‌بررسی توان برخی جدایه‌های باکتریایی به منظور مهار زیستی بیماری لکه زاویه‌ای باکتریایی خیار

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری گروه گیاهپزشکی دانشگاه ولی عصر (ع) رفسنجان-

2 عضو هیات علمی گروه گیاهپزشکی دانشگاه ولی عصر رفسنجان

چکیده

باکتری‌های محرک رشد گیاهان با داشتن توان بالا در جهت افزایش زیست‌توده و کنترل بیماری‌های گیاهی، می‌توانند جانشین مناسبی برای سموم و کودهای شیمیایی در کشاورزی باشند. به‌منظور بررسی مهار زیستی Pseudomonas syringae pv. lachrymans عامل بیماری لکه زاویه‌ای خیار در شرایط آزمایشگاه و گلخانه‌، از باکتری‌های جداسازی شده از خاک ریزوسفر خیار به همراه تعدادی از نمونه‌های مجموعه باکتری‌های گروه گیاه‌پزشکی دانشگاه ولی‌عصر رفسنجان استفاده شد. بعد از غربالگری آزمایشگاهی، چهار جدایه که دارای توان ایجاد هاله بازدارنده بودند برای بررسی‌های بعدی در شرایط آزمایشگاه و گلخانه انتخاب شدند. خصوصیات مهار زیستی جدایه‌ها مانند تولید متابولیت‌های ثانویه و بعضی از ویژگی‌های تحریک‌کنندگی رشد در شرایط آزمایشگاهی مورد ارزیابی قرار گرفت. در ارزیابی‌های گلخانه‌ای، توان جدایه‌های باکتریایی در کنترل بیماری از طریق ممانعت در ایجاد علایم با تعیین سطح زیر منحنی پیشرفت بیماری و افزایش زیست‌توده گیاه خیار با بررسی شاخص‌های رشدی مطالعه شد. نتایج بررسی گلخانه‌ای نشان داد که جدایه Pseudomonas fluorescens VRPF 54 کارآیی مناسبی در مهار زیستی بیماری داشت. در شرایط گلخانه، این جدایه به تنهایی و تواًم با بیمارگر به طور معنی‌داری باعث افزایش زیست‌توده و کنترل بیماری لکه زاویه‌ای خیار گردید. پس از انتخاب این جدایه، برخی از ویژگی‌های آنتاگونیستی، خصوصیات بیوشیمیایی و شناسایی مولکولی آن در آزمایشگاه مورد بررسی قرار گرفت. همچنین نتایج حاصل از اندازه‌گیری آنزیم‌های پراکسیداز، گایاکول‌پراکسیداز، فنیل‌آلانین‌آمونیالیاز، پلی‌فنل‌اکسیداز و محتوی فنل کل روند افزایشی را نشان داد.

