آزمایش مقدماتی انتقال ژن به چمانواش بلند با واسطه‌گری Agrobacterium tumefaciens

Subject Areas : Journal of Ornamental Plants

مصطفی خوشحال سرمست ¹ , Hassan Salehi ² , Mehrnaz Zarei ³

1 - Department of Horticultural Science, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources (GUASNR), Gorgan 49138-43464, Golestan, Iran
2 - دانشگاه شیراز
3 - Department of Horticultural Science, College of Agriculture, Shiraz University, Shiraz, Iran

Received: 2017-10-07 Accepted : 2017-12-28 Published : 2018-06-01

Keywords: تنش خشکی, چمانواش بلند, آزمون Gus, ژن Uida,

Abstract :

تنش­های غیر زیستی ممکن است گیاهان را به دلیل شرایط محیطی نامناسب و یا کیفیت کم آب متاثر کنند. این تنش­های رشد و نمو گیاه را طی چرخه­های رشدی تحت تاثیر قرار می­دهند. به این دلیل گیاهان متحمل به خشکی ارزشمند بوده و سبز فرش­ها متحمل به خشکی می­توانند برای این منظور تولید شوند. این گونه­ها پوششی مناسبی روی سطح زمین ایجاد کرده و مزایایی مختلفی از جمله زیبایی و جلوگیری از فرسایش خاک دارند که زندگی روزانه ما را متاثر می­کنند. هدف این پژوهش یافتن محیط کشت ایده ال برای القاء پینه و باززایی و به دنبال آن بیش بیان ژن P5CS1 کد کننده پرولین برای بهبود مقاومت به خشکی در چمانواش بلند بود. برای این منظور پوشش بذر حذف و سپس به صورت طولی بریده شد. این تیمار اثر قابل ملاحظه­ای در مقایسه با بذوری داشت که  تحت این تیمار قرار نگرفته بودند.  علاوه بر این راندمان باززایی در محیط­های دارای 2,4-D و بنزیل آدنین افزایش یافت. متوسط پینه­های مقاوم به نشانگر گزینش­گر G418 تنها 10% بود. حضور ژن P5CS1 در پینه­ها با استفاده از آزمون هیستوشیمیایی Gus و واکنش زنجیره­ای پلی­مراز با آغازگر­های اختصاصی تایید شد. بیش از 90% پینه ها قادر به بیان ژن uida بودند که این به احتمال و به طور غیر مستقیم بیان کننده القاء تحمل به خشکی در پینه­ها می­باشد.

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