مطالعه‌ی اثر بیوکنترلی سویه‌های سودوموناس فلورسنت مولد 2 و 4- دی استیل فلوروگلوسینول بر نماتد ریشه گرهی در ریزوسفر ارقام گوجه‌فرنگی

نوع مقاله: مقاله علمی-پژوهشی

نویسندگان

1 دانشکده کشاورزی و منابع طبیعی، دانشگاه خلیج فارس، بوشهر

2 دانشگاه خلیج فارس

3 گروه گیاهپزشکی، دانشکده کشاورزی و منابع طبیعی دانشگاه شاهد تهران

4 دانشگاه شاهد

چکیده

نماتد مولد ریشه‌ گرهی Meloidogyne javanica یکی از عوامل مهم بیمارگر گوجه‌فرنگی در کشور به‌شمار می­رود. در این تحقیق کنترل بیولوژیک این نماتد توسط سویه­های UTPF68 و  UTPF101باکتری Pseudomonas fluorescens، مورد ارزیابی قرار گرفت. ابتدا سویه­ها از نظر دارا بودن ژن phlD از طریق واکنش زنجیره­ای پلیمراز (PCR) مورد بررسی قرار گرفتند. سپس توانایی تولید متابولیت‏های ضد میکروبی توسط این باکتری­ها و تأثیر آن‌ها بر تفریخ تخم و فعالیت لارو سن دوم نماتد در شرایط آزمایشگاه و اثر بیوکنترلی آن‌ها بر فعالیت نماتد در ریزوسفر چهار رقم گوجه‌فرنگی در شرایط گلخانه براساس طرح بلوک­های کامل تصادفی با چهار تکرار مورد بررسی قرار گرفت. نتایج واکنش زنجیره­ای پلیمراز نشان داد که هر دو سویه واجد ژن phlD بوده و قطعه DNA با طول 629 جفت باز در آن‌ها تکثیر شد. سویه­های باکتریایی قادر به تولید آنتی بیوتیک DAPG، سیانید هیدروژن و پروتئاز در شرایط آزمایشگاهی بودند. سویه UTPF68 نسبت به سویه UTPF101 توانایی بیشتری در عدم تفریخ تخم نماتد داشت، درحالی‌که میزان مرگ و میر لارو سن دوم در هر دو سویه یکسان بوده است. نتایج گلخانه‌ای نشان داد که سویه­های باکتری موجب افزایش شاخص­های رشدی ارقام و کاهش فعالیت نماتد در ریزوسفر آن‌ها شدند. میزان افزایش رشد یا کنترل نماتد بسته به رقم گوجه‌فرنگی و سویه باکتری متفاوت بود.

 

کلیدواژه‌ها


Ahmadzadeh, M. & Ghasemi, S. 2012. Introduction ofPseudomonas fluorescens as a New Biocontrol Agent in Iran. Biological Control of Pests and Plant Diseases, 1: 49-60.

Bakker, P.A. H.M., Pieterse, C.M.J. & van Loon, L.C. 2007. Induced systemic resistance by fluorescent Pseudomonas spp. Phytopathology, 97:239–243.

Cronin, D., Moënne-Loccoz, Y., Fenton, A., Dunne, C., Dowling, D.N. & O’Gara, F. 1997. Role of 2,4-diacetylphloroglucinol in the interaction of the biocontrol pseudomonad strain F113 with the potato cyst nematode Globodera rostochiensis. Applied and Environmental Microbiology, 63: 1357 –1361.

Delaney, S. M., Mavrodi, D. V., Bonsall, R. F. & Thomashow, L.S. 2001. phzO – a gene for biosynthesis of 2-hydroxylated phenazine compounds in Pseudomonas aureofaciens 30-84. Journal of Bacteriology, 183: 318–327.

Duffy, B.K. & De ´fago, G. 1999. Environmental factors modulating antibiotic and siderophore biosynthesis by Pseudomonas fluorescens biocontrol strains. Applied and Environmental Microbiology, 65: 2429– 2438.

Dwivedi, D., & Johri 2003. Antifungals from fluorescent pseudomonads: biosynthesis and regulation. Current Science. 85: 1693–1703.

Gallagher, L.A. & Manoil, C.B.N. 2001. Pseudomonas aeruginosa PAO1 kills Caenorhabditis elegans by cyanide poisoning. Journal of Bacteriology, 183: 6207-6214.

Ghafele Bashi, S., Jamali, F., & Ahmadzadeh, M. 2014. Study of biological characteristics and evaluation of biocontrol bacteria Pseudomonas fluorescens UTPF68 against Phytophthora drechsleri on cucumber. Biological Control of Pests and Plant Diseases, 3(2): 105-116.

