RESEARCH ARTICLE Assessment of Genetic Fidelity Using Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats Markers of Lawsonia inermis L. Plants Regenerated by Axillary Shoot Proliferation Arpita Moharana 1 • Aradhana Das 2 • Enketeswara Subudhi 2 • Soumendra K. Naik 1 • Durga P. Barik 1 Received: 9 August 2015 / Revised: 21 March 2016 / Accepted: 22 April 2016 Ó The National Academy of Sciences, India 2016 Abstract An efficient, reliable and reproducible plant regeneration protocol was developed for Lawsonia inermis L. using mature nodal explants. Shoot proliferation (81.6 %) with 7.8 shoots/explant was achieved on Mura- shige and Skoog’s (MS) medium supplemented with 1.0 mg l -1 6-benzyladenine. Shoot numbers were up- scaled by inducing multiple shoots from axenic nodal segments derived from the primary shoots on the shoot regeneration medium. Thus the authors could achieve ca. 129–134 shoots from single nodal explant. Ninety-three percent rooting of in vitro regenerated shoots was achieved on growth regulator free half-strength MS medium. Regenerated plantlets were successfully transferred to soil with 85 % survival rate. Genetic stability analyses of the in vitro regenerated plants using random amplified poly- morphic DNA and inter simple sequence repeat markers revealed a homogeneous amplification profile for all micropropagated plants. This is the first report that evalu- ates the use of molecular markers to establish genetic fidelity of micropropagated L. inermis for the rapid clonal multiplication and true-to-type production of plant for attaining the ever increasing demand in pharmaceutical industries. Keywords Inter simple sequence repeats  Mehendi  Nodal segment  Plant propagation  Random amplified polymorphic DNA Introduction Lawsonia inermis L. (Henna or Mehendi) belonging to family Lythraceae has been known for its medical and cos- metic purposes since centuries. The plant is native of Northern Africa and South-West Asia [1]. It is generally cultivated as a cash crop throughout India and particularly in Sojat area of Pali district, Rajasthan [2]. The dye prepared from the leaves of the plant is popular in India for its cosmetic use during wedding and other festivals to decorate palms, fingers, fingernails and soles [3]. It is also used for dyeing hair, leather, silk and wool [4]. Besides, the plant has been used for treatment of various diseases in traditional system of Indian medicine [5]. It has immunomodulatory, antiviral, antibacterial, antifungal, antifertility, hepatoprotective, anticancer, analgesic and anti-inflammatory properties [6, 7]. Leaves are used as a prophylactic against skin diseases, diarrhea and renal lithiase and gastric problems. The bark is used against jaundice, enlargement of spleen and leprosy. The seeds in powdered form are good medicine for dysen- tery, liver disorders and associated problems. The flowers have strong aroma and have been used for manufacturing of perfume since ancient times. The flowers have also medici- nal properties and are used as cardio tonic, refrigerant, soporific and febrifuge. Root of this plant is helpful in the treatment of gonorrhea, herpes, hysteria and nervous disor- ders [7, 8]. There is an increasing interest among people, both men and women, to dye their hair using dyes of herbal origin over synthetic dye due to reduced chance of side effects. & Soumendra K. Naik sknuu@yahoo.com & Durga P. Barik barikdp@gmail.com 1 Department of Botany, Ravenshaw University, Cuttack, Odisha 753 003, India 2 Centre of Biotechnology, Siksha O Anusandhan University, Bhubaneswar, Odisha 751030, India 123 Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. DOI 10.1007/s40011-016-0740-0