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Efficient In Vitro Regeneration, Analysis of Molecular Fidelity and Agrobacterium tumifaciens - Mediated Genetic Transformation of Grewia asiatica L. | Abstract
Journal of Plant Biochemistry & Physiology

Journal of Plant Biochemistry & Physiology
Open Access

ISSN: 2329-9029

+44 20 3868 9735

Abstract

Efficient In Vitro Regeneration, Analysis of Molecular Fidelity and Agrobacterium tumifaciens - Mediated Genetic Transformation of Grewia asiatica L.

Wani TA, Satiander Rana, Bhat WW, Pandith SA, Niha Dhar, Sumeer Razdan, Suresh Chandra, Namrata Sharma and Surrinder K Lattoo

Grewia asiatica is a dietotheraphtically important fruit bearing shrub, indigenous to India. It is a rich resource of triterpinoids and flavonoids and possesses many putative health benefits. Two of the drawbacks which include short shelf life of its fruits and larger seed volume impedes its full exploitation. Seed abortion for developing seedless cultivars through biotechnological interventions is a viable option. One of the prerequisites for such strategy is to develop an efficient plant regeneration and transformation protocols in G. asiatica. Against this backdrop multiple shoot induction was achieved from nodal explants with axillary buds, on culturing in Woody Plant medium (WM) fortified with 3% (w/v) sucrose, 2 × 10-5M Kinetin (Kn) and 1 × 10-5M indole-3-butyric acid (IBA) giving rise to an average of 4.25 ± 0.71 microshoots per explant. More than 90% of the explants formed micro-shoots with mean shoot length of 10.5 ± 1.96 cm leading to whole plant regeneration. Healthy regenerated shoots showed prolific rooting of more than 95% on WM supplemented with 4.8 × 10-6M indole-3-butyric acid (IBA). Following simple hardening procedures, rooted plantlets, were transferred to soil-sand (1:1; v/v) with about 92% success. Genetic fidelity was assessed using random amplified polymorphic DNA (RAPD). Additionally, Agrobacterium-mediated genetic transformation protocol was developed using A. tumefaciens strain GV2260 harboring binary vector p35SGUSINT containing hygromycin phosphotransferase gene (hpt). Transformation was verified by GUS assay and detection of the hygromycin phosphotransferase (hpt) by polymerase chain reaction. In vitro regeneration and ensuing molecular fidelity of regenerated plants and transformation studies are hitherto unreported for G. asiatica.

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