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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Functional Analysis of the Cellulose Synthase-Like Genes CSLD1, CSLD2, and CSLD4 in Tip-Growing Arabidopsis Cells^1,[W]

Department of Biology, University of Copenhagen, Copenhagen Biocentre, 2200 Copenhagen, Denmark (A.J.B., M.M., C.B., I.S., W.G.T.W.); Plant Biology Division, Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401 (C.-M.Y., E.B.B.); Plant Biochemistry Laboratory, University of Copenhagen, DK–1871 Frederiksberg C, Denmark (J.K.J., H.V.S.); Dewel Microscopy Facility, Appalachian State University, Boone, North Carolina 28608 (G.H.); and Departamento de Ciencias Biológicas, Universidad de los Andes, Bogotá, Colombia (A.J.B.)

A reverse genetic approach was used to investigate the functions of three members of the cellulose synthase superfamily in Arabidopsis (Arabidopsis thaliana), CELLULOSE SYNTHASE-LIKE D1 (CSLD1), CSLD2, and CSLD4. CSLD2 is required for normal root hair growth but has a different role from that previously described for CSLD3 (KOJAK). CSLD2 is required during a later stage of hair development than CSLD3, and CSLD2 mutants produce root hairs with a range of abnormalities, with many root hairs rupturing late in development. Remarkably, though, it was often the case that in CSLD2 mutants, tip growth would resume after rupturing of root hairs. In silico, semiquantitative reverse transcription-polymerase chain reaction, and promoter-reporter construct analyses indicated that the expression of both CSLD2 and CSLD3 is elevated at reduced temperatures, and the phenotypes of mutants homozygous for insertions in these genes were partially rescued by reduced temperature growth. However, this was not the case for a double mutant homozygous for insertions in both CSLD2 and CSLD3, suggesting that there may be partial redundancy in the functions of these genes. Mutants in CSLD1 and CSLD4 had a defect in male transmission, and plants heterozygous for insertions in CSLD1 or CSLD4 were defective in their ability to produce pollen tubes, although the number and morphology of pollen grains was normal. We propose that the CSLD family of putative glycosyltransferases synthesize a polysaccharide that has a specialized structural role in the cell walls of tip-growing cells.

² These authors contributed equally to the article.

³ Present address: Max-Planck-Institute of Molecular Plant Physiology, AM Mühlenberg 1, D–14476 Potsdam-Golm, Germany.

⁴ Present address: Joint Bioenergy Institute, 5885 Hollis Street, Emeryville, CA 94608.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: William G.T. Willats (willats{at}bio.ku.dk).

^[W] The online version of this article contains Web-only data.

www.plantphysiol.org/cgi/doi/10.1104/pp.108.121939

^* Corresponding author; e-mail willats{at}bio.ku.dk.

Received April 25, 2008; accepted August 29, 2008; published September 3, 2008.