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PCR-RFLP ANALYSIS OF DIFFERENT FUSARIUM SPECIES ASSOCIATED WITH ONION PRODUCTION

Yıl 2014, Cilt: 29 Sayı: 3, 194 - 198, 29.10.2014
https://doi.org/10.7161/anajas.2014.29.3.194-198

Öz

The study analyzed genetic relationships among different Fusarium species associated with onion production of Turkey using PCR-RFLP technique. ITS region of ribosomal DNA of 42 fungal isolates, representing F. oxysporum, F. solani, F. acuminatum, F. equiseti, F. proliferatum, F. redolens, and F. culmorum was amplified with primer pairs ITS1/4 and digested with HinfI, AluI, MspI, EcoRI enzymes. PCR amplification yielded a single DNA fragment ranging in size from 550-570 bp from all isolates. Restriction digests revealed 17 restriction patterns, providing the differentiation of Fusarium species from each other. All Fusarium species clustered into different groups in the dendogram derived from the data of PCR-RFLP. Similarity index ranged from 0.70 to 0.13 among Fusarium species. The results revealed that restriction patterns were very useful in the discrimination of Fusarium species, causing economically significant losses in onion production areas.

Kaynakça

  • Baayen, R.P., van den Boogert, P.H.J.F., Bonants, P.J.M., Poll, J.T.K., Blok, W.J., Waalwijk, C. 2000. Fusarium redolens f. sp. asparagi, causal agent of asparagus root rot, crown rot and spear rot. Eur. J. Plant Pathol., 106: 907-912.
  • Bayraktar, H., Dolar, F.S. 2011. Molecular identification and genetic diversity of Fusarium species associated with onion fields in Turkey. J. Phytopathol., 159: 28-34.
  • Booth, C. 1977. Fusarium a laboratory guide to the identification of the major species. C.M.I. Kew surrey, p.58. England.
  • Chehri, K., Salleh, B., Yli-Mattila, T., Reddy, K.R.N., Abbasi, S. 2011. Molecular characterization of pathogenic Fusarium species in cucurbit plants from Kermanshah province, Iran. Saudi J Biol Sci., doi: 10.1016/j.sjbs.2011.01.007.
  • Edel, V., Steinberg, C., Gautheron, N., Alabouvette, C. 1996. Evaluation of restriction analysis of polymerase chain reaction (PCR)-amplifies ribosomal DNA for the identification of Fusarium species. Mycol. Res. 101 (2): 179-187.
  • Hartman, G.L., Datnoff, L.E. 1997. Vegetable Crops. In: Soilborne Diseases of Tropical Crops. Eds, R.S. Hollocks and J.M. Waller, CAB Intertional, Wallingford, Oxon, p. 151-170.
  • Kordalı, Ş., Demirci, E., 1998. Fusarium species from various vegetables in Erzincan. J. Turk. Phytopath., 27(2-3): 131-136.
  • Latiffah, Z., Nur Hayati, M.Z., Baharuddin, S., Maziah, Z. 2009. Identification and pathogenicity of Fusarium species associated with root rot and stem rot of dendrobium. Asian J. Plant Pathol., 3(1): 14-21.
  • Leslie, J.F., Summerell, B.A. 2006. The Fusarium laboratory manual. Blackwell, Ames, p.388.
  • Nelson, P.E., Toussoun T.A., Marasas W.F.O. 1983. Fusarium Species. An illustrated manual for identification. Pennsylvania State University Press, pp. 193.
  • Özer, N., Ömeroğlu, M. 1995. Chemical control and determination of fungal causal agents of wilt disease of onion in Tekirdağ province. J. Turk. Phytopath., 24(2): 47-55.
  • Reader, U., Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol., 1: 17-20.
  • Rohlf, F.I. 1998. NTSYS-pc. Numerical taxonomy and multivariate analysis system, version 2.0. Applied Biostatistics, New York, USA.
  • Sambrook, J., Fritsch, E.F., Maniatis, T. 1989. Molecular Cloning: A Laboratory manual, 2nd edn. (pp 1659). New York, Cold Spring Harbor Laboratory Press.
  • Türkkan, M., Karaca, G. 2006. Determination of fungal root rot disease agents associated with onion fields in Amasya province. Tar. Bil. Der., 12(4): 357-363.
  • White, T.J., Bruns, T.D., Lee, S., Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA for phylogentics. In: Innis MA, Gelfland DH, Sninsky JJ, White TJ, eds. PCR Protocols: A Guide to Methods and Applications. San Diego, CA, USA: Academic Press, pp. 315-22.
  • Young-Mi, L., Choi, Y.K., Min, B.R. 2000. PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp. J. Microbiol., 38(2): 66-73.

