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Gene mapping and cloning of a novel narrow-leaf mutant nal24 in rice

Zhao Rui-Tong1,2,Li Yi-Lin1,Zhou Wan-Feng1,2,Fang Cheng1,Zhang Dong-Yang1,Shang Li-Jun1,Cheng Wei-Min1,Tao Liang-Zhi1,Wu Yue-Jin1,Wu Jian-Dong2,Zhang Cong-He3,Ye Ya-Feng1,*,Liu Bin-Mei1,*   

  1. 1 Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, Anhui, China; 2 Anhui Agricultural University, Hefei 230036, Anhui, China; 3 Winall Hi-tech Seed Co., Ltd., Hefei 230088, Anhui, China
  • Received:2026-03-22 Revised:2026-08-21 Accepted:2026-08-21 Published:2026-09-01
  • Supported by:
    This study was supported by the Key R&D Program of Anhui Province (2023n06020006), the Anhui Provincial Science and Technology Innovation Tackling Plan (202523anull050002), the Yangtze River Delta Science and Technology Innovation Community Joint Research (Basic Research) Project (2024CSJZN01100), the Youth Project C of the National Natural Science Foundation of China (32401738), and the Youth Promotion Association Program of the Chinese Academy of Sciences (2022454).

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Abstract: Rice leaf morphology is a key agronomic trait affecting photosynthetic efficiency and population yield. In this study, a stably inherited narrow-leaf mutant nal24 was obtained by heavy-ion beam irradiation mutagenesis of the japonica rice cultivar Wuyunjing 7 (wyj7). Compared with the wild type, the nal24 mutant exhibited a highly significant reduction in leaf width, accompanied by decreased plant height and increased tiller number. Histological analysis revealed that the narrow-leaf phenotype was caused by reduced numbers of large and small veins as well as reduced distances between small veins. Measurements of photosynthetic characteristics showed that the net photosynthetic rate of nal24 was markedly lower than that of the wild type under different light intensities, indicating reduced photosynthetic capacity. Genetic analysis demonstrated that this trait was controlled by a single recessive nuclear gene. The target gene NAL24 was finely mapped to a 77.6 kb physical interval on chromosome 1 via map-based cloning. Sequencing results identified a single base substitution (T→A) in LOC_Os01g02890, which encodes a phosphatidylserine synthase within this interval, resulting in a premature termination codon. Knockout of this gene in wild-type plants using the CRISPR/Cas9 system produced a narrow-leaf phenotype, and genetic complementation restored wild-type leaf width. Transcriptional analysis indicated that the expression levels of multiple NAL family genes were significantly downregulated in the nal24 mutant, whereas the negative regulator of leaf width, WL1, was significantly upregulated. In conclusion, this study cloned the novel gene NAL24 that regulates rice vein development and leaf morphogenesis. The identification of this gene provides a theoretical basis and genetic resources for elucidating the molecular mechanisms underlying rice leaf shape formation.

Key words: narrow leaf, NAL24, gene mapping, rice, leaf vein development

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