Evaluation of different methods of DNA extraction for detection of bacterium Xanthomonas campestris pv. Campestris in cabbage leaves

• Autorzy: Penazova E. , Eichmeier A. , Cechova J. , Baranek M. , Pokluda R.

Warianty tytułu

PL

Ocena różnych metod ekstrakcji dna w celu wykrycia bakterii Xanthomonas campestris pv. Campestris w liściach kapusty

• Języki publikacji: EN

Abstrakty

EN

The bacterium Xanthomonas campestris pathovar campestris (Xcc) as the causal agent of black rot of cruciferous plants causes considerable losses in agricultural yield all over the world. The control of black rot is difficult as well as the determination of Xccon the basis of morphological parameters or by pathogenicity testing. Ten different possibilities for extraction bacterial DNA followed by PCR detection method were tested to optimize PCR protocol. On the basis of ISTA validated method, three sets of primers UBP 1052F-BACR, DLH 120-125 and ZUP 2309-2310 were used. The results of measured concentration and quality of DNA and efficacy for PCR amplification were compared. Finally, three approaches for DNA extraction within Xanthomonas campestris pv. campestris detection protocol were recommended – commercial kits used for isolation from tissues by Macherey-Nagel and MO BIO and kit intended for microbial cultures by MO BIO.

PL

Bakteria Xanthomonas campestris pathovar campestris (Xcc) jako przyczyna czarnej zgnilizny roślin z rodziny krzyżowych powoduje znaczne straty w plonie rolniczym na całym świecie. Zwalczanie czarnej zgnilizny, a także określenie Xcc na podstawie parametrów morfologicznych lub za pomocą testów patogeniczności jest trudne. W celu zoptymalizowania protokołu PCR przetestowano dziesięć różnych możliwości ekstrakcji DNA, a następnie metodę PCR. Na podstawie walidowanej metody ISTA za-stosowano trzy zestawy primerów, mianowicie UBP 1052F-BACR, DLH 120-125 i ZUP 2309-2310. Porównano wyniki zmierzonego stężenia i jakości DNA oraz skuteczności dla amplifikacji PCR. Zalecono trzy sposoby ekstrakcji DNA w ramach protokołu detekcji Xanthomonas campestris pv. Campestris: zestaw komercyjny używany do izolacji z tkanek według Macherey-Nagel, MO BIO oraz zestaw dla hodowli mikrobowych według MO BIO.

• Wydawca: -
• Czasopismo: Acta Scientiarum Polonorum. Hortorum Cultus
• Rocznik: 2015
• Tom: 14
• Numer: 6
• Opis fizyczny: p.140-150,fig.,ref.

Twórcy

autor

Penazova E.

• Department of Vegetable Growing and Floriculture, Faculty of Horticulture, Mendel University in Brno, Valticka 337, 691 44 Lednice, Czech Republic

autor

Eichmeier A.

• Mendel University in Brno, Brno, Czech Republic

autor

Cechova J.

• Mendel University in Brno, Brno, Czech Republic

autor

Baranek M.

• Mendel University in Brno, Brno, Czech Republic

autor

Pokluda R.