کلیدواژه‌ها


Ahemad M, Khan MS (2012) Evaluation of plant growth promoting activities of rhizobacterium Pseudomonas putida under herbicide stress. Annals of Microbiology 62(2): 1531-1540.
Al-aghabary K, Zhu Z, Shi Q (2005) Influence of silicon supply on chlorophyll content, chlorophyll fluorescence, and antioxidative enzyme activities in tomato plants under salt stress. Journal of Plant Nutrition 27: 2101-2115.
Alexander DB, Zuberer DA (1991) Use of chrome azurol S reagents to evaluate siderophore production by rhizosphere bacteria. Biology and Fertility of Soils 2: 39-45.
Aliye N, Fininsa C, Hiskias Y (2008) Evaluation of rhizosphere bacterial antagonists for their potential to
bioprotect potato (Solanum tuberosum) against bacterial wilt (Ralstonia solanacearum). Biological Control 47: 282-288.
Alstrom S, Burns RG (1989) Cyanid production by rhizobacteria as a possible mechanism of plant growth inhibition. Biology and Fertility of Soils 7: 232-238.
Anonymous (2020) Statistics on agriculture (2019 Crop year). Ministry of Jihad-e-Agriculture, Information and Communication Technology Center 3: 59-60. (In Persian).
Bent E, Tuzan S, Chanway CP, Enebak S (2000) Alteration in plant growth and in root hormone levels of lodgeple pines inoculatied with rhizobacteria. Canadian Journal of Microbiology 47: 793-800.
Blokhina O, Virolainen E, Fagerstedt KV (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Annals of Botany 91(2): 179-194.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantites of protein utilizing the principles of protein dyebinding. Analytical Biochemistry 72: 248-254.
Cappuccino JG, Sherman N (1992) Microbiology: A Laboratory Manual. Third ed. Benjamin/Cummings Publishing Companies based in New York. pp. 125-179.
Card SD, Walter M, Jaspers MV, Sztejnberg A, Stewart A (2009) Targeted selection of antagonistic microorganisms for control of Botrytis cinerea of strawberry in New Zealand. Australasian Plant Pathology 38(2): 183-192.
Chen H, Jegadeesh N, Wermers R (2000) The value of active mutual fund management: an examination of the stockholdings and trades of managers. Journal of Financial and Quantitative Analysis 35: 343–368.
Compant S, Duffy B, Nowak J, Clément C, Barka EA (2005) Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Applied and Environmental Microbiology 71(9): 4951-4959.
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: 537-546.
Duncan LW, Ferris H (1983) Validation of a model for prediction of host damage by two nematode species. Journal of Nematology 15: 227-234.
Etebarian HR (2008) Vegetable diseases and methods of control. Tehran University Press, Iran. (In Persian).
Goszczynska T, Serfontein J, Serfontein S (2000) Introduction to practical phytobacteriology; a manual for phytobac-teriology. Safrinet, the Southern African (SADC) Loop of BIONET-International, ARC-Plant Protection Institute, Pretoria, South Africa.
Guo JH, Qi HY, GuoYH, Ge HL, Gong LY, Zhang LX, Sun PH (2004) Biocontrol of tomato wilt by plant growth promoting rhizobacteria. Biological Control 29(1): 66-72.
Harighi B (2007) Angular leaf spot of cucumber caused by Pseudomonas syringae pv. lachrymans in Kurdistan.Plant Disease 91(6): 789.
Hynes RK, Leung GC, Hirkala DL, Nelson LM (2008) Isolation, selection, and characterization of beneficial rhizobacteria from pea, lentil, and chickpea grown in western Canada. Canadian Journal of Microbiology 54(4): 248–258.
Kasana RC, Salwan R, Dhar H, Dutt S, Gulati A (2008) A rapid and easy method for the detection of microbial cellulases on agar plates using Gram’s iodine. Current Microbiology 57: 503-507.
Kritzman G, Zutra D (1983) Systemic movement of Pseudomonas syringae pv. lachrymans in the stem, leaves, fruits, and seeds of cucumber. Canadian Journal of Plant Pathology 5: 273-278.
Lebeda A, Widrlechner M, Staub J, Ezura H, Zalapa J, Kristkova H (2007) Cucurbits (Cucurbitaceae; Cucumis spp., Cucurbita spp., Citrullus spp.). Genetic Resources, Chromosome Engineering and Crop Improvement. Vegetable Crops 3: 271-376.
Maurhofer M, Keel C, Haas D, Defago G (1995) Influence of plant species on disease suppression by Pseudomonas fluorescens strain CHA0 with enhanced production. Plant Pathology 44: 40–50.
Mota MS, Gomes CB, Souza Junior IT, Moura AB (2017) Bacterial selection for biological control of plant disease: criterion determination and validation. Brazilian Journal of Microbiology 48: 62-70.
Nakkeeran S, Kavitha K, Renukadevi P, Chandrasekar G, Fernando WGD (2005) Induction of plant defence compounds by Pseudomonas chlororaphis PA23 and Bacillus subtilis BSCBE4 in controlling damping-off of hot pepper Pythium aphanidermatum . Biocontrol Science and Technology 16(4): 403-416.
