Agrobacterium Protocols Vol. 2, 2nd Ed. (Front Matters Only) Kan Wang Humana Press 2006英文电子书.pdfVIP

METHODS IN MOLECULAR BIOLOGY™ 344 Agrobacterium Agrobacterium Protocols Protocols Second Edition Second Edition Volume 2 Volume 2 Edited by Edited by Kan Wang Kan Wang 1 Carrot (Daucus carota L.) Owen Wally, Jayaraj Jayaraman, and Zamir K. Punja Summary Plants are susceptible to infection by a broad range of fungal pathogens. Many horticul- turally important crop species lack adequate genetic resistance to disease. Studies on poten- tial mechanisms of disease resistance in plants have revealed the importance of a range of pathogenesis-related (PR) proteins with antifungal activity in reducing colonization of plant tissues by pathogens. We are evaluating a range of PR-proteins, through heterologous expression in transgenic carrot tissues, for their effects on fungal disease development. The protocols for carrot transformation with a thaumatin-like protein are described. In addition, the use of herbicide resistance as a selectable marker in carrot transformation is illustrated. In this protocol, petiole segments from carrot seedlings are exposed to Agrobacterium for 10–30 min and co-cultivated for 3 d, after which herbicide selection is imposed until embryogenic calli are produced after 8–12 wk. The transfer of the embryogenic calli to hormone-free medium yields transgenic plantlets. This genetic transformation protocol has suppo