Detection and molecular characterization of some viruses on Cucurbitaceae plants growing in Hatay province by DAS-ELISA and RT-PCR methods

Abstract

Aims: In this study, it was aimed to investigate the occurrence of cucurbit yellow stunting disorder virus (CYSDV), cucurbit chlorotic yellows virus (CCYV) and beet pseudo-yellows virus (BPYV) in cucumber, zucchini and melon plants grown in Hatay province by Double Antibody Sandwich-Enzyme Linked Immuno Sorbent Assay (DAS-ELISA) and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) methods, to analyse the phylogenetic relationships of identified isolates.

Methods and Results: Totally 90 symptomatic and non-symptomatic cucumber, zucchini and melon samples were collected from Hatay province. DAS-ELISA and RT-PCR methods were used to investigate the presence of CCYV and RT-PCR method was used for CYSDV and BPYV. The most common symptoms observed are leaf shrinkage, deformation, leaf wrinkles, curling and vein clearing. According to the DAS-ELISA results, CCYC was not found in the samples tested. As a result of RT-PCR analysis, BPYV and CCYV could not found in the samples tested, but PCR amplicons of 364 bp amplifying the heat shock protein 70h (HSP70h) gene of CYSDV were obtained from 11 samples (9 zucchini, 2 melons). As a result of bidirectional sequence analysis of 10 CYSDV izolates, it was determined that the obtained nucleotide sequences were highly similar (99%) to that of the CYSDV isolates registered in the GeneBank. In this study, CYSDV was reported for the first time in the zucchini and melon plants growing in Hatay province.

Conclusions: CYSDV was detected for the first time in samples collected from zucchini and melon plants in Hatay province. BPYV and CCYV could not be detected in any of the samples tested. The fact that 12.22% of the tested samples were infected with CYSDV highlights the need of using healthy plant materials.

Significance and Impact of the Study: In this study, the presence of CYSDV was proven by RT-PCR analyses for the first time in zucchini and melon plants in Hatay province.

Keywords

• Boron content
• Olive orchard
• soil properties
• Gaziantep

Hatay ilinde yetişen Cucurbitaceae familyasına ait kültür bitkilerinde bazı virüslerin DAS-ELISA ve RT-PCR yöntemleriyle saptanması ve moleküler karakterizasyonu

Öz

Amaç: Bu çalışmada, Hatay ilinde yetiştirilen hıyar, kabak ve kavun bitkilerinde cucurbit yellow stunting disorder virus (CYSDV), cucurbit chlorotic yellows virus (CCYV) ve beet pseudo-yellows virus (BPYV)’lerinin Double Antibody Sandwich-Enzyme Linked Immuno Sorbent Assay (DAS-ELISA) ve Reverse Transcription-Polymerase Chain Reaction (RT-PCR) yöntemleriyle belirlenmesi, elde edilen virüs izolatlarının filogenetik ilişkilerinin ortaya konulması amaçlanmıştır.

Yöntem ve Bulgular: Hatay ilinden simptomlu ve simptomsuz 90 adet hıyar, kabak ve kavun örneği toplanmıştır. CCYV’nin varlığını araştırmak için DAS-ELISA ve RT-PCR yöntemleri, CYSDV ve BPYV’lerinin varlığını araştırmak için ise RT-PCR yöntemi kullanılmıştır. Gözlenen en yaygın simptomlar yapraklarda küçülme, deformasyon, yaprak kırışıklıkları, kıvrılmalar ve damar açılmalarıdır. DAS-ELISA testi sonuçlarına göre testlenen örneklerde CCYC bulunamamıştır. RT-PCR analizleri sonucunda testlenen örneklerde BPYV ve CCYV bulunamamış ancak CYSDV’nin heat shock protein 70h (HSP70h) genini çoğaltan 364 bp büyüklüğünde PCR amplikonları 11 örnekte (9 kabak, 2 kavun) elde edilmiştir. Bu ürünlerden 10 tanesinin doğrudan iki yönlü sekans analizi sonucunda, elde edilen nükleotit dizilerinin gen bankasında kayıtlı CYSDV izolatları ile yüksek oranda (%99) benzerlik gösterdiği saptanmıştır. Bu çalışma ile CYSDV ilk defa Hatay ilinde kabak ve kavun bitkilerinde rapor edilmiştir.

Genel Yorum: Hatay ili kabak ve kavun alanlarından toplanan örneklerde CYSDV ilk kez tespit edilmiştir. Testlenen örneklerinin hiçbirinde BPYV ve CCYV belirlenememiştir. Testlenen örneklerin %12.22 oranında CYSDV ile enfekteli bulunması, sağlıklı bitki materyallerinin kullanılmasının önemini göstermektedir.

Çalışmanın Önemi ve Etkisi: Hatay ilinde ilk kez bu çalışma ile kabak ve kavun bitkilerinde CYSDV’ünün varlığı, RT-PCR analizleri ile kanıtlanmıştır.

Anahtar Kelimeler

• Cucurbitaceae
• virüs
• RT-PCR
• DAS-ELISA
• sekans analizleri

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