4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles

Silver nanoparticles (AgNPs), the most extensively used nanomaterials, are inevitably released into the environment, yet their toxicological effects on plants remain not fully elucidated. To address a gap in the literature regarding the effects of AgNPs on cotton (Gossypium hirsutum L. cv. TM-1), a...

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Main Authors: Yuanhao Ding, Ping Li, Wenjie Lai, Yalan Li, Sihui Huang, Qiuping Zhong, Long Lu, Liyan Yin, Wei Gao, Haiyan Hu
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Plant Stress
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667064X25002386
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author Yuanhao Ding
Ping Li
Wenjie Lai
Yalan Li
Sihui Huang
Qiuping Zhong
Long Lu
Liyan Yin
Wei Gao
Haiyan Hu
author_facet Yuanhao Ding
Ping Li
Wenjie Lai
Yalan Li
Sihui Huang
Qiuping Zhong
Long Lu
Liyan Yin
Wei Gao
Haiyan Hu
author_sort Yuanhao Ding
collection DOAJ
description Silver nanoparticles (AgNPs), the most extensively used nanomaterials, are inevitably released into the environment, yet their toxicological effects on plants remain not fully elucidated. To address a gap in the literature regarding the effects of AgNPs on cotton (Gossypium hirsutum L. cv. TM-1), a 15-days exposure experiment was conducted using AgNPs (10 mg/L), along with Ag+ (0.4 mg/L AgNO3) serving as a mock control and water as the control (CK). Compared to the Ag+and CK groups, the roots of cotton seedlings exposed to AgNPs showed noticeable browning with significant reductions in biomass and root length. To explore the protein-level changes in cotton root response to AgNPs, a 4D label-free proteomics analysis was performed on the AgNPs, Ag+, and CK groups. Finally, 1588 differentially expressed proteins (DEPs) unique expressed in the AgNPs-treated group, which were primarily enriched in lignin and gossypol biosynthesis pathways, as revealed by GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses. Subsequent phloroglucinol staining and quantification confirmed an increased lignin biosynthesis, while LC-MS examination demonstrated elevated gossypol production in cotton roots following AgNPs exposure. Furthermore, roots from a glandless cotton mutant (CCRI12gl, lacking gossypol) were significant shorter than those of its wide-type counterpart (CCRI12, glandular, containing gossypol) after AgNPs treatment, suggesting that gossypol enhances cotton tolerance to AgNPs. Additionally, parallel reaction monitoring (PRM) technology validated that the expression levels of proteins involved in lignin and gossypol biosynthesis were consistent with the findings of the 4D label-free proteomics analysis. This study provides new insights into the mechanisms of plant responses to AgNPs exposure and offers a fundamental reference for investigating the toxicity of nanometal in crops.
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spelling doaj-art-a539010544844aeeba9734a32c5ecb752025-08-23T04:49:42ZengElsevierPlant Stress2667-064X2025-09-011710097010.1016/j.stress.2025.1009704D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticlesYuanhao Ding0Ping Li1Wenjie Lai2Yalan Li3Sihui Huang4Qiuping Zhong5Long Lu6Liyan Yin7Wei Gao8Haiyan Hu9College of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, China; School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, ChinaCollege of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, ChinaCollege of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, ChinaCollege of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, ChinaCollege of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, ChinaCollege of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, ChinaSchool of Life Science, Henan University, Henan 4750004, PR China; National Key Laboratory of Cotton Bio-breeding and Integrated Utilization (Henan University), Henan, 475004 PR ChinaOne Health Institute, Hainan University, Haikou 570228, China; Corresponding authors.School of Life Science, Henan University, Henan 4750004, PR China; National Key Laboratory of Cotton Bio-breeding and Integrated Utilization (Henan University), Henan, 475004 PR China; Corresponding authors.College of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572000, China; School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China; Corresponding authors.Silver nanoparticles (AgNPs), the most extensively used nanomaterials, are inevitably released into the environment, yet their toxicological effects on plants remain not fully elucidated. To address a gap in the literature regarding the effects of AgNPs on cotton (Gossypium hirsutum L. cv. TM-1), a 15-days exposure experiment was conducted using AgNPs (10 mg/L), along with Ag+ (0.4 mg/L AgNO3) serving as a mock control and water as the control (CK). Compared to the Ag+and CK groups, the roots of cotton seedlings exposed to AgNPs showed noticeable browning with significant reductions in biomass and root length. To explore the protein-level changes in cotton root response to AgNPs, a 4D label-free proteomics analysis was performed on the AgNPs, Ag+, and CK groups. Finally, 1588 differentially expressed proteins (DEPs) unique expressed in the AgNPs-treated group, which were primarily enriched in lignin and gossypol biosynthesis pathways, as revealed by GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses. Subsequent phloroglucinol staining and quantification confirmed an increased lignin biosynthesis, while LC-MS examination demonstrated elevated gossypol production in cotton roots following AgNPs exposure. Furthermore, roots from a glandless cotton mutant (CCRI12gl, lacking gossypol) were significant shorter than those of its wide-type counterpart (CCRI12, glandular, containing gossypol) after AgNPs treatment, suggesting that gossypol enhances cotton tolerance to AgNPs. Additionally, parallel reaction monitoring (PRM) technology validated that the expression levels of proteins involved in lignin and gossypol biosynthesis were consistent with the findings of the 4D label-free proteomics analysis. This study provides new insights into the mechanisms of plant responses to AgNPs exposure and offers a fundamental reference for investigating the toxicity of nanometal in crops.http://www.sciencedirect.com/science/article/pii/S2667064X25002386AgNPsCotton4D label-free proteomicsLigninGossypol
spellingShingle Yuanhao Ding
Ping Li
Wenjie Lai
Yalan Li
Sihui Huang
Qiuping Zhong
Long Lu
Liyan Yin
Wei Gao
Haiyan Hu
4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles
Plant Stress
AgNPs
Cotton
4D label-free proteomics
Lignin
Gossypol
title 4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles
title_full 4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles
title_fullStr 4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles
title_full_unstemmed 4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles
title_short 4D label-free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton (Gossypium hirsutum L. cv. TM-1) seedlings after a fifteen-days exposure to silver nanoparticles
title_sort 4d label free proteomics reveals that lignin and gossypol biosynthesis are enhanced in the roots of cotton gossypium hirsutum l cv tm 1 seedlings after a fifteen days exposure to silver nanoparticles
topic AgNPs
Cotton
4D label-free proteomics
Lignin
Gossypol
url http://www.sciencedirect.com/science/article/pii/S2667064X25002386
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