İzmir ili erik bahçelerinde fitoplazmaların PCR
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Tarih
2021
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Yayıncı
Hatay Mustafa Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Yapılan bu çalışmada İzmir ilinde erik ağaçlarındaki fitoplazmaların varlığı belirlenmiş, fitoplazma ile enfekteli erik bahçelerinden toplanan olası fitoplazma vektör böcekler morfolojik olarak teşhis edilmiş, böcek bünyesindeki fitoplazma varlığı moleküler yöntemlerle araştırılmıştır. Erik ağaçlarında görülen en yaygın simptomlar; yapraklarda kloroz ve içe doğru kıvrılma, yapraklarda küçülme, kırmızılaşma ve aşırı sürgün oluşumudur. Yapılan arazi çalışmaları sonucunda 9 farklı erik çeşidinden toplam 110 erik örneği ve yine bu bahçelerden Cicadellidae familyasına ait 30 adet Psammotettix provincialis olarak teşhis edilen böcek toplanmıştır. Nested-PCR analizleriyle testlenen erik örneklerinden 29 tanesinde fitoplazma saptanmış olup bunlardan 8 tanesinin PCR ürünleri RFLP analizleri yapılmak üzere RsaI ve SspI restrüksiyon endonükleaz enzimleri ile kesilmiş ve 4 tanesinin elma çoklu sürgün fitoplazması (Apple Proliferation, Candidatus Phytoplasma mali), diğer 4 tane erik örnekleri ise armut yıkım fitoplazması (Pear decline, Candidatus Phytoplasma pyri) ile enfekteli olduğu tespit edilmiştir. Nested-PCR analizleri ile testlenen 30 adet P. provincialis bireyinden 16 tanesi fitoplazma ile enfekteli bulunmuş ve yapılan filogenetik analizlerde ise bu bireylerin Candidatus Phytoplasma asteris (16SrI) ile enfekteli olduğu belirlenmiştir. Fitoplazma pozitif bulunan 29 izolattan 22 tanesi sekans analizine gönderilmiş ve elde edilen diziler virtual RFLP analizine tabi tutularak 17 restriksiyon enzimiyle [AluI, BamHI, BfaI, BstUI (ThaI), DraI, EcoRI, HaeIII, HhaI, HinfI, HpaI, HpaII, KpnI, Sau3AI (MboI), MseI, RsaI, SspI ve TaqI] kesilmiş ve elde edilen bant profilleri referans izolatlarla karşılaştırılmıştır. Virtual RFLP analizleri sonucunda 11 izolatımız Ca. P. mali, 4 izolatımız Ca. P. prunorum, 3 izolatımız Ca. P. pyri, 2 izolatımız Ca. P. asteris ve diğer 2 izolatımız ise Ca. P. trifolii ile benzer enzim kesim profili vermiştir.
In this study, the presence of phytoplasmas in plum trees in Izmir province was determined, the possible phytoplasma vector insects were collected from phyoplasma infected orchards and morphologically identified and the presence of phytoplasma in the insect was investigated by molecular methods. The most common symptoms observed in plum trees were chlorosis and inward curling in the leaves, small leaves, reddening and shoot proliferation. As a result of field surveys, a total of 110 plum samples from 9 different plum varieties and 30 insects identified as Psammotettix provincialis belonging to the Cicadellidae family were collected from these orchards. Phytoplasma was detected in 29 of the plum samples tested by Nested-PCR analyses and 8 PCR products were cut with RsaI and SspI restriction endonuclease enzymes for RFLP analysis and 4 were found to be infected with Apple proliferation phytoplasma (Candidatus Phytoplasma mali) and 4 other plum samples were infected with Pear decline phytoplasma (Candidatus Phytoplasma pyri). 16 of the 30 P. provincialis individuals tested by Nested-PCR analyses were found to be positive for phytoplasma. In the phylogenetic analyzes performed, it was determined that these individuals were infected with Candidatus Phytoplasma asteris (16SrI). 22 out of 29 phytoplasma positive isolates were sent for sequence analysis and the obtained sequences were subjected to virtual RFLP analysis with 17 restriction enzymes [AluI, BamHI, BfaI, BstUI (ThaI), DraI, EcoRI, HaeIII, HhaI, HinfI, HpaI, HpaII, KpnI, Sau3AI (MboI), MseI, RsaI, SspI and TaqI] were cut and the obtained band profiles were compared with reference isolates. As a result of virtual RFLP analysis, 11, 4, 3, 2 and 2 isolates gave a similar enzyme cut profile with Ca. P. mali, Ca. P. prunorum, Ca. P. pyri, Ca. P. asteris and Ca. P. trifolii, respectively.
In this study, the presence of phytoplasmas in plum trees in Izmir province was determined, the possible phytoplasma vector insects were collected from phyoplasma infected orchards and morphologically identified and the presence of phytoplasma in the insect was investigated by molecular methods. The most common symptoms observed in plum trees were chlorosis and inward curling in the leaves, small leaves, reddening and shoot proliferation. As a result of field surveys, a total of 110 plum samples from 9 different plum varieties and 30 insects identified as Psammotettix provincialis belonging to the Cicadellidae family were collected from these orchards. Phytoplasma was detected in 29 of the plum samples tested by Nested-PCR analyses and 8 PCR products were cut with RsaI and SspI restriction endonuclease enzymes for RFLP analysis and 4 were found to be infected with Apple proliferation phytoplasma (Candidatus Phytoplasma mali) and 4 other plum samples were infected with Pear decline phytoplasma (Candidatus Phytoplasma pyri). 16 of the 30 P. provincialis individuals tested by Nested-PCR analyses were found to be positive for phytoplasma. In the phylogenetic analyzes performed, it was determined that these individuals were infected with Candidatus Phytoplasma asteris (16SrI). 22 out of 29 phytoplasma positive isolates were sent for sequence analysis and the obtained sequences were subjected to virtual RFLP analysis with 17 restriction enzymes [AluI, BamHI, BfaI, BstUI (ThaI), DraI, EcoRI, HaeIII, HhaI, HinfI, HpaI, HpaII, KpnI, Sau3AI (MboI), MseI, RsaI, SspI and TaqI] were cut and the obtained band profiles were compared with reference isolates. As a result of virtual RFLP analysis, 11, 4, 3, 2 and 2 isolates gave a similar enzyme cut profile with Ca. P. mali, Ca. P. prunorum, Ca. P. pyri, Ca. P. asteris and Ca. P. trifolii, respectively.
Açıklama
Anahtar Kelimeler
Ziraat, Agriculture
