Gladys Albina Lori

Fusarium graminearum and deoxynivalenol contamination in the durum wheat area of Argentina

Fusarium graminearum head blight of wheat is a destructive disease of the worlds wheat-growing areas. This work was performed to analyze the distribution and contamination of deoxynivalenol (DON) and its relationship with F. graminearum kernel invasion in Argentina durum wheat area during two consecutive harvests. A total of 147 samples (cultivars and lines) of durum wheat from 5 locations of the major cropping area (Southern Buenos Aires Province) were analyzed. Percentage of F. graminearum kernel infection was evaluated following the blotter test (ISTA method) and fusarotoxins were analyzed by thin layer chromatography. None of the varieties and lines were free of F. graminearum infection. In the first harvest fungal invasion was very low. From 40 samples, 55% showed DON contamination but only 4 samples (10%) were higher than 2 ppm. In the second harvest, a crop year conducive to scab development, the highest level of F. graminearum kernel invasion observed was 42% on a sample from the humid area (eastern Buenos Aires Province) DON was detected in 47 (78.2%) of 60 samples analyzed and 19 (31.6%) showed levels of DON higher than those established in the guidelines in Canada and USA for food and feedstuff. In both years all locations situated in the humid area showed levels ranging from 0 to > 8 ppm. Within the durum wheat area differences among locations were found. This analysis indicates the need for more information on the problem and distribution of Fusarium mycotoxins in durum wheat grown in Argentina.

Stability of maize resistance to the ear rots caused by Fusarium graminearum and F. verticillioides in Argentinian and Canadian environments

Sources of resistance to Fusarium spp. are needed to develop maize hybrids resistant to the accumulation of fungal mycotoxins in the grain. In a search for resistant germplasm in 1999 and 2000, a set of Argentinian maize populations was evaluated in Ottawa, Canada, for resistance to ear rots after inoculation with local isolates of Fusarium verticillioides and F. graminearum. Sixteen of these populations, varying in observed resistance levels, were re-evaluated in 2003 and 2004 in Pergamino, Argentina, using local isolates of the same fungi. Conidial suspensions of each fungal species were inoculated into the silk channel of primary ears. Disease severity was assessed after physiological maturity using a scale based on the percentage of visibly infected kernels. Genotype effect was more important than genotype-by-fungal species or genotype-by-fungal species-by-environment interaction effects. In addition, disease severity levels associated with each fungal species were positively correlated (P < 0.05) (r = 0.90, r = 0.81, r = 0.87 and r = 0.53, in Ottawa 1999 and 2000, and Pergamino 2003 and 2004, respectively). Populations ARZM 01107, ARZM 07138, ARZM 10041, ARZM 13031, ARZM 16002 and Pora INTA exhibited the highest and most stable resistance to both species. Considering that disease resistance exhibited low specificity to the environment and to the fungal species in evaluations conducted in a wide range of environments and with fungal isolates collected from different hemispheres, the most resistant populations are potential sources of genes for stable resistance to these Fusarium spp.

Toxigenic Capacity and Trichothecene Production by Fusarium graminearum Isolates from Argentina and Their Relationship with Aggressiveness and Fungal Expansion in the Wheat Spike

At least 20 epidemics of Fusarium head blight (FHB) of wheat have been registered in the last 50 years in Argentina, with variable intensity. Damage induced by the disease is further aggravated by the presence of mycotoxins in affected grains that may cause health problems to humans and animals. The trichothecene chemotype was analyzed for 112 isolates of Fusarium graminearum from Argentina by polymerase chain reaction and two field trials were conducted to study the aggressiveness of a subsample of 14 representative isolates and to analyze deoxynivalenol (DON) production in planta and in vitro. All isolates belonged to the 15-acetyl-DON chemotype. Significant differences were observed in both the symptom severity induced in wheat spikes and the in vivo DON production, and a close correlation was found between these two variables. However, in vitro toxigenic potential was not correlated with the capacity of F. graminearum isolates to produce DON under natural conditions. The progress of infection in the rachis of inoculated wheat spikes was analyzed and the pathogen presence verified in both symptomatic and symptomless spikes. Even isolates with a limited capacity to induce symptoms were able to colonize the vascular tissue and to produce considerable amounts of DON in planta.

