Research was done to determine if enhanced resistance to potato (Solanum tuberosum L.) late blight could be obtained by combining host plant resistance and engineered resistance. Late blight susceptible cultivars, Atlantic, and Spunta and the partially resistant cultivar Libertas were transformed with a fungal glucose oxidase gene, resulting in lines which ranged in transgene copy number from 1 to 8. Glucose oxidase enzyme activity ranged from 0.00 to 96.74×10-5 units/mg plant tissue. There was no correlation between copy number and level of transgene mRNA, level of transgene mRNA and enzyme activity, or between level of enzyme activity and disease resistance. Field and growth chamber evaluation of late blight response demonstrated little to no effect of the glucose oxidase transgene in either late blight susceptible or partially late blight resistant cultivars. However, enzyme activity levels were much lower than levels reported in previous research, which may account for the lack of effect of glucose oxidase against Phytophthora infestans. Twenty-one percent of the transgenic lines were phenotypically off-type compared to nontransgenic controls. Most of the off-type transgenic lines (four out of seven) were derived from `Libertas'. Because several off-type lines did not express the glucose oxidase protein, this phenomenon could not be attributed solely to the glucose oxidase transgene. Based on these results, transgenic lines produced for this study do not increase resistance to P. infestans even in combination with moderate host plant resistance. However, production of greater numbers of transgenic lines with the current construct or, production of transgenic lines in which a different constitutive promoter drives the expression of the glucose oxidase gene might result in greater disease resistance. However, the usefulness of any small increase in resistance would need to be evaluated against the time and cost required for development of transgenic potato cultivars and the potential for off-type tubers and plants.
A High-Throughput Screening System Based on Droplet Microfluidics for Glucose Oxidase Gene Libraries
