Morphological Physiological and Transcriptional Response to Low Nitrogen Stress in Populus Deltoides Marsh. Clones With Contrasting Nitrogen Use Efficiency

Morphological Physiological and Transcriptional Response to Low Nitrogen Stress in Populus Deltoides Marsh. Clones With Contrasting Nitrogen Use Efficiency | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Morphological Physiological and Transcriptional Response to Low Nitrogen Stress in Populus Deltoides Marsh. Clones With Contrasting Nitrogen Use Efficiency Cun Chen, Yanguang Chu, Qinjun Huang, Weixi Zhang, Changjun Ding, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-149379/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Background: Nitrogen (N) is one of the main factors limiting the wood yield in poplar cultivation. Understanding the molecular mechanism of N utilization could play a guiding role in improving the nitrogen use efficiency (NUE). Results: In this study, three N-efficient genotypes (A) and three N-inefficient genotypes (C) of Populus deltoides were cultured under low N stress (5 μM NH 4 NO 3 ) and normal N supply (750 μM NH 4 NO 3 ). The dry matter mass, leaf morphology, and chlorophyll content of both genotypes decreased under N starvation. Interestingly, N starvation induced fine root growth in A, but not in C. Next, a detailed time-course analysis of enzyme activities and gene expression in leaves identified 2,062 differentially expressed genes (DEGs) in A and 1,118 in C, most of which were up-regulated. Moreover, the sensitivity to N starvation of A was weak, and DEGs related to hormone signal transduction played an important role in the low N response in A. The weighted gene co-expression network analysis identified genes related to membrane, catalytic activity, enzymatic activity, and response to stresses might be critical for poplar’s adaption to N starvation and these genes participated in the negative regulation of various biological processes. Finally, ten influential hub genes and twelve transcription factors were identified in the response to N starvation, among them Podel.19G001200, Podel.19G035300, Podel.02G021400, and Podel.04G076900 were related to programmed cell death, and the defense response, and PodelWRKY41 , PodelWRKY75 , PodelWRKY18 , PodelBHLH25 , PodelBHLH30 , PodelBHLH , and PodelHY5 were involved in plant signal transduction. Conclusions: Under the condition of N starvation, A showed stronger adaptability and a better NUE than C in morphology and physiology. The discovery of hub genes and TFs provided a new information for the analysis of the molecular mechanism of N efficient utilization and the improvement of NUE of poplar. Epigenetics & Genomics nitrogen deficiency nitrogen use efficiency gene expression Populus deltoides Marsh Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Full Text Supplementary Files SupplementaryTables.xlsx Additional file 1: Table S1. Low nitrogen adaptation coefficients of traits of N-efficient and N-inefficient genotypes. Table S2. Results of quality analysis of RNA sequencing data. Table S3. Results of the GO functional enrichment analysis of genes in the ‘magenta’ module. Table S4. Genes in the ‘magenta’ module. Table S5. Annotation description of the top 10 genes for connectivity (hub genes) in the ‘magenta’ module. Table S6. Annotation of transcription factors in the 'magenta' module. Table S7. The reaction system of quantitative real-time reverse transcription PCR (qRT-PCR). Table S8. Primers used for quantitative real-time reverse transcription PCR (qRT-PCR) analysis. SupplementaryFigures.docx Additional file 2: Fig. S1. Transcriptome relationships among three biological replicates. A: N-efficient genotypes; C: N-inefficient genotypes. T0, T2, T4, and T6 represent 0, 5, 20, and 40 days of N treatment, respectively. LN: low N treatment. Fig. S2. Expression pattern analysis of genes in the ‘magenta’ module. Red: upregulated; blue: downregulated. A: N efficient genotypes; C: N-inefficient genotypes. T0, T2, T4, and T6 represent 0, 5, 20, and 40 days of N treatment, respectively. LN: low N treatment. Fig. S3. Expression of key genes in nitrogen metabolism in the leaves of genotypes A and C. (A), (B), (C), (D), (E), (F), (G) and (H) represent the expression trends of NRT1;1, NRT1;2, AMT1;6, AMT2;1, NR, NiR, GS2, and GDH2, respectively. The columns represent the results of RNA sequencing, and the lines show the analysis results of qRT-PCR. Vertical bars indicate SDs (n = 3) in qRT-PCR analysis. A: N-efficient genotypes; C: N-inefficient genotypes. