Ahmet HATIPOĞLU, Ahmet USLU, Galip KAŞKAVALCI, Hatice ÖZAKTAN

Preliminary investigations on suppression of meloidogyne incognita (Kofoid & White) chitwood (Nematoda: Heteroderidae) by antagonistic rhizobacteria

Floresan Pseudomonad ırkları nematod ile bulaşık domates köklerinde bulunan hastalıklı Meloidogyne incognita (Kofoid & White, 1919) Chitwood, 1949 (Nematoda: Heteroderidae) 2. dönem larvalarından elde edilmiştir. In vivo’da 5 farklı ırk izole edilmiş, Kökur nematodlarına antagonist etkilerini test etmek amacıyla bu ırklar içinden patojen olmadığı tespit edilen Pseudomonas fluorescens Migula, 1895 (Proteobacteria: Pseudomonadaceae) Strain Pat1 seçilmiştir. Denemelerde hıyar (Cucumis sativus) cv Sardes test bitkisi olarak kullanılmıştır. Bakteri süspansiyonu (109 cfu/ml) üç farklı şekilde uygulanmıştır: 1) Tohum bakterizasyon; 2) Tohum bakterizasyon ve fide içirme ve 3) Fide içirme. Hıyar bitkilerinin fidelerini kök-ur nematodları ile bulaştırmak için, kök-ur nematodlarının yumurtaları bulaşık domates bitkilerinin köklerinden blender-elek yöntemiyle elde edilmiştir. Şaşırtma esnasında hıyar fideleri 20 000 yumurta/bitki ile bulaştırılmıştır. Bitkiler, kontrollü koşullara sahip olan klimatize edilmiş odalarda belirti gelişimi için 10–11 hafta gözlenmiştir. Belirti gelişimi, bitkilerin toprak üstü kısımları için 0–4 skalası, kökleri içinde 0–10 Zeck Kök-ur skalası kullanılarak değerlendirilmiştir. Tohum kaplama ile tohum kaplama ve fide daldırma, sadece kök-ur nematodları ile bulaşık bitkilerin bulunduğu pozitif kontrol ile karşılaştırıldığında, bitkilerin toprak üstü kısımlarındaki hastalık gelişiminin sırasıyla % 35 ve % 21, köklerdeki ur oluşumunun % 44 ve % 39 oranlarında düştüğü belirlenmiştir. Deneme, köklerdeki antagonist bakterilerin kolonizasyonu ve populasyon dinamiğini gözlemek amacıyla iki kez yinelenmiştir.

Meloidogyne incognita (Kofoid&White) chitwood (Nematoda: Heteroderidae)’nın antagonist kök bakterileri ile mücadelesine yönelik ön çalışmalar

Fluorescent Pseudomonad strains have been obtained from diseased second juveniles from Meloidogyne incognita (Kofoid & White, 1919) Chitwood, 1949 (Nematoda: Heteroderidae) infected tomato roots. After the screening in vivo made by 5 different isolated strains, Strain Pat1 has been selected to test the antagonistic effects on the root-knot nematodes. Strain Pat1 has been identified as non plant pathogenic Pseudomonas fluorescens Migula, 1895 (Proteobacteria: Pseudomonadaceae) according to the test result. Cucumber (Cucumis sativus) cv Sardes has been used as test plant. Bacterial suspension (109 cfu/ml) was applied by three different ways: 1) Seed bacterization; 2) Seed bacterization plus seedling drenching; and 3) Seedling drenching. The eggs of rootknot nematodes have been obtained from diseased tomato roots by blender-sieve method for infection cucumber seedlings with root-knot nematodes. 20 000 eggs/plant have been given to cucumber seedlings during transplanting. The plants have been observed for symptom development for 10-11 weeks in climatized room under controlled conditions. Symptom development has been evaluated by using 0-4 scale for upper parts of plants, and 0-10 Zeck’s root-knot scale for the roots. When seed bacterization and seed bacterization plus seedling drenching applications compared with positive control; plants infected with root-knot nematodes alone, it was determined that disease symptoms on upper parts of the plants were reduced 35 % and 21 %; gal formation on the roots were reduced 44 % and 39 %,respectively. The experiment has been repeated twice to observe the colonization and population dynamic of antagonistic bacteria on the roots.

