C. Iglesias

Species of Fusarium Isolated from River and Sea Water of Southeastern Spain and Pathogenicity on Four Plant Species

Species of Fusarium were isolated from water samples collected from the Andarax River and coastal sea water of the Mediterranean in Granada and Almería provinces of southeastern Spain. In total, 18 water samples were analyzed from the Andarax River, and 10 species of Fusarium were isolated: Fusarium anthophilum, F. acuminatum, F. chlamydosporum, F. culmorum, F. equiseti, F. verticillioides, F. oxysporum, F. proliferatum, F. solani, and F. sambucinum. In addition, five species were isolated from 33 sea water samples from the Mediterranean Sea: F. equiseti, F. verticillioides, F. oxysporum, F. proliferatum, and F. solani. When considering the samples by their origins, 77.8% of the river water samples yielded at least one species of Fusarium, with F. oxysporum comprising 72.2% of the total isolates. In the case of marine water, 45.5% of the samples yielded at least one species of Fusarium, with F. solani comprising 36.3% of the total isolates. The pathogenicity of 41 isolates representing nine of the species collected from river and sea water during the study was evaluated on barley, kohlrabi, melon, and tomato. Inoculation with F. acuminatum, F. chlamydosporum, F. culmorum, F. equiseti, F. verticillioides, F. oxysporum, F. proliferatum F. solani, and F. sambucinum resulted in pre- and post-emergence damping off. Pathogenicity of Fusarium isolates did not seem to be related to the origin of the isolates (sea water or fresh water). However, the presence of pathogenic species of Fusarium in river water flowing to the sea could indicate long-distance dispersal in natural water environments.

Fungal microbiota from rain water and pathogenicity of Fusarium species isolated from atmospheric dust and rainfall dust

In order to determine the presence of Fusarium spp. in atmospheric dust and rainfall dust, samples were collected during September 2007, and July, August, and October 2008. The results reveal the prevalence of airborne Fusarium species coming from the atmosphere of the South East coast of Spain. Five different Fusarium species were isolated from the settling dust: Fusariumoxysporum, F.solani, F.equiseti, F.dimerum, and F.proliferatum. Moreover, rainwater samples were obtained during significant rainfall events in January and February 2009. Using the dilution-plate method, 12 fungal genera were identified from these rainwater samples. Specific analyses of the rainwater revealed the presence of three species of Fusarium: F.oxysporum, F.proliferatum and F.equiseti. A total of 57 isolates of Fusarium spp. obtained from both rainwater and atmospheric rainfall dust sampling were inoculated onto melon (Cucumismelo L.) cv. Piñonet and tomato (Lycopersiconesculentum Mill.) cv. San Pedro. These species were chosen because they are the main herbaceous crops in Almeria province. The results presented in this work indicate strongly that spores or propagules of Fusarium are able to cross the continental barrier carried by winds from the Sahara (Africa) to crop or coastal lands in Europe. Results show differences in the pathogenicity of the isolates tested. Both hosts showed root rot when inoculated with different species of Fusarium, although fresh weight measurements did not bring any information about the pathogenicity. The findings presented above are strong indications that long-distance transmission of Fusarium propagules may occur. Diseases caused by species of Fusarium are common in these areas. They were in the past, and are still today, a problem for greenhouses crops in Almería, and many species have been listed as pathogens on agricultural crops in this region. Saharan air masses dominate the Mediterranean regions. The evidence of long distance dispersal of Fusarium spp. by atmospheric dust and rainwater together with their proved pathogenicity must be taken into account in epidemiological studies.

First Report of Fusarium proliferatum Causing Rot of Garlic Bulbs in Spain

In October of 2008, decayed garlic bulbs (Allium sativum L. cv. Blancomor de Vallelado) were received from a producer in Segovia, Spain. In November of 2009, similar symptoms were observed on stored bulbs (cvs. Blancomor de Vallelado and Garcua) from each of 30 municipalities in northwest Segovia and Valladolid. A minimum of one sample was collected from 12 localities. Pieces of symptomatic bulbs were surface disinfested for 2 to 3 min in 0.5% NaOCl and transferred to potato dextrose agar (PDA) and Komadas media. Colonies had catenate microconidia and curved macroconidia that were usually three to five septate. Microconidia were club shaped with a flattened base, aseptate, and were produced on both mono- and polyphialides. On the basis of morphological features, the fungus was identified as Fusarium proliferatum (T. Matsushima) Nirenberg (2,3). Pathogenicity tests were conducted with 12 isolates of the fungi following the method of Dugan et al. (1). Each assay with an isolate consisted of six cloves (cv. Blancomor de Vallelado) disinfested in 0.5% NaOCl for 45 s, rinsed with sterile water, and injured to a depth of 4.5 mm with a probe 1 mm in diameter. The wound was filled with PDA colonized by the appropriate isolate. Six cloves for each tested isolate received sterile agar as a control. The cloves were incubated at 25°C for 5 weeks. The test was repeated once with cv. Garcua. All isolates produced water-soaked, tan lesions. An isolate of the fungus was deposited in the collection of the Plant Production Department of the University of Almeria. No fungi were recovered from the control cloves. F. proliferatum has been reported on garlic in the northwestern United States (1) and Serbia (4). To our knowledge, this is the first report of a Fusarium sp. in the section Liseola attacking garlic in Spain. The fungus seems to be well established on this host in Spain. References: (1) F. M. Dugan et al. Phytopathology 155:437, 2007. (2) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University Press, University Park, 1983. (3) H. Nirenberg et al. Mycologia 90:434, 1998. (4) S. Stankovic et al. Eur. J. Plant Pathol. 118:165, 2007.