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Major revision 16 May, 2021 Reviews received at journal 13 May, 2021 Reviewers agreed at journal 01 May, 2021 Reviews received at journal 12 Feb, 2021 Reviewers agreed at journal 03 Feb, 2021 Reviewers invited by journal 20 Jan, 2021 Editor assigned by journal 20 Jan, 2021 Editor invited by journal 20 Jan, 2021 Submission checks completed at journal 20 Jan, 2021 First submitted to journal 17 Jan, 2021 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-149379","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":8637579,"identity":"5212ddea-6aca-4e91-8637-b547e22621f1","order_by":0,"name":"Cun Chen","email":"","orcid":"","institution":"State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry","correspondingAuthor":false,"prefix":"","firstName":"Cun","middleName":"","lastName":"Chen","suffix":""},{"id":8637580,"identity":"33078cab-f441-48fe-a2ae-ab8a1b0e7d6b","order_by":1,"name":"Yanguang 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Different letters above the column indicate significant differences between groups (p \u003c 0.05). (A) Height after treatment (Height-n, Hn); (B) Ground diameter after treatment (Ground diameter-n, GDn); (C) Fresh weight of the stem (SFW); (D) Dry weight of the stem (SDW); (E) Fresh weight of the root (RFW); (F) Dry weight of the root (RDW); (G) Fresh weight of the leaf (LFW); (H) Dry weight of the leaf (LDW).","description":"","filename":"OnlineFigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/cf8ba1b77842bf79aefe2fef.png"},{"id":5203305,"identity":"7c20dd53-a965-423b-8168-927df0a165a6","added_by":"auto","created_at":"2021-01-23 00:29:36","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":41948,"visible":true,"origin":"","legend":"Effects of low N stress on leaf morphology and chlorophyll content of N-efficient (A-1, A-2 and A-3) and N-inefficient (C-1, C-2 and C-3) genotypes. Different letters above the column indicate significant differences between the groups (p \u003c 0.05). (A) Chlorophyll a (Chl a); (B) Chlorophyll b (Chl b); (C) Carotenoid (Car); (D) Chlorophyll (a+b) (Chl); (E) Leaf length (LL); (F) Leaf width (LW); (G) Leaf area (LA).","description":"","filename":"OnlineFigure2.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/f052c334727151be911d57dc.png"},{"id":5203306,"identity":"94763f21-b700-4c43-9847-0e5f5f373324","added_by":"auto","created_at":"2021-01-23 00:29:36","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":59038,"visible":true,"origin":"","legend":"The change trends of enzyme activities, total amino acid contents, and soluble sugar contents in leaves during N treatment of N-efficient (A) and N-inefficient (C) genotypes. T0, T1, T2, T3, T4, T5, and T6 represent 0, 3, 5, 10, 20, 30, and 40 days of N treatment, respectively. (A) Nitrate reductase activities (NR); (B) Glutamine synthetase activities (GS); (C) Glutamate dehydrogenase activities (GDH); (D) Glutamic acid synthetase activities (GOGAT); (E) Total amino acid contents (AAs); (F) Soluble sugar contents (SSs).","description":"","filename":"OnlineFigure3.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/46e18b5fe2a602f8c160b138.png"},{"id":5203248,"identity":"e573536a-bdd7-4da4-b4be-31caa40d7ff7","added_by":"auto","created_at":"2021-01-23 00:26:36","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":30229,"visible":true,"origin":"","legend":"(A) Bar chart showing numbers of upregulated and downregulated differentially expressed genes (DEGs) in the four comparison groups (T0-C vs. T0-A, T2-LN-C vs. T2-LN-A, T4-LN-C vs. T4-LN-A and T6-LN-C vs. T6-LN-A). The magenta column shows upregulated DEGs, and the cyan column shows downregulated DEGs. LN: low nitrogen treatment. (B) Venn diagram showing that the distribution of DEGs identified in the comparison of genotypes A and C are common and specific to T0, T2, T4, and T6.","description":"","filename":"OnlineFigure4.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/3ce3186ab78c9018c16b915e.png"},{"id":5203261,"identity":"a2fb7411-2c92-4a45-a523-34563ed9b975","added_by":"auto","created_at":"2021-01-23 00:26:37","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":197744,"visible":true,"origin":"","legend":"Results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the special and common differentially expressed genes (DEGs) between genotypes A and C at different time points during the response to low N stress. T0 special, T2 special, T4 special, and T6 special represent the specific DEGs at T0, T2, T4, and T6 between genotypes A and C, respectively. Common indicates the common DEGs at T0, T2, T4, and T6 between genotypes A and C.","description":"","filename":"OnlineFigure5.