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Türkiye’de turunçgil ağaçlarında moleküler tekniklerle Spiroplasma citri’nin belirlenmesi

Year 2020, Volume: 33 Issue: 1, 37 - 42, 01.04.2020
https://doi.org/10.29136/mediterranean.657937

Abstract

Spiroplasma citri, kurak ve yarı kurak iklimde turunçgiller üzerindeki turunçgil palamutlaşma hastalığı (Citrus Stubborn Disease (CSD))’na neden olan bir etmendir. Bu etmenin turunçgil bahçeleri ve fidanlıklarda bulunan genç fidanlardaki enfeksiyonunun erken aşamada tespiti ve karakterizasyonu, hastalığın Spiroplasma citri’den ari bölgelere yayılmasını önlemek için büyük önem taşımaktadır. Bu çalışma, Türkiye'deki farklı Spiroplasma citri suşlarının muhtemel varlığını araştırmak amacıyla yapılmıştır. Eylül ayında Adana ilinin Kozan ilçesinde farklı köylerde bulunan turunçgil bahçelerindeki Stubborn hastalığına benzer çoğunlukla boğum arası kısalma, yapraklarda kaşıklanma ve leke, sezon dışı çiçeklenme ve tipik küçük palamut tipi meyve simptomları gösteren ağaçlardan yaprak ve meyve örneği toplanmıştır. Türk izolatlarının saptanması ve karakterizasyonu için, polimeraz zincir reaksiyonu (PZR) Spiralin-f/r, P89-r/f ve P58-6f/4r primer çiftleri kullanılarak gerçekleştirilmiştir. Spiralin-f/r primerlerinde pozitif sonuç saptanmamıştır. Buna karşın, testlenen tüm örnekler P89-r/f ve P58-6f/4r primer çiftlerine pozitif sonuç vermiştir. P89-r/f primer çiftinden elde edilen PZR ürünleri sekanslanmış ve filogenetik ağaç oluşturulmuştur. Elde edilen filogenetik ağaç, önemli bir turunçgil üretim merkezi olan Adana ilindeki turunçgil ağaçlarında saptanan Spiroplasma citri izolatlarının, Fas, Mısır ve Amerika Birleşik Devletleri (ABD)'nden izole olanlardan daha ziyade Meksika, İspanyol, Cezayir ve İtalyan izolatlarıyla yakından ilişkili olduğunu kanıtlamıştır.

