Isolation and identification of Pseudomonas savastanoi pv. phaseolicola and Xanthomonas axonopodis pv. phaseoli in Middle Black Sea Region of Turkey

Abstract

In 2013-2014, surveys were conducted in order to investigate the presence of Pseudomonas savastonoi pv. phaseolicola (Psp) halo blight and Xanthomonas axonopodis pv. phaseoli (common bacterial blight) (Xap) in common bean production areas of Samsun, Tokat, Amasya at Middle Black Sea region. Total, 154 common bean fields were surveyed and their leaf and bean samples were collected with typically resembling symptoms using guided sampling method. Total of 72 purified bacterial isolates were obtained. All the isolates assayed in pathogenicity, morphological, biochemical tests. Further identification methods were conducted with LOPAT tests and molecular analyses (PCR), their results have revealed that 30 isolates were Psp and 17 isolates were Xap. Psp isolates were separately inoculated on 8 bean varieties to determine their races according to phenotypic reactions. Their reactions on 8 bean varieties were revealed that all Psp isolates were Psp race 1. This work are proved that races of Psp isolates using universal separation set at Middle Black Sea region in which supplies about 16 % of the total bean production in Turkey.

Keywords

• Halo blight
• Common bacterial blight
• Race
• PCR

Orta Karadeniz Bölgesi’nde Pseudomonas savastanoi pv. phaseolicola ve Xanthomonas axonopodis pv. phaseoli’nin izolasyonu ve tanılanması

Abstract

Orta Karadeniz Bölgesi Samsun, Tokat ve Amasya ili fasulye üretim alanlarında 2013-2014 yıllarında fasulye hale yanıklığı hastalığı etmeni Pseudomonas savastonoi pv. phaseolicola (Psp) ve adi yaprak yanıklığı hastalığı etmeni Xanthomonas axonopodis pv. phaseoli (Xap)’nin belirlenmesi için surveyler yapılmıştır. Güdümlü örnekleme yöntemiyle toplam 154 fasulye tarlası gezilerek şüpheli semptom gösteren yaprak ve meyve örnekleri toplanmıştır. İzolasyon çalışmaları sonucu 72 adet saf bakteri izolatı elde edilmiştir. Patojenisite, morfolojik, biyokimyasal testler, tanıyı destekleyici LOPAT testleri ve moleküler analizler (PCR) sonucunda 30 izolat Psp ve 17 izolat Xap olarak tanılanmıştır. Psp izolatları 8 farklı fasulye çeşidinden oluşan ırk ayrım setindeki bitkilere inokule edilmiş ve çeşitlerin verdiği reaksiyonlara göre Psp’nın ırk/ırkları belirlenmiştir. Çeşit reaksiyonlarına göre Psp izolatlarının tamamının 1 nolu ırk olduğu tespit edilmiştir. Bu çalışmada, Türkiye fasulye üretiminin % 16’sının gerçekleştiği Orta Karadeniz bölgesinde evrensel ırk ayrım seti kullanılarak Psp izolatlarının ırk düzeyinde ayrımı ilk kez yapılmıştır.

Keywords

• hale yanıklığı
• adi yaprak yanıklığı
• ırk
• PCR

References

1. Allen RG, Pereira LS, Raes D, Smith M (1998) Crop evapotranspiration guidelines for computing crop water requirements. FAO irrigation and drainage paper 56. Food and Agriculture Organization, Rome.
2. Balkaya A, Yanmaz R, Bozoğlu H, Gülümser A (1999) Samsun ilinin taze fasulye yetiştiriciliği yönünden durumunun belirlenmesi. Karadeniz Bölgesi Tarım Sempozyumu, Samsun, Cilt 1, pp. 51-62.
3. Baştaş KK, Şahin F (2017) Evaluation of seed borne bacterial pathogens on common bean cultivars grown in central Anatolia region Turkey. European Journal of Plant Pathology 147(2): 239-253.
4. Benlioğlu K, Özakman M, Önceler Z (1994) Bacterial blights of beans in Türkiye and resistance to halo blight and common blight. 9th Congress of the Mediterranean Phytopathological Union, Kuşadası-Aydın, pp. 574-550.
5. Bozkurt IA, Soylu S (2001) Farklı fasulye çeşitlerinin fasulye hale yanıklığı etmeni P.s. pv. phaseolicola ırklarına karşı gösterdiği reaksiyonların belirlenmesi. Türkiye IX. Fitopatoloji Kongresi, Tekirdağ, pp. 506-514.
6. Bozoğlu H (1995) Kuru Fasulyede (Phaseolus vulgaris L.) Bazı Tarımsal Özelliklerin GenotipxÇevre İnteraksiyonu ve Kalıtım Derecelerinin Belirlenmesi Üzerine Bir Araştırma. Doktora Tezi, Ondokuz Mayıs Üniversitesi Fen Bilimleri Enstitüsü, Samsun.
7. Dönmez MF, Şahin F, Elkoca E (2013) Identification of bean genotypes from Turkey resistance to common bacterial blight and halo blight diseases. Acta Scientiarum Polonorum, Hortorum Cultus 12(4): 139-151.
8. FAO (2018) Food and Agriculture Organization of the United Nations, Statistical database. Ferreira CF, Pereira MG, Santos AS, Rodrigues R, Bressan- Smith R.E, PioViana A, Daher RF (2003) Resistance to common bacterial blight in Phaseolus vulgaris L. recombinant inbred lines under natural infection of Xanthomonas axonopodis pv. phaseoli, Euphytica 134: 43-46.

