Stability of transgene expression in reduced allergen peanut (Arachis hypogaea L.) across multiple generations and at different soil sulfur levels.

Transgenic peanut (Arachis hypogaea L.) containing a gene designed for RNA interference (RNAi) showed stable complete silencing of Ara h 2 and partial silencing of Ara h 6, two potent peanut allergens/proteins, along with minimal collateral changes to other allergens, Ara h 1 and Ara h 3, across three generations (T3, T4, and T5) under field conditions. Different soil sulfur levels (0.012, 0.3, and 3.0 mM) differentially impacted sulfur-rich (Ara h 2, Ara h 3, and Ara h 6) versus sulfur-poor (Ara h 1) proteins in non-transgenic versus transgenic peanut. The sulfur level had no effect on Ara h 1, whereas low sulfur led to a significant reduction of Ara h 3 in transgenic and non-transgenic seeds and Ara h 2 and Ara h 6 in non-transgenic but not in transgenic peanuts because these proteins already were reduced by gene silencing. These results demonstrate stability of transgene expression and the potential utility of RNAi in allergen manipulation.