{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,13]],"date-time":"2026-06-13T20:30:41Z","timestamp":1781382641013,"version":"3.54.1"},"reference-count":62,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,12,27]],"date-time":"2019-12-27T00:00:00Z","timestamp":1577404800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,12,27]],"date-time":"2019-12-27T00:00:00Z","timestamp":1577404800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Quantitative real-time PCR (qPCR) is commonly used for deciphering gene functions. For effective qPCR analyses, suitable reference genes are needed for normalization. The objective of this study is to identify the appropriate reference gene(s) for qPCR analyses of the leaves and roots of ramie (<jats:italic>Boehmeria nivea<\/jats:italic> L.), an important natural fiber crop. To accomplish this goal, we investigated the expression patterns of eight common plant qPCR reference genes in ramie leaves and roots under five abiotic stresses, five hormonal treatments, and one biotic stress. The relative expression stabilities of the eight genes were evaluated using four common but different approaches: geNorm, NormFinder, BestKeeper, and RefFinder. Across the 11 tested conditions, <jats:italic>ACT1<\/jats:italic> was the most stably expressed among the eight genes while <jats:italic>GAPDH<\/jats:italic> displayed the biggest variation. Overall, while variations in the suggested reference genes were found for different tissue x treatment combinations, our analyses revealed that together, genes <jats:italic>ACT1<\/jats:italic>, <jats:italic>CYP2<\/jats:italic>, and <jats:italic>UBQ<\/jats:italic> can provide robust references for gene expression studies of ramie leaves under most conditions, while genes <jats:italic>EF-1\u03b1<\/jats:italic>, <jats:italic>TUB<\/jats:italic>, and <jats:italic>ACT1<\/jats:italic> can be used for similar studies of ramie roots. Our results should help future functional studies of the genes in ramie genome across tissues and environmental conditions.<\/jats:p>","DOI":"10.1038\/s41598-019-56640-3","type":"journal-article","created":{"date-parts":[[2019,12,27]],"date-time":"2019-12-27T11:02:42Z","timestamp":1577444562000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":37,"title":["Selection of Reference Genes for qPCR Analyses of Gene Expression in Ramie Leaves and Roots across Eleven Abiotic\/Biotic Treatments"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6818-4094","authenticated-orcid":false,"given":"Yongting","family":"Yu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6763-2254","authenticated-orcid":false,"given":"Gang","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yikun","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingqing","family":"Bai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunsheng","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangbin","family":"Zeng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhimin","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Cheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangping","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Litao","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2915-2780","authenticated-orcid":false,"given":"Jianping","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhun","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,12,27]]},"reference":[{"key":"56640_CR1","first-page":"103","volume":"86","author":"RA Srivastava","year":"1998","unstructured":"Srivastava, R. A. Analysis of RNA by Northern blotting using riboprobes. Methods Mol. Biol. 86, 103\u2013112 (1998).","journal-title":"Methods Mol. Biol."},{"key":"56640_CR2","first-page":"31","volume":"37","author":"MJ Tymms","year":"1995","unstructured":"Tymms, M. J. Quantitative measurement of mRNA using the RNase protection assay. Methods Mol. Biol. 37, 31\u201346 (1995).","journal-title":"Methods Mol. Biol."