Largemouth bass is one of the most important freshwater aquaculture species in China. However, the mechanisms underlying the development of skeleton in the teleosts are not fully comprehensive. In a previous study in our laboratory, it was found that the addition of phospholipids to the diet significantly reduced the malformation rate of largemouth bass larvae. The malformation rate decreased from 91.29 ± 1.05% in the fish fed phospholipid-deficient diet (SP_L) to 11.99 ± 1.19% in the fish fed with diet including 8% phospholipids (SP_H). Therefore, high-throughput RNA-Seq was used to compare the transcriptome of largemouth bass larvae between SP_H and SP_L. Thirty whole body samples originated from 3 tanks in 8% phospholipids inclusion group and 3 tanks in no phospholipids inclusion group were used to reduce inaccuracies. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the differentially expressed genes were involved in the mitogen-activated protein kinase (MAPK) signaling pathway, phosphatidylinositol signaling system, glycosphingolipid biosynthesis-lacto and neolacto series, and fatty acid degradation. Among the 584 differentially expressed genes, we paid attention to 6 genes known to be involved in skeletal development. Twist-related protein 2 (twist2) and dishevelled-associated activator of morphogenesis 1 (daam1), two genes related to osteoblast development, were up-regulated in SP_H. Bone gamma-carboxyglutamate (gla) protein (bglap), gene associated with the skeletal development and osteohormatology, was also up-regulated in SP_H. Procollagen C-endopeptidase enhancer b (pcolce b), a gene related to the development of gristle, was up-regulated in SP_H. Higher expression of secretory calcium-binding phosphoprotein 1 and 7 (scpp1 and scpp7) in SP_H suggests the stimulation of process related with tooth and bone development. The trend changes of the above genes all indicated that the lack of phospholipids might affect the skeletal development through the regulation of the expression of genes involved in several biological processes related to bone formation. In summary, those results provided valuable information about the reduction of deformity rates in largemouth bass and contributed to our understanding of the molecular mechanisms and regulative pathways regulating skeletal growth in larval carnivorous fish.