Copper-based Fungicides Research Articles

Impact of Rhizophagus irregularis MUCL 41833 on disease symptoms caused by Phytophthora infestans in potato grown under field conditions

In organic potato production in Europe, only copper-based fungicides allow to directly control Phytophthora infestans (the causal agent of late blight). Due to environmental concerns caused by the repeated and excessive use of Cu before the nineties, the EU legislation has promoted alternatives approaches such as the use of biocontrol agents (e.g. arbuscular mycorrhizal fungi – AMF). Here, two field trials were conducted over two climatic-contrasting growing seasons. Trial 1 was characterized by a dry and hot cultural season with low pressure of P. infestans, while trial 2 was conducted under high humidity and relatively low temperatures with high pressure of the pathogen. In both trials, sprouted potato tubers were inoculated with AMF in the greenhouse before transplanting to the field. A Real-Time quantitative PCR assay was designed to target the inoculant strain Rhizophagus irregularis MUCL 41833 as well as the native Rhizophagus irregularis strains. In both trials, the inoculated AMF was detected in the roots at harvest, demonstrating the capacity of the inoculated strain to incorporate the microbiome of the potato plants. In the first trial, disease severity in AMF pre-colonized potato plants was markedly decreased and the onset of late blight symptoms was delayed by 10 days. In contrast, in the second trial no differences were noticed between AMF pre-colonized and control plants. In both trials, no mycorrhizal effect was noticed on tuber yield. As a conclusion, disease severity of P. infestans, measured by symptoms development on leaves, was decreased in AMF pre-colonized plants under conditions of low pressure of late blight and over a short period of time, while under conditions more adequate to the pathogen, no reduction in symptoms was noticed.

Zagađenje zemljišta u gradskim baštama prouzrokovano primenom đubriva i fungicida na bazi bakra

Urban gardening is an activity that has been rapidly increasing. The aim of this study was to point out the common mistakes in cultivation practices made by producers on small city areas. Total of 96 individual plots under vegetable crops were sampled (56 from garden plots of the city of Novi Sad, and 40 from field plots in the suburbs). There was no significant difference in the content of organic matter between these two groups of tested soils. The contents of readily available P2O5 and K2O were significantly higher in garden soil than in the field plots for production of vegetables. According to the content of pseudototal copper, 39% of samples belonged to the category of critical concentration and/or above MAC in the urban soil. Obtained results prove the pollution of small area gardens due to high anthropogenic effect of excessive use of mineral fertilizers and copper-based fungicides.

Modeling ecotoxicity impacts in vineyard production: Addressing spatial differentiation for copper fungicides

Application of plant protection products (PPP) is a fundamental practice for viticulture. Life Cycle Assessment (LCA) has proved to be a useful tool to assess the environmental performance of agricultural production, where including toxicity-related impacts for PPP use is still associated with methodological limitations, especially for inorganic (i.e. metal-based) pesticides. Downy mildew is one of the most severe diseases for vineyard production. For disease control, copper-based fungicides are the most effective and used PPP in both conventional and organic viticulture. This study aims to improve the toxicity-related characterization of copper-based fungicides (Cu) for LCA studies. Potential freshwater ecotoxicity impacts of 12 active ingredients used to control downy mildew in European vineyards were quantified and compared. Soil ecotoxicity impacts were calculated for specific soil chemistries and textures. To introduce spatial differentiation for Cu in freshwater and soil ecotoxicity characterization, we used 7 European water archetypes and a set of 15,034 non-calcareous vineyard soils for 4 agricultural scenarios. Cu ranked as the most impacting substance for potential freshwater ecotoxicity among the 12 studied active ingredients. With the inclusion of spatial differentiation, Cu toxicity potentials vary 3 orders of magnitude, making variation according to water archetypes potentially relevant. In the case of non-calcareous soils ecotoxicity characterization, the variability of Cu impacts in different receiving environments is about 2 orders of magnitude. Our results show that Cu potential toxicity depends mainly on its capacity to interact with the emission site, and the dynamics of this interaction (speciation). These results represent a better approximation to understand Cu potential toxicity impact profiles, assisting decision makers to better understand copper behavior concerning the receiving environment and therefore how restrictions on the use of copper-based fungicides should be considered in relation to the emission site.

