ردیابی ژن‌های hcnAB و phlD در سودومونادهای فلورسنت با توان مهار زیستیFusarium graminearum و بررسی توانایی آن‌ها در کلنیزاسیون اکتوریزوسفر گندم

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری، دانشکدة علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

2 دانشیار گروه گیاه‌پزشکی، دانشکدة علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

3 استاد گروه گیاه‌‌پزشکی، دانشکدة علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

چکیده

توانایی کنترل بیولوژیکی سودومونادهای فلورسنت از رقابت بر سر فضا و مواد غذایی، آنتی‌بیوز و کلنیزاسیون ریزوسفر ناشی می‌شود. در این مطالعه، توانایی بیوکنترلی 39 سویةPseudomonas fluorescens  در برابر قارچ Fusarium graminearum  عامل بلایت خوشة گندم در شرایط آزمایشگاهی بررسی شد. بازدارندگی از رشد F. graminearum با استفاده از آزمون کشت متقابل، اثر ضد قارچی ترکیبات فرّار و غیرفرّار تولید‌شده توسط سویه‌ها ارزیابی شد. در تمام این آزمایش‌ها چهار سویة I P13، UTPf127، UTPf125 وUTPf105 در کنترل بیمارگر موفق‌تر بودند. در نتیجه، این سویه‌ها از نظر وجود ژن‌های hcnAB و phlD و توانایی کلنیزاسیون اکتوریزوسفر گندم در شرایط گلخانه‌ای بررسی شدند. ژن hcnABدر تمام سویه‌ها ردیابی شد؛ در حالی که، ژن phlD در تمام باکتری‌ها به جز P13 مشاهده شد. جمعیت باکتری‌ها در اکتوریزوسفر گندم در سه دورة چهارده‌ روزه توسط سری رقت روی محیط کشت KB+++ تعیین شدند. نتایج این تحقیق نشان داد که تفاوت معنی‌داری میان سویه‌های مختلف در کلنیزاسیون اکتوریزوسفر در روزهای مختلف وجود دارد و با گذشت زمان جمعیت باکتری‌ها در ریزوسفر کاهش می‌یابد. سویه‌های P13 وUTPf125  در روز 42‌ام قابل ردیابی نبودند، در حالی که، در روز 28‌ام جمعیت آن‌ها بیش از 104×8/1 سلول در گرم وزن تر ریشه ردیابی شده بود. باکتری UTPf127 با جمعیت 104 × 1/1 سلول در گرم وزن تر ریشه در روز 42‌ام بیشترین میزان کلنیزاسیون و پایداری در ریزوسفر را نشان داد.

کلیدواژه‌ها

موضوعات


Alström S, Burns RG (1989) Cyanide production by rhizobacteria as a possible mechanism of plant growth inhibition. Biology and Fertility of Soils 7(3): 232-238.
Amini J (2006) Identification of Fusarium species associated with root and crown of wheat in Kurdistan province. In 17th Iranian Plant Protection Congress. Karaj, Iran, 42.
Amini J, Ershad D, Torrabi M (1998) A survey on mycoflora of wheat root in Tehran province. In Plant Protection Congress. Karaj, Iran, 45.
Arjmandian A, Rohani H (1998) Fungi associated with root and crown of wheat in Hamedan province. In 13th Iranian Plant Protection Congress. Karaj, Iran, 44.
Babadoost M (1995). Incidence of seed-borne fungal diseases of barley in East Azerbaijan and Ardebil Provinces. Iranian Journal of Plant Pathology 31 (1/4). (in Persian)
Bais HP, Weir TL, Perry LG, Gilroy S, Vivanco JM (2006) The role of root exudates in rhizosphere interactions with plants and other organisms. Annu. Rev. Plant Biol. 57: 233-266.
Bakker AW, Schippers B (1987) Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas spp-mediated plant growth-stimulation. Soil Biology and Biochemistry 19(4): 451-457.
