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====== Molecular Methods in Diagnosis and Detection of Regulated Xanthomonads ======

This data is based on the review by [[https://doi.org/10.3390/microorganisms9040862|Catara et al. (2021]]), focused on the molecular methods for diagnosis, detection, and studies on the diversity of plant pathogenic //Xanthomonas//, concentrating especially on regulated pathogens in the European Union published as a collective effort of the [[https://euroxanth.eu/management/wg1|Working Group 1]] 'Diagnostics & Diversity–Population Structure' from the [[https://euroxanth.eu|EuroXanth]] COST Action CA16107.

**Main protocols described for regulated xanthomonads based on amplification of specific target DNAs.**

^  Disease                                                        ^  Bacteria                                                                                                                  ^  Conventional PCR                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     ^  qPCR<sup>1</sup>                                                                                                                                                                                                                                                                                                                                ^  IA<sup>2</sup>    Methods                                                                                                                                                                                                                                                                                                                                        ^
| Onion bacterial blight                                          | //X. euvesicatoria// pv. //alli//                                                                                          | [[https://doi.org/10.1128/AEM.02697-09|Robène-Soustrade (2010)]]                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      | [[https://doi.org/10.1016/j.mimet.2015.04.017|Robène (2015)]]                                                                                                                                                                                                                                                                                    |  NA<sup>3</sup>                                                                                                                                                                                                                                                                                                                                                   |
| Citrus bacterial canker                                         | //X. citri// pv. //citri// \\ //X. aurantifolii// pathotypes B and C                                                       | [[https://doi.org/10.1094/Phyto-86-95|Hartung (1996)]], \\  [[https://doi.org/10.1128/AEM.68.3.1257-1264.2002|Cubero (2002)]], \\  [[https://doi.org/10.1111/j.1365-2672.2005.02787.x|Coletta-Filho (2006)]], \\  [[https://doi.org/10.1016/j.micres.2005.07.005|Suk Park (2006)]], \\  [[https://doi.org/10.1111/j.1365-3059.2010.02390.x|Kositcharoenkul (2011)]], \\  [[https://doi.org/10.7717/peerj.7676|Fonseca (2019)]]                                                                                                                                        | [[https://doi.org/10.1094/PHYTO.2004.94.1.61|Mavrodieva (2004)]], \\  [[https://doi.org/10.1094/PHYTO-95-1333|Cubero (2005)]], \\  [[https://doi.org/10.1186/s12866-020-01972-8|Robène (2020)]]                                                                                                                                                  | [[https://doi.org/10.1186/1471-2180-10-176|Rigano (2010)]], \\  [[https://doi.org/10.3390/microorganisms10061153|Webster (2022)]]                                                                                                                                                                                                                                 |
| Bacterial spot of stone fruits, walnut blight, hazelnut blight  | //X. arboricola// pvs. //pruni//, //corylina// and //juglandis//                                                           | [[https://doi.org/10.1007/s12275-010-0072-3|Park (2010)]], \\  [[https://doi.org/10.1094/PDIS-10-16-1481-RE|Fernandes (2017)]], \\  [[https://doi.org/10.1016/j.mimet.2019.03.003|Jouen (2019)]], \\  [[https://doi.org/10.1007/s42161-020-00505-6|Webber (2020)]], \\  [[https://doi.org/10.3389/fpls.2023.1254107|Kałużna (2023)]]                                                                                                                                                                                                                                  | [[https://doi.org/10.1128/AEM.01593-10|Palacio-Bielsa (2011)]], \\  [[https://doi.org/10.3389/fmicb.2017.00573|Garita-Cambronero (2017)]], \\  [[https://doi.org/10.1094/PDIS-12-18-2253-RE|Martins (2019)]], \\  [[https://doi.org/10.3389/fpls.2023.1254107|Kałużna (2023)]], \\  [[https://doi.org/10.1094/PDIS-01-24-0012-RE|Panth (2024)]]  | [[https://doi.org/10.1111/j.1365-3059.2012.02654.x|Bühlmann (2013)]], \\  [[https://doi.org/10.5423/PPJ.OA.07.2019.0197|Li (2021)]], \\  [[https://doi.org/10.3389/fpls.2023.1254107|Kałużna (2023)]]                                                                                                                                                             |
| Bacterial leaf blight and bacterial leaf streak of rice         | //X. oryzae// pvs. //oryzae// and //oryzicola//                                                                            | [[https://doi.org/10.1007/s002530100641|Sakthivel (2001)]], \\  [[https://pubmed.ncbi.nlm.nih.gov/18852502/|Kang (2008)]], \\  [[https://doi.org/10.1094/PDIS-94-3-0311|Lang (2010)]], \\  [[https://doi.org/10.1094/PDIS-06-10-0399|Cho (2011)]], \\  [[https://doi.org/10.1016/S2095-3119(12)60047-1|Shen (2012)]], \\  [[https://doi.org/10.3389/fmicb.2015.00791|Singhal (2015)]], \\  [[https://doi.org/10.1111/jam.13094|Cui (2016)]]                                                                                                                           | [[https://doi.org/10.1094/PDIS-06-10-0399|Cho (2011)]], \\  [[https://doi.org/10.1016/j.cropro.2012.04.014|Kang (2012)]], \\  [[https://doi.org/10.18699/VJGB-22-66|Koroleva (2022)]]                                                                                                                                                            | [[https://doi.org/10.1128/AEM.00274-14|Lang (2014)]], \\ [[https://doi.org/10.1016/j.bios.2022.114076|Zhu (2022)]], \\ [[https://doi.org/10.1016/j.cropro.2023.106466|Buddhachat (2024)]]                                                                                                                                                                         |