Hussay, R.S. & Barker, K.R. 1973. A compression of methods of collecting inoculates of Meloidogyne spp. Including a new technique. Plant Disease, 57: 1025-1028.

Jamali, F. 2009. Influence of some biotic factors on the expression of hydrogen cyanide- and 2,4-diacetylphloroglucinol biosynthesis genes in Pseudomonas fluorescens on bean rhizosphere, Ph. D. thesis in Plant pathology, College of Agriculture, Tehran University (In Persian).

Jepson, S.B. 1987. Identification of Root-knot Nematodes (Meloidogyne species). C.A.B. Interactional, 256pp.

Iavicoli, A., Boutet, E., Buchala, A. & Me ´traux, J.P. 2003. Induced systemic resistance in Arabidopsis thaliana in response to root inoculation with Pseudomonas fluorescens CHA0. Molecular Plant Microbe Interactions, 16: 851–858.

Keel, C., Schnider, U., Maurhofer, M., Voisard, C., Burger, D., Haas, D & Défago, G. 1992. Suppression of root diseases by Pseudomonas fluorescens CHA0: importance of the bacterial secondary metabolite 2,4-diacetylphloroglucinol. Molecular Plant-Microbe Interactions, 5: 4–13.

Keel, C. & Defago, G. 1997. Interaction between beneficial soil bacteria and root pathogens, mechanisms and ecological impact. In: Gange, A.C., and Brown, V.K (Eds) Multitrophic in teraction system. Oxford: Blackwell science, 27-47 pp.

Khan, A,. Shaukat, S.S., Islam, S. & Khan, A. 2012. Evaluation of Fluorescent Pseudomonad isolates for their activity against some plant-parasitic nematodesamerican-eurasian. J. Agric. and Environ. Sci, 12 (11): 1496-1506.

Maurhofer, M., Keel, C., Schnider, U., Voisard, C., Haas, D. & De ´fago, G. 1992. Influence of enhanced antibiotic production in Pseudomonas fluorescens strain CHA0 on its disease suppressive capacity. Phytopathology, 82: 190–195.

Maurhofer, M., Keel, C., Haas, D. & Defago, G. 1994. Influence of plant species on disease suppression by Pseudomonas fluorescens strain CHA0 with enhanced production. Plant pathology, 44: 40-50.

Maurhofer, M., Keel, C., Haas, D., & Défago, G. 1995. Influence of plant species on disease suppression by Pseudomonas fluorescens strain CHA0 with enhanced antibiotic production. Plant Pathology, 44: 40-50.

Mazzola, M., Fujimoto, K., Thomashow, L.S., & Cook, R.J. 1995. Variation in sensitivity of Gaeumannomyces graminis to antibiotics produced by fluorescent Pseudomonasspp. and effect on biological control of take-all of wheat. Applied and Environmental Microbiology, 61: 2554–2559.

Meyer, S.L.F., Halbrendt, J.M., Carta, L.K., Skantar, A.M., Liu, T., Abdelnabby, H.M.E., & Vinyard, B.T. 2009. Toxicity of 2, 4-diacetylphloroglucinol (DAPG) to Plant-parasitic and Bacterial-feeding Nematodes. Journal of Nematology, 41(4): 274–280.

Mc Spadden Gardener, B., Mavrodi, D.V., Thomashow, L.S., & Weller, D.M. 2001. A rapid polymerase chain reaction-based assay characterizing rhizosphere populations of 2, 4-diacetylphloroglucinol-producing bacteria. Phytopathology, 91: 44-54.

McSpadden Gardener, B., Benitez, M.S., Camp, A. & Zumpetta, C. 2006a. Evaluation of a seed treatment containing a phlD+ strain of Pseudomonas fluorescens on organic soybeans. Biological and Cultural Tests for Control of Plant Diseases Report, 21: FC046.

McSpadden Gardener, B., Kroon van Diest, C. & Beuerlein, J. 2006b. Evaluation of biological seed treatments containing phlD+ strains of Pseudomonas fluorescens on soybeans grown in Ohio. Biological and Cultural Tests for Control of Plant Diseases Report, 21: FC045.

Nitao, J.K., Meyer, S.L.F. & Chitwood, D.J. 1999. In-vitro assays of M. incognita and Heterodera glicines for detection of nematode-antagonistic fungal compounds. Journal of Nematology, 31:772-783.

Raaijmakers, J.M., Weller, D.M., Thomashow, L.S. 1997. Frequency of antibiotic-producing Pseudomonas spp. in natural environments. Applied Environmental Microbiology, 63: 881-887.

