Alternative products to control late season diseases in soybeans / Produtos alternativos para o controle de doenças de final de ciclo na soja

In the last crop seasons, the complex of late season diseases (CLSD) of soybean (Glycine max L. (Merrill)), has been causing considerable reductions in the crop yield. Currently, there are no cultivars resistant to all pathogens that causes CLSD. The present study evaluated the effect of applying the acibenzolar-S-methyl resistance inducer, alternative products and fungicide on the severity of CLSD in the soybean cultivar BMX Potência RR during the 2013/2014 and 2014/2015 crops, in the field. The treatments for the experiments were: 1 - control (water); 2 - acibenzolar-S-methyl; 3 - calcium; 4 - micronutrients: copper, manganese and zinc; 5 - micronutrients: manganese, zinc and molybdenum; 6 - nitrogen-potassium fertilizer; 7 - Ascophyllum nodosum and 8 - azoxystrobin + cyproconazole with the addition of the adjuvant. Four applications of alternative products and two of fungicide were carried out in both harvests. A diagrammatic scale assessed the severity of CLSD at the phenological stage R7.1. The acibenzolar-S-methyl resistance inducer, alternative products (macro and micronutrients) and A. nodosum had no effect on the severity of CLSD in the two harvests. The fungicide (azoxystrobin + cyproconazole) reduced the severity of CLSD and prevented damage to productivity in both experiments.

Nas últimas safras, o complexo de doenças de final de ciclo (DFC) da soja (Glycine max L. (Merrill)), vem provocando reduções consideráveis no rendimento da cultura. Atualmente, não há variedades resistentes a todos os patógenos causadores das DFC. O objetivo do presente trabalho foi avaliar o efeito da aplicação do indutor de resistência acibenzolar-S-methyl, produtos alternativos e fungicida na severidade de DFC na cultivar de soja BMX Potência RR durante as safras 2013/2014 e 2014/2015, em campo. Os tratamentos para os experimentos foram: 1 - testemunha (água); 2 - acibenzolar-S-methyl; 3 - cálcio; 4 - micronutrientes: cobre, manganês e zinco; 5 - micronutrientes: manganês, zinco e molibdênio; 6 - adubo NK; 7 - Ascophyllum nodosum e 8 - azoxistrobina + ciproconazol com adição do adjuvante. Foram realizadas quatro aplicações dos produtos alternativos e duas do fungicida, nas duas safras. A severidade de DFC foi avaliada por escala diagramática no estádio fenológico R7.1. O indutor de resistência acibenzolar-S-methyl, os produtos alternativos (macro e micronutrientes) e A. nodosum não apresentaram efeito sobre a severidade das DFC nas duas safras. O fungicida (azoxistrobina + ciproconazol) reduziu a severidade das DFC e evitou danos a produtividade nos dois experimentos.

Oregano essential oil vapour prevents Plasmopara viticola infection in grapevine (Vitis Vinifera) and primes plant immunity mechanisms.

