Determination of Sensitivity Level of Hypocotyl Explants to Paromomycin Antibiotic and Optimization of Rooting in Transgenic Medicinal Plant Papaver somniferum

Document Type: Original Article

Authors

1 Faculty of Agriculture, Lorestan University, Khorramabad, Iran

2 Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

Abstract

Poppy (Papaver somniferum) is the most important commercial source of analgesic drugs and of compounds related to them. Because of the excellent medicinal prop- erties of poppy, finding genotypes with changed alkaloid content through breeding or by producing transgenic plants in tissue culture has attracted great interest. The present research was conducted to optimize rooting in somatic embryos of trans- genic poppy plants. Sensitivity of hypocotyl explants to the antibiotic paromomycin was first determined in kill-curve experiments. Transformation of the hypocotyl explants was carried out using the GV3101 strain of Agrobacterium tumefaciens having the vector with the NPT II gene. The explants were then transferred to MS- based medium containing the 2, 4-D plant growth regulator (PGR) at 1 mg/L, 10 mM MES, and paromomycinat 15 mg/L. After the somatic embryos were pro- duced, a factorial experiment was conducted in the rooting stage with two factors including medium (MS and 1/2 MS) and PGR (500 μM IBA PGR and no PGR). Also, confirmation of derived transgenic plantlets was confirmed by PCR tech- nique. Results of ANOVA and comparison of the means indicated the highest root- ing percentage happened in the 1/2 MS medium containing the IBA.