بررسی خصوصیات فنوتیپی و تنوع ژنتیکی جدایه هایPseudomonas syringae pv. syringae عامل شانکر باکتریایی بادام در استان خراسان رضوی با استفاده از BOX-PCR

محورهای موضوعی : دو فصلنامه تحقیقات بیماریهای گیاهی

حمیده احسنی ¹ (دانش آموخته گروه بیماری شناسی گیاهی دانشگاه علوم کشاورزی و منابع طبیعی گرگان)
سعید نصرالله نژاد ² (دانشیار گروه بیماری شناسی گیاهی دانشگاه علوم کشاورزی و منابع طبیعی گرگان)
حشمت ا... رحیمیان ³ (استاد دانشگاه علوم کشاورزی و منابع طبیعی ساری)
اسفندیار ظهور ⁴ (مربی پژوهش مرکز تحقیقات علوم کشاورزی و منابع طبیعی خراسان رضوی)
میثم تقی نسب ⁵ (مربی دانشگاه علوم کشاورزی و منابع طبیعی گرگان)

تاریخ دریافت : 1393/12/25 تاریخ پذیرش : 1393/12/25 تاریخ انتشار : 1394/03/01

کلید واژه: Pseudomonas syringae pv. syringae, شانکر باکتریایی, بادام, تنوع ژنتیکی, BOX-PCR,

چکیده مقاله :

شانکر باکتریایی که بوسیله پاتوار­های Pseudomonas syringae Van Hallایجاد می­شود یکی از مشکلات جدی باغ­های درختان میوه هسته­دار می­باشد. در این تحقیق طی سال­های 1391-1390 نمونه‌های دارای علائم شانکر از باغات بادام در استان خراسان رضوی جمع آوری شده و جداسازی باکتری انجام شد. عامل بیماری براساس آزمون­های گروه LOPAT و GATTa، (Pss)Pseudomonas syringae pv. Syringae  شناسایی شد. شانزده جدایه خالص ­سازی شده و مورد بررسی­های فیزیولوژیکی و بیوشیمیایی قرار گرفتند. جدایه­ها براین اساس اختلافات جزئی نشان دادند. تشخیص مولکولی جدایه ها با واکنش زنجیره ای پلی­مراز با آغازگرهای اختصاصی D21 و D22 صورت گرفت. به منظور ارزیابی دامنه تنوع ژنتیکی جدایه­ها، از روش BOX-PCR استفاده شد. از ضریب تشابه جاکارد (Jaccard) برای تعیین تشابه جدایه­ها و از روش UPGMA و نرم افزار NTYSYS-pc برای آنالیز خوشه­ای استفاده شد. در آنالیز خوشه­ای داده­ها، جدایه­ها با آغازگر BOXA1R در سطح تشابه 68 درصد، در 3 گروه قرار گرفتند. نتایج، بیانگر تنوع ژنتیکی جدایه­های عامل بیماری شانکر درختان بادام در استان خراسان رضوی است.

چکیده انگلیسی:

Bacterial canker caused by pathovars of Pseudomonas syringae Van Hall is one of the serious problems in stone fruit productions. In this research samples with canker symptoms were collected from Khorasane Razavi province during 2011–2012 and bacterial isolation was performed. The bacterial strains were identified as Pseudomonas syringae pv. syringae (Pss) on basis of LOPAT and GATTa tests and were confirmed as the causal agent of the almond bacterial canker based on the pathogenicity tests. A total of 16 isolates were characterized based on physiological and biochemical tests. The isolates showed a few differences in phenotypic characteristics. Genomic DNA from strains was used in PCR with D21 and D22 primers. To assess genetic diversity among the strains, rep-PCR analysis was performed using BOXA1R primer. Similarity matrix of the strains was calculated using the Jaccard's coefficient and cluster analysis was performed by UPGMA method using NTSYS-pc. The strains were divided into 3 clusters with BOXA1R at 68% similarity level. The results demonstrated considerable genetic diversity among strains causing canker in almond trees.
 

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