نوع مقاله : مقاله کامل پژوهشی

نویسندگان

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چکیده

گونه­یPhytophthora erythroseptica ، عامل پوسیدگی صورتی سیب­زمینی، یکی از اُاُمیست­های بیماری­زای گیاهی است که خسارت اقتصادی قابل توجهی را در مزرعه و انبار وارد می­کند. به منظور شناسایی و ردیابی دقیق و حساس P. erythroseptica ، شش توالی هسته­ای و میتوکندریایی برای طراحی آغازگرهای اختصاصی مورد بررسی قرار گرفت. به دلیل شباهت زیاد توالی­های P. erythroseptica به خویشاوندان نزدیکش، تنها یک توالی هسته­ای (TigA) برای طراحی آغازگرهای اختصاصی مناسب تشخیص داده شد. با استفاده از آغازگرهای اختصاصی طراحی شده، شیوه­ای از واکنش زنجیره­ای ساده و تودرتو برای شناسایی و ردیابی P. erythroseptica ابداع شد. اختصاصیت آغازگرهای طراحی شده با استفاده از مجموعه­ای از گونه­های فیتوفتورا متعلق به تبارهای فیلوژنتیکی مختلف و نیز خویشاوندان نزدیک P. erythroseptica ارزیابی شد. ردیابی P. erythroseptica در دی­ان­ای خالص بیمارگر و دی­ان­ای استخراج شده از بافت­های گیاهی آلوده شامل سیب­زمینی، گوجه­فرنگی و اسفناج با استفاده از آغازگرها با موفقیت انجام شد. آغازگرهای اختصاصی10 پیکوگرم از دی­ان­ای خالص P. erythroseptica را در واکنش زنجیره­ای پلیمراز ساده ردیابی کردند، با این حال واکنش زنجیره­ای پلیمراز تودرتو، حساسیت آغازگرها را حداقل صد برابر افزایش داد. همچنین، آغازگرهای اختصاصی قادر به ردیابی P. erythroseptica به عنوان والد پدری یا مادری در جدایه­های دورگ بودند؛ این ویژگی به شناسایی یکی از والدین در دورگ­های P. erythroseptica کمک شایانی می­کند

کلیدواژه‌ها

عنوان مقاله [English]

Identification and detection of Phytophthora erythroseptica and its hybrids using specific primers

نویسندگان [English]

  • B. Safaiefarahani
  • R. Mostowfizadeh-Ghalamfarsa
  • A. Habibi

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چکیده [English]

Phytophthora erythroseptica, the causal agent of potato pink rot, is one of the oomycete plant pathogens that causes significant losses in field and storage. In order to develop a sensitive and rapid method for detection and identification of P. erythroseptica, six nuclear and mitochondrial gene regions were investigated to design species-specific primers. Due to the high similarity of P. erythroseptica sequences to its closely related species, only one nuclear region, TigA, was appropriate to design specific primers. Using specific primers, a simple as well as a nested-PCR based method was developed for the identification and detection of P. erythroseptica. The specificity of designed primers was examined using a collection of Phytophthora species from different phylogenetic clades as well as close relatives of P. erythroseptica. In addition to pure DNA, designed primers detected P. erythroseptica in infected plant tissues including potato, tomato and spinach. Specific primers detected 10 pg of p < /em>. erythroseptica pure DNA, however, nested PCR increased primers sensitivity at least 100 times. Moreover, specific primers designed in this study were able to detect P. erythroseptica as the maternal or paternal parent species in hybrid isolates that would make a significant help to recognize one of the parental species in hybrids of P. erythroseptica.

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