Fernando Diánez

Fungicide resistance inBotrytis cinerea isolates from strawberry crops in Huelva (southwestern Spain)

The severity of disease caused byBotrytis cinerea in strawberries is very high and chemical control is common practice; low residue levels of chemical products are required. Thus, it is important to be aware of the development of fungicide resistance in order to choose the best strategies of chemical control. In the present study we evaluated the response of 36B. cinerea isolates against eight different fungicides. The isolates were sampled twice, at the beginning and the end of the season, in 11 commercial strawberry fields located in the area of Huelva (Spain). In addition, two reference isolates, SAS56 and SAS405, were evaluated. The proportion of isolates resistant to benomyl was very high (86%). Resistance to dicarboximides was detected in 44% of the isolates and resistance to pyrimethanil in 25% of the isolates. Different degrees of sensitivity to captan and dichlofluanid were recorded. No resistance was found to diethofencarb plus carbendazim.

Grape Marc Compost Tea Suppressiveness to Plant Pathogenic Fungi: Role of Siderophores

It is important to know about the mechanisms that suppress plant diseases when compost from vegetable residues and/or their liquid extracts (compost tea) are used in order to improve the efficiency of this suppressing effect on pathogens. In this study, we assessed the presence of siderophores in various grape marc aerated compost teas (ACT) and their suppressing effect on nine pathogens: Rhizoctonia solani, Fusarium oxysporum f. sp. radicis-lycopersici, Fusarium oxysporum f. sp. lycopersici race 0, Fusarium oxysporum f. sp. lycopersici race 1, Fusarium oxysporum f. sp. radicis-cucumerinum, Verticillium dahliae, Pythium aphanidermatum, Phytophthora parasitica and the mycopathogen, Verticillium fungicola. Three concentrations (5, 10 and 15%) filtered, microfiltered and sterilized ACT were added to Petri dishes with a PDA medium, and 1 mM of ferric chloride (FeCy. After adding this mixture, a 0.5 cm disc was placed at the center of each dish containing the vegetative and reproductive body of each of the fungi to be tested. All the dishes were incubated at 25°C for seven days, except R. solani y P. aphanidermatum, which developed after 4 days. The addition of 1 mM of FeCl3 deactivated the siderophores present in the ACT, suppressing their inhibition of fungal development. The results obtained with the microfiltered ACT revealed that the microorganisms present in grape marc compost excreted siderophores into the medium which were responsible for inhibiting the growth of the 9 fungi tested. This activity was annulled by the addition of ferric chloride. The same results were achieved with the ACT obtained from filtering. This inhibition was not 100% after adding FeCl3 due to the fact that the microorganisms present in this tea, exhibited other biocontrol mechanisms.

Possibilities of the use of vinasses in the control of fungi phytopathogens

The purpose of this research was to study the biocide effect of three agroindustrial subproducts, concretely sugar beet, sugar cane and wine vinasse. Results from in vitro testing determined that wine vinasse is what shows a 100% capacity to suppress fungal growth with concentrations between 5% and 7% for Fusarium oxysporum f.sp. melonis race 0 and 1, Sclerotinia sclerotiorum, Pythium aphanidermatum and Phytophthora parasitica and 10-15% for F. oxysporum f.sp. radicis-cucumerinum. On the other hand, sugar cane vinasse produced an increase at high concentrations and sugar beet vinasse showed an approximate 100% suppressor effect on fungal growth for only some of the phytopathogens tested: S. sclerotiorum (15%), P. aphanidermatum (7%), P. parasitica (15%) and F. oxysporum f.sp. radicis-cucumerinum (15%). In the soil samples analyzed none of the three vinasse extracts decreased fusaric microbiota, producing an increase in the three samples tested. This would implicitly convey an improvement in soil quality by producing a potential increase in bacterial and fungal microbiota.

