An Efficient Method for Agrobacterium-Mediated Genetic Transformation and Plant Regeneration in Cumin (Cuminum cyminum L.) Sonika Pandey & Avinash Mishra & Manish Kumar Patel & Bhavanath Jha Received: 20 March 2013 / Accepted: 17 June 2013 / Published online: 30 June 2013 # Springer Science+Business Media New York 2013 Abstract Cumin is an annual herbaceous medicinally important plant having diverse applica- tions. An efficient and reproducible method of Agrobacterium-mediated genetic transformation was herein established for the first time. A direct regeneration method without callus induction was optimised using embryos as explant material in Gamborg’ s B5 medium supplemented with 0.5-μM 6-benzyladenine and 2.0-μM α-naphthalene acetic acid. About 1,020 embryos (a mean of 255 embryos per batch) were used for the optimisation of transformation conditions. These conditions were an Agrobacterium cell suspension of 0.6 OD 600 , a co-cultivation time of 72 h, 300-μM acetosyringone and wounding of explants using a razor blade. Pre-cultured elongated embryos were treated using optimised conditions. About 720 embryos (a mean of 180 embryos per batch) were used for transformation and 95 % embryos showed transient β-glucuronidase expression after co-cultivation. Putative transformed embryos were cultured on B5 medium for shoot proliferation and 21 regenerated plants were obtained after selection and allowed to root. T 0 plantlets showed β-glucuronidase expression and gene integration was confirmed via PCR amplification of 0.96 and 1.28 kb fragments of the hygromycin-phosphotransferase II and β- glucuronidase genes, respectively. In this study, a transformation efficiency of 1.5 % was demonstrated and a total of 11 transgenic plants were obtained at the hardening stage, however, only four plants acclimatised during hardening. Gene copy number was analysed by Southern blot analysis of hardened plants and single-copy gene integration was observed. This is the first successful attempt of Agrobacterium-mediated genetic transformation of cumin. Keywords Agrobacterium . Cumin . Regeneration . Southern . Transformation . Transgenic Appl Biochem Biotechnol (2013) 171:1–9 DOI 10.1007/s12010-013-0349-1 Electronic supplementary material The online version of this article (doi:10.1007/s12010-013-0349-1) contains supplementary material, which is available to authorized users. S. Pandey : A. Mishra (*) : M. K. Patel : B. Jha (*) Discipline of Marine Biotechnology and Ecology, CSIR–Central Salt and Marine Chemicals Research Institute, G. B. Marg, Bhavnagar 364002 Gujarat, India e-mail: avinash@csmcri.org e-mail: bjha@csmcri.org