PorthMaghulyEl-KassabyEtAl2018

Reference

Porth, I., Maghuly, F., El-Kassaby, Y.A. and Mansfield, S. (2018) Localization of gene expression, tissue specificity of Populus xylosyltransferase genes by isolation and functional characterization of their promoters. Plant Cell, Tissue and Organ Culture, 134(3):503-508. (URL )

Abstract

Efficient bioconversion of cellulose into glucose using lignocellulose as the feedstock requires improved cell wall traits. Xylan is part of the matrix covering cellulose and linkages of xylan and lignin inhibit cellulases access. Xylose, an uncommon sugar in the yeast fermentation process, represents 20{\%} of the lignocellulose mass fraction, thus xylan related genes are important targets for biotechnology. Initially, the isolation of candidate genes and their functional characterization is a prerequisite. A common strategy is to perform exhaustive promoter characterizations for candidate genes. Here, we report on the characterization of two xylosyltransferases-related gene promoters that were isolated upstream of the respective candidate genes in poplar, a promising feedstock for bioenergy that is characterized by its short-rotation. The species is known to have undergone recent whole genome duplication, thus finding duplicated gene sequences for xylosyltransferases based on phylogeny reconstruction but with distinct expression patterns is expected. To investigate these two genes closely related to the single, well-characterised Arabidopsis thaliana (At) irx10 gene, we constructed two binary vectors, each studying the full-length promoter of the respective gene. For the poplar (Ptr) gene, most likely the functionally closest to the At irx10 gene, localization of gene expression was studied using the green fluorescence protein (GFP). Poplar's glucuronoxylan glucuronosyltransferase protein, PtrGUT2B, expression was localized in the inflorescence, and specifically in the xylem and along the fibres, suggesting that PtrGUT2B gene expression is associated with fibre production similar to the At irx10 gene. Conversely, the second, duplicated, poplar gene termed PtrGUT2A showed expression in the cambium/phloem, vascular bundles, in the primary xylem and the central cylinder that actively undergo cell divisions and differentiation as evidenced by driving {\ss}-glucuronidase (GUS) expression in these specific tissues. There was no staining detected in the mature secondary xylem or interfascicular fibres. Our study demonstrated that the PtrGUT2B is the true ortholog of irx10 and the two isolated promoters can be used to target tissue specific expression in plant biotechnology assays.

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@ARTICLE { PorthMaghulyEl-KassabyEtAl2018,
    AUTHOR = { Porth, I. and Maghuly, F. and El-Kassaby, Y.A. and Mansfield, S. },
    TITLE = { Localization of gene expression, tissue specificity of Populus xylosyltransferase genes by isolation and functional characterization of their promoters },
    JOURNAL = { Plant Cell, Tissue and Organ Culture },
    YEAR = { 2018 },
    VOLUME = { 134 },
    NUMBER = { 3 },
    PAGES = { 503--508 },
    MONTH = { Sep },
    ISSN = { 1573-5044 },
    ABSTRACT = { Efficient bioconversion of cellulose into glucose using lignocellulose as the feedstock requires improved cell wall traits. Xylan is part of the matrix covering cellulose and linkages of xylan and lignin inhibit cellulases access. Xylose, an uncommon sugar in the yeast fermentation process, represents 20{\%} of the lignocellulose mass fraction, thus xylan related genes are important targets for biotechnology. Initially, the isolation of candidate genes and their functional characterization is a prerequisite. A common strategy is to perform exhaustive promoter characterizations for candidate genes. Here, we report on the characterization of two xylosyltransferases-related gene promoters that were isolated upstream of the respective candidate genes in poplar, a promising feedstock for bioenergy that is characterized by its short-rotation. The species is known to have undergone recent whole genome duplication, thus finding duplicated gene sequences for xylosyltransferases based on phylogeny reconstruction but with distinct expression patterns is expected. To investigate these two genes closely related to the single, well-characterised Arabidopsis thaliana (At) irx10 gene, we constructed two binary vectors, each studying the full-length promoter of the respective gene. For the poplar (Ptr) gene, most likely the functionally closest to the At irx10 gene, localization of gene expression was studied using the green fluorescence protein (GFP). Poplar's glucuronoxylan glucuronosyltransferase protein, PtrGUT2B, expression was localized in the inflorescence, and specifically in the xylem and along the fibres, suggesting that PtrGUT2B gene expression is associated with fibre production similar to the At irx10 gene. Conversely, the second, duplicated, poplar gene termed PtrGUT2A showed expression in the cambium/phloem, vascular bundles, in the primary xylem and the central cylinder that actively undergo cell divisions and differentiation as evidenced by driving {\ss}-glucuronidase (GUS) expression in these specific tissues. There was no staining detected in the mature secondary xylem or interfascicular fibres. Our study demonstrated that the PtrGUT2B is the true ortholog of irx10 and the two isolated promoters can be used to target tissue specific expression in plant biotechnology assays. },
    DAY = { 01 },
    DOI = { 10.1007/s11240-018-1426-5 },
    URL = { https://doi.org/10.1007/s11240-018-1426-5 },
}

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