Çanakkale ilinde Karnabahar mozaik virüsü (Cauliflower mosaic virus; CaMV) izolatlarının tanılanması ve karakterizasyonu

Year 2014, Volume: 27 Issue: 1, 1 - 7, 01.06.2014

Hasan Tuna Tuzlalı Savaş Korkmaz

Abstract

Bu çalışmada Çanakkale ili ve ilçelerinde 2010-2011 üretim yılları içinde bir sörvey çalışması yürütülerek Karnabahar mozaik virüsü (Cauliflower mosaic virus, CaMV)’ne benzer semptom gösteren karnabahar ve lahana bitkilerinden 84 örnek toplanmıştır. Toplanan örnekler CaMV’nin varlığını belirlemek amacıyla DAS-ELISA yöntemiyle test edilmiştir. DAS-ELISA testi sonucunda 84 örnekten 63’ü CaMV ile infekteli bulunmuştur. İnfekteli bulunan örnekler içerisinden seçilen 6 izolatın moleküler özelliklerini belirlemek amacıyla kılıf protein (CP) genleri klonlanarak nükleik asit dizilimleri belirlenmiştir. Klonlama ve sekans analizi yapılan CaMV izolatlarına özgü nükleotid dizilimleri gen bankasında bulunan ve dünyanın farklı üretim bölgelerinden CaMV izolatlarının kılıf protein genleriyle karşılaştırılmıştır. Bu karşılaştırmalar sonucunda Çanakkale izolatlarının CP nükleotid dizilimlerinin kendi içinde % 93-100, dünya izolatları ile % 92-97 oranında bir benzerliğe sahip olduğu bulunmuştur. Yapılan filogenetik analizler sonucunda Çanakkale izolatlarının dünyanın farklı bölgelerindeki izolatlarla farklı düzeylerde ilişki gösterdiği saptanmıştır.

Keywords

Çanakkale, Karnabahar mozaik virüsü, DAS-ELISA, klonlama, sekanslama

Identification and characterization of Cauliflower mosaic virus (CaMV) isolates in Çanakkale province

Abstract

In this study, a survey was conducted in Çanakkale province and sub provinces on cauliflower and cabbage plants that show Cauliflower mosaic virus-like symptoms between the years 2010-2011. A total of 84 samples were collected from these areas and tested by DAS-ELISA for the presence of Cauliflower mosaic virus (CaMV). As a result of the DAS-ELISA analysis 63 out of 84 samples were found infected with CaMV. The coat protein genes of 6 isolates were cloned and sequenced. Isolates were further characterized and the sequences obtained from CaMV isolates of Çanakkale were compared with known sequences from other part of the world to determine the genetic differences and evolutionary relationships among CaMV isolates from Çanakkale and the other parts of the world. Comparison of CP genes revealed that CP gene of CaMV isolates from Çanakkale province and sub provinces showed 93-100 % and isolates with the world 92-97 % identity in their nucleotide sequence, respectively. Phylogenetic analysis of the CP gene sequences showed that CaMV isolates from Çanakkale province and sub provinces displayed different level of genetic relationship with CaMV isolates from those of the world.

