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Updated: Jul 30, 2026

Imaging Spatial Reorganization of a MAPK Signaling Pathway Using the Tobacco Transient Expression System
Published on: March 20, 2016
Methotrexate is a new selectable marker for tobacco immature pollen transformation
Tatiana Aionesei1, Julia Hosp, Viktor Voronin
1Max F. Perutz Laboratories at the Vienna Biocenter, Department of Genetics, Vienna University, Dr. Bohrgasse 9/4, A-1030, Vienna, Austria.
Abstract:
We report here a new selectable marker for tobacco immature pollen transformation based on the expression of dihydrofolate reductase (dhfr) gene which confers resistance to methotrexate (Mtx). Two immature pollen transformation approaches, i.e., male germ line transformation and particle bombardment of embryogenic mid-bicellular pollen have been used for the production of stable transgenic tobacco plants. In the first method, two methotrexate-resistant plants were selected from a total of 7161 seeds recovered after transformation experiments. In the second method, four methotrexate-resistant plants were obtained from 29 bombardments using 3.7 x 10(5) pollen grains per bombardment. Southern analysis confirmed the transgenic nature of T0 and T1 candidate transgenic plants, and a genetic analysis showed that the transgenes are transmitted to subsequent generations.
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