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Purification and characterization of a soluble β-1,4-glucan from bean (Phaseolus vulgaris L.)-cultured cells dehabituated to dichlobenil

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Abstract

Bean cells habituated to grow in the presence of dichlobenil exhibited reduced cellulose and hemicellulose content and an increase in pectic polysaccharides. Furthermore, following the extraction of pectins and hemicelluloses, a large amount of neutral sugars was released. These sugars were found to be part of a soluble β-1,4-glucan in a preliminary characterization, as reported by Encina et al. (Physiol Plant 114:182–191, 2002). When habituated cells were subcultured in the absence of the herbicide (dehabituated cells), the release of neutral sugars after the extraction of pectins and hemicelluloses was maintained. In this study, we have isolated a soluble β-1,4-glucan from dehabituated cells by sonication of the wall residue (cellulose fraction) remaining after fractionation. Gel filtration chromatography revealed that its average molecular size was 14 kDa. Digestion of the sample with endocellulase revealed the presence of cellobiose, cellotriose, and cellotetraose. Methylation analysis showed that 4-linked glucose was the most abundant sugar residue, but 4,6-linked glucose, terminal arabinose and 4-linked galactose for xyloglucan, and arabinogalactan were also identified. NMR analysis showed that this 1,4-glucan may be composed of various kinds of substitutions along the glucan backbone together with acetyl groups linked to the OH group of sugar residues. Thus, despite its relatively high molecular mass, the β-glucan remains soluble because of its unique configuration. This is the first time that a glucan with such characteristics has been isolated and described. The discovery of new molecules, as this β-glucan with unique features, may help understand the composition and arrangement of the polymers within plant cell walls, contributing to a better understanding of this complex structure.

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Abbreviations

DCB:

Dichlobenil

XyG:

Xyloglucan

snCR:

Supernatant of cellulose residue

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Acknowledgments

This study was funded by the Spanish Ministry of Science and Innovation (CGL2008-02470/BOS) and the Castilla y León Regional Government (LE044A10-2), and by predoctoral grants awarded by the Spanish Ministry of Science and Innovation and Leon University to Penélope García-Angulo and Ana Alonso-Simón, respectively.

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Authors and Affiliations

  1. Área de Fisiología Vegetal, Universidad de León, 24071, León, Spain

    Ana Alonso-Simón, Antonio E. Encina, Penélope García-Angulo, Jesús M. Álvarez & Jose L. Acebes

  2. Department of Forest Science, Tokyo University of Agriculture, Setagaya-ku, Tokyo, 156-8502, Japan

    Tomoko Seyama

  3. Biomaterial Design Lab., Bio-Architecture Center (KBAC), Kyushu University, Hakozaki, Fukuoka, 812-8581, Japan

    Tetsuo Kondo

  4. Department of Bio-science, Tokyo University of Agriculture, Setagaya-ku, Tokyo, 156-8502, Japan

    Takahisa Hayashi

Authors
  1. Ana Alonso-Simón
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  2. Antonio E. Encina
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  3. Tomoko Seyama
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  4. Tetsuo Kondo
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  5. Penélope García-Angulo
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  6. Jesús M. Álvarez
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  7. Jose L. Acebes
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  8. Takahisa Hayashi
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Correspondence to Jose L. Acebes.

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Alonso-Simón, A., Encina, A.E., Seyama, T. et al. Purification and characterization of a soluble β-1,4-glucan from bean (Phaseolus vulgaris L.)-cultured cells dehabituated to dichlobenil. Planta 237, 1475–1482 (2013). https://doi.org/10.1007/s00425-013-1861-9

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  • DOI: https://doi.org/10.1007/s00425-013-1861-9

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