Bakker PA, Pieterse CM, Van Loon L (2007) Induced systemic resistance by fluorescent Pseudomonas spp. Phytopathology 97(2): 239-243.
Baligh M (1999) Evaluation of Burkholderia cepacia Strains: root colonization of catharanthus roseus and in-vitro inhibition of selected soil-borne fungal pathogens. Paper read at Proceedings of the Oklahoma Academy of Science.
Bennett R, Lynch J (1981) Bacterial growth and development in the rhizosphere of gnotobiotic cereal plants. Journal of General Microbiology 125(1): 95-102.
Bloemberg GV, Lugtenberg BJ (2003) Phenazines and their role in biocontrol by Pseudomonas bacteria. New Phytologist 157(3): 503-523.
Botelho GR, Mendonça-Hagler LC (2006) Fluorescent Pseudomonads associated with the rhizosphere of crops: an overview. Brazilian Journal of Microbiology 37(4): 401-416.
Brisbane PG, Janik LJ, Tate M, Warren R (1987) Revised structure for the phenazine antibiotic from Pseudomonas fluorescens 2-79 (NRRL B-15132). Antimicrobial agents and chemotherapy 31(12): 1967-1971.
Broukanlo madlo P (2011) The population of Fluorescent pseudomonads in wheat cultivars and their biocontrol activity against Fusarium culmorum the causal agent of wheat crown and root rot, University of Tehran, Tehran, Iran. (in Persian)
Buddrus-Schiemann K, Schmid M, Schreiner K, Welzl G, Hartmann A (2010) Root colonization by Pseudomonas sp. DSMZ 13134 and impact on the indigenous rhizosphere bacterial community of barley. Microbial Ecology 60(2): 381-393.
Castric KF, Castric PA (1983). Method for rapid detection of cyanogenic bacteria. Applied and Environmental Microbiology 45(2): 701-702.
Castric PA (1977) Glycine metabolism by Pseudomonas aeruginosa: hydrogen cyanide biosynthesis. Journal of Bacteriology 130(2): 826-831.
Chancey ST, Wood DW, Pierson EA, Pierson LS (2002) Survival of GacS/GacA mutants of the biological control bacterium Pseudomonas aureofaciens 30-84 in the wheat rhizosphere. Applied and Environmental Microbiology 68(7): 3308-3314.
Chin-A-Woeng TF, de Priester W, van der Bij AJ, Lugtenberg BJ (1997) Description of the colonization of a gnotobiotic tomato rhizosphere by Pseudomonas fluorescens biocontrol strain WCS365, using scanning electron microscopy. Molecular Plant-Microbe Interactions 10(1): 79-86.
Cook R (2010) Fusarium root, crown, and foot rots and associated seedling diseases. Compendium of wheat diseases and pests. 3rd edition. In: WW Bockus, R. Bowden, R. Hunger, W. Morrill, T. Murray, and R. Smiley (eds.). The Pennsylvania State University Press, University Park, USA:37-39.
Cronin D, Moenne-Loccoz Y, Fenton A, Dunne C, Dowling DN, O'gara F (1997) Role of 2, 4-diacetylphloroglucinol in the interactions of the biocontrol pseudomonad strain F113 with the potato cyst nematode Globodera rostochiensis. Applied and Environmental Microbiology 63(4): 1357-1361.
Cuong ND, Nicolaisen MH, Sørensen J, Olsson S (2011) Hyphae-colonizing Burkholderia sp.—a new source of biological control agents against sheath blight disease (Rhizoctonia solani AG1-IA) in rice. Microbial Ecology 62(2): 425-434.
Darvishnia M, Alizadeh A, Mohamadi Goltapeh E (1998) Fusarium sepcies and other fungi associated with crown and root rot of wheat in Lorestan province. In 13th Iranian Plant Protection Congress. Karaj, Iran, 20.