References

  • Ahrens U, Seemüller E (1992) Detection of DNA of plant pathogenic mycoplasma like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82: 828-832.
  • Azeri T (1973) First report of Satsuma Dwarf virus disease on satsuma mandarin in Turkey. Plant Disease Reporter 57(2): 149-153.
  • Bové JM (1984) Wall-less prokaryotes of plants. Annual Review of Phytopathology 22: 361-396.
  • Bové JM (1988) Spiroplasma citri. In: IM Smith, J Dunez, RA Lelliott, DH Phillips, SA Archer (Eds.), European handbook of plant diseases, Blackwell Scientific Publications, Oxford, UK, pp. 129-131.
  • Bové JM, Garnier M (2000) Stubborn. In: LW Timmer, SM Garnsey & JH Graham (eds.) Compendium of Citrus Diseases, American Phytopathological Society Press, St. Paul, MN, pp. 48-50.
  • Calavan EC (1979) Symptoms of stubborn disease and the culture of Spiroplasma citri. in: Proc. R.O.C.-U. S. Coop. Sci. Semin. Mycoplasma Dis. Plants. NSC Symp. Ser. No. 1. National Science Council, Republic of China, pp. 67-72.
  • Calavan EC, Bové JM (1989) Ecology of Spiroplasma citri. in: RF Whitcomb & JG Tully (eds.) The Mycoplasmas, Academic Press, Inc., New York, pp. 425-485.
  • Clark MF, Davies DL, Buss SL, Morton A (1989) Serological discrimination among mycoplasma-like organisms using polyclonal and monoclonal antibodies. ActaHorticulturae 235: 107-113.
  • Çağlayan K (1987) Turunçgil yediverenleşme (Stubborn: palamutlaşma) hastalık etmeni Spiroplasma citri'nin izolasyonu, taşınması ve patojen-vektör ilişkilerinin araştırılması. Doktora Tezi, Çukurova Universitesi Adana, s. 133.
  • Çınar A, Kersting U, Önelge N, Korkmaz S, Sas G (1993) Citrus virus and virus-like diseases in the Eastern Mediterranean region of Turkey. In International Organization of Citrus Virologists Conference Proceedings 12: 397-400.
  • Fletcher J, Slack SA (1986) Latex agglutination as a rapid detection assay for Spiroplasmacitri. Plant Disease 70: 754-756.
  • Gromadka R (1995) Szybkaizolacja DNA z agarozy. In: In Zynieria Genetycznai Biologia Molekularna, Techgen Sp. Z.O.O. (ed.). Metody, Podreczniki laboratoryjne IBB PAN, Warszawa, pp. 6-7.
  • Güllü M (1989) Doğu Akdeniz Bölgesi Navel grubu portakal ve Satsuma mandarin ağaçlarında yaygın virus ve virus benzeri hastalıkların surveyi ve indekslenmesi üzerinde araştırmalar. Doktora Tezi, Çukurova Üniversitesi, Adana, s. 266.
  • Kersting U, Şengonca Q (1992) Detection of insect vectors of the citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the citrus growing area of South Turkey. Journal of Applied Entomology 113(1-5): 356-364.
  • Kersting U, Şengonca Ç, Çınar A (1992) Detection of Spiroplasmacitri in non-citrus host plants and their associated leafhopper vectors in southern Turkey. FAO Plant Protection Bulletein 40(3): 89-44.
  • Kersting U, Baspinar H, Cinar A, Şengonea C, Uygun N (1993) New findings on the epidemiology of Spiroplasmacitri in the Eastern Mediterranean region of Turkey. In XII. International Organization of Citrus Virologists Conference Proceedings 12: 336-341.
  • Kersting U, Başpmar H (1995) Seasonal and diurnal flight activity of Circulifer haematoceps (Hom., Cicadellidae), an important leafhopper vector in the Mediterranean area and the Near East. Journal of Applied Entomology 119(1‐5): 533-537.
  • Kersting U, Başpınar H (1997) Occurrence and Distribution of Spiroplasmacitri and Sesame Phyllody in Sesame in Southern Anatolia. Journal of Turkish Phytopathology 26(1): l-9.
  • Kersting U, Başpinar H, Uygun N, Satar S (1997) Comparison of two sampling methods for leafhoppers (Homoptera, Cicadellidae) associated with sesame in the east Mediterranean region of Turkey. Anzeiger für Schädlingskunde, Pflanzenschutz, Umweltschutz 70(7): 131-135.
  • Korkmaz S, Çınar A, Kersting U (1993) Seasonal changes of citrus stubborn disease pathogen, Spiroplasmacitri, by Budwood Grafting. Journal of Turkish Phytopathology pp. 9-16.
  • Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35: 1547-1549.
  • Lee IM, Bottner KD, Munyaneza JE, Davis RE, Crosslin JM, du Toit LJ, Crosby T (2006) Carrot purple leaf: a new spiroplasmal disease associated with carrots in Washington State. Plant Disease 90: 989-993.
  • Liu HY, Gumpf DJ, Oldfield GN, Calavan EC (1983) The relationship of Spiroplasma citri and Circulifer tenellus. Phytopathology 73: 585-590.
  • Oldfield GN, Kaloostian GH, Pierce HD, Calavan EC, Granett AL, Blue RL, Rana GL, Gumpf DJ (1977) Transmission of Spiroplasma citri from citrus to citrus by Scaphytopius nitridus. Phytopathology 67: 763-765.
  • Saglio P, L'Hospital M, Lafléche D, Dupont G, Bové JM, Tully JG, Freundt EA (1973) Spiroplasma citri gen. and sp. n.: A mycoplasma-like organism associated with stubborn disease of citrus. International Journal of Systematic Bacteriology 23: 191-204.
  • Saillard C, Bové JM (1983) Application of ELISA to spiroplasma detection and classification. In: S Razin, J G Tully, (eds.), Methods in mycoplasmology, Academic Press, New York, USA. Vol. 1, pp. 471-476.
  • Sertkaya G, Çınar A (2002) Detection of same weed species as host of S. citri Saglio et al. and its insect vector Curculifer haematoceps (M.-R.). in the Eastern Mediterranean Region of Turkey. Türkiye Herboloji Dergisi 5: 35-41.
  • Swisher KD, Velásquez-Valle R, Mena-Covarrubias J, Munyaneza JE (2016) Occurrence and molecular detection of Spiroplasma citri in carrots and its insect vector, Circulifer tenellus, in Mexico. Journal of Plant Pathology 98(2): 355-360.
  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) Clustal_X Windows Interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24): 4876-4882.
  • Tully JG (1983) Dark-field microscopy. In. S Razin and JG Tully (eds.) Methods in Mycoplasmology, Academic Press, New York.
  • Uygur S, Uygur FN, Çınar A (1991) Çukurova Bölgesinde Spiroplasma citri Saglio et al.’nin konukçusu olan yabancı ot türlerinin saptanması. VI. Türkiye Fitopatoloji Kongresi, İzmir, pp. 311-314.
  • Yokomi RK, Mello AFS, Saponari M, Fletcher J (2008) Polymerase chain reaction based detection of Spiroplasma citri associated with citrus stubborn disease. Plant Disease 92: 253-260.
  • Yokomi RK, Sisterson M (2011) Validation and comparison of a hierarchal sampling plan for estimating incidence of citrus stubborn disease. In International Organization of Citrus Virologists Conference Proceedings 18(18): 1957-2010.

Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques

Year 2020, Volume: 33 Issue: 1, 37 - 42, 01.04.2020
https://doi.org/10.29136/mediterranean.657937

Abstract

Spiroplasma citri is the causal agent of Citrus Stubborn Disease (CSD) on cultivated citrus in the arid and semi-arid climate. Detection and characterization of this agent at the early stage of infection in citrus orchard and young citrus saplings in the nursery is of utmost importance to prevent the introduction and spread of the disease in Spiroplasma citri free areas. The current study was conducted in order to investigate the possible existence of different Spiroplasma citri strains in Turkey. Leaf and fruit samples were collected from ten years old citrus trees showing mostly short internodes, upright and mottled leaves, off-season blossoming and typical small and acorn-shaped fruits symptoms associated with citrus stubborn diseases in different villages of Kozan town of Adana province in Turkey. Polymerase chain reaction (PCR) was performed using Spiralin-f/r, P89-r/f and P58-6f/4r primer couples for the detection and characterization of the Turkish isolates. No positive sample was detected with the primers Spiralin-f/r. On the contrary, all samples reacted positively to the P89-r/f and P58-6f/4r primer couples. The obtained amplicon from the P89-r/f primers pair was sequenced and the phylogenetic tree was constructed. Phylogenetic tree showed that the Turkish Spiroplasma citri isolates are closely related to the Mexican, Spanish, Algerian, and Italian than those isolated from Morocco, Egypt and USA.

