ردیابی، تحلیل تبارزایی و آنالیز ژنتیک جمعیت ویروس ساقه شیاری سیب بر اساس ژن پروتئین پوششی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران.

2 گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه کردستان، سنندج، ایران.

چکیده

چکیده
     ویروس ساقه شیاری سیب (Apple stem grooving virus (ASGV)) یکی از ویروس­های مهم درختان سیب است. در تحقیق حاضر به منظور ردیابی این ویروس، 87 نمونه مشکوک از برگ و پوست درختان سیب دارای علائم و فاقد علائم از مناطق غربی و جنوب غربی استان زنجان و شرق استان کردستان نمونه برداری انجام شد. پس از استخراج آران­ای کل از 50 نمونه از بین نمونه ­های جمع آوری شده، ساخت دی­ان­ای مکمل با استفاده از آغازگر معکوس طراحی شده منطبق بر ژن کامل پروتئین پوششی انجام شد. در واکنش زنجیره­ای پلی­مراز، قطع ه­ای به طول 728 جفت باز مربوط به ژن کامل پروتئین پوششی ویروس ساقه شیاری سیب در 12 نمونه از باغات مختلف از منطقه زنجان و بیجار استان کردستان تکثیر شد. 9 محصول پی­سی­ار برای تعیین توالی انتخاب شد، از 9 محصول پی­سی­آر انتخاب شده چهار نمونه به­ صورت همسانه ­سازی شده در پلاسمید pTG19 و پنج محصول پی­سی­ آر بصورت مستقیم توالی یابی شدند. نتایج تعیین توالی و مقایسه با پایگاه داده­­های بانک ژن نشان داد قطعات تکثیر یافته در پی­سی­ آر مربوط به ژن پروتئین پوششی ویروس ساقه شیاری سیب است. آنالیز تبارزائی جدایه­ های تعیین توالی شده و توالی­های به­ دست آمده از بانک ژن با استفاده از نرم­افزار MEGA 7 و روش Neighbor Joining (NJ) و مدل Jukes Cantor نشان داد که جدایه­ های ایرانی تازه ردیابی شده با جدایه­ هایی از ایران، هند، کره جنوبی و چین در یک خوشه قرار می­ گیرند. نتایج بررسی جهش ­ها نشان داد dN/dS جدایه­ های ایرانی نسبت به سایر جدایه­ ها کمتر است و بیشترین تبادل ژنی جدایه­ های ایرانی با جدایه­ های کره جنوبی و چین است. این اولین گزارش از این ویروس در منطقه زنجان است.

کلیدواژه‌ها


عنوان مقاله [English]

Detection, phylogenetic and population genetic analysis of Apple stem grooving virus based on coat protein gene

نویسندگان [English]

  • Mahsa Mohammadlou 1
  • Davoud Koolivand 1
  • Mohammad Hajizadeh 2
1 Department of Plant Protection, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.
2 Department of Plant protection, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran.
چکیده [English]

Abstract
Apple stem grooving virus (ASGV) is one of the important viruses infecting apple trees. In this research, 87 symptomatic and symptomless leaf and bark samples were collected from the apple orchards in west and south west of Zanjan and east of Kurdistan provinces to detect ASGV. After total RNA extraction from 50 out of 87 samples, cDNAs were synthesized by reverse primer and PCR optimized using a pair of specific primer corresponding to complete coat protein gene. A DNA segment with 728 bp length corresponding to complete coat protein gene was amplified from twelve samples by RT-PCR. Nine PCR-products including four samples cloned into pTG19 and five samples without cloning were sequenced. The sequencing and blast results revealed that the fragments belong to coat protein gene of Apple stem grooving virus. Phylogenetic analysis of new Iranian isolates and those retrieved from GenBank by MEGA 7 and Neighbor-Joining methods in Jukes-Cantor model showed that the new Iranian isolates were placed in a sub-clade with reported Iranian, Indian, South Korean and Chinese isolates. The result revealed that the least dN/dS ratio belonging to Iranian isolates and the most gene flow was calculated between isolates from Iran, China and South Korean. This is the first report of Apple stem grooving virus from the Zanjan region.  
 

کلیدواژه‌ها [English]

  • Keywords: Kurdistan
  • Phylogentic Analysis
  • Expression
  • Cappilovirus
  • Zanjan
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