},{"key":"56640_CR3","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1080\/09553009514550171","volume":"67","author":"SM de Toledo","year":"1995","unstructured":"de Toledo, S. M., Azzam, E. I., Gasmann, M. K. & Mitchel, R. E. Use of semiquantitative reverse transcription-polymerase chain reaction to study gene expression in normal human skin fibroblasts following low dose-rate irradiation. Int. J. Radiat. Biol. 67, 135\u2013143 (1995).","journal-title":"Int. J. Radiat. Biol."},{"key":"56640_CR4","doi-asserted-by":"publisher","first-page":"986","DOI":"10.1101\/gr.6.10.986","volume":"6","author":"CA Heid","year":"1996","unstructured":"Heid, C. A., Stevens, J., Livak, K. J. & Williams, P. M. Real time quantitative PCR. Genome Res. 6, 986\u2013994 (1996).","journal-title":"Genome Res."},{"key":"56640_CR5","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1586\/14737159.5.4.493","volume":"5","author":"SA Bustin","year":"2005","unstructured":"Bustin, S. A. Real-time, fluorescence-based quantitative PCR: a snapshot of current procedures and preferences. Expert Rev. Mol. Diagn. 5, 493\u2013498 (2005).","journal-title":"Expert Rev. Mol. Diagn."},{"key":"56640_CR6","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1016\/S1471-4914(02)02355-9","volume":"8","author":"D Klein","year":"2002","unstructured":"Klein, D. Quantification using real-time PCR technology: applications and limitations. Trends Mol. Med. 8, 257\u2013260 (2002).","journal-title":"Trends Mol. Med."},{"key":"56640_CR7","doi-asserted-by":"publisher","first-page":"503","DOI":"10.1016\/S0301-472X(02)00806-8","volume":"30","author":"DG Ginzinger","year":"2002","unstructured":"Ginzinger, D. G. Gene quantification using real-time quantitative PCR: an emerging technology hits the mainstream. Exp. Hematol. 30, 503\u2013512 (2002).","journal-title":"Exp. Hematol."},{"key":"56640_CR8","doi-asserted-by":"publisher","first-page":"e0117569","DOI":"10.1371\/journal.pone.0117569","volume":"10","author":"C Tian","year":"2015","unstructured":"Tian, C. et al. Selection of suitable reference genes for qPCR normalization under abiotic stresses and hormone stimuli in carrot leaves. PloS One 10, e0117569 (2015).","journal-title":"PloS One"},{"key":"56640_CR9","doi-asserted-by":"publisher","first-page":"e93724","DOI":"10.1371\/journal.pone.0093724","volume":"9","author":"L Huang","year":"2014","unstructured":"Huang, L. et al. Identification of candidate reference genes in perennial ryegrass for quantitative RT-PCR under various abiotic stress conditions. PloS one 9, e93724 (2014).","journal-title":"PloS one"},{"key":"56640_CR10","doi-asserted-by":"publisher","first-page":"e46487","DOI":"10.1371\/journal.pone.0046487","volume":"7","author":"DT Le","year":"2012","unstructured":"Le, D. T. et al. Evaluation of candidate reference genes for normalization of quantitative RT-PCR in soybean tissues under various abiotic stress conditions. PloS one 7, e46487 (2012).","journal-title":"PloS one"},{"key":"56640_CR11","doi-asserted-by":"crossref","unstructured":"Hong, S. Y., Seo, P. J., Yang, M. S., Xiang, F. & Park, C. M. Exploring valid reference genes for gene expression studies in Brachypodium distachyon by real-time PCR. BMC Plant Biol. 8 (2008).","DOI":"10.1186\/1471-2229-8-112"},{"key":"56640_CR12","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2229-13-130","volume":"13","author":"TM Liu","year":"2013","unstructured":"Liu, T. M., Zhu, S. Y., Tang, Q. M., Yu, Y. T. & Tang, S. W. Identification of drought stress-responsive transcription factors in ramie (Boehmeria nivea L. Gaud). BMC Plant Biol. 13, 130 (2013).","journal-title":"BMC Plant Biol."