Tolerance and translocation of heavy metals in young grapevine (Vitis vinifera) grown in sandy acidic soil with interaction of high doses of copper and zinc

Vineyard soils may have high levels of copper due to frequent applications of Bordeaux mixture for the control of fungal diseases. Therefore, many winegrowers have used zinc-based fungicides instead of copper-based fungicides, resulting in high levels of Cu and Zn in vineyard soils. The aim of the study was to evaluate the tolerance of young grapevines grown in soil with interaction of high contents of Cu and Zn, and the translocation of these heavy metals from roots to shoots. We collected soil of a natural field of Rio Grande do Sul, the southernmost state of Brazil. Soil treatments were addition of 0, 30, and 60mg Cu kg−1, and addition of 0, 15, 30, 60, 120, and 180mg Zn kg−1 for each copper dose. A seedling of grapevine per pot was grown for 60days. After that, we measured root and shoot dry matter, Cu and Zn content in roots and shoots, chlorophyll a fluorescence, photosynthetic pigments, and peroxidase and superoxide dismutase activity. Young grapevines showed tolerance mechanisms to high concentrations of Cu and Zn by retaining these metals in the roots, decreasing translocation to the shoots. However, at the highest doses of Cu and Zn, we observed decrease in plant growth and in photochemical efficiency associated with the decrease in photosynthetic pigment content. Furthermore, the activity of antioxidant enzymes increased in intermediate doses of Zn, which indicates activation of the antioxidant system. However, the stress condition in treatments with high levels of Cu and Zn was not reversed.

Black Oat (Avena strigosa Schreb.) Growth and Root Anatomical Changes in Sandy Soil with Different Copper and Phosphorus Concentrations

Successive applications of copper-based (Cu) fungicides have increased Cu concentration in vineyard soils, inducing Cu toxicity in young vines and cover crops such as black oat, thus inhibiting growth and development. However, increasing soil phosphorus (P) content can reduce Cu toxicity symptoms. In this context, the aim of this study was to evaluate the effect of Cu toxicity and its alleviation by P fertilization in black oat cultivated in sandy soil. For the experiment, Typic Hapludalf soil samples were air-dried, prepared, and subjected to increasing doses of Cu (0, 30, and 60 mg kg−1) and P (0 and 100 mg kg−1). Subsequently, the soil was incubated and stored in pots, where black oat seedlings were grown for 30 days in a greenhouse. Plant roots subjected to Cu, especially with the highest Cu concentration and without P addition decreased the root cap size, showing early tissue differentiation and lateral root formation near the apical region. Decrease in dry matter (DM) production of roots (50 %) and shoots (67 %) was also observed in the highest Cu concentration. Plants without P addition, regardless of Cu concentration, also had lower root (33 %) and shoot (65 %) DM production. P addition in soil and its increased concentration reduced root anatomical changes and stimulated plant DM production. Therefore, we conclude that excessive Cu concentration alters black oat root anatomical structure, affecting plant growth, especially in sandy soils with low organic matter content. However, P supply can reduce root Cu toxicity symptoms, thus increasing plant dry matter production.

Some effects of copper-based fungicides on plant-feeding terrestrial molluscs: A role for repellents in mollusc management

The effects of three copper-based fungicides on feeding by four terrestrial molluscs (two slugs and two snails) were assessed in laboratory choice and no-choice tests. These products functioned effectively as short-term feeding deterrents. However, a copper hydroxide-based fungicide was more persistent than copper octanate- and copper diammonia diacetate-based fungicides under field conditions. Tests with foliage treated in the field and fed to Leidyula floridana and Lissachatina fulica showed that significant feeding suppression persisted for several days. We also determined that copper hydroxide was repellent, affecting movement of L. floridana slugs independent of food. Further, we assessed copper hydroxide effects with and without iron phosphate molluscicide bait. Copper hydroxide alone significantly suppressed foliage consumption, but did not affect slug survival. Iron phosphate bait alone caused slug mortality, and significantly suppressed feeding, but only after 24 h. Significantly improved foliage protection, relative to iron phosphate bait application alone, was attained when copper hydroxide repellent was used simultaneously with iron phosphate bait. Where terrestrial molluscs are a plant protection issue, it appears that ancillary benefit can be derived from using copper hydroxide fungicides. These data provide evidence for the potential application of the “push–pull” or “stimulo-deterrent” concept relative to mollusc management; plant protection may be enhanced when attractant and repellent treatments are applied simultaneously.

Fluorescence spectroscopy to study dissolved organic matter interactions with agrochemicals applied in Swiss vineyards.

