A total of 82 fungal isolates was obtained from wheat kernel samples affected by fusarium head blight collected from 20 locations in southern Brazil. Polymerase chain reaction (PCR) assays were used to characterize trichothecene mycotoxin genotypes [deoxynivalenol (DON), nivalenol (NIV) and two acetylated derivatives of DON]. To identify isolates that producing DON and NIV, portions of the Tri13 gene were amplified. To identify 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) genotypes, portions of Tri3 and Tri12 were amplified. Nearly all of the isolates studied (76/82) were of the DON/15-ADON genotype. Six of the isolates were of the NIV genotype. The DON/3-ADON genotype was not observed. Portions of three genes were sequenced from representative isolates of the NIV and DON/ 15-ADON genotypes and compared with sequences from curated reference isolates of Fusarium in GenBank. blast queries for individual gene sequences and pairwise comparisons of percentage identity and percentage divergence based on 1676 bp of concatenated DNA sequence suggested that the isolates representing the DON/15-ADON genotype were Fusarium graminearum sensu stricto and the isolates representing the NIV genotype were Fusarium meridionale . This is the first detailed report of trichothecene mycotoxin genotypes of F. graminearum and F. meridionale in Brazil.
Plants express different defence mechanisms in response to pathogens. Understanding the recognition of pathogenassociated molecular patterns (PAMPs) by specific receptors, and the role of endogenous signals such as AtPep1 that regulate expression of genes in Arabidopsis thaliana, has aided the understanding of the defence mechanisms in different species. The aim of this study was to identify possible orthologous sequences of AtPROPEPs in tomato (Solanum lycopersicum) and characterize its role in resistance to necrotrophic pathogens. The presence of an orthologue of the A. thaliana AtPROPEP1 gene in S. lycopersicum, SlPROPEP, by in silico analysis, is reported here. This has 96% identity with the C-terminal region of a previously described potato peptide, another possible orthologue of AtPep1. A virus-induced gene silencing (VIGS) system was employed to investigate the role of the SlPROPEP. Silencing of SlPRO-PEP in tomato made plants more susceptible to Pythium dissotocum; approximately 30% of SlPROPEP-silenced plants showed stem constriction compared with 4% in control plants. Furthermore, quantification of P. dissotocum by qPCR revealed that the increase in symptom severity in SlPROPEP-silenced plants was associated with a 15 times increase in growth of the pathogen compared to control plants. Silencing of SlPROPEP also resulted in decreased expression of genes involved in plant defence against pathogens, such as PR-1, PR-5, ERF1, LOX-D and DEF2. These results suggest that SlPROPEP is involved in tomato resistance to P. dissotocum and probably acts as a pathogen-associated molecular pattern through signalling pathways mediated by jasmonic acid/ethylene (JA/ET).
Erwinia carotovora subsp. atroseptica (Eca), E. carotovora subsp. carotovora (Ecc) and E. chrysanthemi (Ech) may cause potato (Solanum tuberosum) blackleg. To determine the occurrence of these pathogens in the conditions found in the State of Rio Grande do Sul (RS), potato plants showing blackleg symptoms were harvested from 22 fields in nine counties in Serra do Nordeste, Planalto, Depressão Central, and Grandes Lagoas, from September to December of 1999 (Spring-Summer season). Green pepper (Capsicum annuum) fruits were used as a host to enrich for pectolytic erwinia from potato stems with blackleg symptoms. Bacteria were subsequently isolated on non-selective medium. Isolates that were Gram-negative, facultatively anaerobic, and pitted crystal-violet-pectate medium were tested for biochemical traits to identify the species and subspecies. Four hundred strains were identified as either Eca, Ecc or Ech. Although the three erwinias were found in RS potato fields, only three strains of Ech were found in one field. Frequencies of Eca and Ecc were 55 and 42%, respectively. Eight strains could not be assigned based on the biochemical characterization.
;Canavalia ensiformis (jackbean) seeds contain the proteins urease and canatoxin, a variant form of the jackbean urease. Here we have cloned a cDNA encoding another isoform of urease, called JBURE-II. This cDNA was obtained by RT-PCR using as template total RNA extracted from C. ensiformis tissues. Nucleotide sequence analysis showed that JBURE-II clones share 86% similarity with known jackbean urease. The presence in C. ensiformis of a family of urease-related genes with at least three members was demonstrated by Southern blot analysis. In order to understand the pattern of expression of the JBURE-II gene, we collected tissue samples from different stages of flower and embryo development. The results of RT-PCR show that JBURE-II is expressed from flower buds throughout seed maturation. Semi-quantitative RT-PCR indicates that expression of urease and JBURE-II genes is induced in seedlings and in leaves treated with abscisic acid, a phytohormone involved in seed maturation and wound response. This work constitutes the first report on the presence of a family of urease genes in jackbean, and provides characterization of a cDNA encoding a new member of this gene family.
Com a finalidade de verificar a ocorrência e a distribuição de biovares de Ralstonia solanacearum no Estado do Rio Grande do Sul, plantas com sintomas de murcha-bacteriana foram coletadas, na primavera de 1999, em 25 lavouras de batata (Solanum tuberosum) em dez municípios das quatro áreas de produção: Serra do Nordeste, Planalto Superior, Depressão Central e Grandes Lagoas. A determinação da espécie foi feita com teste sorológico através de ELISA e da biovar com base no metabolismo oxidativo de açúcares e álcoois. Noventa e quatro porcento dos 490 isolados foram identificados como biovar 2. A ocorrência da biovar 1 foi registrada na Serra do Nordeste e Depressão Central. A ocorrência das biovares não mostrou relação com a temperatura média local ou cultivar de batata.
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