},{"key":"56640_CR13","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1016\/j.indcrop.2014.10.002","volume":"68","author":"E Kipriotis","year":"2015","unstructured":"Kipriotis, E., Xiong, H. P., Vafeiadakis, T., Kiprioti, M. & Alexopoulou, E. Ramie and kenaf as feed crops. Ind. Crop. Prod. 68, 126\u2013130 (2015).","journal-title":"Ind. Crop. Prod."},{"key":"56640_CR14","first-page":"105","volume-title":"J. Northwest A&F Univ. (Nat. Sci. Ed.)","author":"YT Yu","year":"2011","unstructured":"Yu, Y. T. et al. Identification of a nematode isolate from rot root of ramie. J. Northwest A&F Univ. (Nat. Sci. Ed.) 39, (105\u2013109 (2011)."},{"key":"56640_CR15","doi-asserted-by":"publisher","first-page":"358","DOI":"10.1111\/jph.12428","volume":"164","author":"YT Yu","year":"2016","unstructured":"Yu, Y. T. et al. First report of black leaf spot caused by Alternaria alternata on ramie in China. J. Phytopath. 164, 358\u2013361 (2016).","journal-title":"J. Phytopath."},{"key":"56640_CR16","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1080\/07060661.2016.1230150","volume":"38","author":"YT Yu","year":"2016","unstructured":"Yu, Y. T. et al. First report of brown root rot caused by Pythium vexans on ramie in Hunan, China. Can. J. Plant Path. 38, 405\u2013410 (2016).","journal-title":"Can. J. Plant Path."},{"key":"56640_CR17","first-page":"318","volume":"44","author":"M Yang","year":"2014","unstructured":"Yang, M. et al. Identification of the pathogen causing leaf spot disease on Boehmeria nivea. Acta Phytopathol. Sin. 44, 318\u2013322 (2014).","journal-title":"Acta Phytopathol. Sin."},{"key":"56640_CR18","doi-asserted-by":"publisher","first-page":"1508","DOI":"10.1094\/PDIS-08-10-0556","volume":"94","author":"XX Wang","year":"2010","unstructured":"Wang, X. X. et al. First report of anthracnose caused by Colletotrichum gloeosporioides on ramie in China. Plant Dis. 94, 1508\u20131508 (2010).","journal-title":"Plant Dis."},{"key":"56640_CR19","doi-asserted-by":"publisher","first-page":"225","DOI":"10.4149\/av_2010_03_225","volume":"54","author":"J Li","year":"2009","unstructured":"Li, J., Zhang, X. Y. & Qian, Y. J. Molecular characterization of Ramie mosaic virus isolates detected in Jiangsu and Zhejiang provinces, China. Acta virologica 54, 225\u2013228 (2009).","journal-title":"Acta virologica"},{"key":"56640_CR20","first-page":"23","volume":"10","author":"LB Zeng","year":"2013","unstructured":"Zeng, L. B., Xue, Z. D., Yu, Y. T., Yan, Z. Y. & Yang, R. L. Changes of occurrence and control of Cocytodes caerulea Guen\u00e9e. Hunan Agr Sci 10, 23\u201324 (2013).","journal-title":"Hunan Agr Sci"},{"key":"56640_CR21","first-page":"17","volume":"4","author":"DX Peng","year":"1992","unstructured":"Peng, D. X. Effect of draught and flooding on the development of ramie plant. China\u2019s Fiber Crop. 4, 17\u201321 (1992).","journal-title":"China\u2019s Fiber Crop."},{"key":"56640_CR22","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1016\/j.envpol.2009.08.043","volume":"158","author":"B Yang","year":"2010","unstructured":"Yang, B. et al. Constitutional tolerance to heavy metals of a fiber crop, ramie (Boehmeria nivea), and its potential usage. Environ. Poll. 158, 551\u2013558 (2010).","journal-title":"Environ. Poll."},{"key":"56640_CR23","first-page":"246","volume":"22","author":"LY Li","year":"2006","unstructured":"Li, L. Y., Cui, G. X., Zou, X. M. & Liu, F. Root growth, fiber yield and quality for difference potassium efficiency ramie genotypes under lower potassium stress. Chin. Agr. Sci. Bull. 22, 246\u2013249 (2006).","journal-title":"Chin. Agr. Sci. Bull."},{"key":"56640_CR24","doi-asserted-by":"publisher","first-page":"3702789","DOI":"10.1155\/2016\/3702789","volume":"2016","author":"L Zeng","year":"2016","unstructured":"Zeng, L. et al. Transcriptome analysis of ramie (Boehmeria nivea L. Gaud.) in response to ramie moth (Cocytodes coerulea Guenee) infestation. BioMed Res. Int. 2016, 3702789 (2016).","journal-title":"BioMed Res. Int."