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

Document Type : Research Paper

Authors

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.

Abstract

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.

Keywords

Article Title [Persian]

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 [Persian]

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 [Persian]

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

References

Abou-Mandour AA, Fischer S and Czygan FC, 1979. Regeneration of intact plants from haploid and diploid callus cells of Catharanthus roseus. Zeitschrift für Pflanzenphysiologie 91(1): 83-88.
Amiri S, Fotovat R, Tarinejhad A, Panahi B and Mohammadi SA, 2019a. Optimization of hormonal combinations for in vitro regeneration of lesser periwinkle (Vinca minor L.) and assessment of genetic homogeneity. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences doi.org/10.1007/s40011-019-01141-6.
Amiri S, Panahi B, Mohammadi R and Fattahi F, 2019b.  Effect of plant growth regulator combination on direct in vitro regeneration of Persian lilac (Melia azedarach L.). Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. doi: 10.1007/s40011-019-01099-5.
Bagum T and Mathur M, 2014. In vitro regeneration of Catharanthus roseus and Bacopa monnieri and their survey around Jaipur district. International Journal of Pure and Applied Bioscience 2(4): 210-221.
Bakrudeen AAA, Subha Shanthi G, Kavitha MS and Rao MV, 2011. In vitro micropropagation of Catharanthus roseus – an anticancer medicinal plant. Acta Botanica Hungarica 53(1-2): 197-209.
Barrales-Cureño HJ, Andrade-Hoyos P, Luna-Cruz A, Reyes-Reyes C, Chávez-Salinas S and López-Valdez LG, 2017. In Vitro Biotechnological Production and Pharmacological Studies of Antileukemic Alkaloids of Catharanthus roseus. In: Naeem M, Aftab T and Khan M (eds.) Catharanthus roseus. Pp. 17-34. Springer, Cham, Switzerland.
Dhandapani M, Kim DH and Hong SB, 2008. Efficient plant regeneration via somatic embryogenesis and organogenesis from the explants of Catharanthus roseus. In Vitro Cellular & Developmental Biology - Plant 44: 18-25.
Dhruva B, Ramakrishnan T and Vaidyanathan C, 1977. Studies in Catharanthus roseus callus cultures, callus initiation and differentiation. Current Science 46: 364-365.
Ghorbanzadeh Neghab M  and  Panahi B, 2017. Molecular characterization of Iranian black cumin (Nigella sativa L.) accessions using RAPD marker. BioTechnologia 98(2): 97-102.
Khatun MM, Tanny T, Yesmin S, Salimullah MD and Alam I, 2018. Evaluation of genetic fidelity of in vitro-propagated Aloe vera plants using DNA-based markers. ScienceAsia 44: 87-91.
Kumar A,   Mishra P,   Baskaran K,   Shukla AK,  Shasany AK and  Sundaresan V, 2016. Higher efficiency of ISSR markers over plastid psbA‐trnH region in resolving taxonomical status of genus Ocimum L. Ecology and Evolution 6(21): 7671-7682.
Kumar A, Prakash K, Sinha RK and Kumar N, 2013. In vitro plant propagation of Catharanthus roseus and assessment of genetic fidelity of micropropagated plants by RAPD marker assay. Applied Biochemistry and Biotechnology 169(3): 894-900.
Kyozuka J, 2007. Control of shoot and root meristem function by cytokinin. Current Opinion in Plant Biology 10(5): 442-446.
Li Sh, Huang P, Ding G, Zhou L, Tang P, Sun M, Zheng Y and Lin S, 2017. Optimization of hormone combinations for root growth and bud germination in Chinese fir (Cunninghamia lanceolata) clone leaf cuttings. Scientific Reports 7: 5046. doi:10.1038/s41598-017-05295-z.