• Mendel University in Brno, Brno, Czech Republic

Bibliografia

• Ahrends, U., Seeműller, E., (1992). Detection of DNA of plant pathogenic mycoplasmalike organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phy-topathology, 82, 828–832.
• Alvarez, A.M. (2001). Black rot of crucifers. In: Mechanisms of resistance to plant diseases, Slusarenko, A.J., Fraser, R.S.S., van Loon, L.C. (eds). Dordrecht: Kluwer Academic Publishers, 21–44.
• Berg, T., Tesoriero, L., Hailstones, D.L. (2005). PCR-based detection of Xanthomonas campestrispathovars in Brassica seed. Plant Pathol., 54, 416–427. DOI: 10.1111/j.0032-0862.2005.01186.x.
• Bernatzky, R., Tanksley, S.D. (1986). Genetics of actin-related sequences in tomato. Theoret. App. Gen., 72, 314–321.
• Bila, J., Mortensen, C.N., Andersen, M., Vicente, J.G., Wulff, E.G. (2013). Xanthomonas cam-pestris pv. campestris race 1 is the main causal agent of black rot of Brassicas in Southern Mozambique. African J. Biotech., 12(6), 602–610. DOI: 10.5897/AJB12.2455.
• Divashuk, M., Mayer, N., Kroupin, P., Rubets, V., Pylnev, V., Tkhi Tkhu Lin, N., Soloviev, A., Karlov, G. (2012). The association between the allelic state of Vp-1B and pre-harvest sprouting tolerance in red-seeded hexaploid triticale. Open J. Gen., 2, 51–55. DOI: 10.4236/ojgen.2012.21006.
• Fargier, E., Manceau, C. (2007). Pathogenicity assays restrict the species Xanthomonas cam-pestris into three pathovars and reveal nine races within X. campestris pv. campestris. Plant Pathol., 56,805–818.
• Gogorcena, Y., Arulsekar, S., Dandekar, A.M., Parfitt, D.E. (1993). Molecular markers for grape characterization. Vitis, 32, 183–185.
• Grimault, V., Andro, C., Politikou, A. (2012). PCR as a new identification method of Xanthomo-nascampestris pv. campestris on Brassica spp. seed. In: Method Validation Reports on Rules Proposals for the International Rules for Seed Testing 2013 Edition, International Seed Testing Association. Bassersdorf, Switzerland, 2–11.
• Holleinová, V., Čechová, J. (2012). The detection of viruses and phytoplasmas in dwarfed shoots of grapevine varieties Aurelius and Neuburger. Acta Univ. Agricult. Silvicult. Mendel. Brunnen., 60, 8, 73–78. ISTA (2013). 7–019
• Detection of Xanthomonas campestris pv. campestris on Brassica spp. (pre-pared by Roberts, S.J., Koenraadt, H.) International Rules for Seed Testing, Annexe to Chapter 7: Seed Health Testing Methods, Bassersdorf, Switzerland, International Seed Testing Association.
• Klimyuk, V.I, Carroll, B.J., Thomas, C.M., Jones, J.D.G. (1993). Alkali treatment for rapid prepa-ration of plant material for reliable PCR analysis. Plant J., 3, 493–494.
• Krichen, L., Martins, J.M.S., Lambert, P., Daaloul, A., Trifi-Farah, N., Marrakchi M., Audergon, J-M. (2008). Using AFLP markers for the analysis of the genetic diversity of apricot cultivars in Tunisia. J. Am. Soci. Horticult. Sci., 133, 204–212.
• Mynarzová, Z. (2013). Ověření úspěšnosti genetických transformací podnožových odrůd révy vinné za účelem navození jejich rezistence vůči viru GFLV. Diploma thesis. Mendel Univer-sity in Brno. Faculty of Horticulture Lednice, Czech Republic.
• Palacio-Bielsa, A., Cambra, A., López, M.M. (2009). PCR detection and identification of plant-pathogenic bacteria: updated review of protocols (1989–2007). J. Plant Pathol., 91(2), 249–297.
• Pilařová, P., Marandel, G., Decroocq, V., Salava, J., Krška B., Abbott, A.G., (2010). Quantitative trait analysis of resistance to plum pox virus in the apricot F1 progeny ‘Harlayne’ × ‘Vestar’. Tree Gen., 6, 3, 467–475. DOI: 10.1007/s11295-009-0264-3.
• Rijlaarsdam, A., Woudt, B., Simons, G., Koenraadt, H., Oosterhof, J., Asma, M., Buddiger, P., Roorda, P., Grimault, V., De Koning, J. (2004). Development of specific primer for the molecular detection of Xanthomonas campestris pv. campestris. EPPO Conference on Quality of Diagnosis and New Diagnostic Methods for Plant Pests,Noordwijkerhout, NL, 2004-04-09/22.
• Smulders, J.J.M., Noordijk, Y., Rus-Kortekaas, W., Bredemeijer, G.M.M., Vosman, B. (2003). Microsatellite genotyping of carnation varieties. Theoret. App. Gen., 106, 1191–1195.
• Williams, P.H. (2007). Black rot. In: Compendium of Brassica diseases, Rimmer, S.R., Shattuck, V., Buchwaldt, L. (eds). St. Paul, Minn., American Phytopathological Society, 60–62. ISBN 978-0-89054-344-3.