-based plastic bags were used for fresh produce packaging and their fungicidal and/or fungistatic effects on postharvest pathogens of cherry tomatoes, lychee and grapes were investigated. These in vivo experiments have resulted in reduced decay development and significantly extended shelf life. The presented technology holds a great potential for the development of custom-made active packaging for the food and postharvest industries, in a global effort to reduce food loss and waste.

Wild populations of Croatian clary sage (Salvia sclarea L.) were examined for variability to determine cultivation suitability in Croatia and Israel for breeding purposes. Phenotypic variability (coefficient of variation; %) was recorded for inflorescence weight (39.6%), inflorescence yield (52.8%), and essential oil yield (67.6%) when grown in Croatia. Associations were identified between inflorescence yield and essential oil yield (r = 0.9; P < 0.0001), inflorescence weight and inflorescence yield (r = 0.8; P < 0.0001), and inflorescence weight and inflorescence length (r = 0.6; P = 0.0056), suggesting that populations with elongated inflorescence are indirectly associated with higher essential oil yield. In Israel, the populations reached full bloom between the end of May and early June, corresponding on average to 397.5 days post planting. Linalyl acetate, linalool, α–terpineol, sclareol, and geranyl acetate were the leading essential oil components in both Croatia and Israel. The principal compounds in the oil were linalyl acetate (48.5%) and linalool (17.7%), signifying that the Croatian populations were of the linalool chemotype. A two-way ANOVA indicated an interaction between growing location (Croatia vs. Israel) and population for linalool (P = 0.02), α–terpineol (P = 0.007), and linalyl acetate (P = 0.09); evidence of an environmental effect on essential oil composition. The variation observed suggested that the wild population of clary sage in Croatia had the genetic heterogeneity essential for breeding. Nevertheless, the differences in essential oil composition between Croatia and Israel suggest that breeding efforts should be separately focused for each agriculture production system.

The sweet potato whitefly Bemisia tabaci Genn (Hemiptera: Aleyrodidae) has been recorded to differentially prefer rosemary (Rosmarinus officinalis) varieties in commercial fields in Israel. As chemical signaling is a significant component in plant-insect interaction, the present study examined the involvement of rosemary essential oil volatiles in this differential colonization to elucidate the rosemary-whitefly ecological interaction. Thirty-two rosemary varieties with different chemical profiles were used. The average whitefly preference was 25.1% with a significant variation of 51.4%, partitioning the sampled varieties into five preference groups, hence suggesting rosemary as a non-preferred host for the insect. All relations between preference and the major volatiles 1,8-cineole, camphor, linalool, verbenone, bornyl acetate and borneol were significantly (P ≤ 0.05) or notably (0.05 < P ≤ 0.09) negative (r < 1). Therefore, revealing that whitefly preference for rosemary is based on a continuum of repellency rather than attraction. 'Choice' bioassays with a range of the major volatile concentrations and gas chromatography-mass spectrometry (GC-MS) volatile blends (fractions) validated this observation. Principle component analysis of the entire chemical profile of two extreme varieties, representing high and low preferences, identified that approximately 43% of the volatiles in the essential oil were directly associated with repellency. Keeping in mind the remaining 57% of the compounds, this myriad of volatiles exhibit the ecological complexity of the rosemary-whitefly eco-system, explaining that whitefly preference to rosemary is repellency based.

This work describes the fabrication of antimicrobial multilayered polymeric films containing carvacrol (used as a model essential oil) by co-extrusion and multiplication technique. The microlayering process was utilized to produce films, with up to 65 alternating layers, of carvacrol-containing low-density polyethylene (LDPE) and ethylene vinyl alcohol copolymer (EVOH). Carvacrol was melt compounded with LDPE or loaded into halloysite nanotubes (HNTs) in a pre-compounding step prior film production. The detailed nanostructure and composition (in terms of carvacrol content) of the films were characterized and correlated to their barrier properties, carvacrol release rate, and antibacterial and antifungal activity. The resulting films exhibit high carvacrol content despite the harsh processing conditions (temperature of 200 &deg, C and long processing time), regardless of the number of layers or the presence of HNTs. The multilayered films exhibit superior oxygen transmission rates and carvacrol diffusivity values that are more than two orders of magnitude lower in comparison to single-layered carvacrol-containing films (i.e., LDPE/carvacrol and LDPE/(HNTs/carvacrol)) produced by conventional cast extrusion. The (LDPE/carvacrol)/EVOH and (LDPE/[HNTs/carvacrol])/EVOH films demonstrated excellent antimicrobial efficacy against E. coli and Alternaria alternata in in vitro micro-atmosphere assays and against A. alternata and Rhizopus in cherry tomatoes, used as the food model. The results presented here suggest that sensitive essential oils, such as carvacrol, can be incorporated into plastic polymers constructed of tailored multiple layers, without losing their antimicrobial efficacy.