9. Fourie D (1998) Characterization of halo blight races on dry bean in South Africa. Plant Disease 82(3): 307-310.
10. Fourie D, Miklas P, Ariyaranthe H (2004) Genes conditioning halo blight resistance to races 1,7 and 9 Occur in A Tight Cluster. The XL VII Report of The Bean Improvement Cooperation March 2004: 103-104.
11. Goto M (1992) Fundamentals of bacterial plant pathology. Academic Pres, California, Inc., pp. 635.
12. Güven K, Jones JB, Momol MT, Dickstein ER (2004) Phenotypic and genetic diversity among Pseudomonas syringae pv. phaseolicola. Journal of Phytopathology 152: 638-666.
13. Hall R (1994) Compendium of bean diseases. APS Pres, St, Paul, Minessota, pp. 73.
14. Howard RJ, Garland JA, Seaman WL (1994) Diseases and pests of vegatable crops in Canada. The Canadian Phytopathological Society, Canada, pp. 554.
15. King EO, Ward MK, Raney DE (1954) Two simple media for the demonstration of pyocianin and fluoresin. Journal of Laboratory and Clinical Medicine 44: 301-307.
16. Klement Z, Rudolph K, Sands DC (1990) Methods in phytobacteriology, Akademia Kiado, Budapest, pp. 568.
17. Lelliott RA, Stead DE (1987) Methods for the diagnosis of bacterial diseases plants. Blackwell Scientific Publications, Oxford, UK.
18. Nywall RF (1989) Field crop diseases handbook 2nd edn. Van Nostrand Reinhold, New York, pp. 817.
19. Popovic T, Balaz J, Nikolic Z, Starovic M, Gavrilovic V, Aleksic G, Zivkovic S (2010) Detection and identification of Xanthomonas axonopodis pv. phaseoli on bean seed collected in Serbia. African Journal of Agricultural Research 5(19): 2730-2736.
20. Saettler AW (1994) Bacterial brown spot, halo blight and common bacterial blight in compendium of bean diseases (Hall, R., Edt). APS Pres, The American Phytopathological Society, pp. 73.
21. Sambrook J, Fritschi EF, Maniatis T (1989) Molecular cloning: A laboratory manual, cold spring harbor laboratory press, New York.
22. Saygılı H, Şahin F, Aysan Y (2008) Bitki bakteri hastalıkları. Meta Basım, İzmir, s. 317.
23. Schaad NW, Jones JB, Chun W (2001) Laboratory guide for identification of plant pathogenic bacteria.
24. Schwartz HF (1989) Bean production problems in the tropics, The International Center for Tropical Agriculture., Columbia, pp. 285-301.
25. Sigee DC (1993) Bacterial plant pathology cell and molecular aspect. Cambridge University. Pres, pp. 325.
26. Taylor JD, Teverson DM, Davis JHC (1996) Sources of resistance to Pseudomonas savastanoi pv. phaseolicola races in Phaseolus vulgaris. Plant Pathology 45: 479-485.
27. TÜİK (2019) Tarımsal Yapı. T. C. Başkanlık Devlet İstatistik Enstitüsü, biruni.tuik.gov.tr. 10 Temmuz 2020
28. Wortmann CS, Allen DJ (1994) African bean production environments their definition, characteristics and constraints.Network on bean research in Africa, Occasional paper series No.11.
29. Zaiter HZ, Coyne DP, Vidaver AK, Steadman JR (1989) Differantial reaction of tepary bean lines to Xanthomonas campestris pv. phaseoli. American Society for Horticultural Science 24(1): 134-137.