PDF

___

Ahl, P., C Voisard & G. Defago, 1986. Iron bound siderophores, cyanic acid and antibiotics involved in suppression of Thielaviopsis basicola by Pseudomonas fluorescens strain. Journal of Phytopathology, 116: 121-134.
Bakker, P. A. H. M., R. Van Peer & B. Schippers, 1991. Suppression of soil-borne plant pathogens by fluorescent pseudomonads: mechanisms and prospects. In: Beemster,A. B. R., G. J. Bollen, M. Gerlagh, M. A. Ruissen & B. Schippers (eds), Biotic Interactions and Soil-borne Diseases, pp. 217—230, Elsevier, Amsterdam.
De Meyer, G. & M. Hofte, 1997. Salicylic acid produced by the rhizo-bacterium Pseudomonas aeruginosa 7NSK2 induces resistance to leaf infection by Botrytis cinerea on bean. Phytopathology, 87: 588-593.
Elekçioğlu, İ. H., B. Ohnesorge, G. Lung & N. Uygun, 1994. Plant parasitic nematodes in the Mediterranean Region of Turkey. Nematol. Medit., 22: 59-63.
Fahy, P. C. & G. J. Persley, 1983. Plant Bacteria Diseases, A Diagnostic Guide. Academic Pres, New York, 393 pp.
Garima, G., A. Singh & P. C. Trivedi, 2005. Bacteria: A Potential bioagent against Root-knot nematode, Meloidogyne incognita. National Symposium on Recent Advances and Research Priorities in Indian Nematology, 9-10th December 2005, IARI, New Delhi,14 pp.
Hamid, M., I. A. Siddiqui & S. S. Shaukat, 2003. Improvement of Pseudomonas fluorescens CHA0 biocontrol activity against root-knot nematode by the addition of ammonium molybdate. Lett. Appl. Microbiol., 36: 239-244.
Hasky-Günther, K., S. Hoffmann-Hergarten & R. A. Sikora, 1998. Resistance against the potato cyst nematode Globodera pallida systemically induced by the rhizobacteria Agrobacterium radiobacter (G12) and Bacillus sphaericus (B43). Fund.Appl. Nematol., 21: 511-517.
Johnsson, L., M. Hökeberg & B. Gerhardson, 1998. Performance of the Pseudomonas chlororaphis biocontrol agent MA 342 against seed-borne diseases in field experiments. European Journal of Plant Pathology, 104: 701-711.
Jothi, G & M. Sivakumar, 2003. Induced systemic resistance by Pseudomonas fluorescens against root-knot nematode in tomato. 6th International PGPR Workshop, 5-10 October 2003, Calicut, India, 480pp.
Kaşkavalcı, G. & C. Öncüer, 1999. Aydın İli’nin yazlık sebze yetiştirilen önemli bölgelerinde bulunan Meloidogyne Goeldi, 1887 (Tylenchida: Meloidogynidae) türlerinin yayılışları ve ekonomik önemleri üzerinde araştırmalar. Türk. entomol. derg.,23(2): 149-160. (In Turkish with English abstract).
King, E. O., M. K. Ward & D. E. Raney, 1954. Two simple media for the demonstration of pyocyanin and fluorescin. J. Lab. Clin.Med., 44: 301-307.
Klement, Z., K. Rudolph & D. C. Sands, 1990. Methods in phytopathology. Akademiai Kiado, 153-180, Budapest.
Lelliot, R. A. & D. E., Stead, 1987. Methods for The Diagnosis of Bacterial Diseases of Plants. Black Well Scientific Puplication, 157 pp, Oxford, UK.
Leong, J., 1986. Siderophores: their biochemistry and possible role in the biocontrol of plant pathogens. Annual Review of Phytopathology, 24: 187-209.
Mercado-Blanco, J., D. Rodrigez-Jurado, A. Hervas & R. M. Jimenez-Diaz, 2004. Suppression of Verticillium wilt in olive plantings stocks by root-associated fluorescent Pseudomonas spp. Biological Control, 30: 474-486.
