Detection and characterization of phytoplasmas in some cucurbits (Cucurbitaceae) and bindweed (Convolvulaceae) in Hatay Province of Turkey

Abstract

Aims: During a disease survey conducted in 2019, typical symptoms including yellowing of the leaves, stunting, shortening of internodes, proliferation of auxiliary shoots (witches'-broom), the bunchy appearance of growth at the ends of stems (rosetting), virescence of flowers or sterility, phyllody, small and deformed leaves and fasciation were observed on suspected cucurbit plants growing in Hatay province. Therefore, this study was carried out to determine the phytoplasma infection(s) on cucurbit crops and bindweed in Hatay province of Turkey.

Methods and Results: In total; 4 pumpkins (winter squash: Cucurbita moschata Duchesne), 12 summer squash (Cucurbita pepo L.) and 2 bindweed (Convolvulus arvensis L.) samples were collected from suspected plants in cucurbit fields in Hatay province. Some of the plants (2 pumpkins, 10 summer squash and 2 bindweeds) exhibited symptoms related to phytoplasmas. Total nucleic acid was extracted by CTAB method. PCR amplification of 16S rDNA with phytoplasma specific primer pairs; F1/R0 followed by R16F2n/R2 confirmed the phytoplasma presence. Expected size amplicons of ~1.2 kb were obtained from infected samples from symptomatic cucurbits (12/12) and bindweed (1/2), but not from asymptomatic cucurbit plants (0/4). The sequences of the 4 symptomatic samples were deposited in NCBI GenBank (MT163353, MT163393, MT163396, and MT163469).

Conclusions: After sequencing and phylogenetic analyses it was revealed that the squash isolates had 99% sequence identity with “Candidatus phytoplasma trifolii” (16SrVI) and the bindweed isolate had 99% sequence identity with “Candidatus phytoplasma solani” (16SrXII).

Significance and Impact of the Study: There is a lack of knowledge on the status of phytoplasma infections and natural hosts in cucurbits in Turkey. According to our knowledge, this is the first report of a member of 16SrVI group, Candidatus phytoplasma trifolii associated with phytoplasma infection in winter squash-pumpkin (C. moschata) and summer squash (C. pepo) in Turkey.

Keywords

• Ca. phytoplasma solani
• Ca. phytoplasma trifolii
• Phytoplasma
• Summer squash (Cucurbita pepo)
• Turkey.

Hatay İli Bazı Kabak (Cucurbitaceae) ve Tarlasarmaşığı'nda (Convolvuluceae) Fitoplazmaların Belirlenmesi ve Karakterizasyonu

Öz

Amaç: 2019’da bir hastalık surveyinde, şüpheli kabak bitkilerinde yaprakların sararması, bodurluk, boğum aralarının kısalması, çoklu sürgün oluşumu, tepe sürgünlerinde çalı görünümü, çiçeklerin yeşillenmesi ve kısırlık, fillodi, küçük ve şekli bozulmuş yapraklar ve gövde yassılaşması gibi fitoplazmalara özgü simptomlar gözlenmiştir. Böylelikle, bu çalışma Hatay ili kabak bitkilerinde ve tarla sarmaşığında fitoplazma hastalıklarının araştırılması amacıyla yürütülmüştür.

Yöntem ve Bulgular: Hatay ili kabak tarlalarından şüpheli toplam 4 kış kabağı (Cucurbita moschata), 12 yaz kabağı (Cucurbita pepo) ve 2 tarla sarmaşığı (Convolvulus arvensis) örnekleri toplanmıştır. Bitkilerden bazıları (2 kış kabağı, 10 yaz kabağı ve 2 tarla sarmaşığı) fitoplazmalara özgü simptom sergilemiştir. Toplam nükleik asit izolasyonu, CTAB metoduna göre yapılmıştır. Direct ve Nested PCR çalışmalarında sırasıyla F1/R0 ve R16F2n/R2 evrensel primer çiftleri kullanılmıştır. Simptom sergileyen kabak bitkilerinde (12/12) ve tarla sarmaşığında (1/2) beklenen ~1.2 kb büyüklüğünde bant elde edilirken simptomsuz kabaklarda elde edilememiştir. Simptom gösteren örneklerden 4 tanesi NCBI Genbankası’na (MT163353, MT163393, MT163396 ve MT163469) yüklenmiştir.

Genel Yorum: Sekans ve filogenetik analizlerin ardından, kabak izolatlarının “Candidatus phytoplasma trifolii” (16SrVI), tarla sarmaşığı izolatının ise “Candidatus phytoplasma solani” (16SrXII) olduğunu göstermiştir.

Çalışmanın Önemi ve Etkisi: Türkiye'de kabakgillerde fitoplazma enfeksiyonlarının ve doğal konakçıların durumu hakkında bilgi eksikliği bulunmaktadır. Mevcut bilgilerimize göre, Türkiye’de 16SrVI grubunun bir üyesi olan Candidatus phytoplasma trifolii'nin kışlık kabaklardan bal kabağı (C. moschata) ve yazlık kabaklardan sakız kabağı (C. pepo) bitkilerinde enfeksiyon oluşturduğu ilk kez belirlenmiştir.

Anahtar Kelimeler

• Ca. phytoplasma solani
• Ca. phytoplasma trifolii
• Fitoplazma
• Kabak
• Türkiye

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