Keywords

Çanakkale, Cauliflower mosaic virus, DAS-ELISA, Cloning, sequencing

References

• Agama K, Beach J, Schoelz J, Leisner SM (2002) The 5’-third of Cauliflowermosaicvirusgene VI conditions resistance breakage in Arabidopsisecotype Tsu-0.Phytopathology 92:190-196.
• Al-Kaff NS, Covev SN (1995) Biological diversity of Cauliflowermosaicvirus isolates expressed in two Brassicaspecies. Plant Pathology 44: 516-526.
• Alan B (2012) Doğu Akdeniz Bölgesi’nde yetiştirilen bazı kışlık sebzelerde görülen virüslerin tanılanması ve karakterizasyonu. Doktora Tezi, Çukurova Üniversitesi Fen Bilimleri Enstitüsü, Adana.
• Blanc S, Hebrard E, Drucker M,Froissart R (2001) Moleculerbasis of vector transmission Cauliflowermosaicvirus Cabb-S strainand S Delta II hybridbytwospecies of aphid: Myzuspersicae (sulzer) and Brevicorynebrassicae(L.) Res. Virol. 141: 677-683.
• Cheng RH, Olson NH, Baker TS (1992) CauliflowerMosaicVirus: A 420 subunit (T=7), multilayerstructure. Virology186: 655-668.
• Clark MF, Adams AN (1977) Characteristic of themicroplatemethod of enzyme-linked immunosorbent assay for the detection of plantviruses. J. Gen. Virol.34: 475-483.
• Çevik B, Pappu SS, Pappu HR, Benscher D, Lee RF, Futch SH., Rucks P, Niblett CL (1995) Molecular cloning and sequencing of coat protein genes of Citrustristezavirus isolated from Meyerlemon and homelytangortrees in Florida. In: Proc. Intern Org. Citrus Virologist. IOCV University of California Riverside, CA. 47-53.
• Daubert S, Routh G (1990) Point mutations in Cauliflowermosaicvirus gene IV conferhostspesifics symptoms virus gene IV conferhostspesifics symptoms changes. Mol. PlantsMic. Interac. 3: 341-345.
• Erkan S, Eşiyok D, Eser B (1990) A New viralagent affecting cauliflower and cabbageplantsin Turkey Journal of Turkish Phytopathology 19: 95-97.
• Farzadfar S, Pourrahim R, Golnaraghi AR, Ahounmanesh A (2005) Occurrence of Cauliflowermosaicvirus in different Cruciferous plants in Iran. Plant Pathol. 54: 810.
• Farzadfar S, Ahounmanesh A, Mosahebi GH, Oshima K, Koohi-Habibi M, Pourrahim R, Golnaraghi AR (2007) Partialbiolaogical and molecular characterization of Cauliflowermosaicvirus Isolates in Iran. Plant Pathology Journal6: 291-298.
• Hardwick NV, Davies JML, Wright DM (1994) The incidence of three virusdiseases of winteroilseedrape in EnglandandWales in the 1991/92 and 1992/93 growingseason. PlantPathol. 43: 1045-1049.
• Jiang B, Hong N, Wang GP, Hu J, Zhang JK, Wang CX, Liu Y, Fan XD (2008) Characterization of Citrustristezaviruss trains from southern Chinabased on analysis of restrictionpatterns and sequences of theircoat protein genes.VirusGenes 37: 185-192.
• Kennedy JS, Day MF,Eastop VF (1962) A conspectus of aphids as vectors of plantviruses. Common Wealth Institute of Entomol London,The Eastern Press Ltd, London.
• Korkmaz S, Çevik B, Kurtuluş E, Tuzlalı HT (2011) Güneybatı Marmara Bölgesi’nde Brassicaceae ve Alliceaefamilyasına bağlı bitkilerde virüs hastalıklarının teşhisi. Çanakkale Tarım Sempozyumu Dünü, Bugünü Geleceği, Çanakkale s. 460-467.
• Melcher U (1989) Symptoms of Cauliflowermosaicvirus in infection Arabidopsisthaliana and turnip. Bot Gaz.150:137-139.
• Moreno A, Blas CDE, Burrun R, Nebrada M, Palacious I, Duque M, Ferreres A (2004) The incidence and distribution of viruses infecting lettuce, cultivated Brassica and assocatiated natural vegetationin Spain. Ann. Appl. Biol. 144: 339-346.
• Palacios I, Drucker M, Blanc S, Leite S, Moreno A (2002) Cauliflowermosaicvirus is preferentially acquired from the phloem by its aphid vectors. J. Genet. Virol.83: 3163-3171.
• Shahraeen N (2012) An overview of oilseedrape (canola) virus diseases in Iran. International Research Journal of Microbiology Vol. 3: 24-28.
• Schoelz J, Shepherd RJ, Daubert S (1986) Region VI of Cauliflowermosaicvirus encodes a hostrange determinant. Mol. Cell. Biol. 6: 2632-2637.
• Shephard R J (1981) Cauliflowermosaicvirus. AAB Descriptions of Plant Viruses No. 243.
• Suehiro N, Matsuda K, Okuda S, Natsuaki T (2005) A simplified method for obtaining plantviral RNA for RT-PCR. J VirolMethods125: 67-73.
• Wintermantel WM, Anderson EJ, Schoelz JE (1993) Identification of domains within gene VI of Cauliflowermosaicvirus that influence systemic infection of Nicotianabiglovii in a light-dependentmanner. Virology 196: 789-798.