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The cuticle covering the outer surface of anthers is essential for male reproductive development in plants. However, the mechanism underlying the synthesis of these lipidic polymers remains unclear. HOTHEAD (HTH) in Arabidopsis thaliana is a presumptive glucose-methanol-choline (GMC) oxidoreductase involved in the biosynthesis of long-chain α-,ω-dicarboxylic fatty acids. In this study, we characterized the function of an anther-specific gene HTH1 in rice. HTH1 contains a conserved GMC oxidoreductase-like domain, and the sequence of HTH1 was highly similar to that of HTH in A. thaliana. Quantitative real-time PCR (qRT-PCR) and in situ hybridization analyses showed that HTH1 was highly expressed in epidermal cells of anthers. Rice plants with HTH1 suppression through CRISPR (clustered regularly interspaced short palindromic repeats) and RNA interference (RNAi) displayed defective anther wall and aborted pollen. Disorganized cuticle layers in anthers and shriveled pollen grains were observed in HTH1-RNAi lines. The total amounts of long-chain fatty acids and cutin monomers in anthers of HTH1-RNAi lines were significantly reduced compared with the wild type. Our results suggested that HTH1 is involved in cutin biosynthesis and is required for anther development and pollen fertility in rice.

HOTHEAD-Like HTH1 is Involved in Anther Cutin Biosynthesis and is Required for Pollen Fertility in Rice