| Bacterial spot of pepper and tomato                             | //X. euvesicatoria// pvs. //euvesicatoria// and //perforans//, \\ //X. hortorum// pv. //gardneri//, \\ //X. vesicatoria//  | [[https://doi.org/10.1111/j.1472-765X.2009.02690.x|Moretti (2009)]], \\  [[https://doi.org/10.1111/j.1365-2672.2012.05431.x|Araújo (2011)]], \\  [[https://doi.org/10.1007/s10658-013-0225-4|Araújo (2013)]], \\  [[https://doi.org/10.1016/j.cropro.2012.08.008|Beran (2013)]], \\  [[https://doi.org/10.17660/ActaHortic.2009.808.13|Koenraadt (2019)]], \\  [[https://doi.org/10.1016/j.cropro.2019.104978|Pečenka (2020)]], \\  [[https://doi.org/10.5423/PPJ.OA.08.2022.0119|Siddique (2023)]], \\  [[https://doi.org/10.1094/PHYTO-12-22-0479-R|Utami (2023)]]  | [[https://doi.org/10.1094/PDIS-09-15-1085-RE|Strayer (2016)]], \\  [[https://doi.org/10.1016/j.cropro.2019.104978|Pečenka (2020)]], \\  [[https://doi.org/10.5423/PPJ.OA.08.2022.0119|Siddique (2023)]], \\  [[https://doi.org/10.1094/PHYTO-12-22-0479-R|Utami (2023)]]                                                                         | [[https://doi.org/10.1038/s41598-018-32295-4|Larrea-Sarmiento (2018)]], \\  [[https://doi.org/10.1094/PHYTO-03-18-0101-R|Strayer-Scherer (2019)]], \\  [[https://doi.org/10.3390/microorganisms8091301|Stehlíková (2020)]], \\  [[https://doi.org/10.1094/PDIS-05-22-1098-RE|Beran (2023)]], \\  [[https://doi.org/10.5423/PPJ.OA.08.2022.0119|Siddique (2023)]]  |
| Bacterial angular leaf spot of strawberry                       | //X. fragariae//                                                                                                           | [[https://doi.org/10.17660/ActaHortic.1997.439.136|Hartung (1997)]], \\  [[https://agris.fao.org/agris-search/search.do?recordID=IT2001060012|Cruz (1999)]], \\  [[https://doi.org/10.1016/j.mimet.2004.04.002|Stöger (2004)]], \\  [[https://doi.org/10.1111/j.1365-2338.2005.00812.x|Moltmann (2005)]]                                                                                                                                                                                                                                                              | [[https://doi.org/10.1094/PHYTO-98-3-0359|Turechek (2008)]], \\  [[https://doi.org/10.1111/j.1365-3059.2007.01813.x|Vandroemme (2008)]], \\  [[https://doi.org/10.17660/ActaHortic.2009.842.44|Cubero (2009)]]                                                                                                                                   | [[https://doi.org/10.1371/journal.pone.0147122|Wang (2016)]], [[https://doi.org/10.1111/ppa.12665|Gétaz (2017)]], \\  [[https://doi.org/10.1094/PDIS-10-19-2248-RE|Wang (2020)]], \\  [[https://doi.org/10.1111/jph.12900|Immanuel (2020)]]                                                                                                                       |
| Bacterial blight of anthurium and other aroids                  | //X. phaseoli// pv. //dieffenbachiae//                                                                                     | [[https://doi.org/10.1007/s10658-005-7062-z|Khoodoo (2005)]], \\  [[https://doi.org/10.1128/AEM.72.2.1072-1078.2006|Robène-Soustrade (2006)]], \\  [[https://doi.org/10.1111/ppa.12083|Chabirand (2014)]]                                                                                                                                                                                                                                                                                                                                                             | [[https://doi.org/10.1016/j.mimet.2019.03.003|Jouen (2019)]]                                                                                                                                                                                                                                                                                     | [[https://doi.org/10.1007/s10658-015-0653-4|Jun-Hai (2015)]]                                                                                                                                                                                                                                                                                                      |
| Bacterial leaf spot of poinsettia                               | //X. axonopodis// pv. //poinsetticola//                                                                                    | [[https://doi.org/10.5423/PPJ.OA.04.2015.0049|Back (2015)]]                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           |  NA                                                                                                                                                                                                                                                                                                                                              |  NA                                                                                                                                                                                                                                                                                                                                                               |
| Bacterial leaf streak and black chaff of cereals                | //X. translucens// pv. //translucens//                                                                                     | [[https://doi.org/10.1111/j.1365-2338.1995.tb01459.x|Maes (1995)]], \\  [[https://doi.org/10.1094/PHYTO-07-22-0262-SA|Roman-Reyna (2023)]], \\  [[https://doi.org/10.1094/PHYTO-10-22-0381-SA|Hong (2023)]]                                                                                                                                                                                                                                                                                                                                                           | [[https://doi.org/10.1094/PDIS-03-22-0574-RE|Sarkes (2022)]], \\  [[https://doi.org/10.1094/PHYTO-01-23-0022-SA|Tambong (2023)]], \\  [[https://doi.org/10.1094/PDIS-05-23-0887-SR|Fu (2023)]], \\  [[https://doi.org/10.1094/PDIS-09-23-1827-RE|Tambong (2024)]]                                                                                | [[https://doi.org/10.1094/PHYTO-08-16-0286-R|Langlois (2017)]], \\  [[https://doi.org/10.1094/PDIS-03-22-0574-RE|Sarkes (2022)]]                                                                                                                                                                                                                                  |
| Common blight of bean                                           | //X. phaseoli// pv. phaseoli, \\ //X. citri// pv. //fuscans//                                                              | [[https://doi.org/10.1094/Phyto-86-361|Audy (1996)]]                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  |  NA                                                                                                                                                                                                                                                                                                                                              | [[https://doi.org/10.1094/PDIS-02-19-0325-RE|De Paiva (2020)]]                                                                                                                                                                                                                                                                                                    |