The emergence of antibiotic resistance of pathogenic bacteria has led to renewed interest in exploring the potential of plant-derived antimicrobials e.g., essential oils (EOs), as an alternative strategy to reduce microbial contamination. However, the volatile nature of EOs presents a major challenge in their incorporation into polymers by conventional high-temperature processing techniques. Herein, we employ halloysite nanotubes (HNTs) as efficient nano-carriers for carvacrol (a model EO). This pre-compounding encapsulation step imparts enhanced thermal stability to the carvacrol, allowing for its subsequent melt compounding with low-density polyethylene (LDPE). The resulting polymer nanocomposites exhibit outstanding antimicrobial properties with a broad spectrum of inhibitory activity against Escherichia coli, Listeria innocua in biofilms, and Alternaria alternata. Their antimicrobial effectiveness is also successfully demonstrated in complex model food systems (soft cheese and bread). This superior activity, compared to other studied carvacrol containing films, is induced by the significantly higher carvacrol content in the film as well as its slower out-diffusion from the hybrid system. Thus, these new active polymer nanocomposites presents an immense potential in controlling microbial contamination and biofilm related adverse effects, rendering them as excellent candidate materials for a wide range of applications.

Whitefly (Bemisia tabaci) is an important insect pest, causing severe damage to agricultural crops. The pest was recorded in a commercial rosemary (Rosmarinus officinalis, Lamiaceae) field, colonizing rosemary variety (var.) '2', but not '11'. A series of field and controlled laboratory choice bioassays confirmed the observed phenomenon. Mature potted plants of the two varieties were randomly organized in a lemon verbena (Lippia citrodora) and lemon grass (Cymbopogon spp.) fields. Seven days later var. '2' was significantly more colonized by whiteflies than var. '11'. Under lab conditions, whiteflies were significantly more attracted to var. '2' plantlets than to var. '11' following choice bioassays. Furthermore, cotton plants dipped in an essential oil emulsion of var. '2' had significantly greater colonization than cotton plants dipped in the essential oil emulsion of var. '11'. Similar results were obtained in 'plant-plant', 'plant-no plant' as well as, 'essential oil-essential oil' choice bioassay designs. Analyses of the essential oils of the two varieties identified a set of common and unique volatiles in each variety. Among these volatiles were β-caryophyllene and limonene, two compounds known to be associated with plant-insect interactions. The attraction of B. tabaci to pure (>95%) β-caryophyllene and limonene using a range of concentrations was examined in vitro by choice bioassays. The compounds were attractive to the insect at moderate concentration, but not at the lowest or highest concentrations used, where the insect was not attracted or repelled, respectively. Limonene attracted the insects at rates that were 10-fold lower than β-caryophyllene. The results emphasized the role of host plant volatiles in shaping the structure of B. tabaci populations in nature and in agricultural systems, and provided insights into the factors that contribute to the development of insect populations with unique characteristics. The results could also serve for future development of bio-pesticides and in breeding programs.

Active antimicrobial packaging is a promising form of active packaging that can kill or inhibit microorganism growth in order to maintain product quality and safety. One of the most common approaches is based on the release of volatile antimicrobial agents from the packaging material such as essential oils. Due to their highly volatile nature, the challenge is to preserve the essential oils during the high-temperature melt processing of the polymer, while maintaining high antimicrobial activity for a desired shelf life. This study suggests a new approach in order to achieve this goal. Antimicrobial active films are developed based on low-density polyethylene (LDPE), organo-modified montmorillonite clays (MMT) and carvacrol (used as an essential oil model). In order to minimize carvacrol loss throughout the polymer compounding, a pre-compounding step is developed in which clay/carvacrol hybrids are produced. The hybrids exhibit a significant increase in the d-spacing of clay and enhanced thermal stability. The resulting LDPE/(clay/carvacrol) films exhibit superior and prolonged antibacterial activity against Escherichia coli and Listeria innocua, while polymer compounded with pure carvacrol loses the antibacterial properties within days. The films also present an excellent antifungal activity against Alternaria alternata, used as a model plant pathogenic fungus. Furthermore, infrared spectroscopy analysis of the LDPE/(clay/carvacrol) system displayed significantly higher carvacrol content in the film as well as a slower out-diffusion of the carvacrol molecules in comparison to LDPE/carvacrol films. Thus, these new films have a high potential for antimicrobial food packaging applications due to their long-lasting and broad-spectrum antimicrobial efficacy. Copyright © 2014 John Wiley & Sons, Ltd.

