- Tytuł:
-
Molekularna diagnostyka wybranych patogenów z rodzaju Phytophthora w ramach integrowanej ochrony roślin
Molecular diagnostic of Phytophthora pathogens as a tool for Integrated Pest Management - Autorzy:
-
Nowakowska, J.A.
Malewski, T.
Tereba, A.
Borys, M.
Oszako, T. - Powiązania:
- https://bibliotekanauki.pl/articles/989551.pdf
- Data publikacji:
- 2016
- Wydawca:
- Polskie Towarzystwo Leśne
- Tematy:
-
fitopatologia
Phytophthora
wykrywanie
identyfikacja
Phytophthora alni subsp.multiformis
Phytophthora lacustris
Phytophthora taxon hungarica
metody badan
metoda real time PCR
sondy TaqMan
real time pcr
taqman
butt and fine root pathogens
phytophthora - Opis:
- Traditional detection methods such as baiting or direct isolation take a long time and are incapable to handling large volume of material to be tested. The real−time PCR−based techniques are faster, more sensitive, more easily automated, and do not require post−amplification procedures. Species−specific primers for Phytophthora were designed based on the internal transcribed spacer regions (ITS) of rDNA collected from the NCBI DNA database. Primers and probes were designed using the Allele ID 7 at default search criteria. Specific probes were labeled with the reporter dyes JOE (6−carboxy−4,5−dichloro−2,7−dimethoxyfluorescein) at the 5' end and HBQ1 quencher at the 3' end (Sigma−Aldrich). The specificity of primers and fluorogenic probes was tested against genomic DNA of P. alni subsp. multiformis, P. lacustris and P. taxon hungarica. The real−time PCR reactions with the specific probes and primers yielded positive results with five concentrations of standards obtained by standard PCR reaction for corresponding Phytophthora species. The negative control (lack of DNA pathogens) yielded no amplification products. Standard curves showed a linear correlation between input DNA and cycle threshold (Ct) values with R² from 0.994 (P. alni) to 0.998 (P. taxon hungarica). The amplification efficiency of target DNA varied from 94.6% (P. alni) to 100% (P. taxon hungarica). The validation of the primers and probes designed for analysed Phytophthora species was performed on pure cultures, on soil samples from the forest nursery and declining oak stands. The designed probes displayed the high specificity of the detection of investigated species in pure cultures. The presented new molecular TaqMan probes can fully assist the integrated pest management as a powerful tool for a quick detection of above pathogenic organisms in forest nurseries. The molecular detection of harmful phytophthoras and in consequences diminishing of fungicides use for their control in forestry fully support European Union directives as well as the ‘Good plant protection practice measures' elaborated by European and Mediterranean Organisation of Plant Protection.
- Źródło:
-
Sylwan; 2016, 160, 05; 365-370
0039-7660 - Pojawia się w:
- Sylwan
- Dostawca treści:
- Biblioteka Nauki
Artykuł