{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T08:51:48Z","timestamp":1791535908293,"version":"4.3.4"},"publisher-location":"New York, NY","reference-count":25,"publisher":"Springer US","isbn-type":[{"value":"9781071651612","type":"print"},{"value":"9781071651629","type":"electronic"}],"license":[{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.springernature.com\/gp\/researchers\/text-and-data-mining"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.springernature.com\/gp\/researchers\/text-and-data-mining"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026]]},"DOI":"10.1007\/978-1-0716-5162-9_15","type":"book-chapter","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T22:03:29Z","timestamp":1787609009000},"page":"185-194","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Estimation of DNA Methylation in Wheat Using Reverse Phase Ultra-High-Performance Liquid Chromatography Coupled with Mass Spectrometry During Leaf Rust Pathogenesis"],"prefix":"10.1007","author":[{"given":"Uzma","family":"Afreen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aryan","family":"Verma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kunal","family":"Mukhopadhyay","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3023-9018","authenticated-orcid":false,"given":"Manish","family":"Kumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,7,2]]},"reference":[{"issue":"15","key":"15_CR1","doi-asserted-by":"publisher","first-page":"7730","DOI":"10.1021\/acs.analchem.6b01682","volume":"88","author":"J Yin","year":"2016","unstructured":"Yin J, Xu T, Zhang N, Wang H (2016) Three-enzyme cascade bioreactor for rapid digestion of genomic DNA into single nucleosides. Anal Chem 88(15):7730\u20137737. https:\/\/doi.org\/10.1021\/acs.analchem.6b01682","journal-title":"Anal Chem"},{"issue":"26","key":"15_CR2","doi-asserted-by":"publisher","first-page":"21883","DOI":"10.1021\/acsami.8b05399","volume":"10","author":"J Yin","year":"2018","unstructured":"Yin J, Chen S, Zhang N, Wang H (2018) Multienzyme cascade bioreactor for a 10 min digestion of genomic DNA into single nucleosides and quantitative detection of structural DNA modifications in cellular genomic DNA. ACS Appl Mater Interfaces 10(26):21883\u201321890. https:\/\/doi.org\/10.1021\/acsami.8b05399","journal-title":"ACS Appl Mater Interfaces"},{"issue":"12","key":"15_CR3","doi-asserted-by":"publisher","first-page":"1921","DOI":"10.1007\/s12298-024-01539-1","volume":"30","author":"U Afreen","year":"2024","unstructured":"Afreen U, Mukhopadhyay K, Kumar M (2024) DNA methylation in wheat: current understanding and future potential for enhancing biotic and abiotic stress tolerance. Physiol Mol Biol Plants 30(12):1921\u20131933. https:\/\/doi.org\/10.1007\/s12298-024-01539-1","journal-title":"Physiol Mol Biol Plants"},{"key":"15_CR4","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.trac.2013.04.010","volume":"50","author":"A Ga\u0142uszka","year":"2013","unstructured":"Ga\u0142uszka A, Migaszewski Z, Namie\u015anik J (2013) The 12 principles of green analytical chemistry and the SIGNIFICANCE mnemonic of green analytical practices. TrAC, Trends Anal Chem 50:78\u201384. https:\/\/doi.org\/10.1016\/j.trac.2013.04.010","journal-title":"TrAC, Trends Anal Chem"},{"issue":"5","key":"15_CR5","doi-asserted-by":"publisher","first-page":"798","DOI":"10.1016\/j.jpba.2005.02.037","volume":"38","author":"SK