Use of solid phase microextraction coupled to capillary gas chromatography-mass spectrometry for screening Fusarium spp. based on their volatile sesquiterpenes

Solid phase microextraction (SPME) coupled to capillary gas chromatography (CGC) and mass spectrometry (MS) was used to evaluate the use of fungal volatiles to discriminate Fusarium species from wheat cultivars in the Argentina pampa region. Monosporic fungal isolates were grown on rice in sealed containers for 1 week and volatile organic compounds (VOC) were sampled for 30 min from the head space by SPME and analysed by CGC and CGCMS. VOC profiles of Fusarium species F. graminearum, F. poae, F. equiseti, F. verticillioides and F. oxysporum were discriminated by comparison of their profiles in the elution zone corresponding to sesquiterpenes. Trichothecene-producer and non-trichothecene producer Fusarium species were separated by the presence of trichodiene in their VOC fingerprints. Within trichothecene-producers, F. graminearum, F. poae and F. equiseti differed on the structure of their volatile sesquiterpenes. This technique might be also helpful to detect F. graminearum, the major head blight disease-...

Early detection of toxigenic Fusarium graminearum in wheat

Fusarium graminearum (Schwabe) contaminates agricultural crops and commodities with trichothecenes, mostly deoxynivalenol and its acetyl-derivatives. Current techniques available to detect final mycotoxin contamination products usually require an extended time lag between sampling and the corresponding report, and include different clean-up steps and eventually derivatisation. This study was aimed to develop a methodology to detect toxigenic F. graminearum prior to mycotoxin production. Headspace solid-phase microextraction coupled to capillary gas chromatography is shown to be useful to predict the potential of trichothecene mycotoxin formation by detecting the presence of F. graminearum at early stages of fungal growth in wheat cultivars, based on the detection of trichodiene (TRI), the volatile intermediate of trichothecenes. We showed that TRI is a useful marker to detect toxigenic Fusarium in wheat spikes from live plants, regardless of the actual development of Fusarium head blight (FHB). This is th...

Fusarium Wilt of Lettuce Caused by Fusarium oxysporum f. sp. lactucae in Argentina

The green belt area surrounding the city of La Plata, Argentina, produces more than 70% (around 280 ha) of the lettuce (Lactuca sativa L.) cultivated under greenhouse for fresh consumption in the country. In February 2011, April 2012, and December 2013, butterhead lettuce plants from cv. Lores showing wilt and stunted growth symptoms, red-to-brown discoloration of vascular tissues, and yellow leaves were found in greenhouses in La Plata. Sections of tap root, crown and stem from symptomatic plants showing dark-brown streaking of the vascular tissue were surface sterilized and isolations were made. A total of 21 monosporic isolates obtained from different lettuce production fields were identified as Fusarium oxysporum Schltdl. based on morphological characteristics (2). Vegetative compatibility group (VCG) analysis (1) was conducted on 11 of these isolates and all isolates belonged to the same VCG. To fulfil Kochs postulates, two pathogenicity tests were conducted with these 11 isolates in winter (July 2012) and summer (December 2013). Healthy 20-day-old butterhead lettuce seedlings of two cultivars (Reina de Mayo and Lores in the winter and summer tests, respectively) were inoculated by dipping the roots of each plant in a spore suspension (~3 × 105 CFU ml-1), planted in 1-liter pots containing autoclaved soil and grown in a greenhouse with only natural daylight. Control treatments were prepared by dipping the seedling roots in sterilized distilled water. All inoculated plants showed wilt symptoms 15 to 20 days after inoculation (dai) and 45 to 50 dai in the summer and winter pathogenicity tests, respectively. The delay in the appearance of symptoms observed during the winter test is consistent with the effect of planting date on the development and final incidence of Fusarium wilt of lettuce reported by Matheron et al. (3). No symptoms were observed in control treatments. F. oxysporum was re-isolated from vascular tissues of the stems of symptomatic plants and the formae speciales lactucae J.C. Hubb. & Gerik was confirmed by PCR using the specific primer pair GYCF1 and R943 (4). The identification of only one VCG for the tested isolates agrees with the hypothesis of seed transmission of the pathogen, which might explain the dissemination of Fusarium wilt of lettuce in geographically distant areas (2). Studies are being carried out to determine the race of these Argentinian isolates of F. oxysporum f. sp. lactucae. To our knowledge, this is the first report of F. oxysporum f. sp. lactucae infecting lettuce in Argentina. References: (1) J. C. Correll et al. Phytopathology 77:1640, 1987. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) M. E. Matheron et al. Plant Dis. 89:565, 2005. (4) G. C. Y. Mbofung and B. M. Pryor. Plant Dis. 94:860, 2010.