The Interactive Effects of Temperature and Osmotic Potential on the Growth of Aquatic Isolates of Fusarium culmorum

The mycelial growth of 10 Fusarium culmorum strains isolated from water of the Andarax riverbed in the provinces of Granada and Almeria in southeastern Spain was tested on potato-dextrose-agar adjusted to different osmotic potentials with either KCl or NaCl (−1.50 to −144.54 bars) at 10°C intervals ranging from 15° to 35°C. Fungal growth was determined by measuring colony diameter after 4 d of incubation. Mycelial growth was maximal at 25°C. The quantity and capacity of mycelial growth of F. culmorum were similar at 15 and 25°C, with maximal growth occurring at −13.79 bars water potential and a lack of growth at 35°C. The effect of water potential was independent of salt composition. The general growth pattern of Fusarium culmorum growth declined at potentials below −13.79 bars. Fungal growth at 25°C was always greater than growth at 15°C, at all of the water potentials tested. Significant differences were observed in the response of mycelia to water potential and temperature as main and interactive effects. The number of isolates that showed growth was increasingly inhibited as the water potential dropped, but some growth was still observable at −99.56 bars. These findings could indicate that F. culmorum strains isolated from water have a physiological mechanism that permits survival in environments with low water potential. Propagules of Fusarium culmorum are transported long distances by river water, which could explain the severity of diseases caused by F. culmorum on cereal plants irrigated with river water and its interaction under hydric stress or moderate soil salinity. The observed differences in growth magnitude and capacity could indicate that the biological factors governing potential and actual growth are affected by osmotic potential in different ways.

Comparative study of the pathogenicity of seabed isolates of Fusarium equiseti and the effect of the composition of the mineral salt medium and temperature on mycelial growth

The pathogenicity of seven strains of Fusarium equiseti isolated from seabed soil was evaluated on different host plants showing pre and post emergence damage. Radial growth of 27 strains was measured on culture media previously adjusted to different osmotic potentials with either KCl or NaCl (-1.50 to -144.54 bars) at 15o, 25o and 35o C. Significant differences and interactive effects were observed in the response of mycelia to osmotic potential and temperature.

Stemphylium Leaf Blight of Garlic (Allium sativum ) in Spain: Taxonomy and In Vitro Fungicide Response.

The most serious aerial disease of garlic is leaf blight caused by Stemphylium spp. Geographical variation in the causal agent of this disease is indicated. Stemphylium vesicarium has been reported in Spain, whereas S. solani is the most prevalent species recorded in China. In this study, Stemphylium isolates were obtained from symptomatic garlic plants sampled from the main Spanish production areas. Sequence data for the ITS1–5.8S–ITS2 region enabled assignation of the isolates to the Pleospora herbarum complex and clearly distinguished the isolates from S. solani. Conidial morphology of the isolates corresponded to that of S. vesicarium and clearly discriminated them from S. alfalfae and S. herbarum on the basis of the size and septation pattern of mature conidia. Conidial morphology as well as conidial length, width and length:width ratio also allowed the Spanish isolates to be distinguished from S. botryosum and S. herbarum. Control of leaf blight of garlic is not well established. Few studies are available regarding the effectiveness of chemical treatments to reduce Stemphylium spp. incidence on garlic. The effectiveness of nine fungicides of different chemical groups to reduce Stemphylium mycelial growth in vitro was tested. Boscalid + pyraclostrobin (group name, succinate dehydrogenase inhibitors + quinone outside inhibitors), iprodione (dicar-boximide), and prochloraz (demethylation inhibitors) were highly effective at reducing mycelial growth in S. vesicarium with EC50 values less than 5 ppm. In general, the effectiveness of the fungicide was enhanced with increasing dosage.