},{"key":"56640_CR25","doi-asserted-by":"publisher","first-page":"21989","DOI":"10.3390\/ijms160921989","volume":"16","author":"YT Yu","year":"2015","unstructured":"Yu, Y. T. et al. Identification of ramie genes in response to Pratylenchus coffeae infection challenge by digital gene expression analysis. Int. J. Mol. Sci. 16, 21989\u201322007 (2015).","journal-title":"Int. J. Mol. Sci."},{"key":"56640_CR26","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.gene.2014.12.057","volume":"558","author":"TM Liu","year":"2015","unstructured":"Liu, T. M., Zhu, S. Y., Tang, Q. M. & Tang, S. W. Genome-wide transcriptomic profiling of ramie (Boehmeria nivea L. Gaud) in response to cadmium stress. Gene 558, 131\u2013137 (2015).","journal-title":"Gene"},{"key":"56640_CR27","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.gene.2014.09.014","volume":"552","author":"SY Zhu","year":"2014","unstructured":"Zhu, S. Y., Tang, S. W., Tang, Q. M. & Liu, T. M. Genome-wide transcriptional changes of ramie (Boehmeria nivea L. Gaud) in response to root-lesion nematode infection. Gene 552, 67\u201374 (2014).","journal-title":"Gene"},{"key":"56640_CR28","doi-asserted-by":"publisher","first-page":"1175","DOI":"10.1007\/s00425-014-2040-3","volume":"239","author":"G Deng","year":"2014","unstructured":"Deng, G. et al. Comparative proteome analysis of the response of ramie under N, P and K deficiency. Planta 239, 1175\u20131186 (2014).","journal-title":"Planta"},{"key":"56640_CR29","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1007\/s11105-011-0361-y","volume":"30","author":"XX Wang","year":"2012","unstructured":"Wang, X. X. et al. Characterization by suppression subtractive hybridization of transcripts that are differentially expressed in leaves of anthracnose-resistant ramie cultivar. Plant Mol. Biol. Rep. 30, 547\u2013555 (2012).","journal-title":"Plant Mol. Biol. Rep."},{"key":"56640_CR30","doi-asserted-by":"publisher","first-page":"675","DOI":"10.1007\/s00438-014-0842-4","volume":"289","author":"TM Liu","year":"2014","unstructured":"Liu, T. M., Zhu, S. Y., Tang, Q. M. & Tang, S. W. Identification of 32 full-length NAC transcription factors in ramie (Boehmeria nivea L. Gaud) and characterization of the expression pattern of these genes. Mol. Genet. Genomics 289, 675\u2013684 (2014).","journal-title":"Mol. Genet. Genomics"},{"key":"56640_CR31","doi-asserted-by":"publisher","first-page":"101","DOI":"10.3724\/SP.J.1006.2010.00101","volume":"36","author":"XF Ma","year":"2010","unstructured":"Ma, X. F. et al. Cloning and tissue expression of Actin1 gene in different fiber development phases of ramie [Boehmeria nivea (Linn.) Gaud]. Acta Agron. Sin. 36, 101\u2013108 (2010).","journal-title":"Acta Agron. Sin."},{"key":"56640_CR32","first-page":"76","volume":"34","author":"ZJ Tian","year":"2008","unstructured":"Tian, Z. J., Yi, R., Chen, J. R., Guo, Q. Q. & Zhang, X. W. Cloning and expression of cellulose synthase gene in ramie [Boehmeria nivea (Linn.) Gaud.]. Acta Agron. Sin. 34, 76\u201383 (2008).","journal-title":"Acta Agron. Sin."},{"key":"56640_CR33","doi-asserted-by":"publisher","first-page":"3493","DOI":"10.3390\/ijms16023493","volume":"16","author":"X An","year":"2015","unstructured":"An, X. et al. Transcriptome profiling and identification of transcription factors in ramie (Boehmeria nivea L. Gaud) in response to PEG treatment, using illumina paired-end sequencing technology. Int. J. Mol. Sci. 16, 3493\u20133511 (2015).","journal-title":"Int. J. Mol. Sci."