A rapid and simple method of assaying plants transformed with hygromycin or PPT resistance genes.

摘要

A simple leaf assay was developed that rapidly and reliably identifies transgenic plants expressing the hygromycin resistance gene hph or the phosphinothricin [glufosinate] resistance gene bar. Transgenic barley (cv. Schooner) and rice (cv. Taipei 309) plants containing hph or bar were produced by Agrobacterium-mediated transformation. Leaf tips were cut from plants propagated either in the glasshouse or in tissue culture and the cut surface embedded in solid medium containing the appropriate selective agent. Optimum concentrations of hygromycin and bialaphos [bilanafos] were 100 and 3 mg/litre, respectively. Non-transgenic barley or rice leaf tips had noticeable symptoms of either bleaching or necrosis after 3 days growth and were completely bleached or necrotic after 1 week. Transgenic leaf tips remained green and healthy over this period. This gave unambiguous discrimination between transgenic and non-transgenic plants. The leaf assay was also effective for the dicot plants tested (tobacco andpeas).