Basal Rot of Hylocereus undatus Caused by Fusarium oxysporum in Buenos Aires, Argentina

Night-blooming cereus (Hylocereus undatus A. Berger) is generally used as rootstock of ornamental cactus because of its rapid growth and tolerance to humid substrates. Since 2002, in Gran Buenos Aires, a new disease has been observed in grafted crops in all production stages. Incidence was as much as 30% in many greenhouses. Symptoms consisted of soft rot that started near the soil line and developed upward until it affected all the rootstock. The scion was not rotten, but died as a consequence of rootstock infection. All the roots were symptomless. For pathogen isolation, symptomatic tissues were surface disinfected by a 1-min immersion in 0.2% NaOCl, placed on potato dextrose agar (PDA), and incubated at 22 ± 2°C. Only a Fusarium spp. was consistently isolated in pure culture. Twenty healthy potted night-blooming cereus plants, 10 of them previously injured with needles on the rootstock near the soil line, were gently removed from the substrate and inoculated by a 1-min immersion of their base in a suspension of 1.4 × 106 conidia per ml of sterile distilled water, prepared from 15-day-old cultures. Ten control plants, five of them previously injured, were immersed in sterile distilled water. Inoculated and noninoculated plants were replanted in the original substrate, placed in a climatic chamber at 22 ± 2°C, and monitored for disease expression. Basal rot was observed on all injured inoculated plants 12 days after inoculation. Symptoms on undamaged plants appeared 22 days after inoculation. After 72 days of incubation, all inoculated plants were totally rotten. Control plants remained symptomless. The same pathogen was reisolated to fulfill Kochs postulates. For species identification, single-spore cultures were grown on PDA and carnation leaf agar in a climatic chamber at 23 ± 2°C with a 12-h darkness/near ultraviolet light regimen. The micromorphology and culture features, mainly conidial ontogeny, were consistent with descriptions of Fusarium oxysporum Schlechtend.:Fr. (1). The pathogen was able to penetrate undamaged tissues. Needle injuries accelerated infection. To our knowledge, this is the first report of Fusarium oxysporum on H. undatus in Gran Buenos Aires, Argentina. A culture of the pathogen was deposited at the fungal collection of PRHIDEB-CONICET (University of Buenos Aires) as BAFCult 3158. Reference: (1) P. E. Nelson et al. Fusarium species. An Illustrated Manual for Identification. The Pennsylvania State University Press, University Park, PA, 1983.

First report of Fusarium solani causing stunt on lisianthus.

Fusarium solani Mart. (Sacc.) is the causal agent of stem rot and damping-off of lisianthus (Eustoma grandiflorum (Raf.) Shinn.) (1). Since the end of the 1980s, when this flower crop was introduced in Argentina, it has been affected by a basal stem rot (2). A previously undescribed disease was observed in 100% of the greenhouses in the Buenos Aires Province that grow lisianthus. Symptoms that developed after seedlings were transplanted included stunting, shortened internodes with reduced stem diameter, and small narrow leaves that were a dull green color. Some affected plants turned yellow-brownish and died 2 to 3 months after transplanting. Other plants recovered but produced low quality flowers later than normal. A third group of plants remained stunted (5 to 10 cm high) until the last flower harvest (about 8 to 10 months). F. solani was consistently isolated from basal stems and roots of diseased plants. For pathogenicity tests, inoculum was produced by culturing the fungus for 10 days in petri dishes containing sterile moistened rice. Inoculum was air dried, crushed, and mixed with soil that had been autoclaved at 112°C for 40 min on each of two consecutive days. The propagules in the soil were estimated by soil plate dilutions on the Nash & Snyder-PCNB medium at a ratio of about 104 CFU/g soil. Twenty plants of each cultivar Echo White and Echo Blue, whose roots had been pruned, were planted in both infested and noninfested soil. After about 40 days, stunting was observed in 85% of the inoculated plants, while controls remained asymptomatic. F. solani was reisolated from symptomatic plants, thus fulfilling Kochs postulates. A test also was conducted in a commercial greenhouse that produced lisianthus for several years, in which healthy plants were planted in three plots fumigated with methyl bromide and in three nonfumigated plots. The mean cfu/g soil of F. solani in the methyl-bromide treated plots was 5 × 102 and 1.6 × 104 CFU/g in the nontreated plot. After 120 days, the incidence of stunting in the treated plots was 0.6 and about 88% in the control plots. F. solani was recovered from symptomatic plants. Because disinfestation of soil is generally practiced in flower production, stunted plants are limited and can be confused with root problems. This is the first report of F. solani causing stunt on lisianthus. References: (1) J. J. Taubenhaus and W. N. Ezekiel. Phytopathology 24:19, 1934. (2) S. M. Wolcan and G. A. Lori. Invest. Agr. Prot. Veg. 11:465, 1996.

