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OsHXK3 encodes a hexokinase-like protein that positively regulates grain size in rice

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Abstract

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We report the map-based cloning and functional characterization of SNG1, which encodes OsHXK3, a hexokinase-like protein that plays a pivotal role in controlling grain size in rice.

Abstract

Grain size is an important agronomic trait determining grain yield and appearance quality in rice. Here, we report the discovery of rice mutant short and narrow grain1 (sng1) with reduced grain length, width and weight. Map-based cloning revealed that the mutant phenotype was caused by loss of function of gene OsHXK3 that encodes a hexokinase-like (HKL) protein. OsHXK3 was associated with the mitochondria and was ubiquitously distributed in various organs, predominately in younger organs. Analysis of glucose (Glc) phosphorylation activities in young panicles and protoplasts showed that OsHXK3 was a non-catalytic hexokinase (HXK). Overexpression of OsHXK3 could not complement the Arabidopsis glucose insensitive2-1 (gin2-1) mutant, indicating that OsHXK3 lacked Glc signaling activity. Scanning electron microscopy analysis revealed that OsHXK3 affects grain size by promoting spikelet husk cell expansion. Knockout of other nine OsHXK genes except OsHXK3 individually did not change grain size, indicating that functions of OsHXKs have differentiated in rice. OsHXK3 influences gibberellin (GA) biosynthesis and homeostasis. Compared with wild type, OsGA3ox2 was significantly up-regulated and OsGA2ox1 was significantly down-regulated in young panicle of sng1, and concentrations of biologically active GAs were significantly decreased in young panicles of the mutants. The yield per plant of OsHXK3 overexpression lines (OE-4 and OE-35) was increased by 10.91% and 7.62%, respectively, compared to that of wild type. Our results provide evidence that an HXK lacking catalytic and sensory functions plays an important role in grain size and has the potential to increase yield in rice.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

We thank Dr. Hua Zhang and Chennan Ge for Arabidopsis transformation. This work was supported by grants from the National Natural Science Foundation (U21A20211, 91935303, 31901529), the Ministry of Science and Technology (2021YFF1000200), the science and technology major program of Hubei Province (2021ABA011), the earmarked fund for the Agricultural Research System (CARS-01-01) of China, and the Natural Science Foundation of Anhui Province (2008085QC120).

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, National Centre of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, 430070, China

    Peng Yun, Yibo Li, Bian Wu, Yun Zhu, Kaiyue Wang, Pingbo Li, Guanjun Gao, Qinglu Zhang, Xianghua Li & Yuqing He

  2. Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, 230031, China

    Peng Yun & Zefu Li

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  1. Peng Yun
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  2. Yibo Li
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  3. Bian Wu
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Contributions

YP conducted most of the experiments; LYB isolated the sng1 mutant; WB, ZY and WKY carried out part of the phenotyping and expression analysis; LPB, GGJ, ZQL and LXH participated in the field management and logistic work; YP, LYB, LZF and HYQ designed the study; YP and HYQ analyzed the data and wrote the paper.

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Correspondence to Yuqing He.

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Communicated by Joshua N. Cobb.

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Yun, P., Li, Y., Wu, B. et al. OsHXK3 encodes a hexokinase-like protein that positively regulates grain size in rice. Theor Appl Genet 135, 3417–3431 (2022). https://doi.org/10.1007/s00122-022-04189-7

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