Optimization of Hormonal Combinations for In Vitro Regeneration of Lesser Periwinkle ( Vinca minor L.) and Assessment of
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RESEARCH ARTICLE
Optimization of Hormonal Combinations for In Vitro Regeneration of Lesser Periwinkle (Vinca minor L.) and Assessment of Genetic Homogeneity Saleh Amiri1 • Reza Fotovat1 • Alireza Tarinejhad2 • Bahman Panahi3 Seyyed Abolghasem Mohammadi4
•
Received: 29 April 2019 / Revised: 12 August 2019 / Accepted: 3 September 2019 The National Academy of Sciences, India 2019
Abstract Lesser Periwinkle (Vinca minor L.) is an important source of valuable secondary metabolites. To survey the effect of different combinations of plant growth regulators on the direct regeneration of Lesser Periwinkle, axillary and lateral bud segments of V. minor were used as explants. First, for shoot multiplication, Murashige and Skoog (MS) medium were fortified with four concentrations (0, 0.5, 1, and 1.5 mg l-1) of benzylaminopurine (BAP) in combination with three concentrations (0, 0.25, and 0.5 mg l-1) of indol-3-butyric acid (IBA). Then, root induction was assessed in full- and half-strength MS medium supplemented with three concentrations of IBA (0, 0.25, and 0.5). Finally, the genetic homogeneity of the
regenerated plant was subsequently assessed using 10 Inter Simple Sequence Repeat (ISSR) Markers. Higher shoot number per explant (6.50) and shoot length were obtained in MS medium containing the 1.5 mg l-1 BAP with 0.5 mg l-1 IBA and 0.5 mg l-1 BAP with 0.5 mg l-1 IBA, respectively. Moreover, the best root induction rate was observed on half-strength MS containing 1 mg l-1 IBA. PCR profiles with nine ISSR primers on the regenerated and donor plants also verified the genetic homogeneity of the regenerated plantlets. The present study introduced an efficient protocol for future large-scale propagation and production of secondary metabolites in Lesser Periwinkle. Keywords Genetic fidelity Growth regulators Lesser periwinkle Proliferation
Significance Statement Maximum shoot number (6.50 shoots/explant) and shoot length were obtained in MS medium containing the 1.5 mg l-1 BAP with 0.5 mg l-1 IBA and 0.5 mg l-1 BAP with 0.5 mg l-1 IBA, respectively. Moreover, the best root induction was observed on half strength MS containing 1 mg l-1 IBA. PCR profiles with nine ISSR primers in regenerated and donor plants also verified the genetic homogeneity of regenerated plantlets. & Reza Fotovat [email protected]; [email protected] Saleh Amiri [email protected] 1
Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Zanjan, Zanjan, Iran
2
Department of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran
3
Department of Genomics, Branch for Northwest and West Region, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Tabriz, Iran
4
Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
Introduction Lesser Periwinkle (Vinca minor L.) is a perennial subshrub, well known as medicinal and ornamental plant [1, 2]. More than
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