The reduction of synthetic fungicides in agriculture is necessary to guarantee a sustainable production that protects the environment and consumers' health. Downy mildew caused by the oomycete Plasmopara viticola is the major pathogen in viticulture worldwide and responsible for up to 60% of pesticide treatments. Alternatives to reduce fungicides are thus utterly needed to ensure sustainable vineyard-ecosystems, consumer health and public acceptance. Essential oils (EOs) are amongst the most promising natural plant protection alternatives and have shown their antibacterial, antiviral and antifungal properties on several agricultural crops. However, the efficiency of EOs highly depends on timing, application method and the molecular interactions between the host, the pathogen and EO. Despite proven EO efficiency, the underlying processes are still not understood and remain a black box. The objectives of the present study were: a) to evaluate whether a continuous fumigation of a particular EO can control downy mildew in order to circumvent the drawbacks of direct application, b) to decipher molecular mechanisms that could be triggered in the host and the pathogen by EO application and c) to try to differentiate whether essential oils directly repress the oomycete or act as plant resistance primers. To achieve this a custom-made climatic chamber was constructed that enabled a continuous fumigation of potted vines with different EOs during long-term experiments. The grapevine (Vitis vinifera) cv Chasselas was chosen in reason of its high susceptibility to Plasmopara viticola. Grapevine cuttings were infected with P. viticola and subsequently exposed to continuous fumigation of different EOs at different concentrations, during 2 application time spans (24 hours and 10 days). Experiments were stopped when infection symptoms were clearly observed on the leaves of the control plants. Plant physiology (photosynthesis and growth rate parameters) were recorded and leaves were sampled at different time points for subsequent RNA extraction and transcriptomics analysis. Strikingly, the Oregano vulgare EO vapour treatment during 24h post-infection proved to be sufficient to reduce downy mildew development by 95%. Total RNA was extracted from leaves of 24h and 10d treatments and used for whole transcriptome shotgun sequencing (RNA-seq). Sequenced reads were then mapped onto the V. vinifera and P. viticola genomes. Less than 1% of reads could be mapped onto the P. viticola genome from treated samples, whereas up to 30% reads from the controls mapped onto the P. viticola genome, thereby confirming the visual observation of P. viticola absence in the treated plants. On average, 80% of reads could be mapped onto the V. vinifera genome for differential expression analysis, which yielded 4800 modulated genes. Transcriptomic data clearly showed that the treatment triggered the plant's innate immune system with genes involved in salicylic, jasmonic acid and ethylene synthesis and signaling, activating Pathogenesis-Related-proteins as well as phytoalexin synthesis. These results elucidate EO-host-pathogen interactions for the first time and indicate that the antifungal efficiency of EO is mainly due to the triggering of resistance pathways inside the host plants. This is of major importance for the production and research on biopesticides, plant stimulation products and for resistance-breeding strategies.

Fungicide-driven alterations in azole-resistant Aspergillus fumigatus are related to vegetable crops in Colombia, South America.

Aspergillus fumigatus resistant to azole as first-line therapy has been reported in azole-naïve patients. This worldwide resistance phenomenon has been linked to fungicide-driven alterations in the cyp51A gene and its promoter region (such as TR34/L98H and TR46/Y121F/T289A). Azole-resistant A. fumigatus related to the use of triazole fungicides in flower fields was recently reported In Colombia. The purpose of this study was to investigate the presence of azole-resistant A. fumigatus in soil samples from vegetable crops such as carrots, potatoes, maize, strawberries, and pea, and from prepared farming land surrounding the city of Bogotá. Species identification was based on sequencing of the ß-tubulin and calmodulin genes. All A. fumigatus strains were screened for azole resistance on agar supplemented with itraconazole or voriconazole. Among the 60 soil samples, 34 (56.6%) were positive for A. fumigatus and 15 samples exhibited strains (n = 18) that grew on agar supplemented with itraconazole or voriconazole. Triazole-resistant strains were isolated from soil samples associated with carrot, potato, maize, and pea crops. Sequencing of the cyp51A gene and its promoter region indicated polymorphism, mainly with the presence of TR46/Y121F/T289A (n = 8), TR34/L98H, and TR53. Eight resistant isolates exhibited cyp51A wild type without alterations in the promoter region. Our study showed evidence of dissemination of azole-resistant A. fumigatus, with high genetic diversity, in vegetable crops in Colombia. These data underline the need to determine the prevalence of azole resistance in A. fumigatus in clinical and environmental settings for other regions of Colombia as well as Latin America.

Choice of vehicle affects pyraclostrobin toxicity in mice.