SOĞAN ÜRETİMİ İLE İLİŞKİLİ FARKLI FUSARİUM TÜRLERİNİN PCR-RFLP ANALİZİ

Yıl 2014, Cilt: 29 Sayı: 3, 194 - 198, 29.10.2014
https://doi.org/10.7161/anajas.2014.29.3.194-198

Öz

Bu çalışma kapsamında ülkemizde soğan üretimi ile ilişkili farklı Fusarium türleri arasındaki genetik akrabalık PCR-RFLP yöntemi ile araştırılmıştır. F. oxysporum, F. solani, F. acuminatum, F. equiseti, F. proliferatum, F. redolens ve F. culmorum’ u temsil eden 42 fungus izolatının ribozomal DNA ITS bölgesi ITS1/4 primerleri ile çoğaltıldıktan sonra HinfI, AluI, MspI, EcoRI restriksiyon enzimleri ile kesilmiştir. PCR analizi sonucunda tüm izolatlardan 550-570 bp büyüklüğünde tek bir DNA fragmenti çoğaltılmıştır. Enzim kesimleri ise Fusarium türlerinin birbirinden ayrılmasını sağlayan 17 farklı enzim profili oluşturmuştur. ITS bölgesinin RFLP analizi ile oluşturulan dendogramda tüm Fusarium türleri farklı gruplar içerisinde yer almıştır. Fusarium türleri arasındaki benzerlik oranının ise 0.70-0.13 arasında değiştiği tespit edilmiştir. Elde edilen sonuçlar enzim profillerinin soğan üretim alanlarında ekonomik olarak önemli kayıplara yol açan Fusarium türlerinin ayrılmasında oldukça faydalı olduğunu göstermiştir.

Kaynakça

  • Baayen, R.P., van den Boogert, P.H.J.F., Bonants, P.J.M., Poll, J.T.K., Blok, W.J., Waalwijk, C. 2000. Fusarium redolens f. sp. asparagi, causal agent of asparagus root rot, crown rot and spear rot. Eur. J. Plant Pathol., 106: 907-912.
  • Bayraktar, H., Dolar, F.S. 2011. Molecular identification and genetic diversity of Fusarium species associated with onion fields in Turkey. J. Phytopathol., 159: 28-34.
  • Booth, C. 1977. Fusarium a laboratory guide to the identification of the major species. C.M.I. Kew surrey, p.58. England.
  • Chehri, K., Salleh, B., Yli-Mattila, T., Reddy, K.R.N., Abbasi, S. 2011. Molecular characterization of pathogenic Fusarium species in cucurbit plants from Kermanshah province, Iran. Saudi J Biol Sci., doi: 10.1016/j.sjbs.2011.01.007.
  • Edel, V., Steinberg, C., Gautheron, N., Alabouvette, C. 1996. Evaluation of restriction analysis of polymerase chain reaction (PCR)-amplifies ribosomal DNA for the identification of Fusarium species. Mycol. Res. 101 (2): 179-187.
  • Hartman, G.L., Datnoff, L.E. 1997. Vegetable Crops. In: Soilborne Diseases of Tropical Crops. Eds, R.S. Hollocks and J.M. Waller, CAB Intertional, Wallingford, Oxon, p. 151-170.
  • Kordalı, Ş., Demirci, E., 1998. Fusarium species from various vegetables in Erzincan. J. Turk. Phytopath., 27(2-3): 131-136.
  • Latiffah, Z., Nur Hayati, M.Z., Baharuddin, S., Maziah, Z. 2009. Identification and pathogenicity of Fusarium species associated with root rot and stem rot of dendrobium. Asian J. Plant Pathol., 3(1): 14-21.
  • Leslie, J.F., Summerell, B.A. 2006. The Fusarium laboratory manual. Blackwell, Ames, p.388.
  • Nelson, P.E., Toussoun T.A., Marasas W.F.O. 1983. Fusarium Species. An illustrated manual for identification. Pennsylvania State University Press, pp. 193.
  • Özer, N., Ömeroğlu, M. 1995. Chemical control and determination of fungal causal agents of wilt disease of onion in Tekirdağ province. J. Turk. Phytopath., 24(2): 47-55.
  • Reader, U., Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol., 1: 17-20.
  • Rohlf, F.I. 1998. NTSYS-pc. Numerical taxonomy and multivariate analysis system, version 2.0. Applied Biostatistics, New York, USA.
  • Sambrook, J., Fritsch, E.F., Maniatis, T. 1989. Molecular Cloning: A Laboratory manual, 2nd edn. (pp 1659). New York, Cold Spring Harbor Laboratory Press.
  • Türkkan, M., Karaca, G. 2006. Determination of fungal root rot disease agents associated with onion fields in Amasya province. Tar. Bil. Der., 12(4): 357-363.
  • White, T.J., Bruns, T.D., Lee, S., Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA for phylogentics. In: Innis MA, Gelfland DH, Sninsky JJ, White TJ, eds. PCR Protocols: A Guide to Methods and Applications. San Diego, CA, USA: Academic Press, pp. 315-22.
  • Young-Mi, L., Choi, Y.K., Min, B.R. 2000. PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp. J. Microbiol., 38(2): 66-73.
Toplam 17 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Bitki Koruma
Yazarlar

Harun Bayraktar

Kübra Tekin Bu kişi benim

Göksel Özer

Yayımlanma Tarihi 29 Ekim 2014
Yayımlandığı Sayı Yıl 2014 Cilt: 29 Sayı: 3

Kaynak Göster

APA Bayraktar, H., Tekin, K., & Özer, G. (2014). SOĞAN ÜRETİMİ İLE İLİŞKİLİ FARKLI FUSARİUM TÜRLERİNİN PCR-RFLP ANALİZİ. Anadolu Tarım Bilimleri Dergisi, 29(3), 194-198. https://doi.org/10.7161/anajas.2014.29.3.194-198
Online ISSN: 1308-8769