Dataset Information

Living colors in the gray mold pathogen Botrytis cinerea: codon-optimized genes encoding green fluorescent protein and mCherry, which exhibit bright fluorescence.

ABSTRACT: The green fluorescent protein (GFP) and its variants have been widely used in modern biology as reporters that allow a variety of live-cell imaging techniques. So far, GFP has rarely been used in the gray mold fungus Botrytis cinerea because of low fluorescence intensity. The codon usage of B. cinerea genes strongly deviates from that of commonly used GFP-encoding genes and reveals a lower GC content than other fungi. In this study, we report the development and use of a codon-optimized version of the B. cinerea enhanced GFP (eGFP)-encoding gene (Bcgfp) for improved expression in B. cinerea. Both the codon optimization and, to a smaller extent, the insertion of an intron resulted in higher mRNA levels and increased fluorescence. Bcgfp was used for localization of nuclei in germinating spores and for visualizing host penetration. We further demonstrate the use of promoter-Bcgfp fusions for quantitative evaluation of various toxic compounds as inducers of the atrB gene encoding an ABC-type drug efflux transporter of B. cinerea. In addition, a codon-optimized mCherry-encoding gene was constructed which yielded bright red fluorescence in B. cinerea.

SUBMITTER: Leroch M

PROVIDER: S-EPMC3126427 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature

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Living colors in the gray mold pathogen Botrytis cinerea: codon-optimized genes encoding green fluorescent protein and mCherry, which exhibit bright fluorescence.

Leroch Michaela M Mernke Dennis D Koppenhoefer Dieter D Schneider Prisca P Mosbach Andreas A Doehlemann Gunther G Hahn Matthias M

Applied and environmental microbiology 20110304 9

The green fluorescent protein (GFP) and its variants have been widely used in modern biology as reporters that allow a variety of live-cell imaging techniques. So far, GFP has rarely been used in the gray mold fungus Botrytis cinerea because of low fluorescence intensity. The codon usage of B. cinerea genes strongly deviates from that of commonly used GFP-encoding genes and reveals a lower GC content than other fungi. In this study, we report the development and use of a codon-optimized version ...[more]

PMID: 21378036

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Project description:Owing to the remarkable antimicrobial potential of these materials, research into the possible use of nanomaterials as alternatives to fungicides in sustainable agriculture is increasingly progressing. Here, we investigated the potential antifungal properties of chitosan-decorated copper oxide nanocomposite (CH@CuO NPs) to control gray mold diseases of tomato caused by Botrytis cinerea throughout in vitro and in vivo trials. The nanocomposite CH@CuO NPs were chemically prepared, and size and shape were determined using Transmission Electron Microscope (TEM). The chemical functional groups responsible for the interaction of the CH NPs with the CuO NPs were detected using the Fourier Transform Infrared (FTIR) spectrophotometry. The TEM images confirmed that CH NPs have a thin and semitransparent network shape, while CuO NPs were spherically shaped. Furthermore, the nanocomposite CH@CuO NPs ex-habited an irregular shape. The size of CH NPs, CuO NPs and CH@CuO NPs as measured through TEM, were approximately 18.28 ± 2.4 nm, 19.34 ± 2.1 nm, and 32.74 ± 2.3 nm, respectively. The antifungal activity of CH@CuO NPs was tested at three concentrations of 50, 100 and 250 mg/L and the fungicide Teldor 50% SC was applied at recommended dose 1.5 mL/L. In vitro experiments revealed that CH@CuO NPs at different concentrations significantly inhibited the reproductive growth process of B. cinerea by suppressing the development of hyphae, spore germination and formation of sclerotia. Interestingly, a significant control efficacy of CH@CuO NPs against tomato gray mold was observed particularly at concentrations 100 and 250 mg/L on both detached leaves (100%) as well as the whole tomato plants (100%) when compared to the conventional chemical fungicide Teldor 50% SC (97%). In addition, the tested concentration 100 mg/L improved to be sufficient to guarantee a complete reduction in the disease's severity (100%) to tomato fruits from gray mold without any morphological toxicity. In comparison, tomato plants treated with the recommended dose 1.5 mL/L of Teldor 50% SC ensured disease reduction up to 80%. Conclusively, this research enhances the concept of agro-nanotechnology by presenting how a nano materials-based fungicide could be used to protect tomato plants from gray mold under greenhouse conditions and during the postharvest stage.

Dataset Information

Living colors in the gray mold pathogen Botrytis cinerea: codon-optimized genes encoding green fluorescent protein and mCherry, which exhibit bright fluorescence.

Publications

Living colors in the gray mold pathogen Botrytis cinerea: codon-optimized genes encoding green fluorescent protein and mCherry, which exhibit bright fluorescence.

Similar Datasets

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