Suppressive effects of grape marc compost on phytopathogenic oomycetes

Abstract Suppression of soil-borne diseases of horticultural crops by compost has been attributed to the activities of antagonistic micro-organisms. A great diversity of biological control agents naturally colonize compost. This is especially true for biological control agents effective against the soil-borne oomycete pathogens. The purpose of this research was to determine the suppressive capacity of grape marc compost against Pythium aphanidermatum and Phytophthora parasitica. Antagonist in vitro assays were performed with 432 microbial morphologies isolated from grape marc compost. Seven micro-organisms were selected for further bioassay with cucumber – P. aphanidermatum, and two micro-organisms for tomato – P. parasitica. Those experiments indicate that grape marc compost reduces the severity of Pythium dumping-off on cucumber, but do not reduce the severity of Phytophthora root rot on tomato. The enrichment of compost or vermiculite with the selected microbes for compost do not improve the suppressive effects.

First Report of Fusarium oxysporum f. sp. radicis-cucumerinum on Cucumber in Spain

During December 1999, root and stem rot was observed on greenhouse-grown cucumber (cvs. Albatros, Brunex, Acapulco, and Cerrucho) plants in Almería, Spain, using rock wool cultures. The disease caused severe damage, estimated at a loss of up to 75% of the plants, in the first greenhouse affected; afterward, the disease was found in eight additional greenhouses (14 ha) in 1999 and 2000. Stem lesions extended up to 10 to 12 cm above the crown in mature plants, although no fruit damage was observed. In the advanced stages, abundant development of orange sporodochia was evident on crown and stem lesions, without vascular discoloration. Root, crown, and stem pieces that were placed on potato dextrose agar (PDA) after surface-disinfection with 5% sodium hypochlorite, rinsed, and dried resulted in pure fungal colonies. Based on morphological characteristics of conidia, phialides, and chlamydospores from the isolations, the fungus was identified as Fusarium oxysporum Schlechtend.:Fr. Pathogenicity tests were conducted on cucumber (cvs. Marketmore 76 and Cerrucho [F1 hybrid]), melon (cvs. Amarillo oro, Perlita, Piboule, Tania, and Nipper [F1]), watermelon (cvs. Sugar Baby, Sweet Marvel, Jubilee, and Pata Negra and hybrid Crimson sweet), Cucurbita maxima × Cucurbita moschata, zucchini (cv. Senator), and loofah (Luffa aegyptiaca) at several stages: (i) pregermination; (ii) 1 or 2 true leaves; and (iii) more than 10 true leaves. Five fungal isolates were grown on PDA or shaken potato dextrose broth at 25°C for 8 days. Inoculation was performed in pots (10 seeds or plants of each cultivar or hybrid and isolate) by drenching with 100 ml of a fungal suspension (104 to 106 CFU/ml). Sterile water was applied to noninoculated control plants. Tests were repeated in growth chambers at 25°C (night) and 28°C (day) with a 16-h photoperiod. Fifteen to fifty days after inoculation, cucumber and melon plants at all three stages developed symptoms of root and crown rot in 100% of inoculated plants, with no observed vascular discoloration. Fifty days after inoculation, all three stages of C. maxima × C. moschata and zucchini remained symptomless. Loofah and watermelon germinated poorly or not at all when inoculated at the pregermination stage. Fifteen to fifty days after inoculation, 100% of inoculated cucumber and melon plants developed symptoms. Watermelon plants inoculated at the 10 or more true-leaf stage did not develop disease symptoms. No symptoms developed on noninoculated control plants. F. oxysporum was reisolated from infected roots, crowns, and stems of inoculated plants, confirming Kochs postulates. The main symptoms on cucumber infected by F. oxysporum f. sp. cucumerinum are wilt, yellowing, and vascular discoloration. In contrast, based on inoculation of the host differentials and the resulting disease symptoms found in this study, the fungus was identified as F. oxysporum f. sp. radicis-cucumerinum (1). To our knowledge, this is the first report of F. oxysporum f. sp. radicis-cucumerinum causing root and crown rot in cucumber in Spain. Reference: (1) D. J. Vakalounakis. Plant Dis. 80:313, 1996.