UV/Vis fluorescence spectroscopy was used to study the possible interactions of dissolved organic matter (DOM) with the herbicide glyphosate and copper-based fungicide used in vineyards. The study focused on the role of DOM in the transport of these micropollutants from parcels to surface waters (river, lake). Soil solution and river water samples were collected in the Lavaux vineyard area, western Switzerland. Their fluorescence excitation emission matrices (EEM) were decomposed using parallel factor (PARAFAC) analysis, and compared to their content in glyphosate and copper. PARAFAC analysis of EEM of both types of samples showed the contribution of protein-like and humic-like fluorophores. In soil water samples, complexes between fulvic-like and humic-like fluorophores of DOM, copper, and glyphosate were likely formed. In surface water, DOM-copper and glyphosate-copper interactions were observed, but not between glyphosate and DOM.

Effect of copper-based fungicide (bordeaux mixture) spray on the total copper content of areca nut: Implications in increasing prevalence of oral submucous fibrosis.

Background:Potentially malignant disorders like oral submucous fibrosis (OSMF) often precede oral squamous cell carcinoma (OSCC). The rate of transformation of OSMF to OSCC ranges from 3 to 19%. OSMF is etiologically related to chewing of areca nut (betel nut), and the high copper content in areca nut plays an important role in the pathogenesis of the disorder. Even though many studies estimated and confirmed increased copper levels in areca nuts, studies tracing the source of the increased copper content are scarce. Interestingly, on review of agricultural literature, it was found that most of the areca nut plantations in South India commonly use a copper-based fungicide, bordeaux mixture (BM).Aim:The aim of the study was to estimate and compare the copper content in areca nuts from plantations with and without copper-based fungicide usage.Materials and Methods:Four areca nut plantations from Dakshina Kannada district, Karnataka (group A) and four plantations from Ernakulam district, Kerala (group B) were selected for the study. The plantations from Karnataka used copper-based fungicide regularly, whereas the latter were devoid of it. Areca nut samples of three different maturities (unripe, ripe, and exfoliated) obtained from all plantations were dehusked, ground, and subjected to atomic absorption spectrometry (AAS) for copper analysis.Results:There was statistically significant difference in the copper content of areca nuts from both groups. The areca nuts from plantations treated with copper-based fungicide showed significantly higher copper levels in all maturity levels compared to their counterparts in the other group (P < 0.05).Conclusions:The high copper content in areca nut may be related to the copper-based fungicide treatment on the palms. These areca nuts with high copper content used in quid or commercial products may be responsible for the increasing prevalence of OSMF.

Contamination of apple orchard soils and fruit trees with copper-based fungicides: sampling aspects.

Accumulations of copper in orchard soils and fruit trees due to the application of Cu-based fungicides have become research hotspots. However, information about the sampling strategies, which can affect the accuracy of the following research results, is lacking. This study aimed to determine some sampling considerations when Cu accumulations in the soils and fruit trees of apple orchards are studied. The study was conducted in three apple orchards from different sites. Each orchard included two different histories of Cu-based fungicides usage, varying from 3 to 28 years. Soil samples were collected from different locations varying with the distances from tree trunk to the canopy drip line. Fruits and leaves from the middle heights of tree canopy at two locations (outer canopy and inner canopy) were collected. The variation in total soil Cu concentrations between orchards was much greater than the variation within orchards. Total soil Cu concentrations had a tendency to increase with the increasing history of Cu-based fungicides usage. Moreover, total soil Cu concentrations had the lowest values at the canopy drip line, while the highest values were found at the half distances between the trunk and the canopy drip line. Additionally, Cu concentrations of leaves and fruits from the outer parts of the canopy were significantly higher than from the inner parts. Depending on the findings of this study, not only the between-orchard variation but also the within-orchard variation should be taken into consideration when conducting future soil and tree samplings in apple orchards.