},{"key":"56640_CR34","doi-asserted-by":"publisher","first-page":"e0160885","DOI":"10.1371\/journal.pone.0160885","volume":"11","author":"P Wang","year":"2016","unstructured":"Wang, P. et al. Selection of suitable reference genes for RT-qPCR normalization under abiotic stresses and hormone stimulation in Persimmon (Diospyros kaki Thunb). PloS one 11, e0160885 (2016).","journal-title":"PloS one"},{"key":"56640_CR35","doi-asserted-by":"publisher","first-page":"W71","DOI":"10.1093\/nar\/gkm306","volume":"35","author":"A Untergasser","year":"2007","unstructured":"Untergasser, A. et al. Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res. 35, W71\u201374 (2007).","journal-title":"Nucleic Acids Res."},{"issue":"3","key":"56640_CR36","doi-asserted-by":"publisher","first-page":"e0122515","DOI":"10.1371\/journal.pone.0122515","volume":"10","author":"W De Spiegelaere","year":"2015","unstructured":"De Spiegelaere, W. et al. Reference Gene Validation for RT-qPCR, a Note on Different Available Software Packages. PLoS One 10(3), e0122515 (2015).","journal-title":"PLoS One"},{"key":"56640_CR37","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2002-3-7-research0034","volume":"3","author":"J Vandesompele","year":"2002","unstructured":"Vandesompele, J. et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3, RESEARCH0034 (2002).","journal-title":"Genome Biol."},{"key":"56640_CR38","doi-asserted-by":"publisher","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","volume":"64","author":"CL Andersen","year":"2004","unstructured":"Andersen, C. L., Jensen, J. L. & Orntoft, T. F. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 64, 5245\u20135250 (2004).","journal-title":"Cancer Res."},{"key":"56640_CR39","doi-asserted-by":"publisher","first-page":"509","DOI":"10.1023\/B:BILE.0000019559.84305.47","volume":"26","author":"MW Pfaffl","year":"2004","unstructured":"Pfaffl, M. W., Tichopad, A., Prgomet, C. & Neuvians, T. P. Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper\u2013Excel-based tool using pair-wise correlations. Biotechnology Lett. 26, 509\u2013515 (2004).","journal-title":"Biotechnology Lett."},{"key":"56640_CR40","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1007\/s11103-012-9885-2","volume":"80","author":"F Xie","year":"2012","unstructured":"Xie, F., Xiao, P., Chen, D., Xu, L. & Zhang, B. miRDeepFinder: a miRNA analysis tool for deep sequencing of plant small RNAs. Plant Mol Biol. 80, 75\u201384 (2012).","journal-title":"Plant Mol Biol."},{"key":"56640_CR41","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1093\/dnares\/dsx047","volume":"25","author":"C Liu","year":"2018","unstructured":"Liu, C. et al. Draft genome analysis provides insights into the fiber yield, crude protein biosynthesis, and vegetative growth of domesticated ramie (Boehmeria nivea L. Gaud). DNA Res. 25, 173\u2013181 (2018).","journal-title":"DNA Res."},{"key":"56640_CR42","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-09584-5","volume":"7","author":"S Ullah","year":"2017","unstructured":"Ullah, S. et al. Interactive effect of gibberellic acid and NPK fertilizer combinations on ramie yield and bast fibre quality. Sci. Rep. 7, 10647 (2017).","journal-title":"Sci. Rep."},{"key":"56640_CR43","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.mgene.2015.01.003","volume":"3","author":"P Satya","year":"2015","unstructured":"Satya, P. et al. Start codon targeted (SCoT) polymorphism reveals genetic diversity in wild and domesticated populations of ramie (Boehmeria nivea L. Gaudich.), a premium textile fiber producing species. Meta gene 3, 62\u201370 (2015).","journal-title":"Meta gene"},{"key":"56640_CR44","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1007\/s11103-014-0214-9","volume":"86","author":"T Liu","year":"2014","unstructured":"Liu, T., Tang, S., Zhu, S., Tang, Q. & Zheng, X. Transcriptome comparison reveals the patterns of selection in domesticated and wild ramie (Boehmeria nivea L. Gaud). Plant Mol. Biol. 86, 85\u201392, https:\/\/doi.org\/10.1007\/s11103-014-0214-9 (2014).","journal-title":"Plant Mol. Biol."