Pyraclostrobin is a strobilurin fungicide that inhibits mitochondrial complex III of fungal and mammalian cells. In toxicity studies that were used to estimate the safety factor, pyraclostrobin was added to animal feed or to aqueous vehicles. However, foods containing residues of pyraclostrobin and other strobilurin fungicides (azoxystrobin, trifloxystrobin, fluoxastrobin) are frequently prepared in vegetable oil prior to human consumption. The primary objective of this study was to determine if pyraclostrobin dissolved in an oil-based vehicle had adverse health outcomes in mice when compared to aqueous-based vehicles. We found that pyraclostrobin does not fully dissolve in aqueous methyl cellulose (MC) or carboxymethyl cellulose (CMC), two vehicles used in industry-sponsored toxicity studies, but does fully dissolve in corn oil. Moreover, C57BL/6 mice receiving pyraclostrobin in corn oil displayed adverse health outcomes, including loss of body weight, hypothermia and diarrhea at lower doses than when added to feed or to aqueous vehicles. Our data suggest that previous studies underestimated the true toxicity of pyraclostrobin in mammals. Additional toxicity tests using oil-based vehicles are recommended to verify current safety recommendations for strobilurin fungicides.

Positive effects of an oil adjuvant on efficacy, dissipation and safety of pyrimethanil and boscalid on greenhouse strawberry.

Methylated vegetable oil adjuvants can enhance initial deposition and decrease the required dosages of pesticides sprayed on plants, so an oil adjuvant mixed with fungicides were used to prevent and control gray mold in greenhouse strawberry. As the persistence and dietary exposure risks from fungicides on strawberries after using adjuvants have not been assessed, the efficacy, dissipation and safety of pyrimethanil and boscalid in the presence and absence of a methylated vegetable oil adjuvant were evaluated. To better describe the actual use of fungicides in greenhouse strawberry, twice repeated application of fungicides were conducted follower by an optimized QuEChERS pre-treatment method. When applied at 60% of their recommended dosages with the adjuvant, the efficacy of pyrimethanil and boscalid for gray mold was similar to that shown by the treatment of 100% fungicides in absence of the adjuvant based on Duncan's Multiple-Range test, and their average residues increased to 89.0% and 89.3%, respectively. The adjuvant enhanced the accumulation effect of pyrimethanil residue by 31.7% after repeated applications, and the half-lives were similar (5.2 and 4.2 d). The adjuvant had comparable accumulation effects (1.75 and 1.83) and similar half-lives (5.4 and 5.5 d) for boscalid. In absence of adjuvant, the risk quotients (RQs) of pyrimethanil (0.41 and 0.33) and boscalid (0.49 and 0.63) after twice applications at pre-harvest interval were lower than 1. Adding the methylated vegetable oil adjuvant to fungicides would result in unprolonging half-life and acceptably low dietary exposure risk on strawberries, but lower dosage of fungicides were used.

Evaluation of cytotoxic and genotoxic effects of Benodanil by using Allium and Micronucleus assays.

The aim of this study was to evaluate the potential cytotoxic effects of Benodanil fungicide by employing both mitotic index (MI) and mitotic phases on the root meristem cells of Allium cepa and genotoxic effects by using in vitro micronucleus assay (MN) in human peripheral blood lymphocyte. In the Allium root growth inhibition test, the EC50 value was first determined as 25 ppm. Then, 2 × EC50 value (50 ppm), EC50 value (25 ppm), and 1/2 × EC50 value (12.5 ppm) were tested with different treatment periods (24, 48, and 72 h). Both negative and positive controls were also used in parallel experiments. We obtained that mitotic index and prophase index decreased when compared with the control in all concentrations. In the micronucleus assay, lymphocytes were treated with various concentrations (250, 500, 750, and 1000 µg/ml) of Benodanil for 24 and 48 h. The results showed that Benodanil did not induce MN frequency in all concentrations of both treatment periods. Additionally, it was determined that this pesticide decreased nuclear division index (NDI) significantly. It was concluded that Benodanil has a cytotoxic effects depending on decreasing of MI and NDI.

Evaluation of the BlightPro Decision Support System for Management of Potato Late Blight Using Computer Simulation and Field Validation.

