Specific detection of Waitea circinata var. zeae using conventional and real-time PCR
Apstrakt
Waitea circinata var. zeae, a pathogen with a relatively narrow host range, has recently been detected in cabbage and oilseed rape in Europe and worldwide. In this study, we developed specific conventional and real-time PCR protocols for direct detection of W. circinata var. zeae from mycelium and diseased plant tissue. The newly developed primer pair zeaefor1/zeaerew1, used in PCR protocols, specifically amplified only target isolates of W. circinata var. zeae when tested against isolates of 11 different binucleate and multinucleate anastomosis groups of Rhizoctonia spp. including AG-A, AG-G, AG-F, AG-U, AG-2-1, AG-2-2, AG-3, AG-4 HGI, AG-4 HGII, AG-4 HGIII, and AG-6 and common soil-borne pathogens. Total of nine previously published primer pairs designed for the detection of various Rhizoctonia spp. were also tested and did not amplify target isolates of W. circinata var. zeae. The detection limit of conventional and real-time PCR protocols was 10–2 and 10–5 (with starting concentrat...ion 9.5 ng/µl), respectively, and both methods are the first available tools for direct detection and identification of W. circinata var. zeae from mycelium and diseased oilseed rape seedlings. Both conventional and SYBR-Green-based real-time PCR protocols are cost-effective and provide a solid basis for further investigations of W. circinata var. zeae, particularly in relation to distribution, host range, and epidemiology.
Ključne reči:
Waitea circinata var. zeae / Waitea circinata / detection / PCR / real-time PCR / pathogens / pathogen detection / protocolsIzvor:
Journal of Plant Diseases and Protection, 2024Izdavač:
- Springer Nature
Finansiranje / projekti:
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200116 (Univerzitet u Beogradu, Poljoprivredni fakultet) (RS-MESTD-inst-2020-200116)
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200214 (Institut za pesticide i zaštitu životne sredine, Beograd) (RS-MESTD-inst-2020-200214)
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200003 (Institut za proučavanje lekovitog bilja 'Dr Josif Pančić', Beograd) (RS-MESTD-inst-2020-200003)
Kolekcije
Institucija/grupa
FiVeRTY - JOUR AU - Vojvodić, Mira AU - Lazić, Dejan AU - Pešić, Brankica AU - Mitrović, Petar AU - Vico, Ivana AU - Bulajić, Aleksandra PY - 2024 UR - http://fiver.ifvcns.rs/handle/123456789/4557 AB - Waitea circinata var. zeae, a pathogen with a relatively narrow host range, has recently been detected in cabbage and oilseed rape in Europe and worldwide. In this study, we developed specific conventional and real-time PCR protocols for direct detection of W. circinata var. zeae from mycelium and diseased plant tissue. The newly developed primer pair zeaefor1/zeaerew1, used in PCR protocols, specifically amplified only target isolates of W. circinata var. zeae when tested against isolates of 11 different binucleate and multinucleate anastomosis groups of Rhizoctonia spp. including AG-A, AG-G, AG-F, AG-U, AG-2-1, AG-2-2, AG-3, AG-4 HGI, AG-4 HGII, AG-4 HGIII, and AG-6 and common soil-borne pathogens. Total of nine previously published primer pairs designed for the detection of various Rhizoctonia spp. were also tested and did not amplify target isolates of W. circinata var. zeae. The detection limit of conventional and real-time PCR protocols was 10–2 and 10–5 (with starting concentration 9.5 ng/µl), respectively, and both methods are the first available tools for direct detection and identification of W. circinata var. zeae from mycelium and diseased oilseed rape seedlings. Both conventional and SYBR-Green-based real-time PCR protocols are cost-effective and provide a solid basis for further investigations of W. circinata var. zeae, particularly in relation to distribution, host range, and epidemiology. PB - Springer Nature T2 - Journal of Plant Diseases and Protection T1 - Specific detection of Waitea circinata var. zeae using conventional and real-time PCR DO - 10.1007/s41348-024-00926-w ER -
@article{ author = "Vojvodić, Mira and Lazić, Dejan and Pešić, Brankica and Mitrović, Petar and Vico, Ivana and Bulajić, Aleksandra", year = "2024", abstract = "Waitea circinata var. zeae, a pathogen with a relatively narrow host range, has recently been detected in cabbage and oilseed rape in Europe and worldwide. In this study, we developed specific conventional and real-time PCR protocols for direct detection of W. circinata var. zeae from mycelium and diseased plant tissue. The newly developed primer pair zeaefor1/zeaerew1, used in PCR protocols, specifically amplified only target isolates of W. circinata var. zeae when tested against isolates of 11 different binucleate and multinucleate anastomosis groups of Rhizoctonia spp. including AG-A, AG-G, AG-F, AG-U, AG-2-1, AG-2-2, AG-3, AG-4 HGI, AG-4 HGII, AG-4 HGIII, and AG-6 and common soil-borne pathogens. Total of nine previously published primer pairs designed for the detection of various Rhizoctonia spp. were also tested and did not amplify target isolates of W. circinata var. zeae. The detection limit of conventional and real-time PCR protocols was 10–2 and 10–5 (with starting concentration 9.5 ng/µl), respectively, and both methods are the first available tools for direct detection and identification of W. circinata var. zeae from mycelium and diseased oilseed rape seedlings. Both conventional and SYBR-Green-based real-time PCR protocols are cost-effective and provide a solid basis for further investigations of W. circinata var. zeae, particularly in relation to distribution, host range, and epidemiology.", publisher = "Springer Nature", journal = "Journal of Plant Diseases and Protection", title = "Specific detection of Waitea circinata var. zeae using conventional and real-time PCR", doi = "10.1007/s41348-024-00926-w" }
Vojvodić, M., Lazić, D., Pešić, B., Mitrović, P., Vico, I.,& Bulajić, A.. (2024). Specific detection of Waitea circinata var. zeae using conventional and real-time PCR. in Journal of Plant Diseases and Protection Springer Nature.. https://doi.org/10.1007/s41348-024-00926-w
Vojvodić M, Lazić D, Pešić B, Mitrović P, Vico I, Bulajić A. Specific detection of Waitea circinata var. zeae using conventional and real-time PCR. in Journal of Plant Diseases and Protection. 2024;. doi:10.1007/s41348-024-00926-w .
Vojvodić, Mira, Lazić, Dejan, Pešić, Brankica, Mitrović, Petar, Vico, Ivana, Bulajić, Aleksandra, "Specific detection of Waitea circinata var. zeae using conventional and real-time PCR" in Journal of Plant Diseases and Protection (2024), https://doi.org/10.1007/s41348-024-00926-w . .