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Issues Mol. Biol.","CIMB"],"abstract":"<jats:p>Reverse transcription quantitative PCR (RT-qPCR) is one of the most widely used techniques for gene expression analysis in molecular biology. However, the accuracy of relative gene expression quantification largely depends on the stability of the reference genes used for normalization. Sambucus nigra L. (elderberry) is a valuable medicinal and edible plant rich in anthocyanins and other bioactive compounds. Despite its increasing research and application value, no reference genes have been validated for this species. In this study, we applied RT-qPCR alongside four algorithms (GeNorm, NormFinder, BestKeeper, and RefFinder) to assess the expression stability of nine candidate reference genes across ten samples representing five tissue types (including stems, flowers, leaves, roots, fruits) at different developmental stages. The results showed that VAMP and Pol were the most suitable reference gene combination for normalization across different tissues of S. nigra. For studies involving only vegetative tissues (leaves and stems), our results recommend that RPB2 and RPB5 are the most stable and suitable reference genes. This study provides reliable reference genes for accurate RT-qPCR-based gene expression analysis in S. nigra and establishes a useful methodological basis for future functional genomics and molecular breeding studies in this species.<\/jats:p>","DOI":"10.3390\/cimb48080776","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:38:54Z","timestamp":1785411534000},"page":"776","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification of Reference Genes for RT-qPCR Assays in Sambucus nigra L."],"prefix":"10.3390","volume":"48","author":[{"given":"Zhengkun","family":"Cui","sequence":"first","affiliation":[{"name":"Shandong Academy of Forestry, Jinan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shandong Academy of Forestry, Jinan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengyu","family":"Gao","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Shandong Agriculture and Engineering University, Jinan 255300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinyin","family":"Fu","sequence":"additional","affiliation":[{"name":"Shandong Academy of Forestry, Jinan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yin","family":"Sun","sequence":"additional","affiliation":[{"name":"Shandong Academy of Forestry, Jinan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Ren","sequence":"additional","affiliation":[{"name":"Shandong Academy of Forestry, Jinan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junxiu","family":"Yao","sequence":"additional","affiliation":[{"name":"Shandong Academy of Forestry, Jinan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1734","DOI":"10.1105\/tpc.108.059774","article-title":"Towards a systematic validation of references in real-time RT-PCR","volume":"20","author":"Gutierrez","year":"2008","journal-title":"Plant Cell"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Ling, H., Wu, Q., Guo, J., Xu, L., and Que, Y. (2014). Comprehensive selection of reference genes for gene expression normalization in sugarcane by real time quantitative RT-PCR. PLoS ONE, 9, Correction in PLoS ONE 2015, 10, e0118444.","DOI":"10.1371\/journal.pone.0118444"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1093\/jxb\/ern305","article-title":"Normalization of qRT-PCR data: The necessity of adopting a systematic, experimental conditions-specific, validation of references","volume":"60","author":"Guenin","year":"2009","journal-title":"J. Exp. Bot."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"research0034.1","DOI":"10.1186\/gb-2002-3-7-research0034","article-title":"Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes","volume":"3","author":"Vandesompele","year":"2002","journal-title":"Genome Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s00425-010-1158-1","article-title":"Evaluation of candidate reference genes for expression studies in Pisum sativum under different experimental conditions","volume":"232","author":"Die","year":"2010","journal-title":"Planta"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1007\/s10126-010-9277-z","article-title":"Normalizing RT-qPCR data: Are we getting the right answers? An appraisal of normalization approaches and internal reference genes from a case study in the finfish Lates calcarifer","volume":"13","author":"Jerry","year":"2011","journal-title":"Mar. Biotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.ymeth.2009.11.001","article-title":"How to do successful gene expression analysis using real-time PCR","volume":"50","author":"Derveaux","year":"2010","journal-title":"Methods"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1007\/s13353-013-0173-x","article-title":"Reference genes in real-time PCR","volume":"54","author":"Kozera","year":"2013","journal-title":"J. Appl. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1038\/ng1032","article-title":"Microarray data normalization and transformation","volume":"32","author":"Quackenbush","year":"2002","journal-title":"Nat. Genet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S0168-1656(99)00163-7","article-title":"Housekeeping genes as internal standards: Use and limits","volume":"75","author":"Thellin","year":"1999","journal-title":"J. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5070","DOI":"10.1038\/s41598-020-61739-z","article-title":"Selection of reference genes for normalization of RT-qPCR data in gene expression studies in Anthonomus eugenii Cano (Coleoptera: Curculionidae)","volume":"10","author":"Pinheiro","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"22205","DOI":"10.1038\/s41598-020-79213-1","article-title":"Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp","volume":"10","author":"Zhang","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"798","DOI":"10.32615\/bp.2020.154","article-title":"Transcriptome-based screening and the optimal reference genes for real-time quantitative PCR in Rehmannia chingii and R. henryi","volume":"64","author":"Zuo","year":"2020","journal-title":"Biol. Plant."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1002\/2211-5463.12903","article-title":"Selection of reference genes for RT-qPCR normalization in blueberry (Vaccinium corymbosum \u00d7 angustifolium) under various abiotic stresses","volume":"10","author":"Deng","year":"2020","journal-title":"FEBS Open Bio"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Dai, Q., Lu, M., Yang, X., Lei, C., Huang, F., Hu, X., Huang, X., Nie, X., Chen, D., and Huang, S. (2025). qRT-PCR Reference Gene Selection for the Discoloration of Tender Leaves in Hawk Tea (Litsea coreana). Curr. Issues Mol. Biol., 47.","DOI":"10.3390\/cimb47020131"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7331","DOI":"10.1038\/s41598-024-58139-y","article-title":"Validation of reference gene stability for normalization of RT-qPCR in Phytophthora capsici Leonian during its interaction with Piper nigrum L","volume":"14","author":"Vijayakumar","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","article-title":"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","volume":"64","author":"Andersen","year":"2004","journal-title":"Cancer Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1023\/B:BILE.0000019559.84305.47","article-title":"Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper\u2013Excel-based tool using pair-wise correlations","volume":"26","author":"Pfaffl","year":"2004","journal-title":"Biotechnol. Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1007\/s10142-023-01055-7","article-title":"RefFinder: A web-based tool for comprehensively analyzing and identifying reference genes","volume":"23","author":"Xie","year":"2023","journal-title":"Funct. Integr. Genom."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1046\/j.1095-8339.2003.t01-1-00158.x","article-title":"An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG II","volume":"141","author":"Apg","year":"2003","journal-title":"Bot. J. Linn. Soc."},{"key":"ref_21","unstructured":"Enescu, C.M., Houston Durrant, T., and Caudullo, G. (2016). Sambucus nigra in Europe: Distribution, habitat, usage and threats. European Atlas of Forest Tree Species, Publication Office of the European Union."},{"key":"ref_22","unstructured":"Hult\u00e9n, E., and Fries, M. (1986). Atlas of North European Vascular Plants North of the Tropic of Cancer, Koeltz Scientific Books."},{"key":"ref_23","unstructured":"Lid, J. (1979). Norsk og Svensk Flora, Det Norske Samlaget."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1095-8339.1997.tb01785.x","article-title":"The geographical relationships of British and Irish vascular plants","volume":"120","author":"Preston","year":"1997","journal-title":"Bot. J. Linn. Soc."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/j.jff.2017.11.025","article-title":"Bioactive properties of Sambucus nigra L. as a functional ingredient for food and pharmaceutical industry","volume":"40","author":"Mlynarczyk","year":"2018","journal-title":"J. Funct. Foods"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/j.ctim.2018.12.004","article-title":"Black elderberry (Sambucus nigra) supplementation effectively treats upper respiratory symptoms: A meta-analysis of randomized, controlled clinical trials","volume":"42","author":"Hawkins","year":"2019","journal-title":"Complement. Ther. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1016\/j.phytochem.2009.06.003","article-title":"Elderberry flavonoids bind to and prevent H1N1 infection in vitro","volume":"70","author":"Roschek","year":"2009","journal-title":"Phytochemistry"},{"key":"ref_28","first-page":"213","article-title":"Elderberry: Botany, horticulture, potential","volume":"37","author":"Charlebois","year":"2010","journal-title":"Hortic. Rev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4202","DOI":"10.1021\/acs.jafc.2c00010","article-title":"Elderberry (Sambucus nigra L.): Bioactive Compounds, Health Functions, and Applications","volume":"70","author":"Liu","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"W200","DOI":"10.1093\/nar\/gky448","article-title":"HMMER web server: 2018 update","volume":"46","author":"Potter","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Paolacci, A.R., Tanzarella, O.A., Porceddu, E., and Ciaffi, M. (2009). Identification and validation of reference genes for quantitative RT-PCR normalization in wheat. BMC Mol. Biol., 10.","DOI":"10.1186\/1471-2199-10-11"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Niu, K.J., Shi, Y., and Ma, H.L. (2017). Selection of Candidate Reference Genes for Gene Expression Analysis in Kentucky Bluegrass (Poa pratensis L.) under Abiotic Stress. Front. Plant Sci., 8.","DOI":"10.3389\/fpls.2017.00193"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/gkp045","article-title":"Amplification efficiency: Linking baseline and bias in the analysis of quantitative PCR data","volume":"37","author":"Ruijter","year":"2009","journal-title":"Nucleic Acids Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"113641","DOI":"10.1016\/j.ab.2020.113641","article-title":"Bona fide gene expression analysis of samples from the bovine reproductive system by microfluidic platform","volume":"596","author":"Fontes","year":"2020","journal-title":"Anal. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Silver, N., Best, S., Jiang, J., and Thein, S.L. (2006). Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR. BMC Mol. Biol., 7.","DOI":"10.1186\/1471-2199-7-33"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1101\/gr.6.10.986","article-title":"Real time quantitative PCR","volume":"6","author":"Heid","year":"1996","journal-title":"Genome Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"367","DOI":"10.4319\/lom.2006.4.367","article-title":"Housekeeping gene selection for quantitative real-time PCR assays in the seagrass Zostera marina subjected to heat stress","volume":"4","author":"Ransbotyn","year":"2006","journal-title":"Limnol. Oceanogr. Methods"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ab.2004.08.024","article-title":"Equivalence test in quantitative reverse transcription Polymerase chain reaction: Confirmation of reference genes suitable for normalization","volume":"335","author":"Haller","year":"2004","journal-title":"Anal. Biochem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1007\/s11103-010-9666-8","article-title":"Genome-wide identification and evaluation of novel internal control genes for Q-PCR based transcript normalization in wheat","volume":"74","author":"Long","year":"2010","journal-title":"Plant Mol. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Xu, E.Y., Schneper, L.M., and Notterman, D.A. (2023). A novel metric to improve mismatched primer selection and quantification accuracy in amplifying DNA repeats for quantitative Polymerase chain reactions. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0292559"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"609","DOI":"10.12688\/wellcomeopenres.23147.1","article-title":"The genome sequence of black elder, Sambucus nigra Linnaeus, 1753 (Adoxaceae)","volume":"9","author":"Christenhusz","year":"2024","journal-title":"Wellcome Open Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"R112","DOI":"10.1186\/gb-2011-12-11-r112","article-title":"Evaluation of genomic high-throughput sequencing data generated on Illumina HiSeq and genome analyzer systems","volume":"12","author":"Minoche","year":"2011","journal-title":"Genome Biol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S., and Madden, T.L. (2012). Primer-BLAST: A tool to design target-specific primers for Polymerase chain reaction. BMC Bioinform., 13.","DOI":"10.1186\/1471-2105-13-134"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"You, F.M., Huo, N., Gu, Y.Q., Lazo, G.R., Dvorak, J., and Anderson, O.D. (2009). ConservedPrimers 2.0: A high-throughput pipeline for comparative genome referenced intron-flanking PCR primer design and its application in wheat