The objective of this study was to evaluate the utility of the BlightPro decision support system (DSS) for late blight management using computer simulation and field tests. Three fungicide schedules were evaluated: (i) calendar-based (weekly) applications, (ii) applications according to the DSS, or (iii) no fungicide. Simulation experiments utilized 14 years of weather data from 59 locations in potato-producing states. In situations with unfavorable weather for late blight, the DSS recommended fewer fungicide applications with no loss of disease suppression; and, in situations of very favorable weather for late blight, the DSS recommended more fungicide applications but with improved disease suppression. Field evaluation was conducted in 2010, 2011, 2012, and 2013. All experiments involved at least two cultivars with different levels of resistance. DSS-guided and weekly scheduled fungicide treatments were successful at protecting against late blight in all field experiments. As expected, DSS-guided schedules were influenced by prevailing weather (observed and forecast) and host resistance and resulted in schedules that maintained or improved disease suppression and average fungicide use efficiency relative to calendar-based applications. The DSS provides an interactive system that helps users maximize the efficiency of their crop protection strategy by enabling well-informed decisions.

Effects of Oral Exposure to Fungicides on Honey Bee Nutrition and Virus Levels.

Sublethal exposure to fungicides can affect honey bees (Apis mellifera L.) in ways that resemble malnutrition. These include reduced brood rearing, queen loss, and increased pathogen levels. We examined the effects of oral exposure to the fungicides boscalid and pyraclostrobin on factors affecting colony nutrition and immune function including pollen consumption, protein digestion, hemolymph protein titers, and changes in virus levels. Because the fungicides are respiratory inhibitors, we also measured ATP concentrations in flight muscle. The effects were evaluated in 3- and 7-d-old worker bees at high fungicide concentrations in cage studies, and at field-relevant concentrations in colony studies. Though fungicide levels differed greatly between the cage and colony studies, similar effects were observed. Hemolymph protein concentrations were comparable between bees feeding on pollen with and without added fungicides. However, in both cage and colony studies, bees consumed less pollen containing fungicides and digested less of the protein. Bees fed fungicide-treated pollen also had lower ATP concentrations and higher virus titers. The combination of effects we detected could produce symptoms that are similar to those from poor nutrition and weaken colonies making them more vulnerable to loss from additional stressors such as parasites and pathogens.

Controlling Aspergillus flavus and Aspergillus parasiticus growth and aflatoxin production in poultry feed using carvacrol and trans-cinnamaldehyde.

Aflatoxins (AF) are toxic metabolites primarily produced by molds, Aspergillus flavus and Aspergillus parasiticus. Contamination of poultry feed with AF is a major concern to the poultry industry due to severe economic losses stemming from poor performance, reduced egg production, and diminished egg hatchability. This study investigated the inhibitory effect of 2 generally regarded as safe (GRAS), natural plant compounds, namely carvacrol (CR) and trans-cinnamaldehyde (TC), on A. flavus and A. parasiticus growth and AF production in potato dextrose broth (PDB) and in poultry feed. In broth culture, PDB supplemented with CR (0%, 0.02%, 0.04% and 0.08%) or TC (0%, 0.005%, 0.01% and 0.02%) was inoculated with A. flavus or A. parasiticus (6 log CFU/mL), and mold counts and AF production were determined on days 0, 1, 3, and 5. Similarly, 200 g portions of poultry feed supplemented with CR or TC (0%, 0.4%, 0.8%, and 1.0%) were inoculated with each mold, and their counts and AF concentrations in the feed were determined at 0, 1, 2, 3, 4, 8, and 12 weeks of storage. Moreover, the effect of CR and TC on the expression of AF synthesis genes in A. flavus and A. parasiticus (aflC, nor1, norA, and ver1) was determined using real-time quantitative PCR (RT-qPCR). All experiments had duplicate samples and were replicated 3 times. Results indicated that CR and TC reduced A. flavus and A. parasiticus growth and AF production in broth culture and chicken feed (P<0.05). All tested concentrations of CR and TC decreased AF production in broth culture and chicken feed by at least 60% when compared to controls (P<0.05). In addition, CR and TC down-regulated the expression of major genes associated with AF synthesis in the molds (P<0.05). Results suggest the potential use of CR and TC as feed additives to control AF contamination in poultry feed.

