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Wax Crystal-Sparse Leaf1 encodes a β–ketoacyl CoA synthase involved in biosynthesis of cuticular waxes on rice leaf

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Abstract

Cuticular waxes, forming the plant/atmosphere interface of plants colonizing the terrestrial environment, are complex mixtures of very-long chain fatty acids (VLCFAs) and their derivatives. In VLCFAs biosynthesis, β-ketoacyl CoA synthase (E.C.2.3.1.119, KCS) is the key enzyme. Using T-DNA insertional mutagenesis, we identified a cuticle-deficient rice mutant, which displayed a pleiotropic phenotype including reduced growth, leaf fusion, sparse wax crystals, enhanced sensitivity to drought and low fertility. Further analysis indicated that T-DNA was inserted in the 5′-UTR intron of the affected gene, Wax Crystal-Sparse Leaf1 (WSL1), and abnormal transcript caused the loss-of-function of WSL1 gene. Genetic complementation experiment confirmed the function of the candidate gene. WSL1 was predicted to encode a polypeptide containing a conserved FAE1_CUT1_RppA domain typical of the KCS family proteins. Qualitative and quantitative wax composition analyses by gas chromatography–mass spectrometry (GC–MS) demonstrated a marked reduction of total cuticular wax load on wsl1 leaf blades and sheaths, and VLCFA precursors of C20–C24 decreased in both. Moreover, ubiquitous expression of the WSL1 gene gave a hint that WSL1-catalyzed elongation of VLCFAs might participate in a wide range of rice growth and development processes beyond biosynthesis of cuticular waxes.

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Abbreviations

VLCFA:

Very-long chain fatty acid (>C18)

KCS:

β-Ketoacyl CoA synthase

SEM:

Scanning electron microscopy

TEM:

Transmission electron microscopy

GC–MS:

Gas chromatography–mass spectrometry

TAIL-PCR:

Thermal asymmetric interlaced PCR

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Acknowledgments

This work was supported by the Basic Research Program of Ministry of Science and Technology of China (Grant 2006CB101900) and by the Alexander-von-Humboldt-Stiftung (Postdoc grant awarded to K. R).

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Authors and Affiliations

  1. State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, People’s Republic of China

    Dongmei Yu, Zhongyou Pei & Chaozu He

  2. Graduate School of the Chinese Academy of Sciences, 100049, Beijing, People’s Republic of China

    Dongmei Yu

  3. Institute of Cellular and Molecular Botany, Department of Ecophysiology, University of Bonn, 53115, Bonn, Germany

    Kosala Ranathunge, Rochus Franke & Lukas Schreiber

  4. Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, 571737, Hainan, People’s Republic of China

    Huasun Huang

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  1. Dongmei Yu
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  2. Kosala Ranathunge
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  3. Huasun Huang
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  4. Zhongyou Pei
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Correspondence to Chaozu He.

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425_2008_770_MOESM1_ESM.tif

Fig. S1 Morphology of the complemented line HC-8, wild-type (wt) and wsl1 plants. The plant size and vigor of HC-8 was similar to those of the wild-type (TIFF 1,907 kb)

425_2008_770_MOESM2_ESM.tif

Fig. S2 Multiple alignment of WSL1 and six well-characterized KCS family members in Arabidopsis. Arrowheads indicated the conserved Cys223, His391 and Asn424 in all KCS family members (TIFF 1,253 kb)

425_2008_770_MOESM3_ESM.tif

Fig. S3 Composition of the cutin on leaves of wild-type, wsl1 and the complemented line HC-8. No significant difference was found. Cutin was analyzed as described in Jung et al. (2006) (TIFF 1,498 kb)

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Yu, D., Ranathunge, K., Huang, H. et al. Wax Crystal-Sparse Leaf1 encodes a β–ketoacyl CoA synthase involved in biosynthesis of cuticular waxes on rice leaf. Planta 228, 675–685 (2008). https://doi.org/10.1007/s00425-008-0770-9

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