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Plant and Cell Physiology, 2001, Vol. 42, No. 3 251-263
© 2001 Oxford University Press


Paper

Role of the Putative Membrane-Bound Endo-1,4-ß-Glucanase KORRIGAN in Cell Elongation and Cellulose Synthesis in Arabidopsis thaliana

Shigeru Sato ,1,7, Tomohiko Kato ,1,8, Koichi Kakegawa2, Tadashi Ishii2, Yao-Guang Liu3, Tatsuya Awano4, Keiji Takabe4, Yoshiharu Nishiyama5, Shigenori Kuga5, Shusei Sato6, Yasukazu Nakamura6, Satoshi Tabata6 and Daisuke Shibata ,1,8,9

1 Mitsui Plant Biotechnology Research Institute (disbanded in March 1999), Tsukuba, Ibaraki, 305-0047 Japan 2 Forestry and Forest Products Research Institute, P.O. Box 16, Tsukuba Norin Kenkyu Danchi-nai, Ibaraki, 305-8687 Japan 3 Yao-Guang Liu, Genetic Engineering Laboratory, Biotechnology Center, South China Agricultural University, Guangzhou 510642, People’s Republic of China 4 Graduate School of Agriculture, Kyoto University, Sakyo, Kyoto, 606-8502 Japan 5 Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657 Japan 6 Kazusa DNA Research Institute, 1532–3 Yana, Kisarazu, Chiba, 292-0812 Japan

Abstract

A temperature-sensitive, elongation-deficient mutant of Arabidopsis thaliana was isolated. At the non-permissive temperature of 31°C, the mutation impaired tissue elongation; otherwise, tissue development was normal. Hypocotyl cells that had established cell walls at 21°C under light-dark cycles ceased elongation and swelled when the mutant was shifted to 31°C and darkness, indicating that the affected gene is essential for cell elongation. Analysis of the cell walls of mutant plants grown at 31°C revealed that the cellulose content was reduced to 40% and the pectin content was increased to 162% of the corresponding values for the wild type grown at the same temperature. The increased amounts of pectin in the mutant were bound tightly to cellulose microfibrils. No change in the content of hemicellulose was apparent in the 31°C-adapted mutant. Field emission–scanning electron microscopy suggested that the structure of cellulose bundles was affected by the mutation; X-ray diffraction, however, revealed no change in the crystallite size of cellulose microfibrils. The regeneration of cellulose microfibrils from naked mutant protoplasts was substantially delayed at 31°C. The recessive mutation was mapped to chromosome V, and map-based cloning identified it as a single G->A transition (resulting in a Gly429->Arg substitution) in KORRIGAN, which encodes a putative membrane-bound endo-1,4-ß-glucanase. These results demonstrate that the product of this gene is required for cellulose synthesis.

Footnotes

7 Present address: Oji Paper Co. Ltd., Nobono-cho 24-9, Kameyama, Mie, 519-0212 Japan.

8 Present address: Kazusa DNA Research Institute, 1532-3 Yana, Kisarazu, Chiba, 292-0812 Japan.

9 Corresponding author: E-mail, shibata@kazusa.or.jp; Fax, +81-438-52-3948; Phone, +81-438-52-3947.


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