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:: دوره 8، شماره 2 - ( 1400 ) ::
جلد 8 شماره 2 صفحات 22-1 برگشت به فهرست نسخه ها
انتقال ژن به‌روش آگروباکتریوم و تحلیل گیاهان تراریخت در صنوبر سیاه هیبرید (Populus × euromericana Dode Guinier)
شهنوش نیری، بهرام باغبان کهنه‌روز*
چکیده:   (891 مشاهده)
صنوبر سیاه هیبرید (Populus × euramericana)، گیاهی با کاربردهای مهم اقتصادی در صنایع خمیرچوب و کاغذ، سوخت زیستی و مواد نانوسلولزی و حذف آلودگی‌های خاک است. در این پژوهش، شیوه‌نامه انتقال ژن به‌روش آگروباکتریوم و باززایی مستقیم گیاه با استفاده از بافت میانگره ساقه صنوبر سیاه هیبرید ارائه و تحلیل مولکولی گیاهان تراریخت انجام شد. جهت باززایی گیاه کامل از کشت ریزنمونه­های میانگره ساقه در محیط‌کشت القای ساقه‌زایی حاوی محیط موراشیگ و اسکوگ (MS) و غلظت‌های مختلف ترکیباتBAP × IBA  و برای ریشه زایی (RIM) از محیط‌کشت القای ریشه‌زایی حاوی غلظت های IBA × NAA استفاده شد. با ارزیابی فاکتورهای فیتوهورمونی در باززایی مستقیم، بالاترین میزان باززایی ساقه (28.57 ساقه در هر ریزنمونه) از ریزنمونه‌های کشت شده در محیط‌کشت SIM حاوی 0.5 میلی‌گرم بر لیتر BAP و 0.05 میلی‌گرم بر لیتر IBA به‌دست آمد. بهترین ریشه­زایی ساقه­های باززا در محیط‌کشت RIM حاوی 0.1 میلی‌گرم بر لیتر IBA و 0.05 میلی‌گرم بر لیتر NAA تولید شد. 100 درصد گیاهان باززا با محیط سازگار شده و به گلخانه منتقل گردیدند. نتایج مربوط به حساسیت به علف‌کش نشان داد که 0.5 میکرومولار Basta® سیستم گزینش دقیقی جهت مهار سلول‌های غیرتراریخت فراهم می‌آورد. بالاترین فراوانی انتقال ژن (93.33درصد) از طریق پیش‌کشت ریزنمونه‌ها به‌مدت 6 روز و تلقیح آن‌ها با Agrobacterium tumefaciens سویه LBA4404 ( 0.6=OD600 ) همراه 100 میکرومولار استوسرینگان به‌مدت 10 دقیقه به‌دست آمد. آنالیزهای لکه‌گذاری سادرن، RT-PCR و آزمون هیستوشیمیایی GUS به‌ترتیب درج یک تا دو نسخه ژن gus در DNA ژنومی و بیان پایدار آن‌ را در گیاهان نسل T0 منتخب تأیید کرد. یافته‌های این تحقیق می‌تواند به‌عنوان شیوه­نامه باززایی و انتقال ژن به گیاه صنوبر سیاه هیبرید مورد بهره‌برداری قرار گیرد.
واژه‌های کلیدی: باززایی مستقیم، بیان ژن GUS، لکه‌گذاری سادرن، میانگره‌ساقه، Populus × euramericana
متن کامل [PDF 2325 kb]   (211 دریافت)    
نوع مطالعه: پژوهشي | موضوع مقاله: مهندسی ژنتیک
دریافت: 1400/11/5 | پذیرش: 1401/12/24
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Nayeri S, Baghban Kohnehrouz B. Efficient Agrobacterium-Mediated Transformation and Analysis of Transgenic Plants in Hybrid Black Poplar (Populus × euromericana Dode Guinier). pgr. 2022; 8 (2) :1-22
URL: http://pgr.lu.ac.ir/article-1-238-fa.html

نیری شهنوش، باغبان کهنه‌روز بهرام. انتقال ژن به‌روش آگروباکتریوم و تحلیل گیاهان تراریخت در صنوبر سیاه هیبرید (Populus × euromericana Dode Guinier). پژوهش های ژنتیک گیاهی. 1400; 8 (2) :22-1

URL: http://pgr.lu.ac.ir/article-1-238-fa.html



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دوره 8، شماره 2 - ( 1400 ) برگشت به فهرست نسخه ها
پژوهش های ژنتیک گیاهی Plant Genetic Researches
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