خاموشی همزمان ژن‌های تبائین 6- O- دمتیلاز و کدئین O- دمتیلاز در گیاه دارویی شقایق با روش القاء ویروسی

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد

2 دانشیار گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد

3 استاد گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد

چکیده

گیاه شقایق تنها منبع تجاری برخی آلکالوئیدهای مهم دارویی است. دو آنزیم مهم به نام تبائین 6-O-دمتیلاز (Thebaine 6-O-demethylase; T6ODM) و کدئین O-دمتیلاز (Codeine O-demethylase; CODM) در انتهای مسیر بیوشیمیایی آلکالوئیدهای بنزیل ایزوکوینولینی این گیاه وجود دارند. در این مطالعه فعالیت بیانی ژن‌های دو آنزیم مذکور با روش VIGS (Virus-Induced Gene Silencing) مورد بررسی قرار گرفت. برای این منظور، سازه‌ای براساس ناقل TRV2 که دارای توالی حفاظت شده ژن DIOX (که مشترک با بخش انتهایی ژن‌های T6ODM و CODM) می‌باشد، طراحی شد. برگ­های اولیه گیاه دو تا سه هفته­ای کشت شده در گلدان با آگروباکتریوم دارای سازه خاموشی و برای گیاهان کنترل با آگروباکتریوم حاوی ناقل TRV2 خالی مایه‌زنی شدند. بررسی بیان ژن در سطح نسخه‌برداری با فن Real Time PCR به‌ترتیب نشان‌دهنده کاهش 86 و 87 درصدی بیان ژن‌های CODM و T6ODM بود. این مطالعه اثبات کرد که VIGS ابزار موثری برای کاهش سطوح نسخه‌های ژن‌های بیوسنتزی بنزیل ایزوکوینولین‌هاست.

کلیدواژه‌ها


عنوان مقاله [English]

Co-silencing of Thebaine 6-O-demethylase and Codeine O-demethylase in Papaver somniferum L., Using Virus-Induced Gene Silencing

نویسندگان [English]

  • Parvin Norouzi 1
  • Ahmad Ismaili 2
  • Farhad Nazarian Firouz-Abadi 3
1 MSc Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad
2 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad
3 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad
چکیده [English]

Papaver somniferum remains the sole commercial source for narcotic and variety of semisynthetic drugs. Thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM) genes are related the dioxygenase family that occur at the end of biosynthesis of these alkaloids in this plant. For silencing of CODM and T6ODM genes in Papaver somniferum, a TRV2-based construct containing a conserved sequence of DIOX from the coding region of T6ODM and CODM was designed for simultaneously silencing of two mentioned genes. Emerging first leaves of 2 to 3-week-old seedling were infiltrated with Agrobacterium tumefaciens harboring silencing construct, or the empty vector (TRV2) as a control. Analysis of gene expression at the transcript level by real-time quantitative PCR (polymerase chain reaction) technique showed 86 and 87 percent reduction in gene expression of CODM and T6ODM compared with control plants, respectively. This study demonstrated the efficiency of this method for knockdown the transcript levels of specific BIA biosynthetic genes.

کلیدواژه‌ها [English]

  • Benzylisoquinoline
  • Papaver somniferum
  • Gene silencing
  • Virus induced gene silencing

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