Fusarium moniliforme and F. proliferatum Isolated from Crown and Root Rot of Asparagus and Their Association with Asparagus Decline in Argentina

During the summer of 1995-1996, an 80-ha field of 6-year-old asparagus plants (cv. UC 72) in Saladillo (Province of Buenos Aires) was affected by a decline syndrome (1). The plants showed a decline in vigor and approximately 60 to 70% of the plants died. The symptomatic plants were chlorotic, stunted, with stem lesions and crown and root rot. Fusarium moniliforme and F. proliferatum were isolated from vascular and epidermal tissues of roots, crowns, and stems. Identification of Fusarium to species was made by examining conidiogenous cells from colonies cultured on KCl medium (2). Microconidia were born in long and short chains and false heads. The isolates were identified based on the the presence of polyphialides in F. proliferatum and their absence in F. moniliforme, which produces monophialides only (2). In two separate trials, asparagus seeds (cv. UC 72) were surface sterilized and placed in steamed soil infested with a conidial suspension of each species. The viable propagules in the soil (CFU per g) were estimated by soil plate dilutions on Nash & Snyder-PCNB (pentachloronitrobenzene) medium. The F. moniliforme and F. proliferatum soil densities were 19.2 × 103 and 23 × 103 CFU per g of soil, respectively. The pots were placed in the greenhouse on different benches to avoid cross-contamination. After 4 months, inoculated plants showed root and crown discoloration. F. moniliforme and F. proliferatum were reisolated (64 and 75%, respectively) from discolored portions of internal and external root and crown tissues. Although the stems did not show symptoms, F. moniliforme and F. proliferatum were also recovered (27 and 38%, respectively) from asymptomatic tissues. Six months after inoculation the plants developed chlorotic symptoms with crown and root rot, and then wilted. F. moniliforme and F. proliferatum were reisolated from root systems, crowns, and stems of all inoculated plants. F. moniliforme and F. proliferatum are involved in corn stalk and ear rot in Argentina. Corn and asparagus are frequently grown in close proximity and often follow one another at a particular site. Airborne and soil debris carrying F. moniliforme and F proliferatum from corn may be an additional source of inoculum for asparagus in Argentina. The results indicate that the presence of F. moniliforme and F. proliferatum is a factor that contributes to asparagus decline in Argentina. References: (1) W. H. Elmer et al. Plant Dis. 80:117, 1996. (2) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University, University Park, 1983.

Wheat fusarium head blight 2001 epidemic in the southern Argentinian pampas

Centro de Investigaciones de Fitopatologia (CIDEFI), Facultad de Ciencias Agrarias y Forestales, UNLP. 60 y 119, (1900) La Plata. E-mail:fitopato@ceres.agro.unlp. edu.ar. Comision de Investigaciones Cientificas de la Prov. de Buenos Aires (CIC), ArgentinaAutor para correspondencia: Marcelo CarrazaData de chegada:29/03/2005. Aceito para publicacao em:06/11/2007 1189Wheat Head Blight or Scab is frequently a destructive fungal diseasecaused by several