Effect of meadowsweet flower extract-pullulan coatings on rhizopus rot development and postharvest quality of cold-stored red peppers.

The study involved an examination of the antifungal activity on red peppers of pullulan coating (P) and pullulan coating containing either water-ethanol (P + eEMF) or ethanol extract of meadowsweet flowers (P + eEMF). Pullulan was obtained from a culture of Aureobasidium pullulans B-1 mutant. Both non-inoculated peppers and those artificially inoculated with Rhizopus arrhizus were coated and incubated at 24 °C for 5 days. The intensity of the decay caused by Rhizopus arrhizus in the peppers with P and P + eEMF coatings was nearly 3-fold lower, and in the case of P + weEMF 5-fold lower, than that observed in the control peppers. Additionally, the P + weEMF coating decreased, almost two-fold the severity of pepper decay compared to other samples. The influence of coating of pepper postharvest quality was examined after 30 days of storage at 6 °C and 70%-75% RH. All coatings formed a thin and well-attached additional layer of an intensified gloss. During storage, color, total soluble solid content and weight loss of coated peppers were subject to lower changes in comparison with uncoated ones. The results indicate the possibility of the application of pullulan coatings containing MFEs as an alternative to the chemical fungicides used to combat pepper postharvest diseases.

[Species and ecological control of disease on cultivated Gentiana rigescens in Yunnan].

OBJECTIVE: To find out variety of the fungal diseases of cultivated Gentiana rigescens and provide important basis for prevention. METHODS: The diseases were diagnosed based on field investigate, symptoms observation, pathogen isolation, determination the size of morphological and verification following the Koch's Postulate procedures. RESULTS: Anthracnose (Colletotrichum gloeosporioides), grey mould (Botrytis cinerea), brown spot (Alternaria tenuis), rust (Aecidiumpers), circular spot (Pestalotiopsis), leaf blight (Stemphylium, Ascochyta, Pleospora) and nematodes (Heterodera spp., Meloidogyne spp.) were found on Gentiana rigescens. Anthracnose was the first main disease, the diseased plant rate was over 40% and disease severity was 4 - 5 degree and second disease was rust, incidence of rate was less 10% and other diseases rate was not enough 2%. CONCLUSION: All these diseases on Gentiana rigescens are reported for the first time and Gentiana rigescens is the new host plant of the diseases.

Control of postharvest diseases of fruit by heat and fungicides: efficacy, residue levels, and residue persistence. A review.

Extensive research has been done in recent years to reduce the heavy dependence on chemical fungicides to control postharvest diseases and disorders of horticultural crops. Alternative strategies were based on improved cultural practices, biological control, plant-defense promoters, and physical treatments such as UV illumination, radiofrequency treatment, heat therapy, and storage technologies. Among these, postharvest heat treatments such as hot water dips, short hot water rinsing and brushing, and hot air conditioning have reduced rot development and enhanced fruit resistance to chilling injury in sensitive cultivars while retaining fruit quality during cold storage and shelf life. Additive or synergistic increases in effectiveness were observed by integrating heat therapy with various chemical compounds, thus leading to significant reductions in the application of active ingredients to protect produce from decay. This paper highlights the knowledge on this topic with emphasis on heat therapy effects and factors affecting the uptake, persistence, and performance of fungicide residues when they are applied in combination with hot water.

An attempt of postharvest orange fruit rot control using essential oils from Mediterranean plants.

