Detection of the soybean bacterial blight pathogen Pseudomonas savastanoi pv. Glycinea in seeds by the PCR method
R. I. Tarakanov,
I. M. Ignat’eva,
О. О. Beloshapkina,
S. I. Chebanenko,
О. G. Karatayeva,
F. S.-U. Dzhalilov https://doi.org/10.26897/0021-342X-2024-1-41-52
Abstract
This article describes the application of the classical PCR method for the diagnosis of the soybean bacterial blight pathogen Pseudomonas savastanoi pv. glycinea in seeds. Two methods for pathogen isolation from infected soybean seeds, five methods for DNA extraction and two master mixes for preparation of reaction mixture were investigated. The use of a PCR-based test system with oligonucleotides specific for the coronaphacate ligase (cfl) gene allowed the diagnosis of the soybean bacterial blight pathogen in seeds at a concentration of 2 × 103 CFU/ml. The analytical sensitivity of the protocol was 97.4% of 37 closely related and other bacteria tested. Product yield (amplicon peak area) was shown to be highest (645.0 units) when DNA was isolated using the Proba-GS kit, the master mix 5x MasDDDTaqMIX-2025 and the primers PsgFOR-1 and PsgREV-2 were used at 10 pM per reaction (25 μL).
About the Authors
R. I. Tarakanov
https://www.researchgate.net/profile/Rashit-Tarakanov
Russian State Agrarian University – Moscow Timiryazev Agricultural Academy
Russian Federation
Russian State Agrarian University – Moscow Timiryazev Agricultural Academy
Russian Federation
Rashit I. Tarakanov, assistant at the Department of Plant Protection
49 Timiryazevskaya St., Moscow, 127550
I. M. Ignat’eva
All-Russian Plant Quarantine Centre
Russian Federation
Russian Federation
Irina M. Ignat’eva, Research Associate, Bacteriology Laboratory
32 Pogranichnaya St., Bykovo Vlg., Ramenskiy Urban District, Moscow Region, 140150
О. О. Beloshapkina
Russian State Agrarian University – Moscow Timiryazev Agricultural Academy
Russian Federation
Russian Federation
Olga O. Beloshapkina, DSc (Agr), Prosessor, Prosessor at the Department of Plant Protection
49 Timiryazevskaya St., Moscow, 127550
S. I. Chebanenko
Russian State Agrarian University – Moscow Timiryazev Agricultural Academy
Russian Federation
Russian Federation
Svetlana I. Chebanenko, CSc (Agr), Associate Prosessor, Associate Prosessor at the Department of Plant Protection
49 Timiryazevskaya St., Moscow, 127550
О. G. Karatayeva
Russian State Agrarian University – Moscow Timiryazev Agricultural Academy
Russian Federation
Russian Federation
Oksana G. Karatayeva, CSc (Econ), Associate Prosessor, Associate Prosessor at the Department of Pedagogy and Psychology of Professional Education
49 Timiryazevskaya St., Moscow, 127550
F. S.-U. Dzhalilov
Russian State Agrarian University – Moscow Timiryazev Agricultural Academy
Russian Federation
Russian Federation
Fevzi S.-U. Dzhalilov, DSc (Bio), Prosessor, Head of the Department of Plant Protection
49 Timiryazevskaya St., Moscow, 127550
References
1. Bulletin “Sown areas, gross yields and crop yields in the Russian Federation in 2022”. Ministry of Agriculture of the Russian Federation [Electronic source]. URL: https://mcx.gov.ru/ (access date: 09.01.2024) (In Russ.)
2. Bradley C.A., Allen T.W., Sisson A.J., Bergstrom G.C., Bissonnette K.M. et.al. Soybean Yield Loss Estimates Due to Diseases in the United States and Ontario, Canada, from 2015 to 2019. Plant Health Progress. 2021;4:483–495.
3. Koenning S.R., Wrather J.A. Suppression of soybean yield potential in the continental United States by plant diseases from 2006 to 2009. Plant Health Progress. 2010;11(1):5.
4. Wrather J.A., Chambers A.Y., Fox J.A., Moore W.F., Sciumbato G.L. Soybean disease loss estimates for the southern United States, 1974 to 1994. Plant Disease. 1995;79:1076–1079.
5. Ignjatov M., Milošević M., Nikolić Z., Vujaković M., Petrović D. Characterization of Pseudomonas Savastanoi Pv. Glycinea Isolates from Vojvodina. Phytopathol. Pol. 2007;45:43–54.
