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Increasing vitamin E content of canola (Brassica napus L.) by transferring γ-tmt gene | ||
| Iranian Journal of Genetics and Plant Breeding | ||
| مقاله 9، دوره 8، شماره 1 - شماره پیاپی 15، تیر 2019، صفحه 55-63 اصل مقاله (1.25 M) | ||
| نوع مقاله: Research paper | ||
| شناسه دیجیتال (DOI): 10.30479/ijgpb.2020.12410.1260 | ||
| نویسندگان | ||
| Mehran Babaei؛ Ramin Hosseini* | ||
| Biotechnology Department, Faculty of Agriculture and Natural Sciences, Imam Khomeini International University, P. O. Box: 34149-16818, Qazvin, Iran. | ||
| تاریخ دریافت: 23 دی 1398، تاریخ بازنگری: 09 اسفند 1398، تاریخ پذیرش: 22 اسفند 1398 | ||
| چکیده | ||
| Vitamin E is one of the lipid soluble vitamins consisting of several isoforms, including tocopherols and tocotrienols amongst which the alpha tocopherol is the most active one. The conversion of γ tocophrol to α tocopherol takes place by the activity of γ-tmt enzyme. Many plants including canola lack the γ-tmt gene to enable them to convert γ tocophrol into α tocopherol. The aim of this study was to transfer γ-tmt gene into canola plants to enable them to produce α tocopherol and increase their vitamin E content. γ-tmt gene was isolated from tomato (Lycopersicum esculentum L.), Memory1 cultivar. Then, it was amplified using PCR, digested by XbaΙ enzyme, cloned into the pBluescriptΙΙ cloning vector and subcloned into E. coli. The gene was then transferred into pBI121 vector and subsequently the vector containing the γ-tmt gene was transferred into Agrobactearium tumefaciens. Two canola cultivars Zafam and Hayola 401 were used. The cotyledons of canola seeds were inoculated by Agrobactearium tumefaciens. Seven putative transformants from each cultivar (Zarfam 2 to 8 and Hayola 2 to 8) were chosen for further investigations. After the emergence of shoots and roots, the seedlings were assayed for the presence of γ-tmt gene, by PCR. Vitamin E content of the transformed plants was assayed by FTIR spectrophotometer. Results showed several fold increases in vitamin E contents of the transgenic plants compared to the control. The increases in α-tocopherol content were 2.61 and 2.71 times in Zarfam 6 and Hayola 8, respectively. This approach could be considered as a useful method for fortifying oil seed crops with vitamin E. | ||
| کلیدواژه‌ها | ||
| Agrobacterium tumefaciens؛ Brassica napus؛ Transformation؛ Vitamin E؛ γ-tmt | ||
| عنوان مقاله [English] | ||
| افزایش میزان ویتامین E در کلزا (Brassica napus L.) با استفاده از ژن γ-tmt | ||
| نویسندگان [English] | ||
| مهران بابایی؛ رامین حسینی | ||
| گروه بیوتکنولوژی، دانشکده کشاورزی و منابع طبیعی، دانشگاه بین المللی امام خمینی (ره)، قزوین، ایران، کدپستی: 81861-34149. | ||
| چکیده [English] | ||
| ویتامین E یکی از ویتامینهای محلول در چربی و دارای چندین ایزومر است که شامل توکوفرولها و توکوترینولهاست. تبدیل گاما توکوفرول به آلفا توکوفرول با فعالیت آنزیم γ-tmtصورت می پذیرد. بسیاری از گیاهان از جمله کلزا ژن γ-tmt را ندارند تا آنها را از گاما توکوفرول به آلفا توکوفرول تبدیل سازند. هدف این مطالعه انتقال ژن γ-tmt به گیاه کلزا بود تا آن را به تولید آلفا توکوفرول و افزایش ویتامین E قادر سازد. ژن γ-tmt از گیاه گوجه فرنگی (Lycopersicum esculentum L.) رقم Memory 1 جدا سازی شد. سپس با استفاده از PCR تکثیر، با آنزیم XbaI برش داده، در حامل pBluscriptII همسان سازی و به E. coli منتقل گردید. در ادامه، این ژن به حامل pBI121 و حامل به آگروباکتریوم تومفشینس انتقال داده شد. دو رقم کلزا، زرفام و هایولا 401 استفاده شدند. کوتلیدون بذرهای کانولا با آگروباکتریوم تومفشینس تلقیح شدند. هشت گیاه با پتانسیل تراریخت بودن از هر رقم (زرفام 1-8 و هایولا 1-8) برای آزمایشهای بعدی انتخاب شدند. پس از ظاهر شدن ساقه و ریشه، دانه رستها برای وجود ژن γ-tmt سنجیده شدند. میزان ویتامین E گیاهان تراریخت به کمک اسپکتروفتومتر FTIR سنجیده شد. نتایج افزایش چند برابری ویتامین E را در گیاهان تراریخته نسبت به گیاهان شاهد نشان داد. افزایش ویتامین E به ترتیب 41/2 و 71/2 برابر در رقم زرفام و هایولا بود. لذا، این روش میتواند برای غنی سازی روغن گیاهان روغنی با ویتامین E در نظر گرفته شود. | ||
| کلیدواژه‌ها [English] | ||
| Brassica napus, تراریخت کردن, ویتامین E, آگروباکتریوم تومفشینس, γ-tmt | ||
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