Screening of L4 resistance status to pepper mild mottle virus (PMMoV) and characterization by molecular methods

Abstract

Pepper mild mottle virus (PMMoV) is one of the most common Tobamoviruses can be found in worldwide. This virus has potential to cause economic losses and has kept secret infections until harvest time causing great damages on pepper fruits at this period. During 2016-2019 production periods, it has been frequently seen in pepper grown areas. Several isolates reported from different regions of the world regarding resistance with L4 gene fights against this virus disease. In this study, we aimed to investigate how the PMMoV increased and understood its incidence pepper grown areas in Turkey. In order to determine the effectiveness of L4 resistance, PMMoV and other suspected pepper samples were isolated. The collected samples were studied by RT-PCR technique using PMMoV-Kum isolate and its gene bank’s access number (MK806437) was registered. A sensitive (B1) and 5 resistant (L4B1, L4B2, L4B3, L4B4, L4B5) plants were mechanically inoculated with the PMMoV-Kum isolate and their phenotypic observations were recorded. The PMMoV-Kum isolate was confirmed as avirulent in biological tests, the PMMoV could not overcome L4 gene mediated resistance. Furthermore, a successful primer set has determined as AP-7 / AP-8 in L4 gene mediated resistance against occurrence of any PMMoV symptoms on the tested plants. Determining new PMMoV isolates which break the L4 gene mediated resistance, phylogenetic analyzes was performed to determine its place among the world’s isolates.

Keywords

• L4 gene,Resistance Breaking,Pepper,Pepper mild mottle virus,PMMoV

Biber hafif benek virüs’üne (PMMoV) karşı L4 dayanıklılık durumunun taranması ve moleküler yöntemlerle karakterizasyonu

Abstract

Biber hafif benek virüsü (Pepper mild mottle virus- PMMoV) Tobamovirüsler içerisinde yer alan dünya genelinde biber üretim alanlarında en sık karşılaşılan virüslerden biridir. Bu virüs çoğu zaman hasat dönemine kadar gizli kalabilmekte ve en büyük zararlarını hasat döneminde meyveler üzerinde meydana getirebilmektedir. Bölgemizde son yıllarda biber üretim alanlarında sıklıkla karşılaşılan bir problem olarak karşımıza çıkmaktadır. Dünyanın farklı bölgelerinde L4 geni vasıtasıyla sağlanan dayanıklılığı kıran virüs izolatının rapor edilmesi, ülkemizde de PMMoV’nin epidemi durumunun araştırılması gerekliliğini ortaya koymuştur. L4 dayanıklılığının etkinliğini belirleyebilmek için bölgemizden biber üretim alanlarında PMMoV ile şüpheli örnekler toplanmıştır. RT-PCR tekniği ile doğrulanan bu örnek PMMoV-Kum olarak kodlanmıştır ve Gen bankasına kaydı yapılarak erişim numarası (MK806437) alınmıştır. PMMoV-Kum izolatı 1 hassas (B1) ve 5 dayanıklı (L4B1, L4B2, L4B3, L4B4, L4B5) bitkilerine mekanik inokulasyon yöntemi ile bulaştırılmıştır. Yapılan fenotipik gözlemler neticesinde PMMoV-Kum izolatının L4 genine bağlı dayanıklılığı kıramadığı biyolojik testler ile doğrulanmıştır. Dayanıklı olarak seçilen bitkiler üzerinde herhangi bir PMMoV simptomunun oluşmaması üzerine en başarılı çalışan L4 markırlarını belirlemeye yönelik primerlerin belirlenmesi tez çalışmasının seyrini belirlemiş ve en başarılı markırın AP-7/AP-8 ve P118/119primer çiftleri olduğu tespit edilmiştir.

Keywords

• L4 geni,Dayanıklılığın kırılması,Biber hafif benek virüsü,Biber,PMMoV

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