Haemaphysalis longicornis Neumann, a tick of public health and veterinary importance, spend the major part of their life cycle off-host, especially the adult host-seeking period. Thus, they have to contend with prolonged starvation (sometimes for an extended period of 1-2 years). Here, we investigated the underlying molecular mechanism of tick starvation endurance in the salivary gland, midgut, ovary, and Malpighian tubule of starved H. longicornis ticks using the data-independent acquisition (DIA) quantitative proteomic approach to study the proteome changes. “Ribosome” was the most enriched KEGG pathway with the participation of more than thirty differentially expressed ribosomal proteins, indicating ribosomal stress. Essential synthases such as glutamate synthase, citrate synthase, ATP synthase, and ATP-citrate synthase were upregulated probably due to increased proteolysis and amino acid catabolism during starvation. The upregulation of succinate dehydrogenase, ATP synthase, cytochrome c oxidase, and ADP/ATP translocase closely fits with an increased oxidative phosphorylation function during starvation. The differential expression of superoxide dismutase, glutathione reductase, glutathione S-transferase, thioredoxin, thioredoxin reductase, and peroxiredoxin indicated fasting-induced oxidative stress. The upregulation of heat shock proteins could imply the activation of a protective mechanism that checks excessive protein breakdown during starvation stress. The survival of the ticks reduced after silencing of target genes via RNAi but was not significant relative to GFP-dsRNA injected ticks. The results of this study could provide useful information about the vulnerabilities of ticks that could aid in tick control efforts.