Darvishnia M, Alizadeh A, Zare R, Safaee N, Davari M (2006) Report of three new species from Fusarium graminearum clade for Iran. In 17th Iranian Plant Protection Congress. Karaj, Iran, 449.
De La Fuente L, Quagliotto L, Bajsa N, Fabiano E, Altier N, Arias A (2002). Inoculation with Pseudomonas fluorescens biocontrol strains does not affect the symbiosis between rhizobia and forage legumes. Soil Biology and Biochemistry 34(4): 545-548.
DeCoste NJ, Gadkar VJ, Filion M (2010) Verticillium dahliae alters Pseudomonas spp. populations and HCN gene expression in the rhizosphere of strawberry. Canadian journal of microbiology 56(11): 906-915.
Dekkers L, Phoelich C, Lugtenberg B (1999) Bacterial traits and genes involved in rhizosphere colonization. Atlanta Canada Society for Microbial Ecology, Halifax, Canada.
Delany I, Sheehan MM, Fenton A, Bardin S, Aarons S, O’Gara F (2000) Regulation of production of the antifungal metabolite 2, 4-diacetylphloroglucinol in Pseudomonas fluorescens F113: genetic analysis of phlF as a transcriptional repressor. Microbiology 146(2): 537-546.
Duineveld BM, Van Veen J (1999) The number of bacteria in the rhizosphere during plant development: relating colony-forming units to different reference units. Biology and fertility of soils 28(3): 285-291.
Ebrahimi Kazemabad Z, Rouhani H, Jamali F, Mahdikhani Moghadam F (2013) Identification of phID Gene with Fluorescent Pseudomonads from Rhizospheric Zone of Chickpea and its Relation with Biological Control of Chickpea Fusarium wilt Disease Caused by Fusarium oxysporum f. sp. ciceris. Journal of Plant Protection 27(4): 407-416. (in Persian)
Expert J, Digat B (1995) Biocontrol of Sclerotinia wilt of sunflower by Pseudomonas fluorescens and Pseudomonas putida strains. Canadian Journal of Microbiology 41(8): 685-691.
Fiddaman P, Rossall S (1993) The production of antifungal volatiles by Bacillus subtilis. Journal of Applied Bacteriology 74(2): 119-126.
Foroutan A, Bamdadian T, Valipour M, Kianoosh H (1995) Fungi associate with root and crown rot of wheat in Mazandaran province. In 12th Iranian Plant Protection Congress. Karaj, Iran, 46.
Golzar H, Ershad D (1994) Occurrence of pink snow mold in Gorgan area. Iranian Journal of Plant Pathology 30: 83. (in Persian)
Hajmalek  Zanjani M, Ahmadzadeh M, Sharifi Tehrani A, Behboudi K, RA SR (2011) Effects of Rhizoctonia solani on Root Colonization of Canola by Pseudomonas fluorescens Strain UTPF86. Iranian  Journal of Plant Protection Science 42(1): 163-170. (in Persian)
Henkes GJ, Jousset A, Bonkowski M, Thorpe MR, Scheu S, Lanoue A, Schurr U, Röse US (2011) Pseudomonas fluorescens CHA0 maintains carbon delivery to Fusarium graminearum-infected roots and prevents reduction in biomass of barley shoots through systemic interactions. Journal of Experimental Botany 62(12): 4337-4344.
Howell С, Stipanovic R (1980) Suppression of Pythium ultimium induced damping off of cotton seedlings by Pseudomonas fluorescens and its antibiotic pyoluterin. Phytopathology 70(8): 712-715.
Isnansetyo A, Cui L, Hiramatsu K, Kamei Y (2003) Antibacterial activity of 2, 4-diacetylphloroglucinol produced by Pseudomonas sp. AMSN isolated from a marine alga, against vancomycin-resistant Staphylococcus aureus. International Journal of Antimicrobial Agents 22(5): 545-547.
