بررسی کارایی باکتری‏ های اندوفیت جدا شده از خیار در مهار زیستی بیماری پوسیدگی فوزاریومی طوقه و ریشه خیار

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد، گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه شهید مدنی آذربایجان، تبریز، ایران

2 دانشیار، گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه تبریز، ایران

3 دانشیار، گروه گیاه‌پزشکی، دانشکده کشاوری، دانشگاه شهید مدنی آذربایجان، تبریز، ایران

چکیده

پوسیدگی طوقه و ریشه خیار ناشی از قارچ Fusarium oxysporum f. sp . radicis cucumerinum یکی از بیماری‏ های آوندی مهم و جدی در کشت‎ های گلخانه‎ ای ایران محسوب می‎ شود. از استراتژی‏ های جدید مدیریت این بیماری، مهار زیستی آن توسط باکتری‏ های اندوفیت می‎ باشد. پژوهش حاضر به منظور بررسی کارایی برخی باکتری‏ های اندوفیت خیار در کنترل بیماری پوسیدگی فوزاریومی طوقه و ریشه خیار در شرایط آزمایشگاهی و گلخانه انجام گرفت. در این تحقیق، 140 جدایه باکتری اندوفیت از ریشه، ساقه، برگ، گل، میوه و بذر گیاهان سالم چند رقم رایج خیار جدا شد. جهت تعیین کارایی باکتری ‏های اندوفیت در مهار زیستی از روش‏ه ای کشت متقابل و نیز آزمون متابولیت ‎های قابل نفوذ در آگار استفاده شد. چهارده جدایه از باکتری‏ ها که خاصیت آنتاگونیستی قویتری نشان دادند، برای مطالعات بیشتر انتخاب شدند. پس از بررسی مکانیسم‏ های موثر بر خاصیت آنتاگونیستی نظیر توان تولید آنتی‏بیوتیک‎‏‎ ها، مواد فرار، سیدروفور، اکسین، هیدروژن سیانید و آنزیم پروتئاز، هشت جدایه از این باکتری‏ ها که پتانسیل بیشتری برای فعالیت مهار زیستی نشان دادند برای آزمایش در شرایط گلخانه‏ انتخاب شدند. بر اساس نتایج گلخانه‏ ای ومقایسه میانگین داده‏ های حاصل، دو جدایه برتر انتخاب شدند.این دو باکتری بر اساس توالی ژن 16S rRNA و آزمون های بیوشیمیایی با احتمال 98% به عنوان گونه‎ ها‏ی Lysinibacillus mangiferihumi و Lysinibacillus fusiformis شناسایی شدند.

کلیدواژه‌ها

عنوان مقاله [English]

Investigating of efficacy of endophytic bacteria isolated from cucumber in biocontrol of Fusarium crown and root rot disease of cucumber

نویسندگان [English]

  • H. Ghanadi 1
  • R. Khakvar 2
  • A. Shirzad 3
1 Graduated M.Sc. student of Entomology, Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran
2 Associate Professor, Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
3 Associate Professor, Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran
چکیده [English]

Background and objectives
Cucumber crown and root rot, caused by Fusarium oxysporum f. sp. radicis cucumerinum, is one of its major destructive agents in the glasshouse planting in Iran. This disease is also known as the Fusarium damping-off. Several methods, such as chemical control and agronomic methods have been recommended so far to control this disease, but none of them has established effective control over this disease. One of the newly established strategies for the management of this disease is the biological control using endophytic bacteria, which live inside plants and can enhance plant growth by improving nutrient uptake and producing phytohormones. These bacteria can compete and control the population of other bacteria and plant pathogens within plants. This study evaluated the biocontrol capability of some selected cucumber endophytic bacteria over Fusarium crown and root disease.
Materials and methods
Endophytic bacteria were isolated from roots, stems, leaves, flowers, fruits, and seeds of different healthy cucumber cultivars including, Negin, Danje 98, Danje 195, Nagin and, Native Basmang. Bacterial strains were isolated from plant tissues using different culture media. In-vitro biological control assays, such as cross-culture and agar permeable metabolite assays, were performed to determine the biocontrol efficiency of endophytic bacteria against F. oxysporum f.sp. radicis cucumerinum. The bacterial strains with better results were chosen for glasshouse assay. Two isolates with significant biocontrol efficiency were selected for molecular indentification. The 16srDNA barcoding was performed using universal primers for bacterial identification. Then, the sequencing results were analyzed using MEGA-X software, and phylogenic trees were drawn using the neighbor-joining method. The selected biochemical tests confirmed the molecular results.
Results and discussion
A total of 140 endophytic bacterial isolates were isolated and purified from roots, stems, leaves, flowers, fruits, and seeds of some common healthy cucumber cultivars. Based on initial cross-culture, 14 isolates out of 140 isolates showed significant antagonism properties in the cross-culture and agar permeable metabolite assays, which were selected for further studies. After evaluation of the effective mechanisms, their antagonistic properties, such as the production capability of antibiotics, volatile compounds, siderophore, auxins, hydrogen cyanide, and protease enzyme, eight isolates were selected as the most potential bacterial isolates for biocontrol activity in the greenhouse. Finally, two bacterial isolates were selected for molecular identification using 16srDNA barcoding. The sequencing results indicated that the two selected isolates with 98% similarity could belong to Lysinibacillus mangiferihumi and Lyisinibaillus fusiformis species. The biochemical results confirmed the molecular results.

کلیدواژه‌ها [English]

  • Biocontrol
  • Fusarium oxysporum f. sp. radicis cucumerinum
  • Antagonism
  • Glasshouse

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گیاه پزشکی
دوره 44، شماره 1
اردیبهشت 1400
صفحه 95-111

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