Several studies have shown the negative effects of mature plant resistance (MPR) on aphids in sugar beet, which is correlated to the formation of black deposits in their stomach. However, the underlying mechanism of MPR still needs to be elucidated, by understanding the toxicity effects of MPR on aphids and the role of the plant phenological stage and the environment. Here, we report that MPR in sugar beet does not only affect Myzus persicae mortality rate and the formation of a black deposit in the aphid stomach, but also aphid fecundity and behaviour. In addition, experiments in climate-controlled and field settings showed quantitative variation in MPR to M. persicae between six genotypes of sugar beet. Our results indicate that environmental effects, such as temperature, play a major role in MPR and underscore the importance of proper climate-controlled experiments for investigating MPR. In climate-controlled experiments, 83.3% of aphids on old leaves developed a black deposit, in contrast to only 16.8% of aphids on young leaves. This shows that not only plant age, but also leaf age plays a major role in the intensity of MPR. Further research will be needed to identify the underlying mechanism, before MPR can be used as a viable and sustainable solution to aphid pests in sugar beet.