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/4874f72f81ed967ffbfcf06e.png"},{"id":5203250,"identity":"7ad192a1-79a9-4694-8b5b-cb66e4902b7c","added_by":"auto","created_at":"2021-01-23 00:26:36","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":44477,"visible":true,"origin":"","legend":"(A) and (B) bar charts show numbers of upregulated and downregulated differentially expressed genes (DEGs) in the three comparison groups of A (T0-A vs. T2-LN-A, T2-LN-A vs. T4-LN-A, and T4-LN-A vs. T6-LN-A) and C (T0-C vs. T2-LN-C, T2-LN-C vs. T4-LN-C, and T4-LN-C vs. T6-LN-C) genotypes, respectively. The magenta column shows upregulated DEGs, and the cyan column shows downregulated DEGs. LN: low nitrogen treatment. (C) Venn diagrams showing that the distribution of DEGs identified in the comparison of different periods are common and specific to genotypes A and C. DEGs-A and DEGs-C represent all the DEGs identified from genotypes A and C during low N stress treatment, respectively.","description":"","filename":"OnlineFigure6.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/3409be84610297bf5e1a6770.png"},{"id":5203353,"identity":"9ebcfe26-7563-4a6b-8c51-196fd6cce724","added_by":"auto","created_at":"2021-01-23 00:32:37","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":325630,"visible":true,"origin":"","legend":"Results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the special and common differentially expressed genes (DEGs) between genotypes A and C during the response to low N stress. A-special and C-special represent the specific DEGs in genotypes A and C, respectively. AC-common indicates the common DEGs between genotypes A and C.","description":"","filename":"OnlineFigure7.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/9b893bc9a1517ebbf6fcc890.png"},{"id":5203254,"identity":"389ee214-c803-4e4d-bca3-0899fa532ba7","added_by":"auto","created_at":"2021-01-23 00:26:36","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":68039,"visible":true,"origin":"","legend":"Gene expression patterns across four-time points (T0, T2, T4, and T6) in genotypes A and C under low N stress. (A) and (C) indicate the variation trend of differentially expressed genes (DEGs) in genotypes A and C, respectively. Above the box is the ID of the changing trend, and the number in the box indicates the number of DEGs contained in the trend. The grid with color indicates a significantly enrichment trend ( p \u003c 0.05), and the closer the color is, the more similar the changing trend is. (B) and (D) represent the changing trend of genes in profile 12 with genotype A and profile 17 with genotype C, respectively.","description":"","filename":"OnlineFigure8.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/a9840c368541f3cc16f8fbb3.png"},{"id":5203258,"identity":"4521b4db-b1b1-4558-b622-978525326aeb","added_by":"auto","created_at":"2021-01-23 00:26:37","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":297671,"visible":true,"origin":"","legend":"Results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of differentially expressed genes (DEGs) in profile 12 of genotype A (A-profile 12) and profile 17 of genotype C (C-profile 17). The 10 pathways on the left are the top-10 metabolic pathways with significantly enrichment in A-profile 12, and the 10 pathways on the right are the top 10 metabolic pathways with significantly enrichment in C-profile 17 (p \u003c 0.05).","description":"","filename":"OnlineFigure9.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/0d9622d6ea2f647e1385a9a6.png"},{"id":5203307,"identity":"def1b561-e65a-40a4-ba2b-84fe2a127a4c","added_by":"auto","created_at":"2021-01-23 00:29:36","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":71627,"visible":true,"origin":"","legend":"Weighted gene co-expression network analysis (WGCNA) of differentially expressed genes (DEGs) identified in the genotypes A and C over three-time stages under low N stress. (A) Gene cluster dendrogram and 19 gene module divisions of DEGs, in which a major tree branch represents a module, and different colors represent different modules. (B) Correlation heatmap between modules and traits. Each column presents the experimental traits. The number in the oval box represents the correlation coefficient, which ranges from 1 (cyan) to 1 (red). We set an absolute value of the correlation coefficient greater than 0.75 to indicate that there is a strong correlation between gene modules and traits. The number in the rectangular box on the right indicates the number of the genes contained in the corresponding gene module.","description":"","filename":"OnlineFigure10.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/fa08dfcbfb535516e5b157ba.png"},{"id":5203308,"identity":"50adc6e7-f062-40ee-9042-f507888c8e8a","added_by":"auto","created_at":"2021-01-23 00:29:36","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":105482,"visible":true,"origin":"","legend":"Results of the gene ontology (GO) functional enrichment analysis of genes in the ‘magenta’ module. Red filled GO items were significantly enriched (p \u003c 0.05) and yellow ones were not significantly enriched (p \u003e 0.05). The rectangles in each network diagram represent the top 10 GO terms with significance in cellular component, molecular function, and biological process, respectively.","description":"","filename":"OnlineFigure11.