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Glycome profiling and immunohistochemistry uncover spaceflight-induced changes in non-cellulosic cell wall components in Arabidopsis thaliana seedling roots

Jin Nakashima, Sivakumar Pattathil, Utku Avci, Sabrina Chin, J. Alan Sparks, Michael G. Hahn, Simon Gilroy, Elison B. Blancaflor
doi: https://doi.org/10.1101/2023.03.14.532448
Jin Nakashima
1Analytical Instrumentation Facility, North Carolina State University, 2410 Camp, US Shore Drive, Raleigh, NC 27606 USA
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Sivakumar Pattathil
2Mascoma LLC (Lallemand Inc.), 67 Etna Road, Lebanon, NH 03766, USA
3The University of Georgia, Complex Carbohydrate Research Center, 315 Riverbend Rd, Athens, GA, 30602, USA
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Utku Avci
3The University of Georgia, Complex Carbohydrate Research Center, 315 Riverbend Rd, Athens, GA, 30602, USA
4Department of Agricultural Biotechnology, Faculty of Agriculture, Eskisehir Osmangazi University, Eskisehir 26160, TURKEY
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Sabrina Chin
5Department of Botany, 430 Lincoln Drive, University of Wisconsin, Madison, WI 53706, USA
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J. Alan Sparks
6Noble Research Institute LLC, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA
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Michael G. Hahn
4Department of Agricultural Biotechnology, Faculty of Agriculture, Eskisehir Osmangazi University, Eskisehir 26160, TURKEY
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Simon Gilroy
5Department of Botany, 430 Lincoln Drive, University of Wisconsin, Madison, WI 53706, USA
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Elison B. Blancaflor
7Utilization & Life Sciences Office, Exploration Research and Technology Programs, NASA John F. Kennedy Space Center, FL, 32899, USA
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  • For correspondence: elison.b.blancaflor@nasa.gov
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ABSTRACT

A large and diverse library of glycan-directed monoclonal antibodies (mAbs) was used to determine if plant cell walls are modified by low-gravity conditions encountered during spaceflight. This method called glycome profiling (glycomics) revealed global differences in non-cellulosic cell wall epitopes in Arabidopsis thaliana root extracts recovered from RNA purification columns between seedlings grown on the International Space Station-based Vegetable Production System and paired ground (1-g) controls. Immunohistochemistry on 11-day-old seedling primary root sections showed that ten of twenty-two mAbs that exhibited spaceflight-induced increases in binding through glycomics, labeled space-grown roots more intensely than those from the ground. The ten mAbs recognized xyloglucan, xylan, and arabinogalactan epitopes. Notably, three xylem-enriched unsubstituted xylan backbone epitopes were more intensely labeled in space-grown roots than in ground-grown roots, suggesting that the spaceflight environment accelerated root secondary cell wall formation. This study highlights the feasibility of glycomics for high-throughput evaluation of cell wall glycans using only root high alkaline extracts from RNA purification columns, and subsequent validation of these results by immunohistochemistry. This approach will benefit plant space biological studies because it extends the analyses possible from the limited amounts of samples returned from spaceflight and help uncover microgravity-induced tissue-specific changes in plant cell walls.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted March 15, 2023.
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Glycome profiling and immunohistochemistry uncover spaceflight-induced changes in non-cellulosic cell wall components in Arabidopsis thaliana seedling roots
Jin Nakashima, Sivakumar Pattathil, Utku Avci, Sabrina Chin, J. Alan Sparks, Michael G. Hahn, Simon Gilroy, Elison B. Blancaflor
bioRxiv 2023.03.14.532448; doi: https://doi.org/10.1101/2023.03.14.532448
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Glycome profiling and immunohistochemistry uncover spaceflight-induced changes in non-cellulosic cell wall components in Arabidopsis thaliana seedling roots
Jin Nakashima, Sivakumar Pattathil, Utku Avci, Sabrina Chin, J. Alan Sparks, Michael G. Hahn, Simon Gilroy, Elison B. Blancaflor
bioRxiv 2023.03.14.532448; doi: https://doi.org/10.1101/2023.03.14.532448

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