Insect herbivores of different feeding guilds induced sorghum defenses through differential mechanisms, regardless of the order of herbivore arrival on sorghum plants. Sorghum, one of the world's most important cereal crops, suffers severe yield losses due to attack by insects of different feeding guilds. In most instances, the emergence of these pests are not secluded incidents and are followed by another or can also co-infest host plants. Sugarcane aphid (SCA) and fall armyworm (FAW) are the two most important destructive pests of sorghum, which belongs to sap-sucking and chewing feeding guilds, respectively. While the order of the herbivore arriving on the plants has been found to alter the defense response to subsequent herbivores, this is seldom studied with herbivores from different feeding guilds. In this study, we investigated the effects of sequential herbivory of FAW and SCA on sorghum defense responses and their underlying mechanism(s). Sequential feeding on the sorghum RTx430 genotype by either FAW primed-SCA or SCA primed-FAW were monitored to unravel the mechanisms underlying defense priming, and its mode of action. Regardless of the order of herbivore arrival on sorghum RTx430 plants, significant defense induction was observed in the primed state compared to the non-primed condition, irrespective of their feeding guild. Additionally, gene expression and secondary metabolite analysis revealed differential modulation of the phenylpropanoid pathway upon insect attack by different feeding guilds. Our findings suggest that priming in sorghum plants upon sequential herbivory induces defense by the accumulation of the total flavonoids and lignin/salicylic acid in FAW primed-SCA and SCA primed-FAW interaction, respectively.