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/54e3f1eff323a4e61028ef8c.png"},{"id":5203251,"identity":"057e0870-938d-4938-aa66-88af4b12c703","added_by":"auto","created_at":"2021-01-23 00:26:36","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":215701,"visible":true,"origin":"","legend":"Cytoscape representation of the top 150 network relationships related to hub genes that were selected according to the weight value in the ‘magenta’ module. The color of the lines between genes from orange to green to pink indicates that the correlation (weight value) between genes is becoming stronger, and the thicker the lines, the stronger the correlation (weight value). The larger the node, the pinker the color, indicating the greater connectivity of the gene in the module, and TFs represent transcription factors. The heat map next to the central gene shows the expression level of the gene in different samples, and the color from blue to orange indicates that the expression level is increasing. In the upper row, the four samples from left to right are T0-A, T2-LN-A, T4-LN-A, and T6-LN-A, respectively, and in the next row are T0-C, T2-LN-C, T4-LN-C, and T6-LN-C, respectively.","description":"","filename":"OnlineFigure12.png","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/95615adaf6d2c15bd96b6a57.png"},{"id":13578363,"identity":"e9bbc495-9e0c-4e5c-8842-c9585eab6fb6","added_by":"auto","created_at":"2021-09-17 04:15:23","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2637760,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1_covered.pdf"},{"id":5203387,"identity":"e25fbd13-5a46-49da-9420-cbb11fe489cb","added_by":"auto","created_at":"2021-01-23 00:35:40","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2852889,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1_stamped.pdf"},{"id":5203253,"identity":"99f807d7-93d9-4fab-bb85-e8cd6d9a8eed","added_by":"auto","created_at":"2021-01-23 00:26:36","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":102793,"visible":true,"origin":"","legend":"Additional file 1: Table S1. Low nitrogen adaptation coefficients of traits of N-efficient and N-inefficient genotypes. Table S2. Results of quality analysis of RNA sequencing data. Table S3. Results of the GO functional enrichment analysis of genes in the ‘magenta’ module. Table S4. Genes in the ‘magenta’ module. Table S5. Annotation description of the top 10 genes for connectivity (hub genes) in the ‘magenta’ module. Table S6. Annotation of transcription factors in the 'magenta' module. Table S7. The reaction system of quantitative real-time reverse transcription PCR (qRT-PCR). Table S8. Primers used for quantitative real-time reverse transcription PCR (qRT-PCR) analysis.","description":"","filename":"SupplementaryTables.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/267cc216285ba4370499cd08.xlsx"},{"id":5203310,"identity":"5332f607-8339-4df1-b9da-686dadca4d54","added_by":"auto","created_at":"2021-01-23 00:29:37","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":508932,"visible":true,"origin":"","legend":"Additional file 2: Fig. S1. Transcriptome relationships among three biological replicates. A: N-efficient genotypes; C: N-inefficient genotypes. T0, T2, T4, and T6 represent 0, 5, 20, and 40 days of N treatment, respectively. LN: low N treatment. Fig. S2. Expression pattern analysis of genes in the ‘magenta’ module. Red: upregulated; blue: downregulated. A: N efficient genotypes; C: N-inefficient genotypes. T0, T2, T4, and T6 represent 0, 5, 20, and 40 days of N treatment, respectively. LN: low N treatment. Fig. S3. Expression of key genes in nitrogen metabolism in the leaves of genotypes A and C. (A), (B), (C), (D), (E), (F), (G) and (H) represent the expression trends of NRT1;1, NRT1;2, AMT1;6, AMT2;1, NR, NiR, GS2, and GDH2, respectively. The columns represent the results of RNA sequencing, and the lines show the analysis results of qRT-PCR. Vertical bars indicate SDs (n = 3) in qRT-PCR analysis. A: N-efficient genotypes; C: N-inefficient genotypes.","description":"","filename":"SupplementaryFigures.docx","url":"https://assets-eu.researchsquare.com/files/rs-149379/v1/bdedd306c3a2dd550befc4f1.