Narmadhavathy S, Vanitha S, Karthikeyan G, Raguchander T, Ramjegathesh R (2013) Induced systemic resistance and their implications in host resistance to physic nut against leaf blight disease. Molecular Microbiology Research 3(3): 20-29.
Newberry EA, Jardini TM, Rubio I, Roberts PD, Babu B, Koike ST, Bouzar H, Goss EM, Jones JB, Bull CT, Paret ML (2016) Angular leaf spot of cucurbits is associated with genetically diverse Pseudomonas syringae strains. Plant Disease 100(7):1397-1404.
Nicoli MC, Elizable BE, Piotti A, Lerici CR (1991) Effect of sugar and maillard reaction products on polyphenol oxidase and peroxidase activity in food. Journal of Food Biochemistry 15:169-184.
O’Brien JA, Daudi A, Butt VS, Bolwell GP (2012) Reactive oxygen species and their role in plant defence and cell wall metabolism. Planta 236: 765-779.
Olanrewaju OS, Glick BR, Babalola OO (2017) Mechanisms of action of plant growth promoting bacteria. World Journal of Microbiology and Biotechnology 33: 1-16.
Olczak‐Woltman H, Bartoszewski G, Mądry W, Niemirowicz‐Szczytt K (2009) Inheritance of resistance to angular leaf spot (Pseudomonas syringae pv. lachrymans) in cucumber and identification of molecular markers linked to resistance. Plant Pathology 58: 145-151.
Parizi FM, Taghavi SM (2015) Identification of the causal agent of angular leaf spot disease of cucumber and studying its phenotype characteristics and genetic diversity in Fars and Kerman provinces. Iranian Journal of Plant Pathology 51(1): 68-55 (In Persian).
Patten CL, Glick BR (2002) Role of Pseudomonas putida Indoleacetic acid in development of the host plant root system. Applied and Environmental Microbiology 68 (8): 3795-3801.
Pikovskaya RI (1948) Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Microbiology 17: 362-370.
Polle A, Otter T, Seifert F (1994) Apoplastic peroxidases and lignification in needles of Norway Spruce (Picea abies L.). Plant Physiology 106(1): 53-60.
Pugliese M, Liu BP, Gullino ML, Garibaldi A (2008) Selection of antagonists from compost to control soil-borne pathogens. Journal of Plant Diseases and Protection 115(5): 220-228.
Rafiei S, Khodakaramian G (2015) Isolation and identification of cucumber rhizospheric fluorescent pseudomonads and evaluation of their antagonistic potential as biocontrol agents. Biocontrol in Plant Protection 3(2): 59-75 (In Persian).
Rangarajan S, Saleena LM, Vasudevan P, Nair S (2003) Biological suppression of rice disease by Pseudomonas spp. under saline conditions. Plant and Soil 251:73–82.
Renwick A, Campbell R, Coe S (1991) Assessment of in vivo screening systems for potential biocontrol agents of Gaeumannomyces graminis. Plant Pathology 40: 524-532.
Reuveni R (1995) Biochemical marker for disease resistance, In: Singh, RP and Singh US (ed), Molecular Methods in Plant Pathology. CRC Press, Boca Raton, FL. pp. 99-144.
Roland SF, Laima SK (1999) Phenolics and cold tolerance of Brassica napus. Plant Agriculture 1: 1-5.
Shaad NW, Jones JB, Chum w (2001) Labratory guid for identification of plant pathogenic bacteria. Thrid eds. Amer. Phytopathology Society, St, Paul Minnesota, USA.
Sahu B, Singh J, Shankar G, Pradhan A (2018) Pseudomonas fluorescens PGPR bacteria as well as biocontrol agent: A review. International Journal of Chemical Studies 6(2): 01-07.
Sirko A, Brodzik R (2000) Plant ureases: roles and regulation. Acta Biochimica Polonica 47(4): 1189-1195.
Van Loon LC (1997) Induced resistance in plants and the role of pathogenesis-related proteins. Eurpean Journal of Plant Pathology 103: 753- 765.
Vessey JK (2003) Plant growth promoting rhizobacteria as biofertilizers. Plant and Soil 255: 571-586.
Weller DM, Cook RJ (1983) Suppression of take-all of wheat by seed treatments with fluorescent pseudomonads. Phytopathology 73: 463-469.
Wichmann F, Muller BH, Widmer F, Boller B, Studer B, Kolliker R (2011) Phenotypic and molecular genetic characterization indicate no major race‐specific interactions between Xanthomonas translucens pv. graminis and Lolium multiflorum. Plant Pathology 60: 314-324.
Yedidia I, Shoresh M, Kerem Z, Benhamou N, Kapulnik Y, Chet I (2003) Concomitant induction of systemic resistance to Pseudomonas syringae pv. lachrymans in cucumber by Trichoderma asperellum (T-203) and accumulation of phytoalexins. Applied and Environmental Microbiology 69(12): 7343-7353.
Zheng HZ, Cui CI, Zhang YT, Wang D, Jing Y, Kim KY (2005) Active changes of lignifications-related enzymes in pepper response to Glomus intraradices and/or Phytophthora capsici. Journal of Zhejiang University Science 6(8): 778-786.