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Volume 15
Issue 12
10.3390/plants15121826
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

The Kelch-Repeat Superfamily Gene SiNL4 Regulates the Leaf Width in Foxtail Millet

by
Yuqin Zhao
,
Yixuan Ma
,
Yanyu Yang
,
Lejie Yang
,
Lu Chen
,
Tianguo Wang
,
Shiyuan Wang
,
Kai Zhao
,
Xiaorui Li
,
Shuqi Dong
,
Hongzhi Wang
,
Xiaoqian Chu
,
Jiagang Wang
,
Lulu Gao
* and
Guanghui Yang
*
College of Agronomy, Shanxi Agricultural University, Special Orphan Crops Research Center of the Loess Plateau, MARA, Jinzhong 030800, China
*
Authors to whom correspondence should be addressed.
Plants 2026, 15(12), 1826; https://doi.org/10.3390/plants15121826 (registering DOI)
Submission received: 27 May 2026 / Revised: 6 June 2026 / Accepted: 10 June 2026 / Published: 12 June 2026
Versions Notes

Abstract

The Kelch-repeat superfamily genes played important roles in regulating plant growth and development; however, their functions in foxtail millet (Setaria italica) have not yet been characterized. In this study, SiNL4, a homolog of ZmNL4 controlling leaf width in maize, was knocked out using the CRISPR/Cas9 technology, and two homozygous knockout lines (ko1 and ko2) were obtained. Phenotypic analysis showed that compared with the wild-type Ci846, ko1 and ko2 exhibited reduced leaf width and decreased yield related traits (e.g., panicle weight, grain width, and 1000-grain weight). Cytological analysis showed that changes in leaf width of ko1 and ko2 resulted from a decrease in leaf epidermal cell width and the number of small vascular bundles (SVBs) close to the leaf edge, suggesting that SiNL4 might regulate leaf width by influencing cell expansion and the development of SVB. Spatiotemporal expression analysis indicated that the relative expression level of SiNL4 was high in the stem, leaf, and young panicle. Subcellular localization showed that SiNL4 was mainly localized in the mitochondria and plasma membrane. In addition, the T-DNA insertion mutant (Atnl4) of AT5G18590, the ortholog of SiNL4 in Arabidopsis thaliana, exhibited similar phenotypes with reduced rosette leaf width, seed width, and 1000-seed weight. Moreover, complementary expression of SiNL4 in Atnl4 not only restored the phenotypes, but also significantly increased the 1000-seed weight, indicating that the function of these two genes might be conserved. Meanwhile, we found that SiNL4 knockout caused a decrease in chlorophyll content and net photosynthetic rate (Pn), showing that SiNL4 might be involved in regulating photosynthesis. In summary, this study revealed the function of SiNL4 in regulating leaf width in foxtail millet, providing a potential gene for the genetic improvement of foxtail millet.
Keywords: Kelch-repeat superfamily; leaf width; yield; foxtail millet; SiNL4 Kelch-repeat superfamily; leaf width; yield; foxtail millet; SiNL4

Share and Cite

MDPI and ACS Style

Zhao, Y.; Ma, Y.; Yang, Y.; Yang, L.; Chen, L.; Wang, T.; Wang, S.; Zhao, K.; Li, X.; Dong, S.; et al. The Kelch-Repeat Superfamily Gene SiNL4 Regulates the Leaf Width in Foxtail Millet. Plants 2026, 15, 1826. https://doi.org/10.3390/plants15121826

AMA Style

Zhao Y, Ma Y, Yang Y, Yang L, Chen L, Wang T, Wang S, Zhao K, Li X, Dong S, et al. The Kelch-Repeat Superfamily Gene SiNL4 Regulates the Leaf Width in Foxtail Millet. Plants. 2026; 15(12):1826. https://doi.org/10.3390/plants15121826

Chicago/Turabian Style

Zhao, Yuqin, Yixuan Ma, Yanyu Yang, Lejie Yang, Lu Chen, Tianguo Wang, Shiyuan Wang, Kai Zhao, Xiaorui Li, Shuqi Dong, and et al. 2026. "The Kelch-Repeat Superfamily Gene SiNL4 Regulates the Leaf Width in Foxtail Millet" Plants 15, no. 12: 1826. https://doi.org/10.3390/plants15121826

APA Style

Zhao, Y., Ma, Y., Yang, Y., Yang, L., Chen, L., Wang, T., Wang, S., Zhao, K., Li, X., Dong, S., Wang, H., Chu, X., Wang, J., Gao, L., & Yang, G. (2026). The Kelch-Repeat Superfamily Gene SiNL4 Regulates the Leaf Width in Foxtail Millet. Plants, 15(12), 1826. https://doi.org/10.3390/plants15121826

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