Rosemary is an important crop for industrial production of essential oil. However, there is a significant effect of the genetic variation and the production process on the essential oil composition. Therefore, the present study was aimed at elucidating the effect of season and processing method on essential oil composition using the volatile structure of rosemary germplasm that was cultivated at the Newe Ya'ar research center for 30 years. Analysis of the essential oil composition of thirty-three germplasm identified four chemical groups: a '1,8-cineole' type, a 'verbenone' type, a 'camphor' type, and a 'α-pinene' type. Application of fractional factorial analysis of variance to the data indicated interactions of volatile by germplasm, volatile by method (hydro-distillation or solvent extraction), and volatile by season (summer 2017 or winter 2018), but not for germplasm by method, germplasm by season or method by season. This suggests that the germplasm was seasonally stable. Yet, the interaction between volatile by method pointed out that more camphor and 1,8-cineole were obtained by distillation, whereas more α-pinene was obtained by solvent extraction. Using four germplasm that were selected as chemo-type representatives for comparing hydro-distillation by Clevenger modified apparatus and steam-distillation, α-pinene was identified as a minor component in the hydrosol, whereas 1,8-cineole, camphor, borneol and verbenone were major components in the hydrosol. The outcome of the study reflects the importance of evaluating the effects of environment, genetic and processing methods on essential oil composition for accurate industrial breeding and production of essential oils.

Populations of Mentha longifolia, an endangered species in Israel, were tested for essential oil composition and conservational ability. In 2002-2003, 25 wild populations country-wide were tested, indicating population divergence into two chemotypes. Chemotype A was characterized by high levels of menthone and pulegone, and chemotype B by high levels of piperitenone oxide and piperitone oxide. Chemotype A was more abundant (22 of 25 populations) than chemotype B (11 of 25 populations). However, a chemotype/population interaction was not recorded (P > 0.05). In spring 2003, seven of the 25 wild populations were resampled, propagated, and cultivated at the Newe Ya'ar campus. Then, in 2004, the propagated plants were tested for essential oil composition. The propagated plants maintained the essential oil composition as well as the chemotype-frequency distribution of the original wild population from which they were obtained. Since a chemotype/population interaction was not recorded, and the cultivated plants displayed the wild population essential oil composition, it can be concluded that i) the chemotype diversity is genetically based, and ii) the M. longifolia populations sampled can be horticulturally conserved.

Mentha longifolia is produced in and exported from Israel with annual revenue of US$16 million. In 2010, a severe epidemic of unknown etiology reduced growers' returns up to 50%. Disease symptoms included water-soaked lesions, necrosis, and web-like mycelia on plants. Two isolates (JV-1 and BS-1) from randomly selected symptomatic plants were identified as members of Rhizoctonia solani anastomosis groups 1-IB and 4HG-I, respectively. The fitness of JV-1 and BS-1 interacted with temperatures between 17 and 35°C: JV-1 grew faster and was more aggressive (P < 0.05) at lower temperatures (30°C), and the two isolates performed similarly at intermediate temperatures. Disease developed fastest at 24 to 28°C. Yield was reduced between 46 and 100%. In all, 77.5% fewer plants recovered from disease developing at 24 to 28°C than at 17 to 23 or 30 to 35°C. The relationship of disease to relative humidity (RH) fit a quadratic model (P < 0.0015, R2 = 0.98). Disease was most severe at 100% RH, decreasing by 1.3-, 1.9-, 3-, 4.5-, and 10.5-fold with the reduction of RH from 100% to 88, 76, 69, 55, or 49%, respectively. This is the first report of Rhizoctonia web blight in mint in Israel.