Branch","year":"2005","unstructured":"Branch SK (2005) Guidelines from the International Conference on Harmonisation (ICH). J Pharm Biomed Anal 38(5):798\u2013805. https:\/\/doi.org\/10.1016\/j.jpba.2005.02.037","journal-title":"J Pharm Biomed Anal"},{"issue":"1","key":"15_CR6","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/0021-9673(93)80635-l","volume":"645","author":"SA Leonard","year":"1993","unstructured":"Leonard SA, Wong SC, Nyce JW (1993) Quantitation of 5-methylcytosine by one-dimensional high-performance thin-layer chromatography. J Chromatogr A 645(1):189\u2013192. https:\/\/doi.org\/10.1016\/0021-9673(93)80635-l","journal-title":"J Chromatogr A"},{"issue":"2","key":"15_CR7","doi-asserted-by":"publisher","first-page":"378","DOI":"10.1016\/j.ab.2007.11.026","volume":"374","author":"AA Maga\u00f1a","year":"2007","unstructured":"Maga\u00f1a AA, Wrobel K, Caudillo YA, Zaina S, Lund G, Wrobel K (2007) High-performance liquid chromatography determination of 5-methyl-2\u2032-deoxycytidine, 2\u2032-deoxycytidine, and other deoxynucleosides and nucleosides in DNA digests. Anal Biochem 374(2):378\u2013385. https:\/\/doi.org\/10.1016\/j.ab.2007.11.026","journal-title":"Anal Biochem"},{"issue":"7","key":"15_CR8","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1134\/s0003683816070024","volume":"52","author":"SV Antonova","year":"2016","unstructured":"Antonova SV, Demina NG, Lobanov KV, Kuvaev TA, Mironov AS, Pushkov AA, Rumiantseva NF, Tyaglov BV, Shakulov RS, Malakhova II, Krasikov VD (2016) Analysis of natural nucleosides and their derivatives by thin-layer chromatography. Appl Biochem Microbiol 52(7):714\u2013721. https:\/\/doi.org\/10.1134\/s0003683816070024","journal-title":"Appl Biochem Microbiol"},{"issue":"2","key":"15_CR9","doi-asserted-by":"publisher","first-page":"10","DOI":"10.3390\/mps1020010","volume":"1","author":"C Roberts","year":"2018","unstructured":"Roberts C, Raner G, Isaacs G (2018) High performance liquid chromatography separation of epigenetic cytosine variants. Methods Protoc 1(2):10. https:\/\/doi.org\/10.3390\/mps1020010","journal-title":"Methods Protoc"},{"issue":"1","key":"15_CR10","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1007\/s00216-011-5308-5","volume":"402","author":"B Buszewski","year":"2011","unstructured":"Buszewski B, Noga S (2011) Hydrophilic interaction liquid chromatography (HILIC)\u2014a powerful separation technique. Anal Bioanal Chem\u2002402(1):231\u2013247.\u2002https:\/\/doi.org\/10.1007\/s00216-011-5308-5","journal-title":"Anal Bioanal Chem"},{"issue":"10","key":"15_CR11","doi-asserted-by":"publisher","first-page":"7250","DOI":"10.3390\/ijms12107250","volume":"12","author":"ES Souza","year":"2011","unstructured":"Souza ES, Zaramello L, Kuhnen CA, Da Silva Junkes B, Yunes RA, Heinzen VEF (2011) Estimating the octanol\/water partition coefficient for aliphatic organic compounds using semi-empirical electrotopological index. Int J Mol Sci 12(10):7250\u20137264. https:\/\/doi.org\/10.3390\/ijms12107250","journal-title":"Int J Mol Sci"},{"issue":"5","key":"15_CR12","doi-asserted-by":"publisher","first-page":"518","DOI":"10.1080\/15592294.2021.1924970","volume":"17","author":"D Beck","year":"2021","unstructured":"Beck D, Maamar MB, Skinner MK (2021) Genome-wide CpG density and DNA methylation analysis method (MeDIP, RRBS, and WGBS) comparisons.