Keel C, Schnider U, Maurhofer M, Voisard C, Laville J, Burger U, Wirthner P, Haas D, Dfago G (1992) Suppression of root diseases by Pseudomonas fluorescens CHA0: importance of the bacterial secondary metabolite 2, 4-diacetylphloroglucinol. Mol Plant Microbe Interact 5: 413Lodewyckx.
Kluepfel DA (1993) The behavior and tracking of bacteria in the rhizosphere. Annual Review of Phytopathology 31(1): 441-472.
Kraus J, Loper J (1990) Biocontrol of Pythium damping-off of cucumber by Pseudomonas fluorescens pf-5: Mechanistic studies pp. 175-177. Paper read at Plant Growth Promoting Rhizobacteria. The second international workshop on plant growth-promoting rhizobacteria, Interlacen, Switzerlan.
Kraus J, Loper J (1992) Lack of evidence for a role of antifungal metabolite production by Pseudomonas fluorescens Pf-5 in biological control of Pythium damping-off of cucumber. Phytopathology 82(3): 264-271.
Laville J, Blumer C, Von Schroetter C, Gaia V, Défago G, Keel C, Haas D (1998) Characterization of the hcnABC gene cluster encoding hydrogen cyanide synthase and anaerobic regulation by ANR in the strictly aerobic biocontrol agent Pseudomonas fluorescens CHA0. Journal of Bacteriology 180(12): 3187-3196.
Lugtenberg BJ, Kravchenko LV, Simons M (1999) Tomato seed and root exudate sugars: composition, utilization by Pseudomonas biocontrol strains and role in rhizosphere colonization. Environmental Microbiology 1(5): 439-446.
Madloo PB, Behboudi K, Tohidfar M, Jouzani GS, Ahmadzadeh M (2013) Response of some important Iranian wheat cultivars to Fusarium culmorum under genetic diversity of indigenous bio-control agent fluorescent Pseudomonas spp. Australian Journal of Crop Science 7(7): 1003-1009.
Mansoori B (1995) Soil-borne diseases of wheat in Mazandaran province. In 12th Iranian Plant Protection Congress. Karaj, Iran, 58.
Maurhofer M, Keel C, Haas D, Défago G (1995) Influence of plant species on disease suppression by Pseudomonas fluorescens strain CHAO with enhanced antibiotic production. Plant Pathology 44(1): 40-50.
Maurhofer M, Keel C, Schnider U, Voisard C, Haas D, Défago G (1992) Influence of enhanced antibiotic production in Pseudomonas fluorescens strain CHA0 on its disease suppressive capacity. Phytopathology 82(2): 190-195.
Mazzola M, Cook RJ, Thomashow LS, Weller D, Pierson L (1992) Contribution of phenazine antibiotic biosynthesis to the ecological competence of fluorescent pseudomonads in soil habitats. Applied and Environmental Microbiology 58(8): 2616-2624.
Mazzola M, Funnell DL, Raaijmakers J (2004) Wheat cultivar-specific selection of 2, 4-diacetylphloroglucinol-producing fluorescent Pseudomonas species from resident soil populations. Microbial Ecology 48(3): 338-348.
Ministry of Agriculture Statistics (2015). In The first volume of agricultural statistics crop season 1391-92. Economic and Planning Department. Center for Information and Communication Technology, 154.
Moynihan JA, Morrissey JP, Coppoolse ER, Stiekema WJ, O'Gara F, Boyd EF (2009) Evolutionary history of the phl gene cluster in the plant-associated bacterium Pseudomonas fluorescens. Applied and Environmental Microbiology 75(7): 2122-2131.
Nourozian J, Etebarian HR, Khodakaramian G (2006) Biological control of Fusarium graminearum on wheat by antagonistic bacteria. Songklanakarin J Sci Technol 28(1): 29-38.
