, Volume 228, Issue 4, pp 675–685 | Cite as

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

  • Dongmei Yu
  • Kosala Ranathunge
  • Huasun Huang
  • Zhongyou Pei
  • Rochus Franke
  • Lukas Schreiber
  • Chaozu HeEmail author
Original Article


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., 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.


Cuticular waxes β-Ketoacyl CoA synthase Rice (Oryza sativa L.) T-DNA insertional mutagenesis Wax Crystal-Sparse Leaf1 



Very-long chain fatty acid (>C18)


β-Ketoacyl CoA synthase


Scanning electron microscopy


Transmission electron microscopy


Gas chromatography–mass spectrometry


Thermal asymmetric interlaced PCR



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).

Supplementary material

425_2008_770_MOESM1_ESM.tif (1.9 mb)
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 (1.2 mb)
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 (1.5 mb)
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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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Dongmei Yu
    • 1
    • 2
  • Kosala Ranathunge
    • 3
  • Huasun Huang
    • 4
  • Zhongyou Pei
    • 1
  • Rochus Franke
    • 3
  • Lukas Schreiber
    • 3
  • Chaozu He
    • 1
    Email author
  1. 1.State Key Laboratory of Plant Genomics, Institute of MicrobiologyChinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.Graduate School of the Chinese Academy of SciencesBeijingPeople’s Republic of China
  3. 3.Institute of Cellular and Molecular Botany, Department of EcophysiologyUniversity of BonnBonnGermany
  4. 4.Rubber Research InstituteChinese Academy of Tropical Agricultural SciencesHainanPeople’s Republic of China

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