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Plant Cell Reports

, Volume 36, Issue 10, pp 1655–1666 | Cite as

Wax Crystal-Sparse Leaf 4, encoding a β-ketoacyl-coenzyme A synthase 6, is involved in rice cuticular wax accumulation

  • Lu Gan
  • Shanshan Zhu
  • Zhichao Zhao
  • Linglong Liu
  • Xiaole Wang
  • Zhe Zhang
  • Xin Zhang
  • Jie Wang
  • Jiulin Wang
  • Xiuping Guo
  • Jianmin WanEmail author
Original Article

Abstract

Key message

WSL4 encodes a KCS6 protein which is required for cuticular wax accumulation in rice.

Abstract

Very long chain fatty acids (VLCFAs) are essential precursors for cuticular wax biosynthesis. VLCFA biosynthesis occurs in the endoplasmic reticulum and requires the fatty acid elongase (FAE) complex. The β-ketoacyl-coenzyme A synthase (KCS) catalyzes the first step of FAE-mediated VLCFA elongation. Here we characterized the Wax Crystal-Sparse Leaf 4 (WSL4) gene involved in leaf cuticular wax accumulation in rice. The wsl4 mutant displayed a pleiotropic phenotype including dwarfism, less tiller numbers and reduced surface wax load. Map-based cloning and nucleotide sequencing results revealed that wsl4 carried a single nucleotide substitution in the second exon of a putative KCS6 gene, encoding one subunit of the FAE complex for VLCFAs. Genetic complementation confirmed that the mutation in WSL4 was responsible for the phenotype of wsl4. WSL4 was constitutively expressed in various rice tissues and localized in the endoplasmic reticulum. Both WSL4-RNAi transgenic lines and WSL4 knocked-out mutants exhibited wax-deficient phenotypes similar to the wsl4 mutant. These data indicate that WSL4 is required for cuticular wax accumulation in rice.

Keywords

Rice (Oryza sativaCuticular wax Wax Crystal-Sparse Leaf 4 β-Ketoacyl-coenzyme A synthase 6 Fatty acid elongase 

Abbreviations

CER

Eceriferum

CoA

Coenzyme A

ECR

Enoyl-CoA reductase

ER

Endoplasmic reticulum

FAE

Fatty acid elongation

GC–MS

Gas chromatography–mass spectrometry

GFP

Green fluorescent protein

GUS

β-Glucuronidase

HCD

β-Hydroxyacyl-Coa dehydratase

KCR

β-Ketoacyl-CoA reductase

KCS

β-Ketoacyl-CoA synthase

RNAi

RNA interference

SEM

Scanning electron microscope

TEM

Transmission electron microscopy

ORF

Open reading frame

qRT-PCR

Quantitative RT-PCR

UBQ

Ubiquitin

VLCFAs

Very long chain fatty acids

WSL

Wax Crystal-Sparse Leaf

Notes

Acknowledgements

This work was supported by the National Key R&D Program of China (2016YFD0100600, 2016YFD0200700), and the National Special Project of China (2014ZX08001-006).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

299_2017_2181_MOESM1_ESM.pdf (242 kb)
Supplementary Figure 1. SEM analysis of epicuticular wax crystal patterns on the leaf sheaths of WT (A, B, C) and wsl4 (D, E, F), Bar=1μm. (PDF 241 kb)
299_2017_2181_MOESM2_ESM.xlsx (12 kb)
Supplementary Table 1. Primers used in this paper. (XLSX 11 kb)

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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Lu Gan
    • 1
  • Shanshan Zhu
    • 1
  • Zhichao Zhao
    • 1
  • Linglong Liu
    • 2
  • Xiaole Wang
    • 1
  • Zhe Zhang
    • 1
  • Xin Zhang
    • 1
  • Jie Wang
    • 1
  • Jiulin Wang
    • 1
  • Xiuping Guo
    • 1
  • Jianmin Wan
    • 1
    • 2
    Email author
  1. 1.National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop ScienceChinese Academy of Agricultural SciencesBeijingChina
  2. 2.National Key Laboratory for Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjingChina

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