},{"key":"56640_CR45","doi-asserted-by":"publisher","first-page":"172","DOI":"10.3390\/molecules23010172","volume":"23","author":"Q Liu","year":"2018","unstructured":"Liu, Q. et al. Reference gene selection for quantitative real-Time reverse-transcriptase PCR in annual ryegrass (Lolium multiflorum) subjected to various abiotic stresses. Molecules 23, 172 (2018).","journal-title":"Molecules"},{"issue":"10","key":"56640_CR46","doi-asserted-by":"publisher","first-page":"265","DOI":"10.3390\/genes8100265","volume":"8","author":"Yongting Yu","year":"2017","unstructured":"Yu, Y. et al. Molecular cloning, recombinant expression and antifungal activity of BnCPI, a cystatin in ramie (Boehmeria nivea L.). Genes 8 (2017).","journal-title":"Genes"},{"key":"56640_CR47","doi-asserted-by":"publisher","first-page":"636","DOI":"10.1002\/2211-5463.12191","volume":"7","author":"X An","year":"2017","unstructured":"An, X. et al. Senescence is delayed when ramie (Boehmeria nivea L.) is transformed with the isopentyl transferase (ipt) gene under control of the SAG12 promoter. FEBS Open Bio. 7, 636\u2013644 (2017).","journal-title":"FEBS Open Bio."},{"key":"56640_CR48","doi-asserted-by":"publisher","first-page":"776","DOI":"10.1089\/dna.2016.3251","volume":"35","author":"C Huang","year":"2016","unstructured":"Huang, C. et al. A ramie bZIP transcription factor BnbZIP2 is involved in drought, salt, and heavy metal stress response. DNA Cell Biol. 35, 776\u2013786 (2016).","journal-title":"DNA Cell Biol."},{"key":"56640_CR49","doi-asserted-by":"publisher","first-page":"1742","DOI":"10.1007\/s11738-014-1742-2","volume":"37","author":"JS Zheng","year":"2015","unstructured":"Zheng, J. S. et al. Characterization of a glutamine synthetase gene BnGS1-2 from ramie (Boehmeria nivea L. Gaud) and biochemical assays of BnGS1-2-over-expressing transgenic tobacco. Acta Physiol. Plant. 37, 1742 (2015).","journal-title":"Acta Physiol. Plant."},{"key":"56640_CR50","doi-asserted-by":"crossref","unstructured":"Liu, T., Zhu, S., Tang, Q. & Tang, S. Identification of a CONSTANS homologous gene with distinct diurnal expression patterns in varied photoperiods in ramie (Boehmeria nivea L. Gaud). Gene 560 (2015).","DOI":"10.1016\/j.gene.2015.01.045"},{"key":"56640_CR51","doi-asserted-by":"publisher","first-page":"238","DOI":"10.1016\/j.ab.2009.01.024","volume":"387","author":"T Lovdal","year":"2009","unstructured":"Lovdal, T. & Lillo, C. Reference gene selection for quantitative real-time PCR normalization in tomato subjected to nitrogen, cold, and light stress. Analytical Biochem. 387, 238\u2013242 (2009).","journal-title":"Analytical Biochem."},{"key":"56640_CR52","doi-asserted-by":"publisher","first-page":"264","DOI":"10.1080\/14620316.2016.1148372","volume":"91","author":"GL Wang","year":"2016","unstructured":"Wang, G. L. et al. Comparison of nine reference genes for real-time quantitative PCR in roots and leaves during five developmental stages in carrot (Daucus carota L.). J. Hortic. Sci. Biotechnol. 91, 264\u2013270 (2016).","journal-title":"J. Hortic. Sci. Biotechnol."},{"key":"56640_CR53","doi-asserted-by":"publisher","first-page":"e0152356","DOI":"10.1371\/journal.pone.0152356","volume":"11","author":"Y Zhao","year":"2016","unstructured":"Zhao, Y. et al. Selection of reference genes for gene expression normalization in Peucedanum praeruptorum Dunn under abiotic stresses, hormone treatments and different Tissues. PloS one 11, e0152356 (2016).","journal-title":"PloS one"},{"key":"56640_CR54","doi-asserted-by":"publisher","first-page":"e95445","DOI":"10.1371\/journal.pone.0095445","volume":"9","author":"Y Lin","year":"2014","unstructured":"Lin, Y. et al. Validation of potential reference genes for qPCR in maize across abiotic stresses, hormone treatments, and tissue types. PloS one 9, e95445 (2014).","journal-title":"PloS one"},{"key":"56640_CR55","doi-asserted-by":"publisher","first-page":"1005","DOI":"10.1104\/pp.123.3.1005","volume":"123","author":"JS Jeon","year":"2000","unstructured":"Jeon, J. S. et al. Tissue-preferential expression of a rice alpha-tubulin gene, OsTubA1, mediated by the first intron. Plant Physiol. 