Identification and Expression of Some Plant Cell Wall-Degrading Enzymes Present in Three Ontogenetics Stages of Thecaphora frezii, a Peanut (<i>Arachis hypogaea</i>L.) Pathogenic Fungus

Peanuts can be affected by the presence of pathogenic microorganisms. The fungus Thecaphora frezii (T. frezii), which belongs to the taxonomic class Ustilaginomycetes, is the causal agent of the disease known as “peanut smut”. The life cycle of this fungus includes three stages, namely teliospores, basidiospores and hyphae. In the hyphae stage, infection occurs in the peanut plant, which requires the involvement of some enzymes secreted by the fungus. These include the Plant Cell Wall-Degrading Enzymes (PCWDEs), which degrade various polysaccharides. This study aimed to identify the presence of transcript for enzymes belonging to the PCWDEs from three stages of T. frezii. For this, total RNA was extracted from the three ontogenetic stages of T. frezii. These samples were analyzed using an RNA-Seq approach and some transcripts were quantified using Real Time PCR. The analysis of the data provided by the RNA-Seq of the three T. frezii stages, it was possible to identify some transcripts that could encode enzymes compatible with polysaccharides degradation that are part of the plant cell wall. In T. frezii transcriptome, 40 deduced proteins would be enzymes with functions of PCWDEs were identified. They were divided into 27 glycoside hydrolases;two polysaccharide lyases;three carbohydrate esterases and eight enzymes with auxiliary activities. In addition, the fungal SNF1 gene was identified whose activity could be affected by high glucose level, and indirectly influence the levels of some PCWDEs. The analysis of the PCWDEs could help to understand part of the fungal infection process and possibly find substances that can control its development.

Essential Oils from the Leaves and Flowers of Callistemon viminalis:Chemical Characterization and Evaluation of the Insecticide and Antifungal Activities

The chemical compositions of the essential oils from the leaves and flowers of Callistemon viminalis and their insecticide and fungitoxic activities were determined. The essential oil was extracted by the hydrodistillation method using a modified Clevenger apparatus. The chemical characterization was performed by GC-MS and GC-FID. The evaluation of the insecticidal activity was performed with the Myzus persicae aphid, and the antifungal potential was determined via the inhibition of the mycelial growth of Alternaria alternata, Fusarium oxysorum and Botrytis cinérea phytopathogenic fungi. The principal components of the essential oils from the leaves and flowers were eucalyptol (84.60% and 61.47%), α-pinene (10.28% and 21.48%) and α-terpineol (2.59% and 2.79%), respectively. The use of a 0.5% concentration of the essential oil from the flowers influenced the preference of aphids and their reproduction. The number of adult aphids decreased within a period of 48 hours in the presence of the essential oil from the leaves. In the test with no chance of choice, the mean number of adults decreased with both oils within 48 hours. No inhibition of the mycelial growth of the A. alternate microorganism was observed in any of the treatments with the two essential oils. However, the growth of F. oxysporum and B. cinerea fungi was inhibited. The mycelial growth of F. oxysporum was inhibited with the concentration of 100 μL&#183;Lˉ1 of the oil from the flowers and 250 μL&#183;Lˉ1 of the oil from the leaves. The inhibition of the growth of B. cinerea was observed at concentrations of 500 and 100 μL&#183;Lˉ1 for the oils from the flowers and leaves, respectively. The essential oils presented insecticidal and antifungal potentials. However, further studies are needed for these oils to be used in integrated pest management.

Improving ITS sequence data for identification of plant pathogenic fungi

Plant pathogenic fungi are a large and diverse assemblage of eukaryotes with substantial impacts on natural ecosystems and human endeavours.These taxa often have complex and poorly understood life cycles,lack observable,discriminatory morphological characters,and may not be amenable to in vitro culturing.As a result,species identification is frequently difficult.Molecular(DNA sequence)data have emerged as crucial information for the taxonomic identification of plant pathogenic fungi,with the nuclear ribosomal internal transcribed spacer(ITS)region being the most popular marker.However,international nucleotide sequence databases are accumulating numerous sequences of compromised or low-resolution taxonomic annotations and substandard technical quality,making their use in the molecular identification of plant pathogenic fungi problematic.Here we report on a concerted effort to identify high-quality reference sequences for various plant pathogenic fungi and to re-annotate incorrectly or insufficiently annotated public ITS sequences from these fungal lineages.A third objective was to enrich the sequences with geographical and ecological metadata.The results-a total of 31,954 changes-are incorporated in and made available through the UNITE database for molecular identification of fungi(http://unite.ut.ee),including standalone FASTA files of sequence data for local BLAST searches,use in the next-generation sequencing analysis platforms QIIME and mothur,and related applications.The present initiative is just a beginning to cover the wide spectrum of plant pathogenic fungi,and we invite all researchers with pertinent expertise to join the annotation effort.

Design, synthesis and SAR study of novel sulfonylurea derivatives containing arylpyrimidine moieties as potential anti-phytopathogenic fungal agents

Acetohydroxyacid synthase(AHAS) was considered as a promising target for antifungal agents.Herein,three series of novel sulfonylureas(SUs) 9-11 containing aromatic-substituted pyrimidines were designed and synthesized according to pharmacophore-combination and bioisosterism strategy.The in vitro fungicidal activities against ten phytopathogenic fungi indicated that most of the title compounds exhibited broad-spectrum and excellent fungicidal activities.Based on the preliminary fungicidal activities,a CoMFA model was constructed and the 3 D-QSAR analysis indicated that either a bulky group around the 5-position of the pyrimidine ring or electropositive group around the 2-position of the benzene ring would be favour to fungicidal activities.In order to study interaction mechanism,10 k was automatically docked into yeast AHAS and it further indicated that bearing bulky groups-aryl at the pyrimidine ring was critical to enhance antifungal activities.It revealed that the antifungal activity of derivatives 9-11 probably results from the inhibition of fungal AHAS.Thus,the present results strongly showed that SUs should be considered as lead compounds or model molecules to develop novel antiphyt o pathogenic fungal agents.