Copper Accumulations in Soils, Coffee, Banana, and Bean Plants Following Copper-Based Fungicides in Coffee Farms in Arusha and Kilimanjaro Regions, Tanzania

This study was carried out to investigate the levels of copper (Cu) contamination in coffee fields in Kilimanjaro and Arusha regions, Tanzania, to increase the database on the contamination of soils by Cu-based fungicides in coffee fields. Surface (0–20 cm deep) soil samples were collected from different farms in Kilimanjaro and Arusha regions. Coffee, banana, and bean plant samples were collected from the locations of soil sampling. Soil and plant samples were analyzed at the Department of Soil Science, Sokoine University of Agriculture, Morogoro, Tanzania. It was found that the calcium chloride (CaCl2)–extractable Cu was less than the detection limit of flame atomic absorption spectrometry. Diethylenetriaminepentaacetic acid (DTPA)–extractable (24 to 366 mg Cu kg−1 soil) and aqua regia–extractable (80 to 806 mg Cu kg−1 soil) Cu levels were high enough to raise environmental alarm (based on European Union guidelines) in the Cu fungicide–treated soils as compared with natural Cu levels in untreated soils (1 to 12 mg Cu kg−1 soil for DTPA and 22 to 32 mg Cu kg−1 soil for aqua regia–extractable Cu). Coffee, banana, and bean plants grown on soils contaminated by Cu fungicides had varied concentrations of Cu that were greater than the concentrations of Cu in the plants collected from the uncontaminated soils. Stepwise regression analysis carried out to investigate the relationships between the soil properties and the concentrations of Cu in plants revealed a significant (P = 0.01) positive relationship (R2 = 0.4) between organic carbon and the concentration of Cu in banana leaves. Aqua regia–extractable Cu was positively correlated (P = 0.03, R2 = 0.4) with the concentrations of Cu in banana leaves. For bean leaves, electrical conductivity (EC) had a positive significant (P = 0.01) relationship (R2 = 0.56) with the concentrations of Cu in the plants. It is recommended that further research be carried out to investigate the dynamics and bioavailability of Cu for the different crops interplanted in the coffee fields.

Metabolic changes of Vitis vinifera berries and leaves exposed to Bordeaux mixture

Since the development of Bordeaux mixture in the late 1800's, copper-based fungicides have been widely used against grapevine (Vitis vinifera L.) diseases, mainly in organic but also in conventional viticulture; however their intensive use has raised phytotoxicity concerns. In this study, the composition of grape berries and leaves upon Bordeaux mixture treatment was investigated during the fructification season by a metabolomic approach. Four applications of Bordeaux mixture till 3 weeks before harvest were performed following the regular management practices of organic viticulture. Results showed that the copper-based treatment affected the content in sugars, organic acids, lipids and flavan-3-ols of grapes and leaves at specific developmental stages. Nonetheless, the levels of sucrose, glucose and fructose, and of tartaric and malic acids were not significantly affected in mature grapes. In contrast, a sharp decrease in free natural amino acids was observed, together with a reduction in protein content and in mineral nitrogen forms. The treatment with Bordeaux mixture increased by 7-fold the copper levels in tissue extracts from surface-washed mature berries.

Scientific Opinion on the pest categorisation ofPlenodomus tracheiphilus(Petri) Gruyter, Aveskamp &amp; Verkley [syn.Phoma tracheiphila(Petri) L.A. Kantschaveli &amp; Gikashvili

The European Commission requested the EFSA Panel on Plant Health to perform a pest categorisation of Phoma tracheiphila, the fungal pathogen responsible for “mal secco” disease of citrus. This pathogen is listed in Annex IIAII of Directive 2000/29/EC. Recently, the pathogen has been reclassified as Plenodomus tracheiphilus (Petri) Gruyter, Aveskamp & Verkley, based on molecular phylogenetic analysis. Plenodomus tracheiphilus is a single taxonomic entity, and sensitive and specific methods are available for its differentiation from other related Plenodomus species. The main host is lemon (Citrus limon L.), but the pathogen has also been reported on other species of the genera Citrus, Fortunella, Poncirus and Severinia and on their hybrids. Host plants are widely grown in the southern EU Member States (MSs) and climatic conditions are conducive to disease development in both orchards and nurseries. The pathogen is present in part of the risk assessment area, being mainly reported on lemon grown in Italy, Greece, Cyprus and France, where it has a serious impact on the citrus industry. There are no obvious ecological/climatic factors limiting the potential establishment and spread of the pathogen in the, so far, non-infested citrus-producing EU MSs (i.e. Spain, Portugal, Malta and Croatia). Short-distance spread of the pathogen occurs via water splash and wind-driven rain, whereas movement of infected host plants for planting, particularly asymptomatic plants, is considered to be responsible for the introduction of the pathogen into new areas. Cultural practices and copper-based fungicide sprays may reduce inoculum sources and prevent new infections but they cannot eliminate the pathogen. P. tracheiphilus fulfils all of the pest categorisation criteria for having the potential to be a quarantine pest and a regulated non-quarantine pest, as those are defined in the International Standards for Phytosanitary Measures No 11 and 21, respectively. © European Food Safety Authority, 2014

