First Report of Alternaria alternata Causing Decay in Exported Sweet Basil During Freight from Israel to Europe.

Sweet basil (Ocimum basilicum) is an important spice herb grown in Israel for local markets and export. The crop is used as a fresh culinary herb or spice, and the essential oils are used in cosmetics and food flavorings. Due to increased demand, the production area of basil has increased in Israel. Postharvest losses due to fungal disease are a major economic concern for growers. In the summer of 2019, a leaf spot was observed in postharvest shipments of sweet basil destined for Europe; in late winter of 2022, leaf spots were observed on greenhouse-grown sweet basil. Fungal isolates from infected leaves were characterized by morphology in culture as Alternaria spp. PCR amplification of the Alternaria major allergen Alt a1, ITS, and gdp gene regions of the recovered isolates confirmed the presence of A. alternata, a common pathogen of numerous herbs and spice plants. In vitro growth tests demonstrated that 25°C was the optimum temperature for growth of the isolates. The isolates were tested for pathogenicity and found to infect a commonly grown cultivar of basil, cultivar Eli (previously cultivar Perrie). Foliar symptoms in pathogenicity tests were identical to those observed in commercial shipments and in the field, which completed Koch's postulates. Control of the nascent disease by applying fungicides to the plants may be necessary to reduce postharvest losses.

Chemo-Geographic Variations in Wild Population of Asteriscus graveolens in Israel Based on Volatile Composition Analyses.

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.

Agronomic and economic evaluation of Vetiver grass (Vetiveria zizanioides L.) as means for phytoremediation of diesel polluted soils in Israel.

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.

Chemical and morphological diversity in wild populations of Mentha longifolia in Israel.

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.

Rhizoctonia Web Blight-A New Disease on Mint in Israel.

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 (<24°C), BS-1 grew faster and was more aggressive at higher 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.

Antifungal Susceptibility Testing of Aspergillus niger on Silicon Microwells by Intensity-Based Reflectometric Interference Spectroscopy.

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.

Field Resistance to Potato Stem Colonization by the Black Dot Pathogen Colletotrichum coccodes.

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.

Rosemary-Whitefly Interaction: A Continuum of Repellency and Volatile Combinations.

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.

Disease Potential of Soil- and Tuberborne Inocula of Colletotrichum coccodes and Black Dot Severity on Potato.

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.

Resistance to Root Galling Caused by the Powdery Scab Pathogen Spongospora subterranea in Potato.

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, PO94A009-7 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.

Antimicrobial LDPE/EVOH Layered Films Containing Carvacrol Fabricated by Multiplication Extrusion.

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 °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.

Whitefly attraction to rosemary (Rosmarinus officinialis L.) is associated with volatile composition and quantity.

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 ß-caryophyllene and limonene, two compounds known to be associated with plant-insect interactions. The attraction of B. tabaci to pure (>95%) ß-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 ß-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.

Genetic Variability in the Potato Pathogen Colletotrichum coccodes as Determined by Amplified Fragment Length Polymorphism and Vegetative Compatibility Group Analyses.

ABSTRACT 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.

Variation in essential oil composition within individual leaves of sweet basil (Ocimum basilicum L.) is more affected by leaf position than by leaf age.

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.