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بررسی پروفایل متابولیکی گیاه توت فرنگی در تعامل با باکتری اندوفیت Bacillus spp. سویه DM12 با استفاده از دستگاه کروماتوگرافی گازی– طیف سنجی جرمی و تعیین ویژگی های ضدقارچی سویه باکتری | ||
| مهار زیستی در گیاهپزشکی | ||
| دوره 11، شماره 2، اسفند 1402، صفحه 93-110 اصل مقاله (1.14 M) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/bcpp.2024.367321.376 | ||
| نویسندگان | ||
| زهرا علی جانی1؛ جهانشیر امینی* 2؛ علی اکبر مظفری3 | ||
| 1پژوهشگر پسا دکتری گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه کردستان، سنندج، ایران. | ||
| 2استاد، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه کردستان، سنندج، ایران. | ||
| 3استاد، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه کردستان، سنندج، ایران. | ||
| چکیده | ||
| قارچ Colletotrichum nymphaeae عامل بیماری آنتراکنوز توت فرنگی یکی از مهمترین بیماریهای این گیاه است که سبب آلودگی ریشه، برگ و میوه گیاه توت فرنگی در شرایط مختلف رشد میگردد. مشکل اصلی ارقام توت فرنگی در مقابل این بیماری پوسیدگی میوه است که کمیت و کیفیت و بازار پسندی میوه توت فرنگی را کاهش میدهد. در این تحقیق سویه باکتریایی اندوفیت از گیاهچههای سالم توتفرنگی (Fragaria × ananassa) جداسازی و اثرات آنتاگونیستی این سویه علیه قارچ عامل آنتراکنوز توتفرنگی Colletotrichum nymphaeae در شرایط آزمایشگاه، درون شیشه و گلخانه مورد بررسی قرار گرفت. سویه جداسازی شده با استفاده از ویژگیهای ریخت شناسی، بیوشیمیایی و آنالیز مولکولی ژن 16S rDNA در جنس Bacillus spp. قرار گرفت. سویه DM12 در آزمون کشت متقابل باعث کاهش رشد قارچ بیمارگر (%64/03) گردید. در آزمون تاثیر ترکیبات ضدقارچی تولید شده توسط باکتری اندوفیت میزان بازدارندگی از رشد میسلیوم و جوانه زنی اسپور قارچ بیمارگر به ترتیب %32/86 و %73/98 ارزیابی گردید. تاثیر متابولیت های فرار باکتریایی بر رشد میسلیوم قارچ بیمارگر ناچیز بود (%9/82). با توجه به آزمایشات بیوشیمیایی این سویه قادر به تولید آنزیم های پروتئاز، کیتیناز و پکتیناز و همچنین تولید سیدوفور، هورمون ایندول استیک اسید و جیبرلین بوده و توانایی حلالیت فسفات را دارد. سویه باکتری اندوفیت در شرایط درون شیشه قادر به بازدارندگی بیماری بر روی میوه توت فرنگی به میزان %90/87 گردید. همچنین درشرایط گلخانه در روش خیساندن خاک به میزان %72/22 و در روش اسپری اندام های هوایی به میزان %94/44 قادر به جلوگیری از گسترش بیماری بر روی گیاهچه های توت فرنگی شد. همچنین در این تحقیق تغییرات متابولیت های گیاهی در تیمارهای مختلف با باکتری اندوفیت در مقایسه با شاهد بررسی گردید. نتایج نشان داد که در تیمار شاهد منفی میزان ترکیبات استوگلیسرید (%19/418)، استیک اسید بوتیل استر (%4/734) و ریبیتول (%4/349)، در تیمار باکتری اندوفیت به تنهایی میزان ترکیبات تترامتیل دو–هگزامتان (%21/35)، اتیلن گلیکول منوایزوبوتیل اتر (%18/688) و میرتنول (%8/75)، در تیمار قارچ بیمارگر به تنهایی ترکیبات استوگلیسرید (%18/089)، استیک اسید منو گلیسرید (%17/96) و در تیمار قارچ بیمارگر همراه با باکتری اندوفیت ترکیبات ترت–بوتانتیول (%36/153)، اتوکسی تری اتیل سیلان (%14/126)، 5–متیل آمینو–1–2–3–4–تیاتریازول (%9/53) و 2–3–بوتاندیول (%7/795) بیشترین مقدار را داشتند. برخی از این ترکیبات مانند بوتاندیول، لاکتوز و تترادکان که در تیمار های باکتری اندوفیت به تنهایی و قارچ بیمارگر همراه با باکتری اندوفیت تولید شدند دارای خواص ضد قارچی می باشند. نتایج حاصل از این تحقیق نشان داد که سویه DM12 قادر به کاهش رشد میسلیوم، جوانه زنی اسپور، توسعه پوسیدگی میوه و شدت بیماری آنتراکنوز توت فرنگی در شرایط آزمایشگاه، درون شیشه و گلخانه می باشد. به منظور درک تعامل سویه DM12 با گیاه و سایر عوامل بیماریزای گیاهی و اثرات آن بر سیستم دفاعی گیاه، تحقیقات بیشتری مورد نیاز است. | ||
| کلیدواژهها | ||
| آنتراکنوز توت فرنگی؛ متابولیت های ضد قارچی؛ Bacillus spp؛ GC-Mass | ||
| عنوان مقاله [English] | ||