Characters of compost teas from different sources and their suppressive effect on fungal phytopathogens

Compost teas (CT) are fermented watery extracts of composted materials that are used to control plant diseases and on crop fertilization. In this work, aerated (ACT) and non-aerated compost teas (NCT) were obtained from four different composts: spent mushroom substrate compost, grape marc compost, greenhouse horticultural crop residues compost, and vermicompost. Physico-chemical and microbiological analysis were carried out to determine their properties. In vitro assays were performed to assess their suppressive effect on the mycelial growth of eight fungal phytopathogens. In vivo trials aimed to assess their effect on gummy stem blight (Didymella bryonae) and powdery mildew (Podosphaera fusca) in melon plants. Results showed that ACT and NCT filtrates inhibited the in vitro growth of all tested pathogens while autoclaved CT did not completely lose their inhibitory effect, and CT sterilized by microfiltration had no effect on the pathogen growth. The severity of powdery mildew was highly reduced by ACT and NCT from all sources, though in gummy stem blight assay only a delay in disease development was observed. In general, all compost teas showed a high level of microbial populations and nutrients. Results suggest that the efficacy of ACT and NCT firstly depend on the microbiota present in them. We consider compost teas from the four tested sources as a viable way to manage plant diseases and crop fertilization, throughout its integration in pest management programs and fertirrigation systems under different dilution rates.

Erwinia aphidicola isolated from commercial bean seeds ( Phaseolus vulgaris)

In late 2003, a new disease appeared in protected bean crops in southeastern Spain, causing a decrease of over 50% in production. Several samples of affected plants were collected and analyzed and the agent of this disease was identified as the bacterium Erwinia aphidicola, which had never been described as a pathogen previously. We attempted to determine the possible bacterium transmission through seeds, using 120 commercial bean seeds from the same batch as that used in an affected farm, and 120 seeds from the fruiting plants of the same farm. Seed coats, cotyledons and leaves of plants originating from them, were taken and analyzed. Several of the developed symptoms on plants from commercial and fruiting plant seeds were internervial chlorosis, necrotic pits and rough roots and they coincided with those observed on affected crops. Bacteria present in commercial seed cotyledons were isolated and analyzed by biochemical and molecular tests. Results confirmed the presence of Erwinia aphidicola in four analyzed seeds; moreover, Bacillus simplex/Bacillus muralis, Pseudomonas mendocina, Pseudomonas putida and Paenibacillus polymyxa were also identified.

INCIDENCE OF LECANICILLIUM FUNGICOLA IN WHITE-BUTTON MUSHROOM (AGARICUS BISPORUS) CULTIVATED WITH TWO TYPES OF CASING SOIL

The effect of the agent of dry bubble disease Lecanicillium fungicola on mushroom cultivation using two types of casing (mineral soil and peat-based) was evaluated in two artificially infected cropping trials. The incidence of dry bubble and loss of yield caused by the disease were assessed. Biological efficiency (BE) was used to evaluate mushroom productivity for the different casing soils. The results showed that, in the absence of L. fungicola infection, the higher BE obtained using peat-based casing (higher yield of A. bisporus) compensates for the higher cost of the casing. However, when facing serious infection by L. fungicola, the incidence of disease and the yield of healthy mushrooms were statistically similar for both types of casing, although the number of diseased mushrooms was higher when peat-based casing was used. Therefore, the use of peat-based casings cannot be recommended when there is a high risk of outbreaks of dry bubble, since the yield of healthy mushrooms is not sufficient to compensate for the difference in price between both casing types.

Presence of Siderophores on Grape Marc Aerated Compost Tea

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Presence of Fusarium oxysporum f. sp. melonis Race 1 in Soils Cultivated with Melon in the State of Colima (Mexico)

During years 2001, 2002 and 2003 the gravity of the Fusarium wilt in 1000 hectares of melon culture was evaluated in Colima (Mexico). In spite of the soil disinfections with methyl bromide, the losses could reach 25% of the final production. The analysis of 4 soil samples from the fields with ill plants, in a selective medium for Fusarium, allowed to detect the presence of F. oxysporum. By means of the presented technique “soil phytopathometry”, 31 isolates of F. oxysporum f. sp. melonis were obtained from the soil samples. The isolates were inoculated on melon plants to evaluate their pathogenicity. The 31 isolates inoculated, produced the symptoms of chlorosis and wilting, in melon cultivars that allowed us to affirm that all isolates were race 1 of F. oxysporum f. sp. melonis. Being this the first news of the presence of F. oxysporum f. sp. melonis in the state of Colima (Mexico).