Mahmoudi B, Panahi B, Mohammadi SA, Daliri M and Babayev MSh, 2014. Microsatellite based phylogeny and bottleneck studies of Iranian indigenous goat populations. Animal Biotechnology, 25(3): 210-222.
Maqsood M and Abdul M, 2017. Yeast extract elicitation increases vinblastine and vincristine yield in protoplast derived tissues and plantlets in Catharanthus roseus. Brazilian Journal of Pharmacognosy 27(5): 549-556.
Murashige T and Skoog F, 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15(3): 473-497.
Panahi B and Ghorbanzadeh Neghab M, 2013. Genetic characterization of Iranian safflower (Carthamus tinctorius) using inters simple sequence repeats (ISSR) markers. Physiology and Molecular Biology of Plants 19(2): 239-243.
Panahi B, Mohammadi SA and Doulati-Baneh H, 2019. Characterization of Iranian grapevine cultivars using machine learning models. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences doi.org/10.1007/s40011-019-01131-8.
Parry G and Estelle M, 2006. Auxin receptors: a new role for F-box proteins. Current Opinion in Cell Biology 18(2): 152-156.
Pérez-Alonso N and Jiménez E, 2011. Produccion de metabolitos secundarios de plantas mediante el cultivo in vitro. Biotecnología Vegetal 11(4): 195-211.
Pietrosiuk A, Furmanowa M and Łata B, 2007. Catharanthus roseus: micropropagation and in vitro techniques. Phytochemistry Reviews 6: 459-473.
Pop TI, Pamfil DC and Bellini C, 2011. Auxin control in the formation of adventitious roots. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 39(1): 307-316.
Quint M, Barkawi LS, Fan KT, Cohen JD and Gray WM, 2009. Arabidopsis IAR4 modulates auxin response by regulating auxin homeostasis. Plant Physiology 150(2): 748-758.
Ramawat KG, Bhansali RR and Arya HC, 1978. Shoot formation in Catharanthus roseus (L.) G. Don. callus cultures. Current Science 47: 93-94.
Ruzic DV and Vujovic T, 2008. The effects of cytokinin types and their concentration on in vitro multiplication of sweet cherry cv. Lapins (Prunus avium L.). HortScience 35(1): 12-21.
Salvi ND, George L and Eapen S, 2001. Plant regeneration from leaf base callus of turmeric and random amplified polymorphic DNA analysis of regenerated plants. Plant Cell, Tissue and Organ Cultre 66: 113-119.
Singh R, Kharb P and Rani K, 2011. Rapid micropropagation and callus induction of Catharanthus roseus in vitro using different explants. World Journal of Agricultural Sciences 76: 609-704.
Swanberg A and Dai W, 2008. Plant regeneration of periwinkle (Catharanthus roseus) via organogenesis. HortScience 43(3): 832-836.
Tiwari AK, Kumar G, Tiwari B, Kadam GB and Saha TN, 2017. Genetic diversity among turf grasses by ISSR markers. Indian Journal of Agricultural Sciences 87(2): 251-256.
Tank JG and Thaker VS, 2014. Systemic control of cell division and endoreduplication by NAA
         and BAP by modulating CDKs in root tip cells of Allium cepa. BioMed Research International  doi.org/10.1155/2014/453707.
Vidoz ML, Loreti E, Mensuali A, Alpi A and Perata P, 2010. Hormonal interplay during adventitious root formation in flooded tomato plants. Plant Journal 63(4): 551-562.
Werner ET, Soares TCB, Gontijo ABPL, Souza Neto, JD and do Amaral JAT, 2015. Genetic stability of micropropagated plants of  Crambe abyssinica Hochst using ISSR markers. Genetics and Molecular Research 14(4): 16450-16460.
Zhang QX, Shen YK, Shao RX, Fang J, He YQ, Ren JX, Zheng BS and Chen GJ, 2013. Genetic diversity of natural Miscanthus sinensis populations in China revealed by ISSR markers. Biochemical Systematics and Ecology 48: 248-256.
Journal of Plant Physiology and Breeding
Volume 9, Issue 1
June 2019
Pages 129-135

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