RESUMO
A method for simultaneous detection of multi-mycotoxins in ten kinds of common dried fruits was been exploited by using ultra performance liquid chromatography-tandem mass spectrometry. The edible part of dried fruits was ground up with certain quantity of water. The mash was extracted by 10 mmol/L citric acid-acetonitrile, purified by C18 column, and filtered using 0. 22-μm organic filtration. All the sixteen mycotoxins were well separated in 8 min on ACQUITY UPLC BEH C18 column with acetonitrile as phase A and 10 mmol/L ammonium acetate solution containing 0. 1% formic acid as mobile phase B. Result showed that the limits of detection ( LODs) for the sixteen mycotoxins were 0. 01-1. 00 μg/L, with linearity range of 1-200 μg/L and correlation coefficients above 0 . 9981 . The average recoveries of the sixteen mycotoxins in ten matrices were 70. 48% -118. 85%, and the relative standard deviation ( RSD, n=6 ) was 0. 3% -11. 9%. This economical, fast, simple and efficient method could be used for simultaneous detection of multi-mycotoxins in different dried fruits matrixes.
RESUMO
A method for simultaneous detection of multi-mycotoxins in ten kinds of common dried fruits was been exploited by using ultra performance liquid chromatography-tandem mass spectrometry. The edible part of dried fruits was ground up with certain quantity of water. The mash was extracted by 10 mmol/L citric acid-acetonitrile, purified by C18 column, and filtered using 0. 22-μm organic filtration. All the sixteen mycotoxins were well separated in 8 min on ACQUITY UPLC BEH C18 column with acetonitrile as phase A and 10 mmol/L ammonium acetate solution containing 0. 1% formic acid as mobile phase B. Result showed that the limits of detection ( LODs) for the sixteen mycotoxins were 0. 01-1. 00 μg/L, with linearity range of 1-200 μg/L and correlation coefficients above 0 . 9981 . The average recoveries of the sixteen mycotoxins in ten matrices were 70. 48% -118. 85%, and the relative standard deviation ( RSD, n=6 ) was 0. 3% -11. 9%. This economical, fast, simple and efficient method could be used for simultaneous detection of multi-mycotoxins in different dried fruits matrixes.
RESUMO
The peroxisomal matrix proteins involved in many important biological metabolism pathways in eukaryotic cells are encoded by nucleal genes, synthesized in the cytoplasm and then transported into the organelles. Targeting and import of these proteins depend on their two peroxisomal targeting signals (PTS1 and PTS2) in sequence as we have known so far. The vectors of the fluorescent fusions with PTS, i.e., green fluorescence protein (GFP)-PTS1, GFP-PTS2 and red fluorescence protein (RFP)-PTS1, were constructed and introduced into Magnaporthe oryzae Guy11 cells. Transformants containing these fusions emitted fluorescence in a punctate pattern, and the locations of the red and green fluorescence overlapped exactly in RFP-PTS1 and GFP-PTS2 co-transformed strains. These data indicated that both PTS1 and PTS2 fusions were imported into peroxisomes. A probable higher efficiency of PTS1 machinery was revealed by comparing the fluorescence backgrounds in GFP-PTS1 and GFP-PTS2 transformants. By introducing both RFP-PTS1 and GFP-PTS2 into Deltamgpex6 mutants, the involvement of MGPEX6 gene in both PTS1 and PTS2 pathways was proved. In addition, using these transformants, the inducement of peroxisomes and the dynamic of peroxisomal number during the pre-penetration processes were investigated as well. In summary, by the localization and co-localization of PTS1 and PTS2, we provided a useful tool to evaluate the biological roles of the peroxisomes and the related genes.
Assuntos
Sequência de Bases , Primers do DNA , Genética , DNA Fúngico , Genética , Proteínas Fúngicas , Genética , Metabolismo , Genes Fúngicos , Vetores Genéticos , Proteínas de Fluorescência Verde , Genética , Metabolismo , Proteínas Luminescentes , Genética , Metabolismo , Magnaporthe , Genética , Metabolismo , Microscopia de Fluorescência , Mutação , Receptor 2 de Sinal de Orientação para Peroxissomos , Receptor 1 de Sinal de Orientação para Peroxissomos , Peroxissomos , Metabolismo , Receptores Citoplasmáticos e Nucleares , Genética , Metabolismo , Proteínas Recombinantes de Fusão , Genética , Metabolismo , Transformação GenéticaRESUMO
Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M. grisea. A library of 1,077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures of M. grisea. Three transformants grew slowly, 5 transformants decreased in conidiation and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis.
Assuntos
Proteínas Fúngicas , Genética , Regulação Fúngica da Expressão Gênica , Genética , Genes Reporter , Genética , Vetores Genéticos , Genética , Proteínas de Fluorescência Verde , Genética , Metabolismo , Magnaporthe , Genética , Metabolismo , Regiões Promotoras Genéticas , Genética , Engenharia de Proteínas , Métodos , Proteínas Recombinantes , MetabolismoRESUMO
MGTA1, a putative fungal Zn(II)(2)Cys(6) transcriptional activator-encoding gene, was isolated from rice blast pathogen Magnaporthe grisea, which is homologous to CLTA1 from Colletotrichum lindemuthianum with 51% identity at protein level. MGTA1 cassette contains a 2370 bp open reading frame, consisting of 6 exons, coding a 790 amino acid peptide. MGTA1 gene exists as a single copy in genomes of 7 strains of M. grisea, and is expressed in tip hyphae, conidia, and mature appressoria of strain Guy11.