ردیابی ‎و تفکیک دو عارضه اختلال و عدم غلاف‌بندی سویا در مزارع استان‌های گلستان و ‏مازندران

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی مقطع دکتری بیماری شناسی گیاهی، گروه گیاه پزشکی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان گلستان

2 دانشیار گروه گیاه‌پزشکی، دانشکده تولیدات گیاهی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گلستان

3 استادیار گروه گیاه پزشکی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گلستان،

4 دانشیار دانشگاهعلوم کشاورزی و منابع طبیعی گرگان

چکیده

به منظور ردیابی و تفکیک دو عارضه اختلال و عدم غلاف‌بندی در سویا، ضمن بازدید از 185 مزرعه سویا واقع در استان‌های گلستان و مازندران، فقط از کشت تابستانه 17 مزرعه از پنج نقطه مختلف استان گلستان، بوته‌های دارای علایم اختلال و عدم غلافبندی حاد شناسایی و در ادامه هفده نمونه از هر مزرعه انتخاب و نمونه‌برداری از برگ یا ساقه بوته‌های مذکور به منظوراستخراج RNA و DNA انجام شد. سپس PCR برای ردیابی نپو ویروس، با استفاده از پرایمر دژنره نپو ویروس و برای ردیابی فیتوپلاسما با استفاده از جفت آغازگرهای عمومی و یک مرحله آزمون PCR آشیانه‌ای انجام گردید، نتایج الکتروفورز، تکثیر باند 1800 جفت‌بازی را در PCR عمومی، قطعه 1250 جفت‌بازی را در PCR آشیانه‌ای مربوط به فیتوپلاسما و همچنین تکثیر باند 640 جفت بازی مربوط به یک نپو ویروس را تایید کرد. ضمن اینکه هیچ باندی از بوته‌های سالم مشاهده نشد. همزمان پیوند پوست و مایه‌زنی مکانیکی روی گیاه محک سویا جی پی ایکس با استفاده از نمونه‌های مذکور انجام گرفت که منجر به بروز دو نوع علایم شد. از توالی‌یابی نمونه‌های دارای اختلال (تولید تعداد کمی بذر و غلاف ریز)، Tomato ring spot virus استرین ep31_63026 و از توالی‌یابی نمونه‌های دارای عدم غلافبندی (علفی شدن و عدم تشکیل غلاف و بذر)، فیتوپلاسمای Aster yellows phytoplasma از گروه 16SrI-B شناسایی شدند که نتایج گیاه محک را تایید کرد. بررسی فیلوژنتیکی نتایج توالی‌یابی حضور فیتوپلاسما و نپو ویروس را فقط در کشت تابستانه نمونه‌هایی که دارای علائم اختلال و عدم غلافبندی بودند، تائید نمود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Detection and differentiation of two complications of disorder and lack of podding of soybeans in the fields of Golestan and Mazandaran provinces

نویسندگان [English]

  • Son ay Baghdadi 1
  • Abdolhossein taheri 2
  • saeed nasrollahnejad 2
  • Farzad Aliramaji 3
  • leila fahmideh 4
1 PhD student in Plant Pathology, Gorgan University of Agricultural Sciences and Natural Resources, Golestan
2 Associate Professor of Plant Protection, Plant protection department, Gorgan University of Agricultural Sciences and Natural Resources, Golestan
3 Assistant Professor of Plant protection, Gorgan University of Agricultural Sciences and Natural Resources, Golestan,
4 Associate Professor of Plant breeding and biotechnology department, Gorgan University of Agricultural Sciences and Natural Resources, Golestan
چکیده [English]

In order to detection and differentiation of two complications of disorder and lack of podding in soybeans, while visiting 185 soybean fields in Golestan and Mazandaran provinces, only from the summer cultivation of 17 fields from five different places in Golestan province, plants with signs of disorder and the lack of acute podding were identified and 17 samples were selected from each field and sampling was done from the leaves or stems of the mentioned plants in order to extract RNA and DNA. Then, PCR was performed to detect nepovirus using the degenerate primer of nepovirus and to detect phytoplasma using a pair of general primers and a nested PCR test. The results of electrophoresis confirmed the amplification of the 1800 bp band in the general PCR, the 1250 bp fragment in the nested PCR related to phytoplasma, and also the amplification of the 640 bp band related to a nepovirus. Besides, no band of healthy plants was observed. at the same time, tissue grafting and mechanical inoculation were performed on GPX soybean benchmark plant using the mentioned samples and two types of symptoms appeared. From the sequencing of the disordered samples (production of a small number of seeds and small pods), Tomato ring spot virus strain ep31_63026 and from the sequencing of the samples with non-encapsulation (grassiness and no formation of pods and seeds), Aster yellows phytoplasma from the 16SrI-B group were identified, which confirmed the results of the reference plant. The phylogenetic analysis of sequencing results confirmed the presence of Phytoplasma and Nepovirus only in the summer culture of samples that had symptoms of disorder and lack of podding.

کلیدواژه‌ها [English]

  • Nepovirus
  • vector insects
  • phytoplasma
  • soybean
  • soybean pod

مراجع

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