References

  • Ahrens U, Seemüller E (1992) Detection of DNA of plant pathogenic mycoplasma like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82: 828-832.
  • Azeri T (1973) First report of Satsuma Dwarf virus disease on satsuma mandarin in Turkey. Plant Disease Reporter 57(2): 149-153.
  • Bové JM (1984) Wall-less prokaryotes of plants. Annual Review of Phytopathology 22: 361-396.
  • Bové JM (1988) Spiroplasma citri. In: IM Smith, J Dunez, RA Lelliott, DH Phillips, SA Archer (Eds.), European handbook of plant diseases, Blackwell Scientific Publications, Oxford, UK, pp. 129-131.
  • Bové JM, Garnier M (2000) Stubborn. In: LW Timmer, SM Garnsey & JH Graham (eds.) Compendium of Citrus Diseases, American Phytopathological Society Press, St. Paul, MN, pp. 48-50.
  • Calavan EC (1979) Symptoms of stubborn disease and the culture of Spiroplasma citri. in: Proc. R.O.C.-U. S. Coop. Sci. Semin. Mycoplasma Dis. Plants. NSC Symp. Ser. No. 1. National Science Council, Republic of China, pp. 67-72.
  • Calavan EC, Bové JM (1989) Ecology of Spiroplasma citri. in: RF Whitcomb & JG Tully (eds.) The Mycoplasmas, Academic Press, Inc., New York, pp. 425-485.
  • Clark MF, Davies DL, Buss SL, Morton A (1989) Serological discrimination among mycoplasma-like organisms using polyclonal and monoclonal antibodies. ActaHorticulturae 235: 107-113.
  • Çağlayan K (1987) Turunçgil yediverenleşme (Stubborn: palamutlaşma) hastalık etmeni Spiroplasma citri'nin izolasyonu, taşınması ve patojen-vektör ilişkilerinin araştırılması. Doktora Tezi, Çukurova Universitesi Adana, s. 133.
  • Çınar A, Kersting U, Önelge N, Korkmaz S, Sas G (1993) Citrus virus and virus-like diseases in the Eastern Mediterranean region of Turkey. In International Organization of Citrus Virologists Conference Proceedings 12: 397-400.
  • Fletcher J, Slack SA (1986) Latex agglutination as a rapid detection assay for Spiroplasmacitri. Plant Disease 70: 754-756.
  • Gromadka R (1995) Szybkaizolacja DNA z agarozy. In: In Zynieria Genetycznai Biologia Molekularna, Techgen Sp. Z.O.O. (ed.). Metody, Podreczniki laboratoryjne IBB PAN, Warszawa, pp. 6-7.
  • Güllü M (1989) Doğu Akdeniz Bölgesi Navel grubu portakal ve Satsuma mandarin ağaçlarında yaygın virus ve virus benzeri hastalıkların surveyi ve indekslenmesi üzerinde araştırmalar. Doktora Tezi, Çukurova Üniversitesi, Adana, s. 266.
  • Kersting U, Şengonca Q (1992) Detection of insect vectors of the citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the citrus growing area of South Turkey. Journal of Applied Entomology 113(1-5): 356-364.
  • Kersting U, Şengonca Ç, Çınar A (1992) Detection of Spiroplasmacitri in non-citrus host plants and their associated leafhopper vectors in southern Turkey. FAO Plant Protection Bulletein 40(3): 89-44.
  • Kersting U, Baspinar H, Cinar A, Şengonea C, Uygun N (1993) New findings on the epidemiology of Spiroplasmacitri in the Eastern Mediterranean region of Turkey. In XII. International Organization of Citrus Virologists Conference Proceedings 12: 336-341.
  • Kersting U, Başpmar H (1995) Seasonal and diurnal flight activity of Circulifer haematoceps (Hom., Cicadellidae), an important leafhopper vector in the Mediterranean area and the Near East. Journal of Applied Entomology 119(1‐5): 533-537.
  • Kersting U, Başpınar H (1997) Occurrence and Distribution of Spiroplasmacitri and Sesame Phyllody in Sesame in Southern Anatolia. Journal of Turkish Phytopathology 26(1): l-9.
  • Kersting U, Başpinar H, Uygun N, Satar S (1997) Comparison of two sampling methods for leafhoppers (Homoptera, Cicadellidae) associated with sesame in the east Mediterranean region of Turkey. Anzeiger für Schädlingskunde, Pflanzenschutz, Umweltschutz 70(7): 131-135.
  • Korkmaz S, Çınar A, Kersting U (1993) Seasonal changes of citrus stubborn disease pathogen, Spiroplasmacitri, by Budwood Grafting. Journal of Turkish Phytopathology pp. 9-16.
  • Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35: 1547-1549.
  • Lee IM, Bottner KD, Munyaneza JE, Davis RE, Crosslin JM, du Toit LJ, Crosby T (2006) Carrot purple leaf: a new spiroplasmal disease associated with carrots in Washington State. Plant Disease 90: 989-993.
  • Liu HY, Gumpf DJ, Oldfield GN, Calavan EC (1983) The relationship of Spiroplasma citri and Circulifer tenellus. Phytopathology 73: 585-590.
  • Oldfield GN, Kaloostian GH, Pierce HD, Calavan EC, Granett AL, Blue RL, Rana GL, Gumpf DJ (1977) Transmission of Spiroplasma citri from citrus to citrus by Scaphytopius nitridus. Phytopathology 67: 763-765.
  • Saglio P, L'Hospital M, Lafléche D, Dupont G, Bové JM, Tully JG, Freundt EA (1973) Spiroplasma citri gen. and sp. n.: A mycoplasma-like organism associated with stubborn disease of citrus. International Journal of Systematic Bacteriology 23: 191-204.
  • Saillard C, Bové JM (1983) Application of ELISA to spiroplasma detection and classification. In: S Razin, J G Tully, (eds.), Methods in mycoplasmology, Academic Press, New York, USA. Vol. 1, pp. 471-476.
  • Sertkaya G, Çınar A (2002) Detection of same weed species as host of S. citri Saglio et al. and its insect vector Curculifer haematoceps (M.-R.). in the Eastern Mediterranean Region of Turkey. Türkiye Herboloji Dergisi 5: 35-41.
  • Swisher KD, Velásquez-Valle R, Mena-Covarrubias J, Munyaneza JE (2016) Occurrence and molecular detection of Spiroplasma citri in carrots and its insect vector, Circulifer tenellus, in Mexico. Journal of Plant Pathology 98(2): 355-360.
  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) Clustal_X Windows Interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24): 4876-4882.
  • Tully JG (1983) Dark-field microscopy. In. S Razin and JG Tully (eds.) Methods in Mycoplasmology, Academic Press, New York.
  • Uygur S, Uygur FN, Çınar A (1991) Çukurova Bölgesinde Spiroplasma citri Saglio et al.’nin konukçusu olan yabancı ot türlerinin saptanması. VI. Türkiye Fitopatoloji Kongresi, İzmir, pp. 311-314.
  • Yokomi RK, Mello AFS, Saponari M, Fletcher J (2008) Polymerase chain reaction based detection of Spiroplasma citri associated with citrus stubborn disease. Plant Disease 92: 253-260.
  • Yokomi RK, Sisterson M (2011) Validation and comparison of a hierarchal sampling plan for estimating incidence of citrus stubborn disease. In International Organization of Citrus Virologists Conference Proceedings 18(18): 1957-2010.
There are 33 citations in total.