Twelve essential oils from Mediterranean aromatic plants were tested at different doses against four fungi known as causal agents of post-harvest orange fruit rot: Botrytis cinerea, Penicillium italicum, Phytophthora citrophthora, and Rhizopus stolonifer. Essential oils were obtained from Hyssopus officinalis, Lavandula angustifolia, Majorana hortensis, Melissa officinalis, Ocimum basilicum, Origanum vulgare, Salvia officinalis, and Thymus vulgaris (Family Lamiaceae), Verbena officinalis (Family Verbenaceae), and Pimpinella anisum, Foeniculum vulgare, and Carum carvi (Family Apiaceae). Because preliminary in vitro experiments showed that only the oils from V. officinalis, T. vulgaris, and O. vulgare exhibited some fungistatic activity against the above-named fungi, these three essential oils were used in successive in vivo tests carried out to protect healthy "Washington navel" orange fruits from artificial infection by the same micromycetes. The essential oil of T. vulgaris, at a 2,000 ppm dose, controlled fruit rot by B. cinerea, P. citrophthora, and R. stolonifer but was ineffective against P. italicum. Essential oils of V. officinalis and O. vulgare inhibited infection by the first two fungi and only by P. citrophthora, respectively. This finding represents an important result, with the goal of using the essential oils as natural preservatives for food products, due to their positive effect on their safety and shelf life.

Combined exposure to endocrine disrupting pesticides impairs parturition, causes pup mortality and affects sexual differentiation in rats.

Risk assessment is currently based on the no observed adverse effect levels (NOAELs) for single compounds. Humans are exposed to a mixture of chemicals and recent studies in our laboratory have shown that combined exposure to endocrine disrupters can cause adverse effects on male sexual development, even though the doses of the single compounds are below their individual NOAELs for anti-androgenic effects. Consequently, we have initiated a large project where the purpose is to study mixture effects of endocrine disrupting pesticides at low doses. In the initial range-finding mixture studies, rats were gavaged during gestation and lactation with five doses of a mixture of the fungicides procymidone, mancozeb, epoxyconazole, tebuconazole and prochloraz. The mixture ratio was chosen according to the doses of each individual pesticide that produced no observable effects on pregnancy length and pup survival in our laboratory and the dose levels used ranged from 25 to 100% of this mixture. All dose levels caused increased gestation length and dose levels above 25% caused impaired parturition leading to markedly decreased number of live born offspring and high pup perinatal mortality. The sexual differentiation of the pups was affected at 25% and higher as anogenital distance was affected in both male and female offspring at birth and the male offspring exhibited malformations of the genital tubercle, increased nipple retention, and decreased prostate and epididymis weights at pup day 13. The results show that doses of endocrine disrupting pesticides, which appear to induce no effects on gestation length, parturition and pup mortality when judged on their own, induced marked adverse effects on these endpoints in concert with other pesticides. In addition, the sexual differentiation of the offspring was affected. This as well as the predictability of the combination effects based on dose-additivity modelling will be studied further in a large dose-response study.

Effect of bavistin on in vitro plant conversion from encapsulated uninodal microcuttings of micropropagated Bacopa monnieri (L.)--an Ayurvedic herb.

A reliable and reproducible protocol for contamination free plant recovery system from alginated encapsulated uninodal microcuttings of micropropagated Bacopa monnieri L. have been developed after storage at 18 degrees C for 45 days. Node segments excised from freshly micropropagated plants were encapsulated as single explant beads with 3.0% sodium alginate and 80 mM CaCl2 2 H2O. To find out the optimal concentration of fungicide bavistin for efficient plant recovery different concentrations of bavistin (1.0 - 15 mg l(-1)) were incorporated in to the encapsulation medium. 3.0mg l(-1) bavistin showed no reduction in plant conversion and generated maximum number of shoots (45.6 +/- 1.69) at high frequency with out any contamination after storage up to 45 days at 18 degrees C. At high concentrations (13 and 15 mg l(-1)), rupturing of calcium alginate coats after 8 - 9 days and gradual decline in the number of shoots indicates the toxic effect of bavistin on plant conversion. Encapsulated node cuttings stored up to 45 days regenerated shoots (5.2) and multiple shoots (45.6) in MS basal and hormone medium respectively. Maximum shoot length (8.2 +/- 0.37 cm) was observed from encapsulsted node cuttings incorporated with 3.0 mg l(-1) bavistin on MS basal medium. 90% of the recovered plantlets were hardened off and successfully established in soil.