Raaijmakers, J.M., Vlami, M. & de Souza, J.T. 2002. Antibiotic production by bacterial biocontrol agents. Antonie van Leeuwenhoek, 81: 537–547.

Siddiqui, I.A. & Shaukat, S.S. 2003a. Plant species, host age and host genotype effects on Meloidogyne incognita biocontrol by Pseudomonas fluorescens strain CHA0 and its genetically-modified derivatives. Journal of Phytopathology, 151: 231–238.

Siddiqui, I.A. & Shaukat, S.S. 2003b. Suppression of root-knot disease by Pseudomonas fluorescens CHA0 in tomato: Importance of bacterial secondary metabolic 2, 4-diacetylphloroglucinol. Soil Biology and Biochemistry: 1615-1623.

Siddiqui, I.A. & Shaukat, S.S. 2004a. Suppression of Meloidogyne incognita by Pseudomonas fluorescens strain CHA0 and its genetically modified derivatives: II. The influence of sodium chloride. Nematologia Mediterranea, 32: 127–130.

Siddiqui, I.A. & Shaukat, S.S. 2004b. Systemic resistance in tomato induced by biocontrol bacteria against the root-knot nematode, Meloidogyne javanica is independent of salicylic acid production. Journal of Phytopathology, 152: 48–54.

Siddiqui, I.A. & Shaukat, S.S. 2004c. Trichoderma harzianum enhances the production of nematicidal compounds in vitro and improves biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato. Letters in Applied Microbiology, 38: 169-175.

Siddiqui, I.A., Hass, D. & Heeb, S. 2005a. Extracellular protease of Pseudomonas fluorescens CHA0, a biocontrol factor with activity against root-knot nematode, Meloidogyne incognita. Applied and Environmental Microbiology, 71: 5646-5649.

Siddiqui, I.A., Shaukat, S.S., & Dutt, S. 2005 b. Expolysaccharide over producing variant of Pseudomonas fluorescens strain CHA0 enhances tolerance to various environmental stresses in vitro but does not improve Meloidogyne javanica biocontrol in tomato. International Journal of Biology and Biotechnology, 2: 729-736.

Siddigui, I.A., Shaukat, S.S., Sheikh, I.H. & Khan, A. 2006. Role of cyanide production by Pseudomonas fluorescens CHA0 in the suppression of root-knot nematode, Meloidogyne javanica in tomato. World J. Microbiol. Biotech., 22: 641-650.

Smith, K.P., Handelsman, J. & Goodman, R.M. 1997. Modeling dose-response relationships in biological control: partitioning host response to the pathogen and biocontrol agent. Phytopathology, 87: 720-729.

Tian, B.O., Yang, J. & Zhang, K.Q. 2007. Bacteria used in the biological control of plant-parasitic nematodes: populations, mechanisms of action and future prospects. FEMS Microbiology & Ecology, 61: 197-213.

Timper, P., Kone, D., Yin, J., Ji, P. & McSpandden Gardener, B.B. 2009. Evaluation of an Antibiotic-Producing Strain of Pseudomonas fluorescens for suppression of Plant-Parasitic Nematodes. Journal of Nematology, 41(3): 234-240.

Thompson, D.C. 1996. Evaluation of bacterial antagonists for reduction of summer path symptoms in Kentucky blue grass. Plant Disease, 80: 850-862.

Thomashow, L.S. & Weller, D.M. 1996. Current concepts in the use of introduced bacteria for biological disease control mechanisms and antifungal metabolites. In: Plant-microbe Interactions, 1: 187-235. Stacey, G. and Keen, N.J. (eds.) New York, NY, Chapman & Hall.

van Loon, L.C. & Bakker, P.A.H.M. 2005. Induced systemic resistance as a mechanism of disease suppression by rhizobacteria. In PGPR: Biocontrol and Biofertilization (Siddiqui, Z.A., ed.), Dordrecht, The Netherlands: Springer, pp. 39 –66.

Voisard, C., Keel, C., Haas, D. & Défago, G. 1989. Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions. EMBO Journal, 8: 351-358.

Wang, C., Ramette, A., Pungasamarnwong, P., Zala, M., Natsch, A., Moënne-Loccoz, Y. & Défago, G. 2001. Cosmopolitan distribution of phlD-containing dicotyledonous crop-associated biocontrol pseudomonads of worldwide origin. FEMS Microbiology Ecology, 37: 105-116.

Weller, D.M., van Pelt, J.A., Mavrodi, D.V., Pieterse, C.M.J., Bakker, P.A.H.M. & van Loon, L.C. 2004. Induced systemic resistance (ISR) in Arabidopsisagainst Pseudomonas syringae pv. tomatoby 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas fluorescens. Phytopathology, 94: S108.