Micropropagation and DNA delivery studies in onion cultivars of Bellary, CO3
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J. Crop Sci. Biotech. 2015 (March) 18 (1) : 37 ~ 43 DOI No. 10.1007/s12892-014-0101-6 RESEARCH ARTICLE
Micropropagation and DNA Delivery Studies in Onion Cultivars of Bellary, CO3 Ashwini Malla1,2, Balamurugan Srinivasan1, Bala Murugan Shanmugaraj1, Sathishkumar Ramalingam1,2* Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, India Research & Development centre, Bharathiar University, Coimbatore, Tamil Nadu, India
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Received: September 29, 2014 / Revised: January 22, 2015 / Accepted: February 4, 2015 Ⓒ Korean Society of Crop Science and Springer 2015
Abstract Onion is one of the most important vegetables grown all over the world. In the present study, rapidly growing and maintainable callus was accomplished using the basal meristem plate and twin scale leaves as explants in Bellary and CO3 varieties for callus induction and in vitro bulb formation. The two cultivars were transformed with Agrobacterium tumefaciens EHA 105 harboring the binary vector pCAMBIA 1301 containing the GUS gene with different concentrations of acetosyringone. The outcome of this study revealed that the frequency of callus induction was maximum on Murashige and Skoog medium (MS) fortified with B5 vitamins supplemented with 0.5 and 1 mg L-1 picloram. Regeneration of plantlets from the callus was observed on MS supplemented with 0.5 mg L-1 each of BAP and KIN and 0.1 mg L-1 NAA. In vitro bulb response was observed on MS with B5 vitamins supplemented with 2.0 mg L-1 BAP. GUS gene integration in the transgenic plants was confirmed by PCR. The maximum GUS gene expression was observed at a concentration of 150 µM acetosyringone in the transformed plants by histochemical assay. This protocol allows effective genetic transformation and regeneration system of onion via in vitro bulb development. Key words: bulb formation, callus, genetic transformation, onion, regeneration Abbreviations 2,4-D: 2,4-Dichlorophenoxy Acetic Acid, BAP: 6-Benzyl Amino Purine, KIN: Kinetin, NAA: α-Naphthalene Acetic Acid, IAA: Indole Acetic Acid, IBA: Indole Butyric Acid, TDZ: Thidiazuron, Me Ja: Methyl Jasmonic Acid, MS: Murashige and Skoog Medium, B5: Gamborg’s Vitamins
Introduction Nature has an infinite array of gifts to mankind among which herbs occupy an imperative role. Herbal resurgence is in bloom in the world owing to the medicinal substances present that are known to modern and ancient civilizations for their healing properties. Allium cepa belongs to the genus Allium and family Alliaceae or Liliaceae, commonly known as bulb onion or onion which is one of the most important commercial edible herbs consumed all over the world. Herbal medicines have become more popular in recent years because they don’t have many side effects, adverse or toxic effects Sathishkumar Ramalingam(
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E-mail: [email protected]
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when compared to synthetic medicines. Onion has a number of ethnomedical uses such as treatment of cold, allergie
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