fa کشت بافت و تراریزش نارنج (Citrus aurantium) با استفاده از ژن کد کننده ی پوشش پروتئینی ویروس تریستزای مرکبات (CTV) مهندسی ژنتیک گیاهی Plant پژوهشي Research <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">در</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">این</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">پژوهش</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">به</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">منظور</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">به</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">دست</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">آوردن</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">گیاهان</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">تراریخته</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">ی</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">نارنج،</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">از</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">ریزنمونه</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">های</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">اپی</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">کوتیل</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">و هیپوکوتیل</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">نارنج</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">استفاده</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">و</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">بهبود</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">اندام زایی</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">مستقیم</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">و</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">تولید</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">شاخساره</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">در</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">ریزنمونه</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">ها</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">با</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">استفاده</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">از تیمارهای</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">مختلف</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">بررسی</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">شدند</span></span></strong><strong><span dir="LTR"><span style="font-family:bzarbold;"><span style="font-size:14.0pt;">.</span></span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> تراریزش</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">از</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">طریق</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">همکشتی</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">ریزنمونه</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">ها</span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">با</span></span></strong> <strong><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">Agrobacterium</span></span></span></em></strong> <strong><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">tumefaciens</span></span></span></em></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> سویه</span></span></strong> <strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">EHA 105</span></span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> حامل وکتور</span></span></strong> <strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">pFGC 5941</span></span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> و ژن کد کننده ی پوشش پروتئینی ویروس تریستزای مرکبات </span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:12.0pt;">(</span></span></strong><strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">CTV</span></span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:12.0pt;">)</span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> انجام شده. پس از تراریزش، ریزنمونه هابه محیط کشت انتخابی حاوی علف کش بستا با ترکیبی از سطوح تنظیم کننده رشد</span></span></strong> <strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:10.0pt;">BAP</span></span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">(0، 1 و 2 میلی گرم در لیتر) و </span></span></strong><strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">NAA</span></span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> (0، 0/25 و 0/5) منتقل شدند. اثر زخم زدن بر اندام زایی مستقیم و تولید شاخساره و اثر استفاده از وکیوم بر کارایی تراریزش، بررسی شدند. تایید تراریخته </span></span></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">بودن شاخساره ها در محیط انتخابی حاوی علف کش بستا و انجام واکنش پی.سی.آر با ژن های اختصاصی صورت گرفت. بهترین کارایی اندام زایی یا تولید شاخساره (57 درصد) با استفاده از ترکیب تنظیم کننده های رشد به میزان 2 میلی گرم در لیتر</span></span></strong> <strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">BAP</span></span></span><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> و0/25 میلی گرم در لیتر</span></span></strong> <strong><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">NAA</span></span></span></strong> <strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;">حاصل شد ضمن این که، زخم زدن اثر معنی داری بر تولید شاخساره نداشت. بالاترین میزان شاخساره های تراریخته به میزان11/25درصد در تیمار وکیوم به دست آمد. در نهایت شاخساره های تراریخته بر روی دانهال های رشد یافته در شرایط </span></span></strong><strong><em><span dir="LTR"><span style="font-family:times new roman,serif;"><span style="font-size:12.0pt;">in vitro</span></span></span></em></strong><strong><span style="font-family:b mitra;"><span style="font-size:14.0pt;"> ریزپیوند شدند.</span></span></strong> In this study transgenic plants of sour orange (<em>C. aurantium) </em>that is an important citrus rootstock were produced by <em>Agrobacterium</em>-mediated transformation. Epicotyl and hypocotyl segments-derived explants were co-cultured with <em>Agrobacterium </em>strain EHA105 carrying pFGC5941 plasmid containsing CTV coat protein (p25) gene. One of the main objects of present research was to improve the direct <em>in vitro </em>organogenesis efficiency in <em>C. aurantium</em>. Therefore different combination of BAP (0, 1, 2 mg/L) and NAA (0, 0.25, 0.5 mg/L) were used in selective medium to culture transformed explants. The highest regeneration (57%) was obtained from explant treated with 2 mg/L BAP and 0.25 mg/L NAA. Effects of wounding and vacuum infiltration on transformation efficiency were evaluated either. The best transformation efficiency (11.25%) was obtained from explants that were vacuum infiltrated during transformation and subsequently were cultured in medium containing 2 mg/L BAP and 0.25 mg/L NAA. PCR analysis using two different genes were performed to confirm transformation. Micro grafting of transformed shoots were carried out on non-transgenic, in-vitro grown seedlings. Agrobacterium tumefaciens, اپی کوتیل, اندام زایی مستقیم, تنظیم کننده های رشد, زخم زنی Agrobacterium tumefaciens, Direct Organogenesis, Epicotyl, Growth Regulators, Wounding 83 92 http://gebsj.ir/browse.php?a_code=A-10-223-2&slc_lang=fa&sid=1 Banafsheh Fattah بنفشه فتاح 10031947532846001693 10031947532846001693 Yes Mohammad Mehdi Sohani محمدمهدی سوهانی mhdsohani@gmail.com 0047692881 0047692881 No Abdollah Hatamzadeh عبدالله حاتم زاده 10031947532846001695 10031947532846001695 No Alireza Afsharifar علیرضا افشاری فر afshari@shirazu.ac.ir 10031947532846001696 10031947532846001696 No Behrooz Goleyn بهروز گلعین bgoleincitrus@yahoo.com 10031947532846001697 10031947532846001697 No Mohammad Hossein Rezadoost محمدحسین رضا دوست 10031947532846001698 10031947532846001698 No Mohammad Reza Mirzaei محمدرضا میرزایی 10031947532846001699 10031947532846001699 No Amir Hossein Zamani امیرحسین زمانی 10031947532846001700 10031947532846001700 No