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Plant and Cell Physiology Advance Access originally published online on April 27, 2009
Plant and Cell Physiology 2009 50(6):1099-1115; doi:10.1093/pcp/pcp062
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

KORRIGAN1 and its Aspen Homolog PttCel9A1 Decrease Cellulose Crystallinity in Arabidopsis Stems

Junko Takahashi1, Ulla J. Rudsander2, Mattias Hedenström3, Alicja Banasiak1,8, Jesper Harholt5, Nicolas Amelot1,9, Peter Immerzeel1, Peter Ryden6, Satoshi Endo1,10, Farid M. Ibatullin2, Harry Brumer2, Elena del Campillo7, Emma R. Master4, Henrik Vibe Scheller5, Björn Sundberg1, Tuula T. Teeri2 and Ewa J. Mellerowicz1,*

1Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, SLU, 90183, Umeå, Sweden
2Swedish Center for Biomimetic Fiber Engineering, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, 10691, Stockholm, Sweden
3Department of Chemistry, Umeå University, 90187 Umeå, Sweden
4Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, M5S 3E5, Canada
5Molecular Plant Biology Laboratory, Department of Plant Biology, Faculty of Life Sciences, University of Copenhagen, DK-1871 Frederiksberg C, Denmark
6Department of Food Materials Science, Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK
7Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA

*Corresponding author: E-mail, ewa.mellerowicz{at}genfys.slu.se; Fax: +46-90-786-8165.


  Abstract

KORRIGAN1 (KOR1) is a membrane-bound cellulase implicated in cellulose biosynthesis. PttCel9A1 from hybrid aspen (Populus tremula L. x tremuloides Michx.) has high sequence similarity to KOR1 and we demonstrate here that it complements kor1-1 mutants, indicating that it is a KOR1 ortholog. We investigated the function of PttCel9A1/KOR1 in Arabidopsis secondary growth using transgenic lines expressing 35S::PttCel9A1 and the KOR1 mutant line irx2-2. The presence of elevated levels of PttCel9A1/KOR1 in secondary walls of 35S::PttCel9A1 lines was confirmed by in muro visualization of cellulase activity. Compared with the wild type, 35S::PttCel9A1 lines had higher trifluoroacetic acid (TFA)-hydrolyzable glucan contents, similar Updegraff cellulose contents and lower cellulose crystallinity indices, as determined by 13C solid-state nuclear magnetic resonance (NMR) spectroscopy. irx2-2 mutants had wild-type TFA-hydrolyzable glucan contents, but reduced Updegraff cellulose contents and higher than wild-type cellulose crystallinity indices. The data support the hypothesis that PttCel9A1/KOR1 activity is present in cell walls, where it facilitates cellulose biosynthesis in a way that increases the amount of non-crystalline cellulose.

Keywords: Aspen - Cellulase - Cellulose - Cell wall - Populus - Wood formation

Abbreviations: Ara, arabinose; CaMV, cauliflower mosaic virus; CBM, carbohydrate-binding module; CDS, coding sequence; Col, Columbia; CPMAS, cross-polarization magic angle spinning; EST, expressed sequence tag; Fuc, fucose; Gal, galactose; GalUA, galacturonic acid; GH9, glycoside hydrolase 9; Glc, glucose; Glc-Res, resorufinyl β-D-glucopyranoside; Glc, ; 2, ; -Res, resorufinyl β-D-cellobioside; GlcUA, glucuronic acid; GUS, β-glucuronidase; KOR1, KORRIGAN1; LD, long day; NMR, nuclear magnetic resonance; OE, overexpressing; PLS-DA, partial least squares-discriminant analysis; PSRE, proton spin relaxation editing; Rha, rhamnose; RT–PCR, reverse transcription–PCR; SA, solvent-accessible; SD, short day; SI, solvent-inaccessible; TFA, trifluoroacetic acid; Ws, Wassilewskija; WT, wild type; Xyl, xylose; YFP, yellow fluorescent protein.

8Present address: Institute of Plant Biology, University of Wroclaw, Kanonia 6/8, 50-328, Wroclaw, Poland.

9Present address: UMR 5546 Paul Sabatier University/CNRS, 31326 Castanet-Tolosan, France.

10Present address: Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 703-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

(Received March 4, 2009; Accepted April 23, 2009)
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