Details

Primary Language English
Subjects Agricultural Engineering
Journal Section Makaleler
Authors

Behçet Kemal Çağlar 0000-0001-6618-1658

Gül Satar 0000-0002-5120-3313

Saadettin Baloğlu 0000-0001-9482-9247

Mounira İnas Drais 0000-0002-9934-5898

Khaled Djelouah 0000-0002-4806-122X

Publication Date April 1, 2020
Submission Date December 11, 2019
Published in Issue Year 2020 Volume: 33 Issue: 1

Cite

APA Çağlar, B. K., Satar, G., Baloğlu, S., Drais, M. İ., et al. (2020). Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques. Mediterranean Agricultural Sciences, 33(1), 37-42. https://doi.org/10.29136/mediterranean.657937
AMA Çağlar BK, Satar G, Baloğlu S, Drais Mİ, Djelouah K. Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques. Mediterranean Agricultural Sciences. April 2020;33(1):37-42. doi:10.29136/mediterranean.657937
Chicago Çağlar, Behçet Kemal, Gül Satar, Saadettin Baloğlu, Mounira İnas Drais, and Khaled Djelouah. “Detection of Spiroplasma Citri from Citrus Trees in Turkey by Molecular Techniques”. Mediterranean Agricultural Sciences 33, no. 1 (April 2020): 37-42. https://doi.org/10.29136/mediterranean.657937.
EndNote Çağlar BK, Satar G, Baloğlu S, Drais Mİ, Djelouah K (April 1, 2020) Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques. Mediterranean Agricultural Sciences 33 1 37–42.
IEEE B. K. Çağlar, G. Satar, S. Baloğlu, M. İ. Drais, and K. Djelouah, “Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques”, Mediterranean Agricultural Sciences, vol. 33, no. 1, pp. 37–42, 2020, doi: 10.29136/mediterranean.657937.
ISNAD Çağlar, Behçet Kemal et al. “Detection of Spiroplasma Citri from Citrus Trees in Turkey by Molecular Techniques”. Mediterranean Agricultural Sciences 33/1 (April 2020), 37-42. https://doi.org/10.29136/mediterranean.657937.
JAMA Çağlar BK, Satar G, Baloğlu S, Drais Mİ, Djelouah K. Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques. Mediterranean Agricultural Sciences. 2020;33:37–42.
MLA Çağlar, Behçet Kemal et al. “Detection of Spiroplasma Citri from Citrus Trees in Turkey by Molecular Techniques”. Mediterranean Agricultural Sciences, vol. 33, no. 1, 2020, pp. 37-42, doi:10.29136/mediterranean.657937.
Vancouver Çağlar BK, Satar G, Baloğlu S, Drais Mİ, Djelouah K. Detection of Spiroplasma citri from citrus trees in Turkey by molecular techniques. Mediterranean Agricultural Sciences. 2020;33(1):37-42.

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