American Journal of Plant Sciences, 2014, 5, 1-6 Published Online January 2014 (http://www.scirp.org/journal/ajps ) http://dx.doi.org/10.4236/ajps.2014.51001 Efficient Regeneration System for Genetic Transformation of Mulberry (Morus indica L. Cultivar S-36) Using in Vitro Derived Shoot Meristems D. S. Vijaya Chitra 1* , Bhaskarrao Chinthapalli 1 , G. Padmaja 2 1 Department of Biology, College of Natural Sciences, Arba Minch University, Arba Minch, Ethiopia; 2 Department of Plant Sciences, School of Life Sciences, India University of Hyderabad, Hyderabad, India. Email: * ds_chitra@fastmail.fm Received November 13 th , 2013; revised December 14 th , 2013; accepted December 25 th , 2013 Copyright © 2014 D. S. Vijaya Chitra et al. This is an open access article distributed under the Creative Commons Attribution Li- cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In accordance of the Creative Commons Attribution License all Copyrights © 2014 are reserved for SCIRP and the owner of the intel- lectual property D. S. Vijaya Chitra et al. All Copyright © 2014 are guarded by law and by SCIRP as a guardian. ABSTRACT Shoot meristems used for the study were exercised from the in vitro regenerated shoots cultured on MS medium supplemented with 0.5 mg/L of BAP for multiplication. The sensitivity of the in vitro regenerated was studied using shoot meristems of 0.5 cm. Shoot meristems were cultured on medium containing 10 - 100 mg/l kanamycin to determine the concentration that was lethal for multiple shoot induction and root induction. The response of shoot multiplication decreased (66.2% - 6.2%) as the concentration of kanamycin increased (10.0 - 70.0 mg/L) with complete inhibition of shoot proliferation at 100 mg/L kanamycin. The rooting phase was very sensitive to kanamycin compared to shoot multiplication. The percentage of shoots that rooted decreased (53.8% - 4.8%) with increase in the concentration of kanamycin (10.0 - 70.0 mg/l) on IBA and 2,4-D supplemented medium. For transformation studies, the shoot tips that were infected with Agrobacterium strain were placed on selection me- dium containing MS medium with 0.5 mg/L BAP and 100 mg/L kanamycin and scored for the putative trans- formed shoots. An average of 62.2% of shoot tips developed shoot buds from the base and the shoots reached a length of 0.5 - 1.0 cm at the end of 30 days of culture on the selective medium in comparison to control which showed no response. An average of 66.7% of the regenerated plants showed GUS expression on selection me- dium where 43.2% and 65% of GUS expression was recorded in the leaves and callus. Leaves and callus induced from the controls did not show GUS activity. Stable integration of nptII gene with the genomic DNA from these transformed plants was confirmed through PCR analysis. Our result presents an efficient regeneration system using in vitro derived shoot meristems for Agrobacterium mediated gene transfer. KEYWORDS Morus indica L. Cultivar S-36; In Vitro Regeneration; Shoot Meristems; Kanamycin; Genetic Transformation 1. Introduction Mulberry (Morus spp.), a woody perennial tree plant plays a significant role in sericulture, as its foliage con- stitutes the main diet for silkworm (Bombyx mori L.). The most important factor in the management of sericul- ture is the improvement of mulberry cultivation for achieving higher leaf yields. Mulberry leaves are essen- tial for the survival of silkworms, since silkworms are monophagus insects, which grow only by feeding on mulberry leaves. Thus the cultivation of mulberry is the most important factor in the production of silkworm eggs, rearing of silkworm eggs, rearing of silkworm cocoons and on the whole in the entire operation of sericulture. Mulberry cultivation is fraught with many problems in the form of biotic and abiotic stresses. Among the vari- ous biotic stresses, fungal diseases cause a major damage to this crop [1]. Mulberry is improved qualitatively and quantitatively by conventional genetic approaches. How- * Corresponding author. OPEN ACCESS AJPS