Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean

Soybean (<i>Glycine max</i> (L.) Merr.) is a critical crop in China, serving as a primary source of food, oil, and animal feed. Drought stress significantly impacts soybean growth and yield. MicroRNAs (miRNAs) play crucial roles in plant drought responses. The miR164 family is highly con...

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Main Authors: Wentao Hu, Man Zhang, Jie Lu, Miaomiao Zhang, Reqing He, Youlin Zhu, Dong Wang, Liyun Jiang
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Agronomy
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Online Access:https://www.mdpi.com/2073-4395/15/6/1450
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author Wentao Hu
Man Zhang
Jie Lu
Miaomiao Zhang
Reqing He
Youlin Zhu
Dong Wang
Liyun Jiang
author_facet Wentao Hu
Man Zhang
Jie Lu
Miaomiao Zhang
Reqing He
Youlin Zhu
Dong Wang
Liyun Jiang
author_sort Wentao Hu
collection DOAJ
description Soybean (<i>Glycine max</i> (L.) Merr.) is a critical crop in China, serving as a primary source of food, oil, and animal feed. Drought stress significantly impacts soybean growth and yield. MicroRNAs (miRNAs) play crucial roles in plant drought responses. The miR164 family is highly conserved across plant species and has been shown to participate in drought responses in a range of plants, yet the function of miR164 in soybean remains unclear. In this study, we identified <i>GmNAC35</i> as a direct target of miR164 through published degradome sequencing data and 5′ RLM-RACE assays. Under drought stress, miR164 members (e.g., <i>MIR164a</i>, <i>MIR164f</i>, and <i>MIR164k</i>) rapidly down-regulated, reaching their lowest expression at 2 h and returning to basal levels within 6 h. Conversely, <i>GmNAC35</i> showed an inverse pattern, indicating negative regulation by miR164. Overexpression of <i>GmNAC35</i> enhanced drought tolerance in transgenic soybean plants, as evidenced by higher survival rates and reduced water loss. Transcriptomic analysis revealed that <i>GmNAC35</i> modulates stress-responsive pathways, including ABA signaling and phenylpropanoid biosynthesis. Our findings indicate that miR164 negatively regulates <i>GmNAC35</i>, a positive regulator of drought tolerance. This enhances our understanding of the molecular mechanisms of drought tolerance in soybean and may inform strategies for breeding drought-resistant varieties.
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spelling doaj-art-a0167e87043c40f2a6ea289aad0a487f2025-08-20T03:26:14ZengMDPI AGAgronomy2073-43952025-06-01156145010.3390/agronomy15061450Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in SoybeanWentao Hu0Man Zhang1Jie Lu2Miaomiao Zhang3Reqing He4Youlin Zhu5Dong Wang6Liyun Jiang7Institute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaInstitute of Advanced Agricultural Sciences, College of Life Science, Nanchang University, Nanchang 330031, ChinaSoybean (<i>Glycine max</i> (L.) Merr.) is a critical crop in China, serving as a primary source of food, oil, and animal feed. Drought stress significantly impacts soybean growth and yield. MicroRNAs (miRNAs) play crucial roles in plant drought responses. The miR164 family is highly conserved across plant species and has been shown to participate in drought responses in a range of plants, yet the function of miR164 in soybean remains unclear. In this study, we identified <i>GmNAC35</i> as a direct target of miR164 through published degradome sequencing data and 5′ RLM-RACE assays. Under drought stress, miR164 members (e.g., <i>MIR164a</i>, <i>MIR164f</i>, and <i>MIR164k</i>) rapidly down-regulated, reaching their lowest expression at 2 h and returning to basal levels within 6 h. Conversely, <i>GmNAC35</i> showed an inverse pattern, indicating negative regulation by miR164. Overexpression of <i>GmNAC35</i> enhanced drought tolerance in transgenic soybean plants, as evidenced by higher survival rates and reduced water loss. Transcriptomic analysis revealed that <i>GmNAC35</i> modulates stress-responsive pathways, including ABA signaling and phenylpropanoid biosynthesis. Our findings indicate that miR164 negatively regulates <i>GmNAC35</i>, a positive regulator of drought tolerance. This enhances our understanding of the molecular mechanisms of drought tolerance in soybean and may inform strategies for breeding drought-resistant varieties.https://www.mdpi.com/2073-4395/15/6/1450soybeandrought responsemiR164<i>GmNAC35</i>transcriptomics
spellingShingle Wentao Hu
Man Zhang
Jie Lu
Miaomiao Zhang
Reqing He
Youlin Zhu
Dong Wang
Liyun Jiang
Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean
Agronomy
soybean
drought response
miR164
<i>GmNAC35</i>
transcriptomics
title Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean
title_full Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean
title_fullStr Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean
title_full_unstemmed Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean
title_short Negative Regulation of <i>GmNAC35</i> by miR164 Enhances Drought Tolerance in Soybean
title_sort negative regulation of i gmnac35 i by mir164 enhances drought tolerance in soybean
topic soybean
drought response
miR164
<i>GmNAC35</i>
transcriptomics
url https://www.mdpi.com/2073-4395/15/6/1450
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