SNP discovery. BMC Bioinform., 10.","DOI":"10.1186\/1471-2105-10-331"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Piriyapongsa, J., Ngamphiw, C., Assawamakin, A., Wangkumhang, P., Suwannasri, P., Ruangrit, U., Agavatpanitch, G., and Tongsima, S. (2009). RExPrimer: An integrated primer designing tool increases PCR effectiveness by avoiding 3\u2032 SNP-in-primer and mis-priming from structural variation. BMC Genom., 10.","DOI":"10.1186\/1471-2164-10-S3-S4"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Mallona, I., Weiss, J., and Egea-Cortines, M. (2011). pcrEfficiency: A Web tool for PCR amplification efficiency prediction. BMC Bioinform., 12.","DOI":"10.1186\/1471-2105-12-404"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1002","DOI":"10.1183\/09031936.05.00050205","article-title":"Choice of endogenous control for gene expression in non-small cell lung cancer","volume":"26","author":"Liu","year":"2005","journal-title":"Eur. Respir. J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1016\/j.bbrc.2003.11.177","article-title":"Guideline to reference gene selection for quantitative real-time PCR","volume":"313","author":"Thulke","year":"2004","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1186\/s13007-017-0238-7","article-title":"Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress","volume":"13","author":"Tang","year":"2017","journal-title":"Plant Methods"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Tu, Z., Hao, Z., Zhong, W., and Li, H. (2019). Identification of suitable reference genes for RT-qPCR assays in Liriodendron chinense (Hemsl.) Sarg. Forests, 10.","DOI":"10.3390\/f10050441"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s13353-016-0374-1","article-title":"Evaluation of stability and validation of reference genes for RT-qPCR expression studies in rice plants under water deficit","volume":"58","author":"Auler","year":"2017","journal-title":"J. Appl. Genet."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1186\/s13007-018-0354-z","article-title":"Cowpea and abiotic stresses: Identification of reference genes for transcriptional profiling by qPCR","volume":"14","author":"Amorim","year":"2018","journal-title":"Plant Methods"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/s11103-012-9885-2","article-title":"miRDeepFinder: A miRNA analysis tool for deep sequencing of plant small RNAs","volume":"80","author":"Xie","year":"2012","journal-title":"Plant Mol. Biol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.plaphy.2016.02.011","article-title":"Identification and validation of reference genes for accurate normalization of real-time quantitative PCR data in kiwifruit","volume":"102","author":"Rey","year":"2016","journal-title":"Plant Physiol. Biochem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"7042","DOI":"10.1038\/srep07042","article-title":"The choice of reference genes for assessing gene expression in sugarcane under salinity and drought stresses","volume":"4","author":"Guo","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Ma, L., Wu, J., Qi, W., Coulter, J.A., Fang, Y., Li, X., Liu, L., Jin, J., Niu, Z., and Yue, J. (2020). Screening and verification of reference genes for analysis of gene expression in winter rapeseed (Brassica rapa L.) under abiotic stress. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0236577"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"16640","DOI":"10.1038\/s41598-019-51040-z","article-title":"Stable reference genes for RT-qPCR analysis of gene expression in the Musa acuminata\u2013Pseudocercospora musae interaction","volume":"9","author":"Rego","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1104\/pp.106.092973","article-title":"Increases in the number of SNARE genes parallels the rise of multicellularity among the green plants","volume":"144","author":"Sanderfoot","year":"2007","journal-title":"Plant Physiol."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Zhang, L., Zhang, H., Liu, P., Hao, H., Jin, J.B., and Lin, J. (2011). Arabidopsis R-SNARE Proteins VAMP721 and VAMP722 Are Required for Cell Plate Formation. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0026129"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1915","DOI":"10.1007\/s11095-026-04123-2","article-title":"Targeting VAMPs in Aging-Related Diseases: From Mechanisms to Pharmacotherapy","volume":"43","author":"Jia","year":"2026","journal-title":"Pharm. Res."