123, 1005\u20131014 (2000).","journal-title":"Plant Physiol."},{"key":"56640_CR56","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1104\/pp.005538","volume":"130","author":"XB Li","year":"2002","unstructured":"Li, X. B., Cai, L., Cheng, N. H. & Liu, J. W. Molecular characterization of the cotton GhTUB1 gene that is preferentially expressed in fiber. Plant Physiol. 130, 666\u2013674 (2002).","journal-title":"Plant Physiol."},{"key":"56640_CR57","doi-asserted-by":"publisher","first-page":"961","DOI":"10.1104\/pp.107.107086","volume":"145","author":"RV Oakley","year":"2007","unstructured":"Oakley, R. V., Wang, Y. S., Ramakrishna, W., Harding, S. A. & Tsai, C. J. Differential expansion and expression of alpha- and beta-tubulin gene families in. Populus. Plant Physiol. 145, 961\u2013973 (2007).","journal-title":"Populus. Plant Physiol."},{"key":"56640_CR58","first-page":"10","volume":"21","author":"CH Zhang","year":"2011","unstructured":"Zhang, C. H., Wang, X. M., Wang, L. B., Wu, W. L. & Li, W. L. cDNA fragment cloning and expression analysis of actin gene in Rubus spp. Biotechnology 21, 10\u201315 (2011).","journal-title":"Biotechnology"},{"key":"56640_CR59","first-page":"27","volume":"22","author":"CH Zhang","year":"2012","unstructured":"Zhang, C. H. et al. cDNA fragment cloning and expression analysis of two actin genes in Beach plum (Prunus maritima Marshall). Biotechnology 22, 27\u201331 (2012).","journal-title":"Biotechnology"},{"key":"56640_CR60","doi-asserted-by":"publisher","first-page":"e31263","DOI":"10.1371\/journal.pone.0031263","volume":"7","author":"V Mafra","year":"2012","unstructured":"Mafra, V. et al. Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions. PloS one 7, e31263 (2012).","journal-title":"PloS one"},{"key":"56640_CR61","doi-asserted-by":"publisher","first-page":"286","DOI":"10.1016\/j.plaphy.2016.07.022","volume":"108","author":"HP Sun","year":"2016","unstructured":"Sun, H. P., Li, F., Ruan, Q. M. & Zhong, X. H. Identification and validation of reference genes for quantitative real-time PCR studies in Hedera helix L. Plant Physiol. Biochem. 108, 286\u2013294 (2016).","journal-title":"Plant Physiol. Biochem."},{"key":"56640_CR62","first-page":"55","volume":"22","author":"X Jiang","year":"2014","unstructured":"Jiang, X. et al. Reference gene selection for Real-time quantitative PCR normalization in switchgrass (Panicum virgatum L.) root tissue. J. Agr. Biotechnol. 22, 55\u201363 (2014).","journal-title":"J. Agr. Biotechnol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56640-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56640-3","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56640-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T17:57:36Z","timestamp":1671299856000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56640-3"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12,27]]},"references-count":62,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["56640"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-56640-3","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12,27]]},"assertion":[{"value":"10 August 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"16 December 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"27 December 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"20004"}}