The aroma in sweet basil is a factor affecting the commercial value of the crop. In previous studies leaf age was considered to be a factor that influences the composition of essential oil (EO). In this study it was hypothesized that a single observation of the EO content in leaves from different positions on the main stem (young vs old) could predict the developmental changes in the plant during its life cycle. Plants harvested at week 16 demonstrated an exponential increase (R(2) = 0.92) in EO concentration in leaves on the main stem and lateral shoots, indicating higher EO concentrations in younger than in older leaves. Eugenol and methyleugenol predominated (28-77%) in the extract. Eugenol levels were higher in younger leaves (∼53%), and methyl-eugenol levels predominated in older leaves (∼68%). Linalool was lower in mature leaves than in younger leaves. This suggested that eugenol converted into methyleugenol and linalool decreased as leaf mature. However, in weekly monitored plants, the levels of these compounds in the EO had limited variation in the maturing leaf regardless of its position on the stem. This proposed that the EO composition in an individual leaf is mostly affected by the leaf position on the stem and not by its maturation process. Because leaf position is related to plant development, it is probable that the plant's physiological age at the time of leaf formation from the primordial tissue is the factor affecting the EO composition. It was concluded that interpretation of scientific observations should be carried out with caution and that hypotheses should be tested utilizing multifaceted approaches.

Black dot is caused by the fungus Colletotrichum coccodes (Wallr.) S.J. Hughes. The disease is prevalent in potato fields and can be of economic concern by itself, or as a part of the potato early dying syndrome. Little is known about resistance to this disease. In the present study resistance to potato black dot was tested in Solanum tuberosum Group Andigena. Forty accessions were chosen randomly from the core collection and screened. The accessions originated from Peru, Mexico, Bolivia, Columbia, Ecuador, Argentina and Costa Rica. Five accessions had less (P ≤ 0.05) disease on roots and stems than a set of commercial standards. These accessions were: PIs 189473, 230475, 161683, 243367 and 230470. They were retested to validate their partial resistance. Fifteen plants, each a unique genotype, were selected from each accession and were propagated into multiple clones that were inoculated with the pathogen. Three genotypes were identified with less (P < 0.05) disease on roots and stems than the industry standard. These genotypes originated from the accessions PI-243367 and PI-230475. Clones of the resistant genotypes were added to the potato germplasm collection located at the Potato Improvement Laboratory of the USDA-ARS near Prosser, WA, to be used in the development of black dot resistant commercial varieties.

Spongospora subteranea, the causal agent of potato powdery scab is becoming increasingly important worldwide. Little is known about the genetic basis of resistance to this disease. The present study tested the hypothesis that potato genotypes with stable genetic resistance to “Spongospora root galling” were present in potato germplasm. Root galling index values of 24 genotypes screened for resistance in four field trials (environments) in 2004 and 2005 in Washington State and Idaho were analyzed. Genotypes tested included five resistant, four industry standards and advanced selections from the USDA-ARS, Prosser, WA program. Broad-sense heritability was calculated as 0.76 with a 95% confidence interval of 0.55–0.89, indicating a fairly high genetic component of the trait. Of the 24 genotypes that were tested, eight showed no genotype*environment interactions while six of the remainder had significant variance (i.e., they were unstable) after removal of genotype*environment variance. Among the five resistant genotypes, PA95B2-4 was stable, and PA98N5-2, PA98NM38-1, PO94A009-7 and POR00HG5-1 were stable after the removal of environmental heterogeneity. Among the four industry standards, Shepody was unstable, whereas Ranger Russet, Russet Burbank and Umatilla Russet were stable after the removal of genotype*environment variance. Stable resistance to “Spongospora root galling” was identified. A large portion of the variation was genetic, which will enable breeders to use resistant and stable potato genotypes as parents in future breeding to develop superior commercial potato cultivars with resistance to “Spongospora root galling”.

