Wax crystal-sparse leaf 3 encoding a β-ketoacyl-CoA reductase is involved in cuticular wax biosynthesis in rice
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WSL3 encodes β-ketoacyl-CoA reductase (KCR) in rice, in a similar way to YBR159w in yeast, and is essential for VLCFA biosynthesis and leaf wax accumulation.
Cuticular waxes on plant surfaces limit non-stomatal water loss, protect plants against deposits of dust and impose a physical barrier to pathogen infection. We identified a wax-deficient mutant of rice, wax crystal-sparse leaf 3 (wsl3), which exhibits a pleiotropic phenotype that includes reduced epicuticular wax crystals on the leaf surface and altered wax composition. Map-based cloning demonstrated that defects in the mutant were caused by two adjacent single-nucleotide changes in a gene encoding β-ketoacyl-CoA reductase (KCR) that catalyzes the second step of the fatty acid elongation reaction. The identity of WSL3 was further confirmed by genetic complementation. Transient assays of fluorescent protein-tagged WSL3 in tobacco protoplasts showed that WSL3 localizes to the endoplasmic reticulum, the compartment of fatty acid elongation in cells. Quantitative PCR and histochemical staining indicated that WSL3 is universally expressed in tissues. RNA interference of WSL3 caused a phenotype that mimicked the wsl3 mutant. Very long-chain fatty acids (VLCFAs) 20:0 and 22:0, or 20:1Δ11 and 22:1Δ13, were detected when WSL3 and Arabidopsis fatty acid elongation 1 (FAE1) were co-expressed in a yeast ybr159wΔ mutant strain. Our results indicated that WSL3 affects rice cuticular wax production by participating in VLCFA elongation.
KeywordsWax crystal-sparse leaf 3 β-Ketoacyl-CoA reductase Cuticular wax Fatty acid elongase Rice (Oryza sativa)
Bulked segregant analysis
Bis-N, N-(trimethylsilyl) trifluoroacetamide
Fatty acid elongase
Fatty acid methyl ester
Gas chromatography–mass spectrometry
Green fluorescent protein
Insertion and deletion
Scanning electron microscope
Short-chain alcohol dehydrogenase reductase
Transmission electron microscopy
Open reading frame
Very long-chain fatty acids
Wax crystal-sparse leaf
The authors thank Dr. F. Beaudoin, Department of Biological Chemistry, Rothamsted Research, UK, for providing the yeast ybr159wΔ mutant strain. This work was supported by the National Special Project (2014ZX08009-003-003) and the National Natural Science Foundation of China (91535302, 31571629). We also acknowledge the support of the Jiangsu Collaborative Innovation Center for Modern Crop Production.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interests.
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