شناسایی، خصوصیات مورفوفیزیولوژیک و تفاوت های ژنتیکی گونه قارچی عامل بیماری سرخشکیدگی شاخه درختان پسته استان کرمان بر اساس توالی یابی rDNA-ITS
محورهای موضوعی : میکروب شناسی گیاهی
کلید واژه: بیماری سرخشکیدگی, واکنش زنجیره ای پلی مراز, ITS1, ITS4, درخت فیلوژنتیک.,
چکیده مقاله :
سابقه و هدف: بیماری سرخشکیدگی شاخه یکی از بیماری های مهم درختان پسته استان کرمان است که توسط قارچ های مختلفی ازجمله P. variotii بوجود می آید. هدف از انجام این تحقیق شناسایی قارچ عامل این بیماری در استان کرمان بر اساس خصوصیات مرفولوژیکی و توالی یابی ناحیه rDNA-ITS بود. امکان وجود تفاوت های ژنتیکی در این ناحیه و وجود ارتباطاتی بین تفاوت های ژنتیکی با تفاوت های مرفولوژیکی و سرعت رشد قارچ در شرایط آزمایشگاهی بررسی شد. مواد و روش ها: در طی سال زراعی 1400، نمونه برداری از شاخه های بیمار ارقام مختلف پسته باغات استان کرمان انجام گرفت. مراحل جداسازی، کشت، خالص سازی و شناسایی گونه قارچ عامل بیماری بر اساس خصوصیات مرفولوژیک انجام شد. تکثیر و توالی یابی ناحیه ITS با استفاده از آغازگرهای ITS1 و ITS4 انجام شد. سرعت رشد جدایه های قارچ روی محیط کشت ثبت و میانگین رشد آن ها محاسبه و مقایسه گردید. با بکارگیری نرم افزار مگا10، درخت فیلوژنتیکی ترسیم شد. یافته ها: بر اساس خصوصیات مورفولوژیکی و توالی یابی ITS، تمامی جدایه ها، گونه P. variotii شناسایی شدند. درخت فیلوژنتیکی، وجود تفاوت ژنتیکی در ناحیه ITS جدایه ها را نشان داد. ارتباط معنی داری بین گروه بندی فیلوژنتیکی جدایه ها با تفاوت های مورفولوژیک و میانگین سرعت رشد مشاهده نشد. نتیجه گیری: تفاوت هایی در توالی های ناحیه ITS در سطح درون گونه ای وجود دارد. توالی یابی ITS، پشتیبان ارزشمندی برای تأیید مشاهدات مورفولوژیکی در جهت شناسایی P. voraitii است. ارتباط مستقیمی بین گروه بندی فیلوژنتیکی با تفاوت های مورفولوژیکی و فیزیولوژیکی جدایه ها وجود نداشت.
Background & Objectives: Dieback disease is a major problem in pistachio trees in Kerman province. It is caused by various fungi, including P. variotii. This study aimed to identify this fungal species based on morphology and rDNA-ITS sequencing. The study also investigated genetic differences in this region and correlations between genetic variations, morphology, and fungal growth rate in lab conditions. Materials & Methods: During 2021, diseased branches from different pistachio cultivars in Kerman were sampled. Isolation, cultivation and purification of the isolates were carried out and identification of the fungal species were performed based on morphological characteristics. The ITS region was amplified and sequenced using specific primers ITS1 and ITS4. The growth rate of fungal isolates on culture media was measured and compared. A phylogenetic tree was constructed using MEGA10 software. Results: According to morphological characteristics and the ITS sequences of the isolates, all isolates were identified as P. variotii. The phylogenetic tree demonstrated genetic differences in the ITS region among the isolates. No significant correlation was observed between the phylogenetic grouping of the isolates and the morphological differences or the average growth rate. Conclusion: There were differences in the sequences of the ITS region among the isolates. Sequencing the ITS region provided valuable support to confirm the morphological observations to identify P. variotii, although it could not establish a direct correlation between phylogenetic grouping and the morphological and physiological differences among the isolates.
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