Potato (Solanum tuberosum) germplasm was tested for resistance to stem colonization by the black dot pathogen Colletotrichum coccodes. Forty-six potato selections were tested in three field trials from 2006 to 2008. Resistance was determined by comparing disease severity on aboveground stems to the mean disease severity of the industry standards Russet Burbank, Ranger Russet, and Umatilla Russet. The potato selections were also tested for genotype*environment interaction to determine their genetic stability. Heritability of resistance was calculated to be 0.13 with confidence intervals between 0.00 and 0.68. The selections A0012-5, PA95B2-4, PA98NM38-1, and PO94A009-7 had less black dot than the standards in all years, and also demonstrated genetic stability. These selections also possess resistance to the root galling stage of the powdery scab pathogen Spongospora subterranea f. sp. subterranea. PA95B2-4, PA98NM38-1, and PO94009-7 were derived from an introgression program to incorporate resistance to the Columbia root-knot nematode Meloidogyne chitwoodi from the Mexican wild species Solanum bulbocastanum, and also have the commercial cultivar Summit Russet in their ancestry. These selections are promising steps toward sustainable management of black dot and powdery scab and will be further tested and used for breeding purposes.

Root galls possibly caused by Spongospora subterranea were observed on hairy nightshade (Solanum sarrachoide; HNS). HNS galls and galls from potato were used to artificially inoculate potato and HNS. Eighty-three and 52% potato and HNS plants inoculated with potato inoculum had root galls, respectively. Ten and 31% potato and HNS plants inoculated with HNS inoculum had root galls, respectively. A S. subterranea-specific PCR significantly correlated (P < 0.0001, Phi Coefficient = 0.69) with visual assessment of gall incidence in 34 of 40 samples. Five of 40 samples had no galls, but gave positive PCRs. One of 40 samples had root galls, but gave a negative PCR. More sporosori were produced on potato than on HNS. Therefore, the root galls on HNS were caused by S. subterranea that is able to complete its life cycle on HNS and produce new generations of sporosori that are infectious on potato.

Nitzan, N., Cummings, T. F., Johnson, D. A., Miller, J. S., Batchelor, D. L., Olsen, C., Quick, R. A., and Brown, C. R. 2008. Resistance to root galling caused by the powdery scab pathogen Spongospora subterranea in potato. Plant Dis. 92:1643-1649. Potato (Solanum tuberosum) selections (clones and commercial cultivars) were examined for resistance to root galling, caused by the powdery scab pathogen Spongospora subterranea f. sp. subterranea in seven field trials conducted between 2003 and 2007 in the states of Washington and Idaho. Four industry reference cultivars—Shepody, Russet Burbank, Russet Ranger, and Umatilla Russet—were used as susceptible standards. Every year, selections less susceptible than the standards were considered resistant and progressed to the next season. Selections that did not demonstrate resistance in at least two consecutive trials were discarded. Eight potato selections were more resistant to root galling than the susceptible standards in two or more trials: PA98NM38-1 was more resistant than the susceptible standards in 5 of 5 trials, PO94A009-10 in 4 of 5 trials, PA95B2-4 and PA98N5-2 in 3 of 5 trials, POR00HG5-1 in 2 of 5 trials, PO94A0097 in 3 of 4 trials, PO94A012-2 in 2 of 3 trials, and Summit Russet in 2 of 2 trials. POR00HG5-1 has Solanum hougasii in its ancestry, while the other selections have the Mexican wild species Solanum bulbocastanum and the commercial cultivar Summit Russet appearing in their ancestry. Summit Russet is the most plausible source of resistance.

Inoculum of Colletotrichum coccodes, the cause of potato black dot, is soil- or tuberborne. Understanding the disease potential of sources of inocula is crucial for developing disease management strategies and resistance screening techniques. Two hypotheses were tested in this study: (i) soilborne inoculum causes more disease than tuberborne inoculum and (ii) black dot severity is related to the concentration of soilborne inoculum. Trials were conducted in the greenhouse with standardized inoculum. Plants grown in infested soil had more sclerotia on roots than plants grown from infected tubers in three of four trials. In general, plants grown in infested soil produced fewer tubers and lower yields than the noninoculated plants. Plants grown from infected tubers produced similar numbers of tubers as the noninoculated plants in all trials, and had reduced yields in one of the four trials. Increasing concentrations of soilborne inoculum had a nonlinear association with disease development. Foliar symptoms, sclerotial density on roots, and sclerotial development on stems did not increase when soil inoculum exceeded 0.5 or 1.7 g/liters of soil. In this study, soilborne inoculum caused more disease than tuberborne inoculum and disease severity remained constant above a threshold of soilborne inoculum.

