V. VENKATARAVANAPPA, K. V. ASHWATHAPPA, H. D. VINAYKUMAR, M. NANDAN, M. KRISHNA REDDY, C. N. LAKSHMINARAYANA REDDY

Güney Hindistan'da Çakal Hünnabı (Zizyphus oenoplia L.)'nın Cadı Süpürgesi Hastalığı "Candidatus Phytoplasma balanitae" ile İlişkisi

Araştırma sürecinde (2017-18), Hindistan’ın Karnataka ilçesinde fitoplazmalara özgü cadı süpürgesi (WB: Witches Broom) simptomu gösteren on iki (12) Çakal hünnap (Jackal jujube) bitkisinden örnek toplanmıştır. Örnekler fitoplazmaların 16S rRNA ve SecA genlerine sipesifik primerler kullanılarak Polymerase Chain Reaction (PCR) ile testlenmiş, amplifikasyon bölgeleri klonlanarak sekans dizilimleri çıkarılmış ve takiben NCBI veri tabanında yer alan fitoplazmaların sekans dizilimleri ile mukayese edilmiştir. Yapılan karşılaştırmada JJWB fitoplazma izolatlarının 16S rRNA gen bölgesinin Candidatus Phytoplasma balanitae gurubu (16SrV) ile %97,8-%98,2, SecA gen bölgesinin ise %89,4-%99 oranında Jujube witches broom (JWB) phytoplasma ile benzer nükleotit dizilimlerine sahip oldukları tespit edilmiş ve sonuç filogenetik analiz ile desteklenmiştir. Diğer taraftan, JJWB izolatları Alu I, Hae III ve Mse I enzimleri ile Restriction Fragmenth Length Polymorphism (RFLP) analizine tabi tutulduğunda kontrol olarak kullanılan Ca. P. balanitae (16SrV) gurubu ile 0,91 oranında benzerlik gösterdiği saptanmıştır. Bu oran yeni alt guruplar için eşik benzerlik oranını olan 0,98’den düşük bulunmuş ve bulgular bu veriler çerçevesinde tartışılmıştır.

Anahtar Kelimeler:

Candidatus Phytoplasma balanitae, çakal hünhabı, PCR, fillodi, filogenetik analiz

“Candidatus Phytoplasma balanitae” Associated with Witches’ Broom Disease of Jackal Jujube (Zizyphus oenoplia L.) in South India

During survey, 12 Jackal jujube plant samples showing the symptoms of witches’ broom disease were collected from the Shivamogga district of Karnataka, India, between 2017 and 2018. The causal agent associated with Jackal jujube witches’ broom disease was identified through polymerase chain reaction using phytoplasma 16S rRNA-encoding and SecA gene-specific universal primers. All 12 Jackal jujube plant samples gave positive amplification for the phytoplasmaspecific primers. The amplified polymerase chain reaction products (16S rRNA-encoding gene and SecA gene) were cloned and sequenced. The nucleotide sequence 16S rRNA-encoding and SecA genes comparisons were made with the available phytoplasmas from an NCBI database. The Jackal jujube witches’ broom phytoplasma isolates shared the highest nucleotide identity of 97.8–98.2% (16S rRNA gene) with Candidatus Phytoplasma balanitae group (16SrV) and 89.4–99% (SecA gene) nucleotide identity with Jujube witches’ broom phytoplasma. This was well supported by the close clustering of Jackal jujube witches’ broom phytoplasma isolates in the current study with Candidatus Phytoplasma balanitae (16S rRNA gene) and Jujube witches’ broom-phytoplasma (SecA gene) in phylogenetic analysis. The virtual Restriction fragment length polymorphism (RFLP) pattern generated by Jackal jujube witches’ broom isolates was different (similarity coefficient of 0.91) to the reference pattern of Candidatus Phytoplasma balanitae (16SrV) group with respect to three enzymes (Alu I, Hae III, and Mse I). Based on the threshold similarity coefficient for the new subgroup, delineation should be set at 0.98. The significance of the research is discussed.

Keywords:

Candidatus Phytoplasma balanitae, jackal jujube, PCR, phyllody, phylogenetic analysis,

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