6. Jagtap D. Bio-efficacy of different antibacterial antibiotic, plant extracts and bioagents against bacterial blight of soybean caused by Pseudomonas savastanoi pv. glycinea. Scientific Journal of Microbiology. 2012;1(1):1–9.
7. Tarakanov R.I., Ignatov A.N., Dzhalilov F.S. Isolation of specific bacteriophages – Pseudomonas savastanoi pv. glycinea – and their use in soybean bacterial blight control. Izvestiya of Timiryazev Agricultural Academy (TAA). 2020;(4):43–53. (In Russ.) https://doi.org/10.26897/0021-342X-2020-4-43-53
8. Maslennikova V.S., Cvetkova V.P., Bedareva E.V., Krugovyh A.A. Efficiency of soybean seed treatment with biopreparation in Western Siberia. Agrarnaya nauka – 2022: Proceedings of the All-Russian Conference of Young Researchers, Moscow, November 22–24, 2022. Moscow, Russia: Russian State Agrarian University-Moscow Agricultural Academy named after K.A. Timiryazev, 2022:60–63. (In Russ.)
9. Ignat’eva I.M., Karimova E.V. Study of bacteriosis pathogens of grain legume crops and the development of methods for their diagnosis. Sovremennye podkhody i metody v zashchite rasteniy: Proceedings of the All-Russian Scientific and Practical Conference With International Participation, Ekaterinburg, November 12–14, 2018. Ekaterinburg, Russia: Ural Federal University named after the first President of Russia B.N. Yeltsin, 2018:194–198. (In Russ.)
10. Shepherd L.M., Block C.C. Detection of Pseudomonas savastanoi pv. glycinea in soybean seeds. In: M. Fatmi,R.R. Walcott andN.W. Schaad, eds.Detection of plant-pathogenic bacteria in seed and other planting material, 2nd ed. St. Paul: The American Phytopathological Society, 2017.
11. Ciampi-Guillardi M., Ramiro J., Moraes M.H.D., Barbieri M.C.G., Massola Junior N.S. Multiplex qPCR assay for direct detection and quantification of Colletotrichum truncatum, Corynespora cassiicola, and Sclerotinia sclerotiorum in soybean seeds. Plant Disease. 2020;104(11):3002–3009.
12. Bereswill S., Bugert P., Völksch B., Ullrich M., Bender C.L., Geider K. Identification and relatedness of coronatine-producing Pseudomonas syringae pathovars by PCR analysis and sequence determination of the amplification products. Appl Environ Microbiol. 1994;60(8):2924–2930.
13. Tarakanov R., Shagdarova B., Varlamov V. Dzhalilov, F. Biocidal and resistance-inducing effects of chitosan on phytopathogens. E3S Web Conf. 2021;254:05007.
14. Rooney W., Laird J., Chowdhury M., MacIntosh C., Deng X., McBride P., Milner J. Pseudomonas Syringae Seed Infections [Electronic source]. URL: https://www. protocols.io/view/pseudomonas-syringae-seed-infections-ewov18beygr2/v1 (access date: 09.01.2024)
15. Curtobacterium flaccumfaciens pv. flaccumfaciens. EPPO Global Database. 2011;41. URL: https://gd.eppo.int/taxon/CORBFL/datasheet
16. Javan mard A., Khaledi K., Asadzadeh N., Solimanifarjam A.R. Detection of polymorphisms in the bovine leptin (LEP) gene: association of single nucleotide polymorphism with breeding value of milk traits in Iranian Holstein Cattle. Journal of Molecular Genetics. 2010;2:10–14.
17. Tsygankova S.V., Ignatov A.N., Boulygina E.S., Kuznetsov B.B., Korotkov E.V. Genetic intraspecies relationships in Xanthomonas campestris pv. campestris revealed by novel rep-PCR primers. European J. Plant Pathol. 2004;1(10):845–853.
18. Rueden C.T., Schindelin J., Hiner M.C., DeZonia B.E., Walter A.E., Arena E.T., Eliceiri K.W. ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinformatics. 2017;18:529. https://doi.org/10.1186/s12859-017-1934-z
Review
For citations:
Tarakanov R.I., Ignat’eva I.M., Beloshapkina О.О., Chebanenko S.I., Karatayeva О.G., Dzhalilov F.S. Detection of the soybean bacterial blight pathogen Pseudomonas savastanoi pv. Glycinea in seeds by the PCR method. IZVESTIYA OF TIMIRYAZEV AGRICULTURAL ACADEMY. 2024;1(1):41-52. (In Russ.) https://doi.org/10.26897/0021-342X-2024-1-41-52
Views:
233
Email this article 