The rotation crops wheat, barley, oat, maize, soybean, rye, yellow mustard, alfalfa, and spring canola and weeds eastern black nightshade, velvetleaf, timothy grass, orchard grass, and Giant foxtail common to potato-growing areas in North America were used to study the host range ofColletotrichum coccodes, the causal agent of potato black dot. The fungus was isolated from nine of 14 rotation crops and weeds that were inoculated: yellow mustard, soybean, spring canola, alfalfa, oat, eastern black nightshade, velvetleaf, giant foxtail, and timothy grass. In all, colonization was highest in black nightshades (87%) and velvetleaf (80%). Among the rotation crops, colonization was highest on yellow mustard (59%) followed by spring canola (33%) and soybean (30%).Colletotrichum coccodes was not isolated from wheat, barley, rye, maize, or orchard grass. The results indicated that crops used for rotation with potato should be selected carefully to prevent the increase ofC. coccodes inoculum in the soil and that weeds may help maintain viable inoculum ofC. coccodes in the absence of potato. Based on these results we recommend that wheat, barley, maize, or rye be used in rotation with potato in areas whereC. coccodes is present in high levels in the soil.

Heilmann, L. J., Nitzan, N., Johnson, D. A., Pasche, J. S., Doetkott, C., and Gudmestad, N. C. 2006. Genetic variability in the potato pathogen Colletotrichum coccodes as determined by amplified fragment length polymorphism and vegetative compatibility group analyses. Phytopathology 96:1097-1107. Amplified fragment length polymorphism (AFLP) using three primer sets was used to characterize 211 Colletotrichum coccodes isolates from North America, 112 of which were assigned to six vegetative compatibility groups (VCGs) using nitrate nonutilizing (nit) mutants. These isolates clustered into five corresponding groups by unweighted pairgroup method with arithmetic means-based cluster analysis of AFLP banding patterns. Isolates of C. coccodes belonging to NA-VCG1 and NA-VCG3 were closely related, as were isolates belonging to NA-VCG2 and NA-VCG5. Based on bootstrap analysis of AFLP data, the two isolates originally assigned to NA-VCG4 clustered with isolates belonging to NA-VCG2 and NA-VCG5. C. coccodes isolates that clustered with two isolates belonging to NA-VCG6 were the most diverged from other groups, including seven isolates collected from hosts other than potato. As opposed to the bootstrap analysis, a quadratic discriminant analysis (QDA) of AFLP data correctly categorized the two isolates of NA-VCG4. Furthermore, in isolates where VCG determinations had been made, this model correctly classified isolates of all VCGs. QDA classifications were identical to those made by the bootstrap analysis, with the exception of VCG4. Overall, classifications made by the QDA model were strongly correlated (r = 0.970, P < 0.001) to the VCGs assigned by traditional methods. All 99 C. coccodes isolates evaluated only by AFLP also were subjected to QDA, leading to the assignment of a presumptive VCG for each isolate. No isolates of VCG4 or VCG6 were identified by QDA within this population. Symptoms of black dot developed in plants inoculated with isolates collected from both potato and non-potato hosts. However, total yield was not significantly reduced by infection with non-potato isolates. The lack of any additional groups identified by AFLP analysis may be an indicator of a limited level of genetic variation among North American C. coccodes isolates. AFLP is a much more efficient technique for subspecific characterization in C. coccodes than VCG analysis utilizing nit mutants and will provide an effective means by which the population biology of this pathogen can be further investigated worldwide.

The radial growth rate and sclerotial density ofColletotrichum coccodes isolates representing four major vegetative compatibility groups (VCGs) were determined at three temperatures (21,25, and 30 C) and three pH levels (pH 5, 6, and 7). This is the first report indicating thatC. coccodes isolates characterized to the same VCG share common physiological traits. The optimal temperature for growth of isolates from all four VCGs was 25 C, except for VCG1 isolates, for which growth at 21 and 25 C was equivalent. The growth rate of all isolates, of all VCG groups was decreased at 30 C, whereas sclerotia development was enhanced. The optimal pH level for growth of all isolates was pH 6 and pH 7. Interactions between VCGs and temperature or pH were observed in relation to radial growth rate and sclerotia density. At 25 C, isolates from VCG1 and 2 demonstrated the most rapid growth rate. At 21 and 30 C, VCG1 isolates demonstrated the fastest growth compared with all other VCGs. In response to pH, VCG1 and 2 demonstrated the fastest growth rates at all pH levels. VCG3 and 4 demonstrated the highest sclerotial density at 25 C and at pH 6; and VCG2 and 4 demonstrated the highest sclerotial density at 30 C.