Verónica Fabiana Consolo

Mating-type Distribution and Fertility Status in Magnaporthe grisea Populations from Argentina

Isolates of Magnaporthe grisea causing gray leaf spot on rice were collected in Argentina and analyzed for mating distribution and fertility. One hundred and twenty-five isolates of M.grisea were collected from rice plants between 2000 and 2003. Each isolate was tested for mating type through a polymerase chain reaction based assay. All M. grisea isolates from Argentina belonged to a single mating type, MAT1.1. The fertility status of isolates was determined using controlled crosses in vitro, pairing each isolate with GUY11 and KA9 (MAT1.2 standard hermaphroditic testers). Production of perithecia was scarce among isolates of the blast pathogen since a low percentage of them (7.2%) developed perithecia with only one of the fertile tester (KA9); all crosses failed with the other tester strain. Asci and ascospores were not observed. The presence of only one mating type and the absence of female fertile isolates indicate that sexual reproduction is rare or absent in M. grisea populations associated with rice in Argentina.

Occurrence of Different Species of Fusarium from Wheat in Relation to Disease Levels Predicted by a Weather-Based Model in Argentina Pampas Region

Fusarium head blight (FHB) is an important disease throughout many of the world wheat-growing areas that have humid to semi-humid climate. The infection happens mainly during the anthesis of the wheat, when there have been favorable conditions of moisture and temperature. The direct relation of the infection to environmental factors makes possible the formulation of mathematical models that predict the disease. The causal agent of the FHB of the spike of wheat is attributed principally to Fusarium graminearum. High economic losses due yield decrease have been recorded in Argentina. In the present work, 67 isolates of Fusarium spp. were obtained from samples of wheat grains from Pampas region from 15 locations distributed in Buenos Aires, Entre Ríos, Santa Fe and Córboba provinces during 2006 and 2007 wheat-growing seasons. The identification of species from monosporic isolates was carried out by morphological characterization and use of species-specific PCR-based assays. Both identification criteria were necessary and complementary for the species determination, since in some cases the molecular identification was not specific. Scanty presence of F. graminearum was observed in 2006 wheat-growing season coinciding with the lack of favorable meteorological conditions for producing FHB infection events. High presence of F. graminearum isolates was observed in 2007 wheat-growing season, in accordance with moderate incidence of the disease according to spatial distribution of FHB incidence values. The aim of this report was to identify the causal agent of the FHB disease by different taxonomic criteria and to relate its occurrence with disease incidence values predicted by a weather-based model in Argentina.

Characterization of novel Trichoderma spp. isolates as a search for effective biocontrollers of fungal diseases of economically important crops in Argentina

Monoconidial cultures of 33 isolates of Trichoderma from Buenos Aires Province, Argentina were characterized on the basis of twenty eight morphological, physiological and biochemical features. All of them were screened for proteinase, endochitinase and β-1,3 glucanase activity. Universally primed PCR (UP-PCR) and inter-simple sequence repeat (ISSR) techniques were used to examine the genetic variability among isolates, which resulted in 127 bands for the total number of isolates. These results were subjected to numerical analysis revealing 20 haplotypes grouped in five clusters. The ability of Trichoderma isolates to antogonize soil-borne fungal plant pathogens using a dual culture assay was done against five fungal species: Alternaria sp., Bipolaris sorokiniana, Fusarium graminearum, F. solani, and Pyricularia oryzae. The highest inhibition values (85% RI) were obtained against B. sorokiniana and P. oryzae. Three isolates of T. harzianum named as FCCT2, FCCT3 and FCCT9 were capable of causing a high growth inhibition on four of the fungal species assayed, which was in agreement with their higher extracellular hydrolytic activity. Our results suggest that these isolates have the potential to be effective agents for biocontrol of cereal and tomato fungal pathogens.

Pathogenicity, formulation and storage of insect pathogenic hyphomycetous fungi tested against Diabrotica speciosa

Studies were conducted tosearch for fungal strains with potentialpathogenicity against Diabrotica speciosa(Germar) (Coleoptera: Chrysomelidae).Among sixteen fungal isolates screenedthe most virulent was a Beauveria bassiana(Balsamo) Vuillemin isolate (FHD13) thatcaused 70% mortality of D. speciosathird instar larvae. The LC50 value ofB. bassiana isolate FHD13 was3.48 × 1010 conidia/ml.Different temperatures (4, 17 and 26 °C)and vegetable oils (corn, sunflower and canola)used for storage did not significantly affectviability of conidia. A pathogenicity trialagainst D. speciosa larvae performed withthe corn oil formulation (1 × 108 conidia/mlof oil) caused 65% of mortality.

A conventional PCR technique to detect Septoria tritici in wheat seeds

A PCR assay was developed for detection of wheat seed naturally contaminated with Septoria tritici. S. tritici specific primers were derived from strict alignment of ITS and α-tubulin sequences of the pathogen. The specificity of four sets of synthesised oligonucleotide pairs (A, B, C and D) were tested using isolates from S. tritici, other selected fungi and wheat seeds. A single DNA fragment was amplified from S. tritici isolates with all primer pairs, whereas no product was generated from other DNA sources. S. tritici was also detected in wheat seed lots collected from plants with variable pycnidial coverage on the upper two leaves. PCR detection of as little as 0.5 pg of S. tritici genomic DNAwas possible. This is the first report on the detection of S. tritici DNA in naturally infested wheat seeds. This PCR based assay is simple, rapid, specific, sensitive and suitable for routine detection of the wheat pathogen in infested wheat seeds.

DNA fingerprint and pathotype diversity of Pyricularia oryzae populations from Argentina

The genetic diversity of the rice blast pathogen, Pyricularia oryzae, was analysed in rice-growing provinces of Argentina. A total of 161 isolates of the fungus was collected from 15 rice cultivars at nine locations during 2000-05 and characterised using Pot2-DNA fingerprinting. Based on DNA analysis (isolates with ≥70% band similarity), five lineages were identified and designated A, B, C,Dand E, with 11, 22, 4, 1 and 4 haplotypes identified, respectively. The predominant lineage, B, representing 38% of the collected isolates, was recovered from four cultivars in five locations. In contrast to lineages A and B, which did not contain a dominant haplotype, a single haplotype predominated in lineages C and E. Isolates representing all haplotypes were examined for virulence on a set of differential rice cultivars, near-isogenic lines and commercial cultivars commonly grown in Argentina, revealing 41 pathotypes and 24 international races. There was no significant association between DNA fingerprint similarities and pathotypes. Overall, these data indicated that populations of P. oryzae in Argentina are genetically simple and predominantly clonal yet have a high pathotype diversity.