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eMorphological Physiological and Transcriptional Response to Low Nitrogen Stress in Populus Deltoides Marsh. Clones With Contrasting Nitrogen Use Efficiency\u003c/p\u003e","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-149379/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gics","sideBox":"Learn more about [BMC Genomics](http://bmcgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/gics","title":"BMC Genomics","twitterHandle":"#BMCGenomics","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"nitrogen deficiency, nitrogen use efficiency, gene expression, Populus deltoides Marsh","lastPublishedDoi":"10.21203/rs.3.rs-149379/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-149379/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eNitrogen (N) is one of the main factors limiting the wood yield in poplar cultivation. Understanding the molecular mechanism of N utilization could play a guiding role in improving the nitrogen use efficiency (NUE). \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eIn this study, three N-efficient genotypes (A) and three N-inefficient genotypes (C) of \u003cem\u003ePopulus deltoides\u003c/em\u003e were cultured under low N stress (5 μM NH\u003csub\u003e4\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e) and normal N supply (750 μM NH\u003csub\u003e4\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e). The dry matter mass, leaf morphology, and chlorophyll content of both genotypes decreased under N starvation. Interestingly, N starvation induced fine root growth in A, but not in C. Next, a detailed time-course analysis of enzyme activities and gene expression in leaves identified 2,062 differentially expressed genes (DEGs) in A and 1,118 in C, most of which were up-regulated. Moreover, the sensitivity to N starvation of A was weak, and DEGs related to hormone signal transduction played an important role in the low N response in A. The weighted gene co-expression network analysis identified genes related to membrane, catalytic activity, enzymatic activity, and response to stresses might be critical for poplar’s adaption to N starvation and these genes participated in the negative regulation of various biological processes. Finally, ten influential hub genes and twelve transcription factors were identified in the response to N starvation, among them Podel.19G001200, Podel.19G035300, Podel.02G021400, and Podel.04G076900 were related to programmed cell death, and the defense response, and \u003cem\u003ePodelWRKY41\u003c/em\u003e, \u003cem\u003ePodelWRKY75\u003c/em\u003e, \u003cem\u003ePodelWRKY18\u003c/em\u003e, \u003cem\u003ePodelBHLH25\u003c/em\u003e, \u003cem\u003ePodelBHLH30\u003c/em\u003e, \u003cem\u003ePodelBHLH\u003c/em\u003e, and \u003cem\u003ePodelHY5\u003c/em\u003e were involved in plant signal transduction.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e Under the condition of N starvation, A showed stronger adaptability and a better NUE than C in morphology and physiology. The discovery of hub genes and TFs provided a new information for the analysis of the molecular mechanism of N efficient utilization and the improvement of NUE of poplar.\u003c/p\u003e","manuscriptTitle":"Morphological Physiological and Transcriptional Response to Low Nitrogen Stress in Populus Deltoides Marsh. Clones With Contrasting Nitrogen Use Efficiency","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-01-23 00:26:34","doi":"10.21203/rs.3.rs-149379/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-05-17T03:33:31+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-05-13T12:50:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"fc745473-a312-4e17-b1e7-09d24c45a311","date":"2021-05-01T15:17:21+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-02-12T13:41:32+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"32536766-8a62-4daf-95f5-6842bbf41e89","date":"2021-02-04T04:50:55+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-01-20T15:00:48+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-01-20T14:45:56+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2021-01-20T13:12:15+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2021-01-20T13:09:10+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Genomics","date":"2021-01-17T12:55:06+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gics","sideBox":"Learn more about [BMC Genomics](http://bmcgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/gics","title":"BMC Genomics","twitterHandle":"#BMCGenomics","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f74030dd-9894-4a09-b9e0-00021ee6bbe7","owner":[],"postedDate":"January 23rd, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":2031491,"name":"Epigenetics \u0026 Genomics"}],"tags":[],"updatedAt":"2021-09-07T11:14:11+00:00","versionOfRecord":[],"versionCreatedAt":"2021-01-23 00:26:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-149379","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-149379","identity":"rs-149379","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}