Progress in Plant Protection

The use of quantitative PCR with high resolution melting (qPCR-HRM) analysis to distinguish the Polish mild and yellowing pathotypes of Pepino mosaic virus (PepMV)
Zastosowanie techniki ilościowego PCR z analizą krzywych topnienia (qPCR-HRM) do rozróżniania polskich łagodnych i żółtaczkowych patotypów wirusa mozaiki pepino (Pepino mosaic virus, PepMV)

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

Instytut Ochrony Roślin – Państwowy Instytut Badawczy, Zakład Wirusologii i Bakteriologii, Władysława Węgorka 20, 60-318 Poznań, Polska

Beata Komorowska, e-mail: Beata.Komorowska@inhort.pl

Instytut Ogrodnictwa, Zakład Fitopatologii, Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Polska

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

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

Pepino mosaic virus (PepMV) is a highly infectious virus causing significant losses in tomato greenhouse production worldwide. One of the characteristic symptoms is yellowing discolorations of leaf blades, which is often accompanied by damage of the fruits. It has been found that, two independent point mutations in the gene encoding the coat protein (CP) are responsible for the formation of yellowing symptoms: G463AA/A463AA (155 aa) and GA497U/GG497U (166 aa), respectively. In our study, qPCR-HRM was used to distinguish mild and yellowing pathotypes. Appropriate primers, amplifying a fragment of the CP-encoding gene in which both yellowing mutations are located were designed. The analysis was carried out using 18 isolates originated from greenhouses in different region of Poland. Our experiments confirmed the effectiveness of the designed primers to differentiate the analyzed pathotypes. This technique is a valuable tool for detecting the diversity of PepMV.

 

Wirus mozaiki pepino (PepMV) jest groźnym patogenem powodującym znaczne straty w uprawie szklarniowej pomidora w wielu regionach świata. Jednym z charakterystycznych objawów obecności wirusa w roślinie są żółtaczkowe przebarwienia blaszek, którym często towarzyszy uszkodzenie owoców. Za ich powstawanie odpowiedzialne są dwie niezależne mutacje punktowe w genie kodującym białko płaszcza (CP): G463AA/A463AA (155 aa) i GA497U/GG497U (166 aa). W przeprowadzonych badaniach wykorzystano qPCR-HRM do rozróżniania łagodnych i żółtaczkowych patotypów. Zaprojektowano startery, amplifikujące fragment genu kodującego CP, w którym znajdują się obie mutacje żółtaczkowe. Przeanalizowano 18 izolatów, pochodzących z obiektów szklarniowych na terenie całego kraju. Wykonane doświadczenia potwierdziły skuteczność zaprojektowanych starterów do różnicowania analizowanych patotypów. Technika ta może być z powodzeniem stosowana do rozróżniania łagodnych i żółtaczkowych patotypów PepMV na terenie naszego kraju.

Słowa kluczowe
Pepino mosaic virus; CH2 strain; yellowing pathotype; Solanum lycopersicum; szczep CH2; patotyp żółtaczkowy
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Progress in Plant Protection (2020) 60: 5-10
Data pierwszej publikacji on-line: 2020-02-07 11:13:55
http://dx.doi.org/10.14199/ppp-2020-001
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