شناسایی مخمر Cryptococcus magnus به عنوان عامل شانکر درختان میوه هسته دار در برخی از استان های مرکزی ایران

نوع مقاله: بیماری‌شناسی گیاهی

نویسندگان

1 دانش آموخته کارشناسی ارشد، گروه گیاهپزشکی، دانشگاه علوم کشاورزی و منابع طبیعی ساری

2 استاد، گروه گیاهپزشکی، دانشگاه علوم کشاورزی و منابع طبیعی ساری

3 دانشیار، گروه گیاهپزشکی، دانشگاه علوم کشاورزی و منابع طبیعی ساری

چکیده

شانکر تنه و شاخه یکی از مهمترین بیماری‌های درختان میوه هسته‌دار است که بارزترین علائم آن ایجاد زخم‌های فرورفته روی شاخه‌ها همراه با تراوش صمغ می‌باشد. دو باکتری Xanthomonas arboricola pv. pruni و Pseudomonas syringae pv. syringae به عنوان عوامل اصلی این بیماری قبلا از مناطقی از ایران گزارش شده‌اند. به منظور شناسایی و تعیین پراکنش عوامل مولد شانکر از نواحی مرکزی ایران، نمونه‌های دارای علایم شانکر از باغ‌های این مناطق جمع آوری گردید. پس از کشت نمونه‌ها روی محیط آگار غذایی حاوی سوکرز (NAS) کلنی‌های برجسته، کرم متمایل به صورتی و خمیری مانند جدا شد. بیماری‌زایی نماینده‌هایی از جدایه‌ها روی نهال‌های هلو (Prunus persica) به اثبات رسید. بررسی‌های میکروسکوپی، فنوتیپی و مولکولی برای شناسایی جدایه‌ها انجام شد. با استفاده از آغازگرهای ITS4 و ITS5 این قطعه در واکنش‌های زنجیره‌ای پلی‌مراز (PCR) در جدایه‌های نماینده تکثیر و توالی یابی شد. توالی‌های به دست آمده با استفاده از نرم‌افزار بلاست در ژن بانک (NCBI) با توالی‌های موجود در این پایگاه مقایسه و میزان شباهت تعیین گردید. از ژن 26S و بخشی از ژن‌های RPB1 و RPB2 نیز برای تأیید شناسایی استفاده شد. در بررسی میکروسکوپی جدایه‌ها سلول‌هایی کروی شکل که برخی از آن‌ها در حال جوانه زدن بودند مشاهده شد. در مقایسه توالی قطعه ITS در ژن بانک، جدایه‌‌ها بیشترین شباهت را به گونه‌ Cryptococcus magnus نشان دادند. توالی ژن‌های 26S، RPB1 و RPB2 نیز این شباهت‌ را تایید کردند.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Cryptococcus magnus as the causative agent of stem and branch canker of stone fruit trees in the some central provinces of Iran

نویسندگان [English]

  • M. DEHGHAN-NIRI 1
  • H. RAHIMIAN 2
  • V. BABAEIZAD 3
چکیده [English]

canker is one of the most damaging diseases of stone fruit trees which incites depressed brown to black lesions, often accompanied by exudation of gum, on twigs, branches and trunk of affected trees. The most common causal agents of bacterial canker in the world as well as in Iran are Pseudomonas syringae pv. syringae and Xanthomonas arboricola pv. pruni. To determine the predominant causative microorganism in some major fruit growing areas of the central provinces of Iran, samples were taken from the canker-affected stone fruit trees in these regions and macerates of the affected bark tissues were plated on sucrose nutrient agar (NAS). The predominant colonies appearing on NAS were whitish and yeast-like in appearance. Pathogenicity of representative isolates on peach (Prunus persica) seedlings were confirmed. Microscopic, phenotypic and genotypic characteristics of pure cultures of the isolates were determined. DNAs of representative isolates were extracted and the rDNA ITS regions were amplified and sequenced. The sequences were compared with those deposited in GenBank. Partial sequences of the 26S rRNA, RPB1 and RPB2 genes were determined and compared with those existing in GenBank. Budding globose cells were predominant in cell suspensions studied under the microscope. The isolates showed the highest identity with Cryptococcus magnus based on the sequences of the ITS regions. The identity of the isolates as C. magnus was further verified by comparison of the nucleotide sequences of their 26S, RPB1 and RPB2 genes with the homologous regions in Cryptococcus species.

کلیدواژه‌ها [English]

  • canker
  • homologous
  • ITS
  • sequence
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