\u2002Epigenetics\u200217(5):518\u2013530.https:\/\/doi.org\/10.1080\/15592294.2021.1924970","journal-title":"Epigenetics"},{"key":"15_CR13","doi-asserted-by":"publisher","first-page":"4704","DOI":"10.1016\/j.csbj.2022.08.051","volume":"20","author":"W Gong","year":"2022","unstructured":"Gong W, Pan X, Xu D, Ji G, Wang Y, Tian Y, Cai J, Li J, Zhang Z, Yuan X (2022) Benchmarking DNA methylation analysis of 14 alignment algorithms for whole genome bisulfite sequencing in mammals. Comput Struct Biotechnol J 20:4704\u20134716. https:\/\/doi.org\/10.1016\/j.csbj.2022.08.051","journal-title":"Comput Struct Biotechnol J"},{"key":"15_CR14","doi-asserted-by":"publisher","unstructured":"Schmidt M, Van Bel M, Woloszynska M, Slabbinck B, Martens C, De Block M, Coppens F, Van Lijsebettens M (2017) Plant-RRBS, a bisulfite and next-generation sequencing-based methylome profiling method enriching for coverage of cytosine positions. BMC Plant Biol\u200217(115):1\u201314.\u2002https:\/\/doi.org\/10.1186\/s12870-017-1070-y","DOI":"10.1186\/s12870-017-1070-y"},{"issue":"4","key":"15_CR15","doi-asserted-by":"publisher","first-page":"3105","DOI":"10.3390\/ijms24043105","volume":"24","author":"H Li","year":"2023","unstructured":"Li H, Qamar MTU, Yang L, Liang J, You J, Wang L (2023) Current progress, applications and challenges of multi-omics approaches in sesame genetic improvement. Int J Mol Sci 24(4):3105. https:\/\/doi.org\/10.3390\/ijms24043105","journal-title":"Int J Mol Sci"},{"issue":"9","key":"15_CR16","doi-asserted-by":"publisher","first-page":"1580","DOI":"10.1093\/bioinformatics\/btx830","volume":"34","author":"Y Gao","year":"2017","unstructured":"Gao Y, Wang H, Zhang H, Wang Y, Chen J, Gu L (2017) PRAPI: post-transcriptional regulation analysis pipeline for Iso-Seq. Bioinformatics 34(9):1580\u20131582. https:\/\/doi.org\/10.1093\/bioinformatics\/btx830","journal-title":"Bioinformatics"},{"key":"15_CR17","doi-asserted-by":"publisher","unstructured":"Wardenaar R, Liu H, Colot V, Colom\u00e9-Tatch\u00e9 M, Johannes F (2013) Evaluation of MEDIP-CHIP in the context of whole-genome bisulfite sequencing (WGBS-SEQ) in Arabidopsis. In: Methods in molecular biology, vol. 1067. Humana Press, New York, NY, pp 203\u2013224. https:\/\/doi.org\/10.1007\/978-1-62703-607-8_13","DOI":"10.1007\/978-1-62703-607-8_13"},{"issue":"1","key":"15_CR18","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1093\/jxb\/erv449","volume":"67","author":"C H\u00e9brard","year":"2015","unstructured":"H\u00e9brard C, Peterson DG, Willems G, Delaunay A, Jesson B, Lef\u00e8bvre M, Barnes S, Maury S (2015) Epigenomics and bolting tolerance in sugar beet genotypes. J Exp Bot 67(1):207\u2013225. https:\/\/doi.org\/10.1093\/jxb\/erv449","journal-title":"J Exp Bot"},{"key":"15_CR19","doi-asserted-by":"publisher","unstructured":"Lephatsi M, Nephali L, Meyer V, Piater LA, Buthelezi N, Dubery IA, Opperman H, Brand M, Huyser J, Tugizimana F (2022) Molecular mechanisms associated with microbial biostimulant-mediated growth enhancement, priming and drought stress tolerance in maize plants. Sci Rep 12(10450):1\u201318. https:\/\/doi.org\/10.1038\/s41598-022-14570-7","DOI":"10.1038\/s41598-022-14570-7"},{"issue":"5","key":"15_CR20","doi-asserted-by":"publisher","first-page":"1888","DOI":"10.1111\/nph.18395","volume":"236","author":"AC Silva","year":"2022","unstructured":"Silva AC, Ruiz-Ferrer V, M\u00fcller SY, Pellegrin C, Abril-Ur\u00edas P, Mart\u00ednez-G\u00f3mez \u00c1, G\u00f3mez-Rojas A, Berenguer