In vitro regeneration of periwinkle (Catharanthus roseus L.) and fidelity analysis of regenerated plants with ISSR Markers

Document Type : Research Paper


1 Department of Plant Production and Genetics, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.

2 Department of Plant Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran.

3 Department of Genomics, Northwest and West Region Branch, Agricultural Biotechnology Research Institute, Tabriz, Iran.

4 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran.


Catharanthus roseus is an important multipurpose medicinal plant. In this study, in vitro proliferation and root induction of periwinkle were optimized and regenerated plants were subsequently surveyed for genetic homogeneity using the inter simple sequence repeat (ISSR) markers. Shoot tips and nodal segments were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations of benzylamino-purine (BAP), gibberellic acid (GA3), and indol-3-butyric acid (IBA) hormones. ISSR profiling of regenerated plants as well as the mother plant were surveyed with five primers. The highest establishment rate (80.67%) was obtained in the MS medium containing 1.0 mg L-1 GA3 and 1.0 mg L-1 BAP. Highest proliferation rate (5.20 shoots/explant) and average shoot length (6.30 cm) were observed in 1.5 mg L-1 BAP + 0.5 mg L-1 IBA. Moreover, the best rooting response (85.30%) was observed on half strength MS containing 1.0 mg L-1 IBA. Genetic fidelity analysis using ISSR markers showed the monomorphic banding pattern of the micro-propagated plants and the mother plant, which highlighted their genetic uniformity. This implies that periwinkle micropropagation through shoot tip is the most reliable method for true-to-type production of C. roseus in a large scale.


Article Title [فارسی]

باززایی درون شیشه ای گیاه پروانش (.Catharanthus roseus L) و بررسی پایداری ژنتیک گیاهچه های حاصل از کشت بافت با استفاده از نشانگرهای ISSR

Abstract [فارسی]

پروانش با نام علمی Catharanthus roseus به عنوان یک گیاه دارویی مهم و پرکاربرد از نظر خواص دارویی می­ باشد. در این تحقیق، پس از بهینه­ سازی شاخه ­زایی و ریشه ­زایی گیاه پروانش، یکنواختی ژنتیکی گیاهان باززایی شده با استفاده از پنج جفت نشانگر ISSR مورد ارزیابی قرار گرفت. ریزنمونه­ ها پس از استریل روی محیط کشت موراشیک و اسکوگ (MS) حاوی غلظت­ های متفاوت از هورمون­ های گیاهی بنزیل آمینو پورین (BAP) و اسید ژیبرلیک (GA3) و ایندول بوتیریک اسید (IBA) کشت شدند. تجزیه داده ­ها نشان داد که بیشترین میزان استقرار (80/67 درصد) در یک میلی­گرم در لیتر BAP و GA3 حاصل شد. بیشترین ضریب شاخه­ زایی (5/20 شاخه­ تولید شده بر حسب ریزنمونه) و میانگین طول شاخه (6/30 سانتی­متر) در تیمار حاوی 1/5 میلی­گرم در لیتر BAP +  0/5 میلی­گرم در لیتر IBA حاصل شد. همچنین، بهترین پاسخ ریشه­ زایی(80/30 درصد) در محیط کشت MS­1/2 حاوی یک میلی ­گرم در لیتر IBA به دست آمد. برای بررسی یکنواختی ژنتیکی گیاهان باززایی شده در مقایسه با گیاه مادری از نشانگر ISSR استفاده شد و نتایج نشان داد که ریزازدیادی گیاه پروانش از طریق نوک شاخه­ ها یک روش قابل اطمینان برای تولید مقادیر زیادی گیاهچه با میزان تولید یکنواخت از متابویت های ثانویه می­باشد.

Keywords [فارسی]

  • جوانه جانبی
  • شاخه زایی
  • ضد سرطان
  • نشانگر های ISSR
  • یکنواختی ژنتیکی
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