با همکاری انجمن علمی گیاهان دارویی ایران

نوع مقاله : مقاله علمی پژوهشی

نویسنده

استادیار، موسسه تحقیقات ثبت و گواهی بذر و نهال، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

چکیده

هدف از این مطالعه شناسایی گونه‌های Fusarium همراه با ریشه و بنه زعفران و ارزیابی فعالیت آنزیم‌های مخرب دیواره سلولی تولید شده توسط آن­ها می باشد. به منظور بررسی گونه‌های Fusarium، طی فصل رشدی 1397 در مجموع 53 نمونه ریشه و بنه به طور تصادفی از مزارع زعفران استان خراسان جنوبی جمع­آوری شد. پس از شستشو نمونه­ها با آب مقطر استریل و ضدعفونی سطحی با هیپوکلریت سدیم یک درصد، قطعات کوچک ریشه و بنه روی محیط­های کشت‌ عمومی و اختصاصی کشت داده شدند. تهیه کشت خالص قارچ­ها به روش تک اسپور و نوک ریسه کردن روی محیط کشت‌ آب - آگار دو درصد انجام شد. شناسایی قارچ­ها بر اساس مشخصات میکروسکوپی و ریخت­شناختی با توجه به کلیدهای معتبر شناسایی انجام گرفت. با توجه به خصوصیات ریخت­شناختی، گونه های F. oxysporum، F. culmorum،F. proliferatum و F. graminearum شناسایی شدند. نتایج به دست آمده از تجزیه و تحلیل مولکولی با استفاده از آغازگرهای اختصاصی گونه، شناسایی ریخت­شناختی را تأیید کرد. نتایج نشان داد که 6/22 درصد از نمونه­های زعفران توسط گونه­های Fusarium آلوده بودند. بر اساس مشاهدات ریخت­شناختی و مولکولی، F. oxysporumبه­عنوان گونه‌ غالب با بیشترین درصد فراوانی در نمونه­ها به میزان 4/9 درصد پس از آن F. culmorum (7/5 درصد)،F. proliferatum (8/3 درصد) و F. graminearum (8/3 درصد) بود. نتایج نشان داد که تمام جدایه‌های Fusarium قادر به تولید آنزیم‌های مخرب دیواره سلولی و به طور عمده سلولاز و زایلاناز می‌باشند. سطوح فعالیت آنزیم­های سلولز، زایلاناز و پکتیناز در این مطالعه بالاتر از فعالیت لیپاز در همه جدایه­های مورد بررسی بود. تجزیه و تحلیل روند فعالیت آنزیم‌های مخرب دیواره سلولی نشان داد که برای بسیاری از جدایه‌های مورد بررسی حداکثر سطح فعالیت سلولاز، زایلاناز، پکتیناز و لیپاز به ترتیب در 72، 96، 144 و 192 ساعت پس از کشت در محیط مایع مشاهده شد. سپس فعالیت­های آنزیمی به تدریج با گذشت زمان کاهش یافت، تا اینکه در سطوح ثابتی باقی ماند. بر اساس منابع موجود F. graminearum، F. culmorum و F. proliferatum برای اولین بار در ایران از روی زعفران گزارش می شوند.

کلیدواژه‌ها

عنوان مقاله [English]

Evaluation of cell wall degrading enzymes of Fusarium species associated with root and corm of saffron in South Khorasan province

نویسنده [English]

  • Nima Khaledi

Assistant Professor, Seed and Plant Certification and Registration Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

چکیده [English]

The aim of this study is to identify  Fusarium species associated with root and corm of saffron and evaluate extracellular enzyme activity secreted by them. In order to study the species of Fusarium, a total of 53 root and corm samples were randomly collected from saffron fields of the South Khorasan province during 2017 growing season. After washing the samples with sterile water and disinfected the surface with 1% sodium hypochlorite, small pieces of root and corm were cultured on general and specific culture media. Purification of fungal colonies was performed using single spore and hyphal tip methods on 2% water agar medium. Identification of fungi based on the microscopic and morphological characteristics was performed with valid identification keys. The F. oxysporum, F. graminearum, F. culmorum and F. proliferatum species were identified based on morphological characters. The results obtained from molecular analysis using species-specific primers confirm morphological identification. The results reveal that 22.6% of saffron samples are contaminated by Fusarium species. F. oxysporum is the predominant species with the highest isolation frequency in samples of 9.4% followed by F. culmorum (5.7%), F. proliferatum (3.8%) and F. graminearum (3.8%) based on morphological and molecular observations,

کلیدواژه‌ها [English]

  • Cellulase
  • Identification
  • Molecular
  • Morphological
  • Xylanase
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