Copper reducing strategies for late blight (Phytophthora infestans) control in organic potato (Solanum tuberosum) production

In organic potato ( Solanum tuberosum ) production in Europe,solely copper-based fungicides allow to directly control late blight (caused by Phytophthora infestans ). Due to environmental concerns, the use of copper (Cu) fungicides has to be as efficiently as possible to reduce annual consumption. This can be achieved either by reducing the dose per application or by decreasing the numbers of sprays. Between 2005 to 2009, six field trials were conducted at two sites(3 each) in northern Germany in order to (a) determine theefficacy of a copper fungicide (copper hydroxide) at reducedrates compared to the common practise of the most impor­tant German organic farmers’ associations (3 kg Cu ha-1),and (b) to evaluate the copper reduction potential of anapplication strategy based on a decision support system (DSS). Aclear dose-response relationship with reduced rates of copperhydroxide was not consistent over trials and strategiesalthough there was clear evidence for a considerable potentialto reduce Cu rates compared to the amount of 3 kg Cu ha-1. However, different degrees of late blight pressure stronglyaffected the efficacy of the Cu fungicide in individual years.Under less late blight pressure, a reduction to 1.25 kg Cu ha-1 applied throughout the season was possible without significantloss in efficacy compared to the typical spraying of 2.5 or 3.0 kg Cu ha-1. Due to a higher risk of decreasing efficacy,reducing Cu hydroxide supply was only possible to an amountof 2.0-2.5 kg Cu ha-1 under moderate disease pressure whilehigh late blight pressure in 2007 did not allow a Cu reductionto this extent. Even the highest amount of 3.0 kg Cu ha-1 only decreased the disease by 18% in one single case. Consequently, yields were not consistent, but depended on year, site and late blight pressure. On average over rates and strategies,Cu application increased yields by 15%. In addition, the resultsrevealed the clear tendency that reducing Cu amounts did not impair tuber yield. A spray schedule based on the new GermanDSS OKOSIMPHYT increased effectiveness of Cu sprays,reduced numbers of sprays and the total amount of Cu peryear, even under moderate and high late blight pressure.Therefore, we conclude that the DSS has the potential for further optimization. Knowledge of actual late blight pressure, as provided by OKOSIMPHYT, is necessary to optimize the use of Cu fungicides allowing both to ensure the site-specific tuber yield potential and to reduce the CU application amounts.

Copper content and distribution in vineyard soils of central Serbia

This research studied the copper content of vineyard soils as affected by the long-term use of copper-based fungicides. The soil samples were taken from individual vineyards located in the central region of Serbia, from two depths: 0-30 and 30-60 cm. At the same time, at each site, control samples were collected from a nearby forest in order to determine the background concentrations. The pseudototal (Cu T ) and available (Cu EDTA ) copper content were analysed in 60 soil samples in total, 46 of which represented vineyard soils and 14 control samples. The maximum value of copper was 200.1 mg/kg of pseudototal and 82.1 mg/kg of available copper. Comparison of the copper content in vineyards to the background concentrations of control samples clearly confirmed anthropogenic influence. Out of 46 vineyard soil samples, about one half (22 of them) had the Cu T concentration above the critical level of 60 mg/kg. Eleven samples had the pseudototal content over the MAC of 100 mg/kg. Anthropogenic influence was also confirmed on the basis of copper bioavailability and copper distribution along the soil profile. Available content of over 50 mg/kg was found in 8 out of 46 analysed samples of vineyard soils. According to the percentage contribution of available Cu EDTA to pseudototalCu T , half of the samples were above 36%, which is potentially phytotoxic. The concentration of copper was the highest in the surface layer in the vineyard soil samples. A check of the background Cu levels has shown that the distribution of Cu T and Cu EDTA is uniform throughout the soil profile. Data from some of the analysed plots indicate that the process of erosion is under way at the site. The soil on lower-lying terrain has been found to be more exposed to copper pollution than the soil of higher terrain. Since copper at the surveyed sites is very persistent and accumulates in a short period of time, focus should be placed on the preventive measures of reducing the use of copper-based fungicides to an optimal level.

