Plant and Cell Physiology Advance Access originally published online on October 30, 2007
Plant and Cell Physiology 2007 48(11):1524-1533; doi:10.1093/pcp/pcm139
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Cytological and Biochemical Analysis of COF1, an Arabidopsis Mutant of an ABC Transporter Gene
1International Graduate School of Arts and Science, Yokohama City University, Yokohama, Kanagawa, 230-0045 Japan
2Gene Discovery Research Group, RIKEN Plant Science Center, Yokohama, Kanagawa, 230-0045 Japan
3Graduate School of Agriculture, Kyoto University, Sakyo, Kyoto, 606-8502 Japan and
4Environmental Molecular Biology Laboratory, RIKEN Discovery Research Institute, Wako, Saitama, 351-0198 Japan120
*Corresponding author: E-mail, sinozaki{at}rtc.riken.jp; Fax, +81-45-503-9580.
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Abstract |
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In transposon-tagged lines of Arabidopsis we found two new mutants, cof1-1 and cof1-2 (cuticular defect and organ fusion), that show the phenotype of wilting when grown in soil, organ fusion of rosette leaves and infertility. Toluidine blue testing and scanning electron microscopy observation revealed that these mutants had cuticular defects in the stems and adult leaves, but not in cotyledones. Transmission electron microscopy observation revealed thinner cuticle layers in the mutants, and cuticular materials interspersed between the two fused epidermal layers were observed in the mutant rosette leaves. These two mutants had a transposon insertion in the coding regions of WBC11, which was classified as a member of ABC transporter genes in Arabidopsis. WBC11 showed high sequence similarity to CER5 (also called WBC12), which was involved in cuticular lipid export. Gas chromatographic analysis revealed that C29 alkane extracted from the stem surface of cof1 mutants was reduced whereas C29 ketone was accumulated, which was different from the case of cer5 mutants. While cer5 mutants had fairly normal morphology, cof1 mutants had pleiotropic phenotypes so that COF1/WBC11 could have important roles different from those of CER5/WBC12. We also found that C29 alkane was accumulated in the intracellular extract of cof1 mutants, suggesting a function for WBC11 in the direct transport of lipid molecules. Pollen observation showed that mutant pollen grains were irregularly shaped. The function of COF1/WBC11 in lipid transport for the construction of cuticle layers and pollen coats for normal organ formation is discussed.
Keywords: ABC transporter - Arabidopsis thaliana - Cuticle - Organ fusion - Transposon-tagged line
Abbreviations: Ac/Ds, Activator/Dissociation; ABC transporters, TP-binding cassette transporters; CaMV, cauliflower mosaic virus; cer, eceriferum; cof1, cuticular defect and organ fusions 1; GFP, green fluorescent protein; RT–PCR, reverse transcription–PCR; SEM, scanning electron microscopy; TB, toluidine blue; TEM, transmission electron microscopy; YFP, yellow fluorescent protein; WBC, White–Brown Complex homolog.
5These authors contributed equally to this work.
6Present address: Faculty of Agriculture, Kyusyu University, Fukuoka, Fukuoka, 812-8581 Japan.
(Received September 30, 2007; Accepted October 10, 2007)
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