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Yang, Y., Xu, X., Jing, Z., Ye, J., Li, H., Li, X., Shi, L., Chen, M., Wang, T., and Xie, B. (2022). Genome-Wide Screening and Stability Verification of the Robust Internal Control Genes for RT-qPCR in Filamentous Fungi. J. Fungi, 8.","DOI":"10.3390\/jof8090952"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/j.bbagrm.2016.08.004","article-title":"The RNAs of RNA-directed DNA methylation","volume":"1860","author":"Wendte","year":"2017","journal-title":"Biochim. Biophys. Acta-Gene Regul. Mech."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Rocha, M.A., Gowda, B.S., and Fleischmann, J. (2022). RNAP II produces capped 18S and 25S ribosomal RNAs resistant to 5\u2032-monophosphate dependent processive 5\u2032 to 3\u2032 exonuclease in Polymerase switched Saccharomyces cerevisiae. BMC Mol. Cell Biol., 23.","DOI":"10.1186\/s12860-022-00417-6"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1093\/pcp\/pcr177","article-title":"Regulation of Pol I-Transcribed 45S rDNA and Pol III-Transcribed 5S rDNA in Arabidopsis","volume":"53","author":"Layat","year":"2012","journal-title":"Plant Cell Physiol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1007\/s00343-019-9053-5","article-title":"Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii","volume":"38","author":"Chen","year":"2020","journal-title":"J. Oceanol. Limnol."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Jarosov\u00e1, J., and Kundu, J.K. (2010). Validation of reference genes as internal control for studying viral infections in cereals by quantitative real-time RT-PCR. BMC Plant Biol., 10.","DOI":"10.1186\/1471-2229-10-146"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/nsmb.1539","article-title":"An atypical RNA Polymerase involved in RNA silencing shares small subunits with RNA Polymerase II","volume":"16","author":"Huang","year":"2009","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_68","first-page":"1600","article-title":"NRPB3, the third largest subunit of RNA Polymerase II, is essential for stomatal patterning and differentiation in Arabidopsis","volume":"143","author":"Chen","year":"2016","journal-title":"Development"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"5","DOI":"10.15252\/embj.2020105565","article-title":"The RNA Polymerase II subunit RPB-9 recruits the integrator complex to terminate Caenorhabditis elegans piRNA transcription","volume":"40","author":"Berkyurek","year":"2021","journal-title":"Embo J."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"e00320","DOI":"10.1002\/pld3.320","article-title":"High resolution RNA-seq profiling of genes encoding ribosomal proteins across different organs and developmental stages in Arabidopsis thaliana","volume":"5","author":"Xiong","year":"2021","journal-title":"Plant Direct"},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Zhang, D., Li, J.J., Li, B.Y., Li, C.M., Chen, X.Y., and Ouyang, K.X. (2020). Internal Reference Gene Selection under Different Hormone Stresses in Multipurpose Timber Yielding Tree Neolamarckia cadamba. Forests, 11.","DOI":"10.3390\/f11091014"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1080\/14620316.2021.1879686","article-title":"Selection and validation of reference genes in all-red Amaranth (Amaranthus tricolor L.) seedlings under different culture conditions","volume":"96","author":"Xiao","year":"2021","journal-title":"J. Hortic. Sci. Biotechnol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"3030","DOI":"10.1111\/pce.14916","article-title":"Cytosolic ABA Receptor Kinases phosphorylate the D6 PROTEIN KINASE leading to its stabilization which promotes Arabidopsis growth","volume":"47","author":"He","year":"2024","journal-title":"Plant Cell Environ."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"1674","DOI":"10.1105\/tpc.113.111484","article-title":"D6PK AGCVIII Kinases Are Required for Auxin Transport and Phototropic Hypocotyl Bending in Arabidopsis","volume":"25","author":"Willige","year":"2013","journal-title":"Plant Cell"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1023\/A:1005896711321","article-title":"Morphological changes and hypomethylation of DNA in transgenic tobacco expressing antisense RNA of the S-adenosyl-L-homocysteine hydrolase gene","volume":"35","author":"Tanaka","year":"1997","journal-title":"Plant Mol. Biol."}],"container-title":["Current Issues in Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1467-3045\/48\/8\/776\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:02:38Z","timestamp":1785412958000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1467-3045\/48\/8\/776"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,30]]},"references-count":75,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["cimb48080776"],"URL":"https:\/\/doi.org\/10.3390\/cimb48080776","relation":{},"ISSN":["1467-3045"],"issn-type":[{"value":"1467-3045","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,30]]}}}