Industrial Crops and Products 32 (2010) 366–374 Contents lists available at ScienceDirect Industrial Crops and Products journal homepage: www.elsevier.com/locate/indcrop Rapid plant regeneration, analysis of genetic fidelity and essential aromatic oil content of micropropagated plants of Patchouli, Pogostemon cablin (Blanco) Benth. – An industrially important aromatic plant Anamika Paul a,c , Ganesh Thapa a , Adreeja Basu b , Purabi Mazumdar b , Mohan Chandra Kalita c , Lingaraj Sahoo a,b,∗ a Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India b Center for Energy, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India c Department of Biotechnology, Gauhati University, Guwahati 781014, Assam, India article info Article history: Received 31 December 2009 Received in revised form 26 May 2010 Accepted 28 May 2010 Keywords: Essential oil Leaf explant Organogenesis Pogostemon cablin RAPD abstract Rapid and prolific shoot regeneration via direct organogenesis was induced from leaf explants of Patchouli, Pogostemon cablin (Blanco) Benth., an aromatic plant of immense industrial value, on Murashige and Skoog (MS) medium with benzylaminopurine (BAP) and -naphthaleneacetic acid (NAA) within 4 weeks. The adventitious shoot bud induction and plant regeneration were greatly influenced by the origin, age of donor plant, and leaf position on stem. Leaf explants prepared from in vivo plants of different ages showed higher regeneration response as compared to the explants from in vitro plantlets of respective age. The shoot regeneration ability of explants was significantly related to the age of the donor plants as well as the leaf position on the stem. The highest number of shoots (94.6/explants) was obtained from 96.2% of leaf explants derived from the leaves located on the second node of the 3-month- old in vivo plants on Murashige and Skoog’s (MS) medium containing 2.5 M benzylaminopurine (BAP) and 0.5 M naphthaleneacetic acid (NAA). Incorporation of 1.0 M gibberellic acid (GA 3 ) in MS medium significantly improved the shoot elongation (1.8-fold) within 2 weeks in 95% of the cultures. Regenerated shoots rooted spontaneously on growth regulator-free half strength MS medium and were successfully hardened and transferred to nursery with 96–100% survival rate. Genetic fidelity of the in vitro derived plants was assessed using random amplified polymorphic DNA (RAPD). Fourteen arbitrary decamers displayed same banding profile within all the micropropagated plants and in vivo explant donor plant. The molecular analysis complemented and compared well and showed genetic stability in the plants regenerated through direct shoot bud differentiation from leaf explants. The gas chromatogram of the extracted oils from in vitro derived plants and the mother stock plant showed similar essential oil profiles. Rapid and high multiplication frequency, molecular, genetic and essential oil content stability ensure the efficacy of the protocol developed for the production of this industrially important aromatic plant. © 2010 Elsevier B.V. All rights reserved. 1. Introduction Essential oil produced by Pogostemon cablin (Blanco) Benth. is one of the most important naturally occurring base materials used in the perfume industry which contains various sesquiterpenes and hydrocarbons such as: Patchouli alcohol (patchoulol), patchou- lene, bulnesene, guaiene, caryophyllene, elemene and copaene (Hasegawa et al., 1992). While 92% of the oil is non-odoriferous, the rest is made up of mixture of sesquiterpenes of which nor- ∗ Corresponding author at: Department of Biotechnology, Indian Institute of Tech- nology Guwahati, Guwahati 781039, Assam, India. Tel.: +91 361 2582204; fax: +91 361 2582249. E-mail address: ls@iitg.ernet.in (L. Sahoo). patchoulenol and (-patchoulene and -guaislene are major aroma compounds making it a complex contributing to its characteris- tic odor. The oil has strong fixative properties and blends very well with oils of sandalwood, geranium, vetiver, clove, etc. giv- ing strength and tenacity for making heavy perfumes of lasting odor. The absence of synthetic substitute for Patchouli oil, which has its characteristic woody fragrance and fixative properties, fur- ther enhances its prominent position in the perfume industry, and hence natural oil from cultivation remains the only source of this oil. In addition, Patchouli oil also possesses anti-insecticidal activi- ties (Sharma et al., 1992), anti-fungal and bacteriostatic properties (Kukreja et al., 1990) which are attributed to a number of dehy- droacetic acids. Due to its perfumery uses, the demand for Patchouli oil is increasing dramatically worldwide. The P. cablin plant is a tropical crop and areas of commercial cultivation are located mainly 0926-6690/$ – see front matter © 2010 Elsevier B.V. All rights reserved. doi:10.1016/j.indcrop.2010.05.020