Adıyaman ili badem ağaçlarında önemli prunus virüslerinin DAS-ELISA ve RT-PCR analizleri ile saptanması ve karakterizasyonu
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Dosyalar
Tarih
2020
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Yayıncı
Hatay Mustafa Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Dünyada ve ülkemizde ekonomik önemi yıldan yıla artan badem ağaçlarında ekonomik kayıplara yol açan virüsler içerisinde elma mozaik virüsü (ApMV), prunus nekrotik halkalı leke virüsü (PNRSV), erik cücelik virüsü (PDV) şarka virüsü (PPV) ve elma klorotik yaprak leke virüsü (ACLSV) en yaygın görülenlerdir. Ticari amaçla yapılan badem yetiştiriciliği Adıyaman ili için yeni bir ürün olmakla birlikte kapama bahçelerin sayısı her geçen gün artmakta ve üretim de teşvik edilmektedir. Bu çalışma kapsamında Adıyaman ilinde badem bahçelerinde ApMV, ACLSV, PDV, PNRSV ve PPV'nin varlığı Double Antibody Sandwich–Enzyme Linked Immuno Sorbent Assay (DAS-ELISA) ve Ters Traskripsiyon-Polimeraz Zincir Reaksiyonu (RT-PCR) yöntemleri ile araştırılmış, belirlenen bazı izolatların biyolojik ve moleküler karakterizasyonu yapılmıştır. Badem ağaçlarında görülen bu 5 virüsün varlığını belirlemek amacıyla yapılan DAS-ELISA testleri sonucunda 34 tane örnek PDV ile enfekteli bulunmuş ancak ACLSV, ApMV, PNRSV ve PPV tespit edilememiştir. RT-PCR analizleri ile testlenen 110 badem örneğinde sadece PDV'üne ait beklenen uzunluktaki (756 bp) DNA fragmenti 34 örnekte çoğaltılırken, testlenen örneklerde yine ApMV, ACLSV, PNRSV ve PPV saptanamamıştır. Testlenen badem örneklerinin PDV ile enfeksyion oranı %30.90 olarak saptanmıştır. RT-PCR yöntemiyle çoğaltılan PDV izolatları doğrudan dizilenerek nükleotid ve kodladıkları amino asit dizileri analiz edilerek Adıyaman'dan elde edilen PDV izolatlarının birbirleriyle ve Gen Bankasına kayıtlı diğer ülkelerin izolatlarıyla karşılaştırması yapılmış ve benzerlik oranları ile filogenetik ilişkileri ortaya konulmuştur. PDV izolatlarına ait NJ (Neighbor Joining) dendogramı incelendiğinde Adıyaman PDV badem izolatlarının birbirine çok benzediği ve filogenetik ağaçta aynı grup içinde yer aldığı, gen bankasına kayıtlı diğer PDV izolatlarından ayrı bir grupta kümelendiği belirlenmiştir. Yapılan bu çalışma ile PDV ilk kez Adıyaman ili badem plantasyonlarında saptanmıştır.
There are viral diseases causing economic losses on almond trees whose economic importance is increasing by year in the world and also in our country. Among these viruses, apple mosaic virus (ApMV), prunus necrotic ring spot virus (PNRSV), prune dwarf virus (PDV), plum pox virus (PPV) and apple chlorotic leaf spot virus (ACLSV) are the most common viruses. Although commercial almond cultivation is a new product for Adıyaman province, the number of enclosed orchards is increasing day by day and its production is also encouraged. Within the scope of this study, the presence of ApMV, ACLSV, PDV, PNRSV and PPV on almond orchards in Adıyaman province were investigated by Double Antibody Sandwich-Enzyme Linked Iimmuno Sorbent Assay (DAS-ELISA), Reverse Transcription-Polymerase Chain Reaction (RT-PCR) analysis and molecular characterization of some isolates has been made. As a result of the DAS-ELISA tests performed to determine the presence of these 5 viruses seen in almond trees, 34 samples were found to be infected with PDV and ACLSV, ApMV, PNRSV and PPV were never found in tested samples. In 110 almond samples tested with RT-PCR analysis, only the expected length (756 bp) DNA fragment of PDV was replicated in 34 samples, while ApMV, ACLSV, PNRSV and PPV were not detected in the tested samples. The infection rate of PDV in tested almond samples was found to be 30.90%. PDV isolates amplified by RT-PCR method were directly sequenced and nucleotide and amino acid sequences they encoded were analyzed, PDV isolates obtained from Adıyaman with each other and with different countries isolates deposited in Gen Bank were compared and their similarity rates and phylogenetic relationships were revealed. When the NJ (Neighbor Joining) dendogram of PDV isolates was examined, it was determined that Adıyaman PDV almond isolates were very similar to each other and included in the same group in the phylogenetic tree, clustered in a separate group from other PDV isolates registered in the gene bank. PDV was detected for the first time in almond plantations in Adıyaman province by this study.
There are viral diseases causing economic losses on almond trees whose economic importance is increasing by year in the world and also in our country. Among these viruses, apple mosaic virus (ApMV), prunus necrotic ring spot virus (PNRSV), prune dwarf virus (PDV), plum pox virus (PPV) and apple chlorotic leaf spot virus (ACLSV) are the most common viruses. Although commercial almond cultivation is a new product for Adıyaman province, the number of enclosed orchards is increasing day by day and its production is also encouraged. Within the scope of this study, the presence of ApMV, ACLSV, PDV, PNRSV and PPV on almond orchards in Adıyaman province were investigated by Double Antibody Sandwich-Enzyme Linked Iimmuno Sorbent Assay (DAS-ELISA), Reverse Transcription-Polymerase Chain Reaction (RT-PCR) analysis and molecular characterization of some isolates has been made. As a result of the DAS-ELISA tests performed to determine the presence of these 5 viruses seen in almond trees, 34 samples were found to be infected with PDV and ACLSV, ApMV, PNRSV and PPV were never found in tested samples. In 110 almond samples tested with RT-PCR analysis, only the expected length (756 bp) DNA fragment of PDV was replicated in 34 samples, while ApMV, ACLSV, PNRSV and PPV were not detected in the tested samples. The infection rate of PDV in tested almond samples was found to be 30.90%. PDV isolates amplified by RT-PCR method were directly sequenced and nucleotide and amino acid sequences they encoded were analyzed, PDV isolates obtained from Adıyaman with each other and with different countries isolates deposited in Gen Bank were compared and their similarity rates and phylogenetic relationships were revealed. When the NJ (Neighbor Joining) dendogram of PDV isolates was examined, it was determined that Adıyaman PDV almond isolates were very similar to each other and included in the same group in the phylogenetic tree, clustered in a separate group from other PDV isolates registered in the gene bank. PDV was detected for the first time in almond plantations in Adıyaman province by this study.
Açıklama
Anahtar Kelimeler
Ziraat, Agriculture, Adıyaman, badem, DAS-ELISA, RT-PCR, sekans analizi, virüs, Adıyaman, almond, DAS-ELISA, RT-PCR, sequencing, virus