<sup>1</sup>  qPCR: quantitative real-time PCR; <sup>2</sup>  IA: isothermal amplification: <sup>3</sup>  NA: not applicable.
===== References =====

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Jouen E, Chiroleu F, Maillot-Lebon V, Chabirand A, Merion S, Boyer C, Pruvost O, Robène I (2019). A duplex quantitative real-time PCR assay for the detection and quantification of //Xanthomonas phaseoli// pv. //dieffenbachiae //from diseased and latently infected anthurium tissue. J. Microbiol. Methods 161: 74-83. DOI: [[https://doi.org/10.1016/j.mimet.2019.03.003|j.mimet.2019.03.003]]

Jun-Hai N, Yue-Rong G, Jun-Mei Y, Qing-Yun L, Guang-Sui Y, Cun W, Yu R (2015). Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of bacterial blight pathogen (//Xanthomonas axonopodis// pv. //dieffenbachiae//) in anthurium. Eur. J. Plant Pathol. 142: 801-813. DOI: [[https://doi.org/10.1007/s10658-015-0653-4|10.1007/s10658-015-0653-4]]

Kałużna M, Prokić A, Obradović A, Weldon WA, Stockwell VO, Pothier JF (2023). Specific and sensitive detection tools for //Xanthomonas arboricola// pv. corylina, the causal agent of bacterial blight of hazelnut, developed with comparative genomics. Front. Plant Sci. 14: 1254107. DOI: [[https://doi.org/10.3389/fpls.2023.1254107|10.3389/fpls.2023.1254107]]

Kang MJ, Kim MH, Hwang DJ, Cho MS, Seol Y, Hahn JH, Ryu S, Park DS (2012). Quantitative in planta PCR assay for specific detection of //Xanthomonas oryzae// pv. //oryzicola //using putative membrane protein based primer set. Crop. Prot. 40: 22-27. DOI: [[https://doi.org/10.1016/j.cropro.2012.04.014|10.1016/j.cropro.2012.04.014]]

Kang MJ, Shim JK, Cho MS, Seol YJ, Hahn JH, Hwang DJ, Park DS (2008). Specific detection of //Xanthomonas oryzae// pv. //oryzicola //in infected rice plant by use of PCR assay targeting a membrane fusion protein gene. J. Microbiol. Biotechnol. 18: 1492-1495. PMID: [[https://pubmed.ncbi.nlm.nih.gov/18852502/|18852502]]

Khoodoo MHR, Sahin F, Jaufeerally-Fakim Y (2005). Sensitive detection of //Xanthomonas axonopodis// pv. //dieffenbachiae// on //Anthurium andreanum// by immunocapture-PCR (IC-PCR) using primers designed from sequence characterized amplified regions (SCAR) of the blight pathogen. Eur. J. Plant Pathol. 112: 379-390. DOI: [[https://doi.org/10.1007/s10658-005-7062-z|10.1007/s10658-005-7062-z]]

Koenraadt H, van Betteray B, Germain R, Hiddink G, Jones JB, Oosterhof J (2009). Development of specific primers for the molecular detection of bacterial spot of pepper and tomato. Acta Hortic. 808: 99-102. DOI: [[https://doi.org/10.17660/ActaHortic.2009.808.13|10.17660/ActaHortic.2009.808.13]]

Koroleva ML, Blinova SA, Shvartsev AA, Kurochkin VE, Alekseev YI (2022). Molecular genetic detection and differentiation of //Xanthomonas oryzae// pv. //oryzicola//, bacterial leaf streak agents of rice. Vavilovskii Zhurnal Genet. Selektsii. 26: 544-552. DOI: [[https://doi.org/10.18699/VJGB-22-66|10.18699/VJGB-22-66]]

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