Copper homeostasis in grapevine: functional characterization of the Vitis vinifera copper transporter 1

The Vitis vinifera copper transporter 1 is capable of self-interaction and mediates intracellular copper transport. An understanding of copper homeostasis in grapevine (Vitis vinifera L.) is particularly relevant to viticulture in which copper-based fungicides are intensively used. In the present study, the Vitis vinifera copper transporter 1 (VvCTr1), belonging to the Ctr family of copper transporters, was cloned and functionally characterized. Amino acid sequence analysis showed that VvCTr1 monomers are small peptides composed of 148 amino acids with 3 transmembrane domains and several amino acid residues typical of Ctr transporters. Bimolecular fluorescence complementation (BiFC) demonstrated that Ctr monomers are self-interacting and subcellular localization studies revealed that VvCTr1 is mobilized via the trans-Golgi network, through the pre-vacuolar compartment and located to the vacuolar membrane. The heterologous expression of VvCTr1 in a yeast strain lacking all Ctr transporters fully rescued the phenotype, while a deficient complementation was observed in a strain lacking only plasma membrane-bound Ctrs. Given the common subcellular localization of VvCTr1 and AtCOPT5 and the highest amino acid sequence similarity in comparison to the remaining AtCOPT proteins, Arabidopsis copt5 plants were stably transformed with VvCTr1. The impairment in root growth observed in copt5 seedlings in copper-deficient conditions was fully rescued by VvCTr1, further supporting its involvement in intracellular copper transport. Expression studies in V. vinifera showed that VvCTr1 is mostly expressed in the root system, but transcripts were also present in leaves and stems. The functional characterization of VvCTr-mediated copper transport provides the first step towards understanding the physiological and molecular responses of grapevines to copper-based fungicides.

Effects of elevated copper levels on biological nitrogen fixation and occurrence of rhizobia in a Tanzanian coffee-cropped soil

A study was conducted to investigate the effects of increasing copper concentration in soil on rhizobial occurrence and the process of biological nitrogen fixation. Two slow-growing bradyrhizobial strains CP1 and GM8 and two fast-growing rhizobial strains PV1 and PV2 were studied by comparing their performance under increasing copper concentrations in greenhouse-based assays involving modified Leonard jar assemblies and potted-soil experiments. Additionally, field samples from soils grown to coffee and subjected to long-term use of copper-based fungicides were analyzed for their total indigenous rhizobial populations using the most probable number-plant infection technique. Results indicated that elevated copper levels in the growth medium had inhibitory effects on nodulation, biological N2 fixation and overall rhizobial numbers in soil. Significant (p= 0.05%) reductions in fresh nodule mass, fresh nodule volume and total shoot nitrogen were recorded when copper concentration was increased from 0 to 100 ppm in both modified Leonard jar assemblies and potted-soil trials. Effective decrements in all the three parameters of fresh nodule mass, fresh nodule volume and total shoot nitrogen were more pronounced with the slow-growing bradyrhizobial strains of CP1 and GM8 than with the fast-growing PV1 and PV2. The MPN-plant infection technique results showed a non-significant (p=0.05) but substantial decrement in rhizobial and bradyrhizobial numbers when the copper-contaminated field soil (82.5 mgCu/g soil) was compared to a control soil (1.8 mgCu/g soil). The study concludes, therefore, that elevated levels of copper in soil could be harmful to free-living rhizobia and their abilities to fix N2 in respective symbiotic associations with legume species. Such negative effects were more pronounced in the slow-growing bradyrhizobial than rhizobial species used.

Influence of the farming system on the epiphytic yeasts and yeast-like fungi colonizing grape berries during the ripening process

Grape berries are colonized by a wide array of epiphytic microorganisms such as yeast and filamentous fungi. This microbiota plays a major role in crop health and also interferes with the winemaking process. In this study, culture-dependent and -independent methods were used to investigate the dynamics and diversity of the yeast and yeast-like microorganisms on the grape berry surface during maturation and the influence of cropping systems in this microflora. The results showed a significant impact of both the farming system and the maturity stage on the epiphytic yeast and yeast-like community. A quantitative approach based on counting cultivable populations indicated an increase in the yeast and yeast-like population during the grape ripening process, reaching a maximum when the berries became overripe. The cultivable yeast and yeast-like population also varied significantly depending on the farming system. Microorganism counts were significantly higher for organically- than conventionally-farmed grapes. The yeast and yeast-like community structures were analysed by culture independent methods, using CE-SSCP. The results revealed changes in the genetic structure of the yeast and yeast-like community throughout the ripening process, as well as the impact of the farming system. Copper-based fungicide treatments were revealed as the main factor responsible for the differences in microbial population densities between samples of different farming systems. The results showed a negative correlation between copper levels and yeast and yeast-like populations, providing evidence that copper inhibited this epiphytic community. Taken together, our results showed that shifts in the microbial community were related to changes in the composition of the grape–berry surface, particularly sugar exudation and the occurrence of copper residues from pesticide treatments.

