Wednesday, November 7, 2007
259-7

Transformation of Taro (Colocasia esculenta) with a Wheat Oxalate Oxidase Gene Increased Resistance to Taro Leaf Blight.

Xiaoling He1, Susan Miyasaka1, and Y. Judy Zhu2. (1) University of Hawaii, 3190 Maile Way, St. John 102, Honolulu, HI 96822, (2) Hawaii Agriculture Research Center, 99-193 Aiea Heights Dr., Aiea, HI 96701

Taro (Colocasia esculenta) is one of the most important crops in the Pacific Islands; however, taro yields have been declining in Hawaii over the past 30 years partly due to diseases caused by oomycete and fungal pathogens. To engineer taro for increased disease resistance, methods for an efficient regeneration and transformation system of cv. Bun Long were established. Regenerative calli were initiated after shoot tip explants were grown on a Murashige and Skoog (MS) medium with 2 mg L-1 benzyl adenine (BA) and 1 mg L-1 napthalene acetic acid (NAA). Multiple shoots from these calli were induced on MS medium supplemented with 4 mg L-1 BA. Transformation of cv. Bun Long plants with a wheat oxalate oxidase gene (gf2.8) was achieved via Agrobacterium tumefaciens. Transformation was conducted using EHA105 harboring pBI121:gf2.8 that contains the genes neomycin phosphotransferase II (nptII), ß-glucuronidase (gus) and the intact gf2.8. Thirty putative shoot lines were regenerated from 200 calli under geneticin (G418) selection. Nine out of the ten lines that were examined showed strong GUS expression. Eight of these nine GUS positive lines also were confirmed to be transgenic using polymerase chain reaction (PCR), Southern blot, and reverse transcription (RT)-PCR analysis of the specific gf2.8 gene fragment, indicating both the presence and transcription of the gf2.8 gene in these transformed lines. In a laboratory-based bioassay, one transgenic line with the wheat oxalate oxidase gene showed increased resistance to taro leaf blight pathogen Phytophthora colocasiae. To our knowledge, this is the first report of a transformed taro line that exhibited increased disease resistance.