Oostendorp, M. & R. A. Sikora, 1990. In-vitro interrelationships between rhizosphere bacteria and Heterodera schachtii. Review de Nematologie, 13:269-274.
Racke, J. & R. A. Sikora, 1992. Isolation, formulation and antagonistic activity of rhizobacteria toward the potato cyst nematode Globodera pallida. Soil Biology and Biochemistry, 24: 521-526.
Ramakrishnan, S., C. V. Sivakumar & K. Poornima, 1998. Management of rice root nematode, Hirschmanniella gracilis (de Man) with Pseudomonas fluorescens. J. Biol. Control., 9:135-141.
Reitz, M., K. Rudolph, I. Schroder, S. Hoffmann-Hergarten, J. Hallmann & R. A. Sikora,2000. Lipopolysaccharides of Rhizobium etli strain G12 act in potato roots as an inducing agent of systemic resistance to infection by the cyst nematode Globodera pallida. Appl. Environ. Microbiol., 66: 3515-3518.
Schroth, M. N., J. G. Hancock, 1981. Selected topics in biological control. Annual Review of Microbiology, 35: 453-476.
Shanthi, A. & C. V. Sivakumar, 1995. Biocontrol potential of Pseudomonas fluorescens(Migula) against root-knot nematode, Meloidogyne incognita infecting tomato. J. Biol.Control., 9: 113-115.
Siddiqui, I.A. & S. Ehteshamul-Haque, 2000. Use of Pseudomonas aeruginosa for the control of root rot-root knot disease complex in tomato. Nematologia Mediterranea, 28: 189-192.
Siddiqui, I. A. & S. S. Shaukat, 2002 a. Resistance against the damping-off fungus Rhizoctonia solani systemically induced by the plant growth promoting rhizobacteria Pseudomonas aeruginosa (IE-6S+) and P. fluorescens (CHA0). J. Phytopathol., 150: 500-506.
Siddiqui, I. A. & S. S. Shaukat, 2002 b. Rhizobacteria-mediated induction of systemic resistance (ISR) in tomato against Meloidogyne javanica. J. Phytopathol., 150:469-473.
Siddiqui, I. A., S. A. Qureshi, V. Sultana, S. Ehteshamul-Haque & A. Ghaffar, 2000.Biological control of root rot-root knot disease complex of tomato. Plant Soil, 227:163-169.
Stockwell, V. O., K. B. Johnson & J. E. Loper, 1998. Establishment of bacterial antagonists of E. amylovora on pear and apple blossoms as influenced by inoculum preparation. Phytopathology, 88: 506-513.
Verma, K. K., 2005. Management of Meloidogyne javanica by bacterial antagonist, Pseudomonas fluorescens as seedling root dip in tomato. National Symposium on Recent Advances and Research Priorities in Indian Nematology, 9-10th December 2005, IARI, New Delhi, 22 pp.
Wei, G., J. W. Kloepper & S. Tuzun, 1996. Induced systemic resistance to cucumber diseases and increased plant growth by plant growth-promoting rhizobacteria under field conditions. Phytopathology, 86: 221-24.
Weller, D. M., 1988. Biocontrol of soilborne plant pathogens in the rhizosphere with bacteria. Annual Review of Phytopathology, 26: 379-407.
Yüksel, H., 1974. Kök-ur nematodlarının (Meloidogyne spp.) Türkiye’deki durumu ve bunların populasyon problemleri üzerinde düşünceler. Atatürk Üni. Zir. Fak. Zir.Derg., 5 (1): 83-105 (Ayrı Baskı).
Zeck, W. M., 1971. A rating scheme for field evaluation of Root-knot nematode infestation.Pflanzenschutz Nachrichten Bayer, 10: 141–144.