Modeling losses of copper-based fungicide foliar sprays in wash-off under simulated rain

Wash-off experiments of three Cu-based fun- gicides were conducted with a single raindrop simulator with known drop size and fall height. Losses were quan- tified as total Cu (CuT), in solution (CuL), and particulate (CuP). Cu wash-off time course was modeled for two dif- ferent drop sizes using a stochastic model based on the cumulative detachment by random scattered raindrop impacts. In other set of experiments, the influence of raindrop size, fall height, and fungicide dose was analyzed statistically by means of a full factorial design. Most Cu was lost as particles sized from 0.3 to 1 lm. The stochastic model gave good estimations with two detachment per- formance levels. The best-fitting model parameters were as follows: the single element area covered by one impact (1.7 and 0.38 cm 2 for the large and small raindrop, respec- tively), the average number of repeated drop impacts on one single element area that exhaust non-rainfast fungicide (4.2 ± 3.0 small drops and 2.5 ± 0.5 large drops for the high performance level, low performance level needed 30 ± 10 large drops and 40 ± 88 small drops), and the mass washed-off by a single-drop impact (from 1.27 ± 0.2 l gC u to 3± 1 ng Cu per impact). Factorial design showed the dosage was the most influential factor in the three fungicide formulations. The model can help to estimate fungicide losses in field from rainfall and canopy properties. However, the particulate/soluble loss ratio cannot be predicted by the model since both the particle detachment and solubilization were not clearly related with the raindrop energy.

Compatibility of Isaria fumosorosea (Hypocreales: Cordycipitaceae) Blastospores with Agricultural Chemicals Used for Management of the Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae).

Biorational insecticides are being increasingly emphasized for inclusion in integrated pest management programs for invasive insects. The entomopathogenic fungus, Isaria fumosorosea, can be used to help manage the Asian citrus psyllid with minimal impact on beneficial arthropods, but its effectiveness may be compromised by agrochemicals used to control concurrent arthropod pests and diseases. We evaluated the compatibility of I. fumosorosea blastospores with a range of spray oils and copper-based fungicides registered for use in citrus groves. Results of laboratory and greenhouse tests showed a range of responses of the fungus to the different materials, including compatibility and incompatibility. Overall, I. fumosorosea growth in vitro was reduced least by petroleum-based materials and most by botanical oils and borax, and some of the copper-based fungicides, suggesting that tank mixing of I. fumosorosea with these latter products should be avoided. However, equivalent negative effects of test materials on fungal pathogenicity were not always observed in tests with adult psyllids. We hypothesize that some oils enhanced adherence of blastospores to the insect cuticle, overcoming negative impacts on germination. Our data show that care should be taken in selecting appropriate agrochemicals for tank-mixing with commercial formulations of entomopathogenic fungi for management of citrus pests. The prospects of using I. fumosorosea for managing the invasive Asian citrus psyllid and other citrus pests are discussed.

Horticultural Use of Copper-Based Fungicides Has Not Increased Copper Concentrations in Sediments in the Mid- and Upper Yarra Valley

The use of Cu-based fungicide can pose a risk to nearby surface water bodies due to the run-off of accumulated Cu from agricultural soils. In 2008, we conducted a reconnaissance survey of the presence and concentration of copper in sediments at 18 sites within the Yarra River Catchment, an important horticultural production system in south-eastern Australia. Observed Cu concentrations in sediment samples from the study sites (mean (95 % confidence interval) 12.0 (10.6–13.6) mg/kg dry weight) were similar to the concentrations present in the samples from the reference sites (mean (95 % confidence interval) 12.0 (6.7–16.8) mg/kg dry weight). The data on Cu and other metals in the sediments suggest that that there is unlikely to have been wide spread, diffuse, off-site transport of Cu from the soils of horticultural properties to nearby surface waterways in the Yarra River Catchment and that that observed sediment metal concentrations are unlikely to pose an ecological risk to sediment-dwelling organisms at the study sites.