Although it is widely recognized that nutritional symbionts can manipulate host reproduction, the underlying molecular and cellular mechanisms are largely unclear. The facultative symbiont in bacteriocyte induces female-biased sex ratio of whiteflies. Here, we demonstrate that a maternal gene () and its encoded protein have lower expression levels in ovaries of cured whiteflies. Tud family proteins can interlink the various stages of biosynthesis of PIWI-interacting RNA (piRNA), a class of small noncoding RNAs. We find that affects the abundance of a piRNA through the maternal gene , thereby regulating the expression of the () (), which is the switch of activity of the vacuolar (H+)-ATPase that plays a crucial role in maintaining the homeostasis of intracellular energy and supporting mitochondrial respiration. This regulation adjusts the ATP level in ovaries of whiteflies. The ATP level shapes the F-actin pattern in ovaries and eggs of whiteflies, ultimately manipulating whitefly fertilization. Silencing inhibited whitefly fertilization by impairing ATP levels and F-actin patterns in ovaries and eggs. This study reveals that symbiont and maternal protein associations can regulate host fertilization by piRNA biosynthesis.