Endocrine disruptors and hypospadias: role of genistein and the fungicide vinclozolin.

OBJECTIVES: The phytoestrogen (plant estrogen) genistein, present in soy products, is of interest because in utero exposure to genistein can cause hypospadias in our mouse model and maternal consumption of soy is prevalent in human populations. Another compound of interest is the fungicide vinclozolin, which also causes hypospadias in the mouse and rat and can occur concurrently with genistein in the diet as a residue on exposed foods. A study in the United Kingdom found no relationship between a maternal organic vegetarian diet and hypospadias frequency, but women who consumed nonorganic vegetarian diets had a greater percentage of sons with hypospadias. Because nonorganic diets can include residues of pesticides such as vinclozolin, we sought to assess the interaction of realistic daily exposures to genistein and vinclozolin and their effects on the incidence of hypospadias. METHODS: Pregnant mice were fed a soy-free diet and orally gavaged from gestational days 13 to 17 with 0.17 mg/kg/day of genistein, 10 mg/kg/day of vinclozolin, or genistein and vinclozolin together at the same doses, all in 100 microL of corn oil. The controls received the corn oil vehicle. The male fetuses were examined at gestational day 19 for hypospadias, both macroscopically and histologically. RESULTS: We identified no hypospadias in the corn oil group. The incidence of hypospadias was 25% with genistein alone, 42% with vinclozolin alone, and 41% with genistein and vinclozolin together. CONCLUSIONS: These findings support the idea that exposure to these compounds during gestation could contribute to the development of hypospadias.

Dynamics of pesticides in potato crops.

This study investigated the behavior of pesticides commonly used on potato in a part of northwestern Spain with a large area devoted to this crop. Experimental potato plots were treated with commercial formulations of two insecticides, a nematicide, a herbicide, and a combination of two fungicides, and the concentrations of the active ingredients were monitored throughout the growing season in the 0-1 and 1-15 cm soil layers, in water dripping off the potato plants, and in the potatoes themselves. The technique used for pesticide determination was gas chromatography with mass selective detection. For potato analyses, pesticide extraction was optimized. The only pesticide ever detected in potato tubers was metalaxyl, the concentration of which never exceeded half the maximum residual limit even when it was applied several times more often than is officially recommended for potato crops. Metalaxyl was also the only pesticide detected in wash-off, apparently due to its being more soluble in water than the other pesticides applied to the growing plants. A mathematical model of pesticide transport in soil was fitted to the experimental data using the program HYDRUS-1D.

Large-scale preparation of the phytoalexin elicitor glucohexatose and its application as a green pesticide.