Ravanlou A, Banihashemi Z (1999) Taxonomy and pathogenicity of Fusarium spp. associated with root and crown rot of wheat in Fars Province. Iranian Journal of Plant Pathology 35: 37-45. (in Persian)
Reddy T, Khudiakov I, Borovkov A (1969) Pseudomonas fluorescens strain 26-o--producer of phytotoxic substances. Mikrobiologiia 38(5): 909.
Safaee D (2004) Fungi associated with root and Crown rot of wheat in Kermanshah province. In 16th Iranian Plant Protection Congress. Tabriz, Iran, 38.
Saremi H (2004) Crown rot and root rot diseases on wheat caused by Fusarium graminearum as new species in Zanjan, East Azarbyjan and Ardabil provinces. In 16th Iranian Plant Protection Congress Tabriz, Iran, 44.
Savchuk S, Fernando WGD, Parks PS (2001) Potential for biocontrol of Sclerotinia sclerotiorum on canola. Canadian journal of plant pathology 23(2): 205.
Schippers B, Bakker AW, Bakker PA (1987) Interactions of deleterious and beneficial rhizosphere microorganisms and the effect of cropping practices. Annual review of phytopathology 25(1): 339-358.
Shanahan P, O'Sullivan DJ, Simpson P, Glennon JD, O'Gara F (1992) Isolation of 2, 4-diacetylphloroglucinol from a fluorescent pseudomonad and investigation of physiological parameters influencing its production. Applied and Environmental Microbiology 58 (1):353-358.
Siddiqui IA, Shaukat SS (2003) Suppression of root-knot disease by Pseudomonas fluorescens CHA0 in tomato: importance of bacterial secondary metabolite, 2, 4-diacetylpholoroglucinol. Soil Biology and Biochemistry 35(12): 1615-1623.
Sobhanipoor A (2003) Biological control of Fusarium wilt disease by bacteria Pseudomonas fluorescens in cumin, University of Tehran, Tehran, Iran. (in Persian)
Stockwell C, Bergstrom G, Luz Wd (1999). Selection of microbial antagonists for biological control of Fusarium head blight of wheat. Paper read at Proceedings of the 1999 National Fusarium Head Blight Forum, Michigan State University, University Printing, East Lasting, MI.
Stockwell C, Luz Wd, Bergstrom G (1997) Biocontrol of wheat scab with microbial antagonists. Phytopathology 87: S94.
Thomashow LS, Weller DM (1996) Current concepts in the use of introduced bacteria for biological disease control: mechanisms and antifungal metabolites. In Plant-microbe interactions: Springer, 187-235.
Velusamy P, Immanuel JE, Gnanamanickam SS, Thomashow L (2006) Biological control of rice bacterial blight by plant-associated bacteria producing 2, 4-diacetylphloroglucinol. Canadian journal of microbiology 52(1): 56-65.
Vincent MN, Harrison L, Brackin J, Kovacevich P, Mukerji P, Weller D, Pierson E (1991) Genetic analysis of the antifungal activity of a soilborne Pseudomonas aureofaciens strain. Applied and Environmental Microbiology 57(10): 2928-2934.
Weller D (1983) Colonization of wheat roots by a fluorescent pseudomonad suppressive to take-all. Phytopathology 73(11): 1548-1553.
Weller DM (1994) Current challenges in introducing beneficial microorganisms into the rhizosphere. Molecular ecology of rhizosphere microorganisms 1-18.
Weller DM (2007) Pseudomonas biocontrol agents of soilborne pathogens: looking back over 30 years. Phytopathology 97(2): 250-256.
Weller DM, Raaijmakers JM, Gardener BBM, Thomashow LS (2002) Microbial populations responsible for specific soil suppressiveness to plant pathogens 1. Annual Review of Phytopathology 40(1): 309-348.
Yan Z, Reddy M, Kloepper JW (2003) Survival and colonization of rhizobacteria in a tomato transplant system. Canadian Journal of Microbiology 49(6): 383-389.