تنوع ژنوتیپی جدایه‎های tumefaciens Agrobacterium و A. vitis از میزبان‌های مختلف در برخی از استان‌های غربی و جنوبی ایران با استفاده از نشانگر RAPD

نوع مقاله: مقاله کامل پژوهشی

نویسندگان

1 به ترتیب دانش آموخته کارشناسی ارشد بیماری شناسی گیاهی ،استادو استاد بیماری شناسی گیاهی دانشکده کشاورزی، دانشگاه شیراز

2 دانشیار پژوهشکده بیوتکنولوژی دانشکده کشاورزی، دانشگاه شیراز

چکیده

به منظور بررسی تنوع ژنتیکی گونه­های  tumefaciens Agrobacterium و A.vitis در سال­های 93-1392 از باغ‎های مو، گلخانه‌های پرورش گل رز و مزارع چغندر قند در استان­ جنوبی (فارس) ، جنوب غربی(کهکیلویه و بویراحمد) و غربی (کردستان و کرمانشاه)، از گیاهان دارای علائم گال طوقه و ریشه نمونه برداری صورت گرفت. بر اساس ویژگی‎های فنوتیپی، آزمون­های بیو‌شیمیایی، بیماری­زایی و همچنین با استفاده از آغازگرهای عمومی جنس (virD2A/virD2C) و اختصاصی در سطح گونه (VCF/VCR و PGF/PGR) بیمارگر به عنوان، tumefaciens A. و A.vitis  شناسایی شد. علاوه بر این با استفاده از جفت آغازگرهای  VisF/VisRو VisFF1/R2 نوع اوپین جدایه‌های A.vitis تشخیص داده شد. جهت بررسی تنوع ژنتیکی بین جدایه­های دو گونه tumefaciens. A و A.vitis در آزمون چندشکلی تصادفی قطعات تکثیر شده­ی دی­ان­اِی (RAPD) از 11 آغازگر تصادفی استفاده شد. در گونه tumefaciens. A جدایه‌های متعلق به میزبان‌های مختلف در سطح تشابه 73 درصد به سه گروه و جدایه‌های A.vitis به دو گروه  تقسیم شدند. نتایج نشان داد که جدایه‌های متعلق به دو گونه tumefaciens. A و A.vitis دارای تنوع زیادی بودند و جدایه­های به دست آمده از یک منطقه­ جغرافیایی در چند گروه مختلف قرار گرفتند. همچنین بین تنوع ژنتیکی جدایه­های مورد بررسی و منطقه­ جغرافیایی یا میزبان آن­ها ارتباطی وجود نداشت .با استفاده از آغازگرهای VisF/VisRو VisFF1/R2 مشخص شد  که 9/57 درصد از جدایه­های A.vitis دارای ژن کد کننده ویتوپین، 3/26 درصد از جدایه دارای ژن کد کننده اوپین اکتوپین / نوپالین  بودند و 8/15 درصد جدایه­ها قادر به تکثیر هیچ یک از قطعات ذکر شده نبودند

کلیدواژه‌ها


عنوان مقاله [English]

Genotypic Diversity among Agrobacterium tumefaciens and A. vitis Isolates From Different hosts in Some of Western and Southern Provinces of Iran using RAPD Marker*

نویسندگان [English]

  • H. Mafakheri 1
  • S. M. Taghavi 1
  • Z. Banihashemi 1
  • A. Niazi 2
1 Former M.Sc. Student, Prof and Prof of Plant Phathol., respectively, School of Agric., Shiraz University., Shiraz, Iran
2 Associate Prof. of Institute of Biotechnology, School of Agric., Shiraz Univ., Shiraz, Iran.
چکیده [English]

To assess the genetic diversity of Agrobacterium tumefaciens and A.vitis , during 2013 and 2014, samples of grapevine, rose and sugar beet with gall symptoms on crown and roots were collected from different areas of southern (Fars and kohgiluye and Boyerahmad) and western (Kurdistan and Kermanshah) provinces of  Iran. Based on phenotypic characteristics, biochemical and pathogenicity tests as well as using the generic primers virD2A/virD2C the isolates were identified as Agrobacterium. Furthermore, species-specific primer pairs VCF/VCR for A. tumefaciens, as well as PGF/PGR for A. vitis were used to determine the identity of the causal agent.For detection of opine type among A. vitis isolates, a PCR assay was carried out using primer pairs VisF/VisR and VisFF1/ R2. Genetic diversity of A. tumefaciens and A.vitis isolates were investigated by random amplified polymorphic DNA technique using 11 random primers. The A.tumefaciens isolates clustered in three groups at 73% level of similarity and A.vitis clustered in two  groups. Based on the results,  A. tumefaciens and A.vitis isolates showed high genetic diversity even in a same geographical region. In conclusion, no correlation was observed between genetic diversity and geographical origins or their host plants. For detection of opine genes among A. vitis isolates, a PCR assay was carried out using primer pairs VisF/VisR and VisFF1/R2. Our results revealed that 57.9% of A. vitis isolates produced vitopine and 26.3% of them produced octopine or nopaline, whereas 15.8% of isolates were unable to amplify specific fragment.

کلیدواژه‌ها [English]

  • Crown gall
  • Ti plasmid
  • Molecular marker
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