Large-scale preparation of the phytoalexin elicitor was achieved through a highly regio- and sterereoselective synthesis using 2,3,4,6-tetra-O-benzoyl-d-glucopyranosyl trichloroacetimidate (1), 1,2:5,6-di-O-isopropylidene-alpha-d-glucofuranose (2), and 6-O-acetyl-2,3,4-tri-O-benzoyl-alpha-d-glucopyranosyl trichloroacetimidate (3) as the synthons. Coupling of 1 with 2 gave the 1-->3-linked disaccharide; subsequent selective removal of 5,6-O-isopropylidene to give 5 followed by selective 6-O-glycosylation with 1 afforded the trisaccharide 6. Hydrolysis to remove the 1,2-O-isopropylidene was accompanied by ring expansion, giving 3,6-branched pyranosyl trisaccharide. Acetylation, selective 1-O-deacetylation, and activation with trichloroacetonitrile gave the trisaccharide donor 7. The trisaccharide acceptor 9 was prepared from condensation of the disaccharide 5 with 3 and subsequent 6-O-deacetylation. Coupling of the trisaccharide donor 7 with the trisaccharide acceptor 9 and subsequent deprotection afforded the glucohexatose elicitor. The cost of the produced glucohexatose should be low enough to allow its applications in agriculture as a green pesticide. At a concentration of 5-10 mg/L, the resultant elicitor was used to treat growing orange trees and harvested oranges, giving very encouraging results, comparable with those obtained using commercial pesticides at a concentration of 1400 mg/L (Topsin-M) for growing trees and 900 mg/L (Tecto) for harvested oranges, respectively. Treatment of tomato leaves against Botrytis cinerea with the synthetic elicitor at a concentration of 10 mg/L gave 82% inhibition, comparable with the inhibition of 84% by Wanmeiling at a concentration of 1000 mg/L. Treatment of tea leaves also showed promising results.

Fungicide sprays can injure the stigmatic surface during receptivity in almond flowers.

Fungicides can be detrimental to flower development, pollen function and fruit set in a number of crops. Almond is a self-incompatible nut crop that has a fruit set of only approx. 30 % of the total number of flowers. Thus, interference of pollination and fertilization by fungicide sprays is of concern, and identification of chemicals having the least detrimental effects would be desirable. The objective of this study was to evaluate the effect of fungicide sprays on stigma morphology in almond using a laboratory spray apparatus that simulated field applications. Four fungicides (azoxystrobin, myclobutanil, iprodione and cyprodinil) were applied, and fresh, unfixed stigmatic surfaces were observed using a scanning electron microscope at 4 and 24 h after spraying. Increased exudate accumulation was induced by azoxystrobin at both time periods, and localized damage and collapse of stigmatic cells were observed after 24 h. Damaged stigmatic papillae exhibited wrinkling, surface distortion or collapse. Likewise, myclobutanil caused significant damage to and collapse of papillae; these were more extensive at later observations. Iprodione had no effect on exudate accumulation but caused marked and severe collapse of stigmatic papillae which was pronounced at 24 h. Cyprodinil promoted a copious increase in exudate secretion and caused the most severe collapse of stigmatic cells of all the fungicides evaluated. Damage was somewhat localized at 4 h but more global at 24 h. This study has verified that certain fungicide sprays have direct detrimental effects on stigma morphology and enhance exudate production in almond flowers.

Residue uptake and storage responses of Tarocco blood oranges after preharvest thiabendazole spray and postharvest heat treatment.

Tarocco blood oranges (Citrus sinensis Linn. Obsek) were subjected to a single preharvest spray with thiabendazole (TBZ) at a concentration of 1% active ingredient (ai) in water and harvested 2 weeks after fungicide application or heated at 37 degrees C for 48 h under saturated humidity after harvest. The two treatments were also combined before cold quarantine (3 weeks at 2 degrees C), subsequent storage (3 weeks at 8 degrees C), and simulated marketing period (SMP) (1 week at 20 degrees C). Fruit not treated with TBZ and unheated were used as controls. The residue levels of TBZ (active ingredient, whole fruit basis) after spray were approximately 6.3 and 5.4 mg x kg(-1) before fruit storage respectively, a level close to the tolerance limit set by the European Community. TBZ showed a high persistence during quarantine, storage, and SMP. TBZ spray significantly reduced the incidence and severity of chilling injury (CI) and decay during the postquarantine period and SMP. Heat treatment (HT) produced beneficial effects in controlling CI, especially during SMP, when applied in combination with TBZ. However, HT remarkably promoted the development of secondary fungal infections such as Phytophthora rots and adversely affected fruit flavor and taste. The occurrence of off-flavor and off-taste was found to be perceptible after heating.