| Analysis of strawberry metabolic profiling in interaction with Bacillus spp. strain DM12 using Gas Chromatography-Mass Spectrometry and Determination of its Antifungal Activity | ||
| نویسندگان [English] | ||
| Zahra Alijani1؛ Jahanshir Amini2؛ Ali Akbar Mozafari3 | ||
| 1Post- Doctora research, Department of Plant Protection, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran | ||
| 2Professor, Department of Plant Protection, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran | ||
| 3Professor, Department of Horticultural Sciences, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran. | ||
| چکیده [English] | ||
| Colletotrichum nymphaeae infects the roots, leaves and fruits of the plant at different developmental stages and is one of the most important diseases of strawberry. The principal problems to strawberry cultivation are fruit rot caused by anthracnose that affect the quality and quantity of production and marketing in the field, and post–harvest. An endophytic bacterial strain, isolated from Fragaria × ananassa, and antifungal properties against Colletotrichum nymphaeae was assayed under in vitro, in vivo, and greenhouse experiments. Bacterial strain was identified as Bacillus spp. DM12 (MH161581) using phenotypic, biochemical and molecular phylogenetic analysis of the 16S rDNA gene. DM12 strain inhibited mycelial growth of fungal pathogen (64.03%) using dual–culture test. The cell–free culture compounds produced by DM12 prevented mycelial growth and conidial germination of C. nymphaeae by 32.86% and 73.98%, respectively but, inhibition percentage of mycelial growth of pathogen by volatile compounds was less (9.82%). As well as, protease, chitinase, pectinase, siderophore, IAA, gibberellin, and phosphate solubilization tests for this strain were positive. Anthracnose disease at post–harvest on fruit suppressed by the strain DM12 (90.87%). Also, biocontrol efficacy on strawberry plants by drenching soil and spraying methods were 72.22% and 94.44%, respectively, 60 days after inoculation. PCR amplification represented the presence of genes of surfactin. In addition, metabolite profile of strawberry was changed on presence of bacterial strain that a number of metabolites in control treatment with maximum area percent were Acetoglyceride (19.418%), Acetic acid, butyl ester (4.734%) and Ribitol (4.349%), in treatment inoculated with DM12 strain alone were Tetramethyl–2–hexadecen (21.35%), Ethylene glycol monoisobutyl ether (18.688%) and Myrtenol (8.75%), in treatment inoculated with fungal pathogen alone were Acetoglyceride (18.089%) and Acetic acid, monoglyceride (17.96%) and in treatment inoculated with C. nymphaeae and DM12 strain together were tert–Butanethiol (36.153%), Ethoxy triethyl silane (14.126%), 5–(Methylamino)–1,2,3,4–thiatriazole (9.53%) and 2,3–Butanediol (7.795%). Some of these compounds, such as Butanediol, Lactose and Tetradecane, which were produced in DM12 strain alone and C. nymphaeae and DM12 strain together treatments, have antifungal properties. These results showed that strain DM12 has able to decrease mycelial growth, conidial germination, fruit decay development and disease severity of strawberry anthracnose under in vitro, in vivo and greenhouse conditions. Further works are required in order to understanding the interaction of DM12 with the plant and others phytopathogens and its effects on the plant defense system. . | ||
| کلیدواژهها [English] | ||
| Antifungal metabolites, Bacillus spp, GC-Mass, Strawberry anthracnose | ||
| مراجع | ||
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