RESUMO
Fermentation is an important industrial process for microbial metabolite development and has wide applications in various fields. Aspergillus is the most important genus of fungi used for the production of microbial enzymes such as cellulase. The Aspergillus genome encodes various cellulolytic enzymes. In this study, we assayed the gene expression and cellulolytic enzyme production of three isolates: A. niger (KSU009), A. terreus (KC462061), and A. flavus (KSU014). Two fermentation systems, submerged fermentation and biofilm fermentation (BF), were used for this purpose. Gene expression analysis by RT-PCR showed that cbhB, exo, eglA, eglB, eglC, and ß-actin genes were differentially expressed in the two fermentation systems for these three isolates during enzyme production. Furthermore, the expression of all genes was found to be higher in the BF system. The six genes were not expressed in the isolates with no cellulolytic enzyme production. The isolates were identified by morphological and molecular methods, which were based on macroscopic characteristics and sequence analysis of ITS1, ITS2, and the 5.8S regions of rDNA.
Assuntos
Aspergillus/enzimologia , Aspergillus/genética , Celulase/genética , Regulação Fúngica da Expressão Gênica , Genes Fúngicos , Aspergillus/isolamento & purificação , Sequência de Bases , Celulase/metabolismo , Fermentação/genética , Filogenia , Especificidade da Espécie , Transcrição Gênica , beta-Galactosidase/metabolismoRESUMO
Twelve species from six fungal genera were found to be associated with corn (Zea mays L.) grain samples collected from three main regions of Saudi Arabia. The average frequencies of the most common genera were Aspergillus (11.4%), Fusarium (9.5%), Penicillium (5.1%), and Alternaria (5.8%). Fifteen isolates of Aspergillus flavus were screened by HPLC for their ability to produce aflatoxins (AF). The percentage of aflatoxigenic A. flavus isolates was 53%. Eight isolates produced AF, at concentrations ranging 0.7-2.9 ppb. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular markers were used to genetically characterize isolates of A. flavus and to discriminate between the aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, which was useful for genetic characterization. The clustering in the RAPD and ISSR dendrograms obtained was unrelated to geographic origin. The RAPD and ISSR markers could not discriminate between aflatoxigenic and non-aflatoxigenic isolates, but the ISSR primers were somewhat better.
Assuntos
Aflatoxinas/toxicidade , Aspergillus flavus/genética , Aspergillus flavus/isolamento & purificação , Sementes/microbiologia , Manejo de Espécimes , Zea mays/microbiologia , Aspergillus flavus/efeitos da radiação , Primers do DNA/metabolismo , Geografia , Filogenia , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA Polimórfico , Arábia Saudita , Raios UltravioletaRESUMO
Twelve species belonging to six fungal genera were found to be associated with wheat (Triticum aestivum L.) grain samples collected from three main regions in Saudi Arabia. The most common genera (average frequency) were Aspergillus (14.3%), Fusarium (29.1%), Penicillium (9.3%), and Alternaria (8.2%). Nineteen isolates of Aspergillus flavus were screened for their ability to produce aflatoxins using HPLC. Thirteen isolates produced aflatoxins ranging from 0.5 to 2.6 µg/kg. Inter-simple sequence repeats (ISSR), and random amplified polymorphic DNA (RAPD) molecular markers were used, with the aim of genetically characterizing strains of A. flavus to discriminate between aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, useful for genetic characterization. Clustering based on RAPD and ISSR dendograms was unrelated to geographic origin. RAPD and ISSR markers were not suitable to discriminate aflatoxigenic and non-aflatoxigenic isolates, but ISSR primers were better compared to RAPD.
Assuntos
Aflatoxinas/genética , Aspergillus flavus/genética , Variação Genética , Aflatoxinas/classificação , DNA Fúngico/genética , Grão Comestível/genética , Grão Comestível/parasitologia , Repetições de Microssatélites , Técnica de Amplificação ao Acaso de DNA Polimórfico , Arábia Saudita , Triticum/genética , Triticum/parasitologiaRESUMO
Twenty-one isolates of Rhizoctonia solani were categorized into three anastomosis groups consisting of AG-4-HG-I (eight isolates), AG-2-2 (nine isolates) and AG-5 (four isolates). Their pathogenic capacities were tested on cotton cultivar Giza 86. Pre-emergence damping-off varied in response to the different isolates; however, the differences were not significant. Soluble proteins of the fungal isolates were electrophoresed using SDS-PAGE and gel electrophoreses. A dendrogram of the protein banding patterns by the UPGMA of arithmetic means placed the fungal isolates into distinct groups. There was no evidence of a relationship between protein dendrogram, anastomosis grouping or level of virulence or geographic origin. The dendrogram generated from these isolates based on PCR analysis with five RAPD-PCR primers showed high levels of genetic similarity among the isolates from the same geographical locations. There was partially relationship between the genetic similarity and AGs or level of virulence or geographic origin based on RAPD dendrogram. These results demonstrate that RAPD technique is a useful tool in determining the genetic characterization among isolates of R. solani.
