Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize

Nitrogen use efficiency remains the primary bottleneck for sustainable maize production. This study elucidates the functional mechanisms of the amino acid transporter <i>ZmAAP1</i> in nitrogen absorption and stress resilience. Through systematic evolutionary analysis of 55 maize inbred l...

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Main Authors: Mo Zhu, Ziyu Wang, Shijie Li, Siping Han
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/14/2242
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author Mo Zhu
Ziyu Wang
Shijie Li
Siping Han
author_facet Mo Zhu
Ziyu Wang
Shijie Li
Siping Han
author_sort Mo Zhu
collection DOAJ
description Nitrogen use efficiency remains the primary bottleneck for sustainable maize production. This study elucidates the functional mechanisms of the amino acid transporter <i>ZmAAP1</i> in nitrogen absorption and stress resilience. Through systematic evolutionary analysis of 55 maize inbred lines, we discovered that the <i>ZmAAP1</i> gene family exhibits distinct chromosomal localization (Chr7 and Chr9) and functional domain diversification (e.g., group 10-specific motifs 11/12), indicating species-specific adaptive evolution. Integrative analysis of promoter cis-elements and multi-omics data confirmed the root-preferential expression of ZmAAP1 under drought stress, mediated via the ABA-DRE signaling pathway. To validate its biological role, we generated transgenic maize lines expressing Arabidopsis thaliana <i>AtAAP1</i> via Agrobacterium-mediated transformation. Three generations of genetic stability screening confirmed the stable genomic integration and root-specific accumulation of the AtAAP1 protein (Southern blot/Western blot). Field trials demonstrated that low-N conditions enhanced the following transgenic traits: the chlorophyll content increased by 13.5%, and the aboveground biomass improved by 7.2%. Under high-N regimes, the gene-pyramided hybrid ZD958 (AAP1 + AAP1) achieved a 12.3% yield advantage over conventional varieties. Our findings reveal <i>ZmAAP1</i>’s dual role in root development and long-distance nitrogen transport, establishing it as a pivotal target for molecular breeding. This study provides actionable genetic resources for enhancing NUE in maize production systems.
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spelling doaj-art-652919ba6d974da0b26d3e95625bf98f2025-08-20T03:56:47ZengMDPI AGPlants2223-77472025-07-011414224210.3390/plants14142242Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in MaizeMo Zhu0Ziyu Wang1Shijie Li2Siping Han3Institute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, ChinaInstitute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, ChinaInstitute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, ChinaInstitute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, ChinaNitrogen use efficiency remains the primary bottleneck for sustainable maize production. This study elucidates the functional mechanisms of the amino acid transporter <i>ZmAAP1</i> in nitrogen absorption and stress resilience. Through systematic evolutionary analysis of 55 maize inbred lines, we discovered that the <i>ZmAAP1</i> gene family exhibits distinct chromosomal localization (Chr7 and Chr9) and functional domain diversification (e.g., group 10-specific motifs 11/12), indicating species-specific adaptive evolution. Integrative analysis of promoter cis-elements and multi-omics data confirmed the root-preferential expression of ZmAAP1 under drought stress, mediated via the ABA-DRE signaling pathway. To validate its biological role, we generated transgenic maize lines expressing Arabidopsis thaliana <i>AtAAP1</i> via Agrobacterium-mediated transformation. Three generations of genetic stability screening confirmed the stable genomic integration and root-specific accumulation of the AtAAP1 protein (Southern blot/Western blot). Field trials demonstrated that low-N conditions enhanced the following transgenic traits: the chlorophyll content increased by 13.5%, and the aboveground biomass improved by 7.2%. Under high-N regimes, the gene-pyramided hybrid ZD958 (AAP1 + AAP1) achieved a 12.3% yield advantage over conventional varieties. Our findings reveal <i>ZmAAP1</i>’s dual role in root development and long-distance nitrogen transport, establishing it as a pivotal target for molecular breeding. This study provides actionable genetic resources for enhancing NUE in maize production systems.https://www.mdpi.com/2223-7747/14/14/2242NUEAAPsmaizephylogenetic analysisexpression analysis
spellingShingle Mo Zhu
Ziyu Wang
Shijie Li
Siping Han
Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize
Plants
NUE
AAPs
maize
phylogenetic analysis
expression analysis
title Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize
title_full Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize
title_fullStr Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize
title_full_unstemmed Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize
title_short Genetic Improvement and Functional Characterization of AAP1 Gene for Enhancing Nitrogen Use Efficiency in Maize
title_sort genetic improvement and functional characterization of aap1 gene for enhancing nitrogen use efficiency in maize
topic NUE
AAPs
maize
phylogenetic analysis
expression analysis
url https://www.mdpi.com/2223-7747/14/14/2242
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AT ziyuwang geneticimprovementandfunctionalcharacterizationofaap1geneforenhancingnitrogenuseefficiencyinmaize
AT shijieli geneticimprovementandfunctionalcharacterizationofaap1geneforenhancingnitrogenuseefficiencyinmaize
AT sipinghan geneticimprovementandfunctionalcharacterizationofaap1geneforenhancingnitrogenuseefficiencyinmaize