Progress in Plant Protection

Jednoczesne wykrywanie wirusa mozaiki arbuza (watermelon mosaic virus, WMV) oraz wirusa żółtej mozaiki cukinii (zucchini yellow mosaic virus, ZYMV) za pomocą reakcji dupleks RT-PCR

Simultaneous detection of watermelon mosaic virus (WMV) and zucchini yellow mosaic virus (ZYMV) using duplex RT-PCR reaction

Julia Minicka, e-mail: j.minicka@iorpib.poznan.pl

Instytut Ochrony Roślin – Państwowy Instytut Badawczy, Władysława Węgorka 20, 60-318 Poznań, Polska

Agnieszka Taberska, e-mail: a.taberska@iorpib.poznan.pl

Instytut Ochrony Roślin – Państwowy Instytut Badawczy, Władysława Węgorka 20, 60-318 Poznań, Polska

Beata Hasiów-Jaroszewska, e-mail: B.Hasiow@iorpib.poznan.pl

Instytut Ochrony Roślin – Państwowy Instytut Badawczy, Władysława Węgorka 20, 60-318 Poznań, Polska
Streszczenie

Wirus mozaiki arbuza (watermelon mosaic virus, WMV) oraz wirus żółtej mozaiki cukinii (zucchini yellow mosaic virus, ZYMV) są groźnymi patogenami infekującymi uprawy roślin dyniowatych na całym świecie. W Polsce występują powszechnie, szczególnie w uprawach cukinii, powodując straty w jakości i ilości plonów. Celem pracy było opracowanie i zoptymalizowanie reakcji dupleks RT-PCR do jednoczesnego wykrywania obu wirusów często stwierdzanych w infekcji mieszanej. Zaprojektowane startery amplifikowały gen kodujący białko płaszcza (CP) obu wirusów odpowiednio wielkości: 977 pz WMV oraz 415 pz ZYMV. Czułość wykrywania reakcji dupleks RT-PCR wyniosła 500 pg/μl całkowitego RNA. Specyficzność wykrywania poszczególnych produktów była sprawdzana za pomocą sekwencjonowania metodą Sangera. Opracowana technika może być wykorzystywana do rutynowego wykrywania obu wirusów w zainfekowanych próbkach.

 

Watermelon mosaic virus (WMV) and zucchini yellow mosaic virus (ZYMV) are dangerous pathogens infecting cucurbit crops worldwide. In Poland, both viruses are common, especially in the cultivation of zucchini, causing losses in the quality and quantity of crops. The aim of the study was to develop and optimize the duplex RT-PCR reaction for simultaneous detection of both viruses often occurring in mixed infection. The designed primers amplified the gene encoding the coat protein (CP) of both viruses with a size of 977 bp for WMV and 415 bp for ZYMV. The duplex RT-PCR detection limit was 500 pg/μl of total RNA. The specificity of detection of individual products was checked by Sanger sequencing. The developed assay can be used for routine detection of both viruses in infected samples.

Słowa kluczowe
dupleks RT-PCR; wykrywanie; wirus mozaiki arbuza; wirus żółtej mozaiki cukinii; koinfekcja; duplex RT-PCR; detection; watermelon mosaic virus; zucchini yellow mosaic virus; co-infection
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Progress in Plant Protection (2021) : 0-0
Data pierwszej publikacji on-line: 2021-03-22 10:34:51
http://dx.doi.org/10.14199/ppp-2021-008
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