Transgenic Plants from Ri-Transformed Callus of Datura innoxia. Alkaloid Content of Normal and Transformed Tissues PDF

Keywords

hairy roots, transgenic plants, transgenic callus

Abstract

Hairy roots were induced by inoculation of sterile plantlets of Datura innoxia with Agrobacterium rhizogenes. The hairy roots were separated and maintained on hormone-free liquid MS medium. They proliferated 30 fold, based on the initial fresh weight, after 4 weeks of subculturing. Some segments of the hairy roots proliferated into undifferentiated callus (transgenic callus), which in turn differentiated into transgenic shoots, upon transfer to the light on the same basal MS medium. The shoots were rooted by subculturing on either hormone-free medium or on MS medium supplemented with 0.3 mg/l IAA to yield transgenic plantlets. Genetic transformation into hairy roots, transgenic callus and plantlets was confirmed by chromatographic detection of opines in these tissues. The hairy roots gave the highest total alkaloids and hyoscyamine contents and was second to the normal plant in scopolamine production. The alkaloid content of the normal plant was higher than that of the tested transgenic plants, transgenic callus and normal callus.