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Genome-wide identification and functional validation of asparagine synthetase genes (NtASNs) in Nicotiana tabacum.

Yan-Qun Zhang·Xin-Yao Huang·Wu-Xia Xi·Pei-Song Yang·Jia-Rong Wu·Jian-Cheng Wu·Shao-Ming Zhao·Hang Li·Rong Zhang·Shi-Ping Zhou·Wei-Guang Wang·Qiu-Fen Hu·Hua-Yin Liu
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Abstract

Asparagine (Asn) is pivotal for plant nitrogen (N) metabolism and plays indispensable roles in plant growth, development, and stress tolerance. However, the systematic characteristics and core functions of asparagine synthetase genes (NtASNs) in tobacco remain unclear. Through a comprehensive genome-wide investigation, nine members of the NtASN gene family were identified. Subsequent CRISPR/Cas9-mediated knockout and overexpression assays of these NtASN genes revealed that NtASN1e, NtASN2a, and NtASN2b are the core genes responsible for Asn biosynthesis in tobacco. Their knockout reduced asparagine synthetase activity and Asn content, delayed seed germination by 2-3 days, and displayed elevated oxidative injury when exposed to salinity conditions. In contrast, overexpression of these genes elevated Asn accumulation. Subcellular localization analysis indicated that NtASN1e was localized to both the cytoplasm and chloroplasts, whereas NtASN2a exhibited dual localization in the cytoplasm and endoplasmic reticulum, and NtASN2b was mainly localized in the cytoplasm. This study systematically clarifies the evolutionary characteristics and core functions of the NtASN gene family and provides candidate genes for optimizing nitrogen metabolism and improving salt-stress adaptation in tobacco. These findings hold important practical significance for molecular breeding and product quality improvement in industrial crops.

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NicotianaAspartate-Ammonia LigaseAsparaginePlant ProteinsGenes, PlantPlants, Genetically ModifiedGenome, PlantGene Expression Regulation, PlantMultigene FamilyAsparagine synthetaseBioinformaticsGene editingGene familyNicotiana tabacumJournal Article

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Yan-Qun Zhang, Xin-Yao Huang, Wu-Xia Xi, Pei-Song Yang, Jia-Rong Wu, Jian-Cheng Wu, Shao-Ming Zhao, Hang Li, Rong Zhang, Shi-Ping Zhou, Wei-Guang Wang, Qiu-Fen Hu, Hua-Yin Liu. 2026-08-19. Genome-wide identification and functional validation of asparagine synthetase genes (NtASNs) in Nicotiana tabacum.. https://doi.org/10.1016/j.plantsci.2026.113380

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