ABSTRACT
Background: In Saccharomyces cerevisiae, Msn2, which acts as a key transcription factor downstream the MAPKHOG cascade pathway, also regulates the expression of genes related to stress responses. However, little is known about the regulation mechanisms of the transcription factor in Setosphaeria turcica. Results: In this study, a zinc finger DNA-binding protein, designated as StMSN2, was cloned from S. turcica. Sequencing results showed that StMSN2 had a 1752 bp open reading frame (ORF), which was interrupted by an intron (135 bp) and encoded a putative 538-amino acid protein. Phylogenetic analysis further revealed that StMsn2 was more closely related to Msn2 of Aspergillus parasiticus. StMSN2 was cloned into the pET-28a vector with His (Histidine) tags and induced with 1 mM IPTG (isopropyl-ß-D-thiogalactoside) at 37°C. The recombinant His-tagged StMsn2 was purified, and a band of size approximately 58.8 kDa was obtained. The high specificity of the polyclonal antibody Msn2-2 was detected with the StMsn2 protein from S. turcica and prokaryotic expression system, respectively. Conclusions: A new gene, named StMSN2, with 1617 bp ORF was cloned from S. turcica and characterized using bioinformatics methods. StMsn2 was expressed and purified in a prokaryotic system. A polyclonal antibody, named Msn2-2, against StMsn2 with high specificity was identified.
Subject(s)
Plant Diseases , Ascomycota/genetics , Ascomycota/pathogenicity , Transcription Factors/isolation & purification , Ascomycota/metabolism , Stress, Physiological , Transcription Factors/genetics , Transcription Factors/metabolism , Carrier Proteins , Gene Expression , Blotting, Western , Open Reading Frames , Zinc Fingers , Cloning, Molecular , Zea mays , Escherichia coli , Helminthosporium , EpitopesABSTRACT
To study the chemical components and the antifungal activity of extraction from conidia of Trichoderma viride LTR-2.The extraction were obtained by distilling with Methylene dichloride from conidia of Trichoderma viride LTR-2 cultured on wheat bran solid matrix.Antifungal activity were determined by mycelium growth method.The chemical components of the extraction were analysed by GC-MS,the relative components in the extraction were determined by area normalization.The extraction not only have broad-spectrum control,showed antibiosis against eleven different plant fungal pathogens in PDA dish,such as Rhizoctonia solani,Alternaria brassica,Verticillium dahliae,Macrophoma kawatsukai,Fusarium moniliforme,Botrytis cinerea,Rhizoctonia cerealis,Fusarium oxysporum f.sp.vasinfectum,Bipolaris sorokinana,Fusarium graminearum,Alternaria.mali,but also have high inhibitory effect,and had 89.3% suppressive rate to Rhizoctonia cerealis.About sixty components were separated and identified by GC-MS,majority components were Hydrocarbon,the number of the Hydrocarbon were fourty-three kinds.Ergosterol was the major chemical components of the extract,and has 41.90% content.Other components comprised:Ketone,Organic acid,Alcohol,Ene,et al.Conclusion:The extraction from conidia of Trichoderma viride LTR-2 have antifungal activity.The extration comprised 2H-Pyran-2-one,5,6-dihydro-6-pentyl,it has 2.35% content.reference others literature,2H-Pyran-2-one,5,6-dihydro-6-pentyl may be the suppressive component of the extration.