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Plant and Cell Physiology Advance Access published online on April 27, 2009
Plant and Cell Physiology, 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 e-mail: journals.permissions@oxfordjournals.org

KORRIGAN1 and its aspen homologue PttCel9A1 decrease cellulose crystallinity in Arabidopsis stems

Junko Takahashi1, Ulla J. Rudsander2, Mattias Hedenström3, Alicja Banasiak1,7, Jesper Harholt4, Nicolas Amelot1,8, Peter Immerzeel1, Peter Ryden5, Satoshi Endo1,10, Farid M. Ibatullin2, Harry Brumer2, Elena del Campillo6, Emma R. Master7, Henrik Vibe Scheller4, Björn Sundberg1, Tuula T. Teeri2 and Ewa J. Mellerowicz1

1 Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, SLU, 90183, Umeå, Sweden
2 Swedish Center for Biomimetic Fiber Engineering, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, 10691, Stockholm, Sweden
3 Department of Chemistry, Umeå University, 90187 Umeå, Sweden
4 Molecular Plant Biology Laboratory, Department of Plant Biology, Faculty of Life Sciences, University of Copenhagen, DK-1871 Frederiksberg C, Denmark
5 Department of Food Materials Science, Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom
6 Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD20742, Maryland, USA
7 Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, M5S 3E5, Canada

Corresponding author: Dr. Ewa J. Mellerowicz, Address: Department of Forest Genetics and Plant Physiology, SLU, Umeå Plant Science Centre (UPSC), 90183, Umeå, Sweden. Tel: 46 90 786 8367, Fax: 46 90 786 8165, E-mail: ewa.mellerowicz{at}genfys.slu.se


  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 to WT, 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 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: cellulase - cellulose - cell wall - aspen - Populus - wood formation

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

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

10 Present address: Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113-0033, Tokyo, Japan.

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