E, Testillano PS, Andr\u00e9s MF, Fenoll C, Akker SED, Escobar C (2022) The DNA methylation landscape of the root-knot nematode-induced pseudo-organ, the gall, in Arabidopsis, is dynamic, contrasting over time, and critically important for successful parasitism. New Phytol 236(5):1888\u20131907. https:\/\/doi.org\/10.1111\/nph.18395","journal-title":"New Phytol"},{"key":"15_CR21","doi-asserted-by":"publisher","unstructured":"Starczak M, Abakir A, Ruzov A, Gackowski D(2022)\u2002Detection\u2002and\u2002quantification\u2002of RNA modifications on RNA\u2013DNA hybrids using\u2002SID-UPLC-MS\/MS.\u2002In:\u2002Methods in molecular\u2002biology, vol. 2528.\u2002Humana\u2002Press, New York, NY, pp 127\u2013143. https:\/\/doi.org\/10.1007\/978-1-0716-2477-7_9","DOI":"10.1007\/978-1-0716-2477-7_9"},{"key":"15_CR22","doi-asserted-by":"publisher","unstructured":"White LK, Hesselberth JR (2022) Modification mapping by nanopore sequencing. Front Genet 13(1037134):1\u201314.\u2002https:\/\/doi.org\/10.3389\/fgene.2022.1037134","DOI":"10.3389\/fgene.2022.1037134"},{"issue":"10","key":"15_CR23","doi-asserted-by":"publisher","first-page":"9373","DOI":"10.1007\/s11033-012-1801-y","volume":"39","author":"D Singh","year":"2012","unstructured":"Singh D, Bhaganagare G, Bandopadhyay R,Prabhu KV, Gupta PK, Mukhopadhyay K (2012) Targeted spatio-temporal expression-based characterization of state of infection and time-point of maximum defense in wheat NILs during leaf rust infection. Mol Biol Rep 39(10):9373\u20139382. https:\/\/doi.org\/10.1007\/s11033-012-1801-y","journal-title":"Mol Biol Rep"},{"issue":"1","key":"15_CR24","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1007\/s00425-016-2600-9","volume":"245","author":"D Kumar","year":"2016","unstructured":"Kumar D, Dutta S, Singh D, Prabhu KV, Kumar M, Mukhopadhyay K (2016) Uncovering leaf rust responsive miRNAs in wheat (Triticum aestivum L.) using high-throughput sequencing and prediction of their targets through degradome analysis. Planta 245(1):161\u2013182. https:\/\/doi.org\/10.1007\/s00425-016-2600-9","journal-title":"Planta"},{"key":"15_CR25","doi-asserted-by":"publisher","unstructured":"Li XL, Yuan J, Dong YS, Fu CH, Li MT, Yu LJ (2016) Optimization of an HPLC method for determining the genomic methylation levels of Taxus Cells. J Chromatogr Sci  54(2):200\u2013205. https:\/\/doi.org\/10.1093\/chromsci\/bmv129","DOI":"10.1093\/chromsci\/bmv129"}],"container-title":["Methods in Molecular Biology","Plant Biotic Stress"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/978-1-0716-5162-9_15","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T08:25:05Z","timestamp":1791534305000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/978-1-0716-5162-9_15"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026]]},"ISBN":["9781071651612","9781071651629"],"references-count":25,"URL":"https:\/\/doi.org\/10.1007\/978-1-0716-5162-9_15","relation":{},"ISSN":["1064-3745","1940-6029"],"issn-type":[{"value":"1064-3745","type":"print"},{"value":"1940-6029","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026]]},"assertion":[{"value":"2 July 2026","order":1,"name":"first_online","label":"First Online","group":{"name":"ChapterHistory","label":"Chapter History"}},{"value":"The authors have no conflicts of interest to declare that are relevant to the content of this chapter.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of Interest"}}]}}