Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the Alkaline Protease () Gene.

is a fungal pathogen that infects maize and produces aflatoxins. Host-Induced Gene Silencing (HIGS) has been shown to reduce host infection by various fungal pathogens. Here, the alkaline protease () gene was targeted for silencing through HIGS. An RNAi vector carrying a portion of the gene was incorporated into the B104 maize genome. Four out of eight transformation events containing the gene, Alk-3, Alk-4, Alk-7 and Alk-9, were self-pollinated to T4/T6 generations. At T3, the Alk-transgenic lines showed up to 87% reduction in aflatoxin accumulation under laboratory conditions. T4 transgenic Alk-3 and Alk-7 lines, and T5 and T6 Alk-4 and Alk-9 showed an average of 84% reduction in aflatoxin accumulation compared to their null controls under field inoculations ( < 0.05). F hybrids of three elite maize inbred lines and the transgenic lines also showed significant improvement in aflatoxin resistance ( < 0.006 to < 0.045). Reduced growth and levels of fungal DNA were observed in transgenic kernels during in vitro inoculation. Alk-4 transgenic leaf and immature kernel tissues also contained about 1000-fold higher levels of -specific small RNAs compared to null controls, indicating that the enhanced aflatoxin resistance in the transgenic maize kernels is due to suppression of infection through HIGS of gene.