Light-harvesting chlorophyll a/b binding (Lhc) family proteins play a significant role in photosynthetic processes. Our objective was systematic identification and analysis of the Lhc family in cotton, as well as the relationship between Lhc family genes and chlorophyll synthesis during photosynthetic processes. We used genome-wide identification, phylogenetic analysis, chromosomal distribution and collinearity to examine potential functions of Lhc superfamily genes in upland cotton. Subcellular localization, qRT-PCR, a yeast two hybrid (Y2H) , and Virus-induced gene silencing (VIGS) experiment were used to explore function of GhLhcb2.3. Focusing on GhLhc family, gene structural analysis of G. hirsutum Lhc family genes (GhLhc) indicated the conservation of selected Lhc family members. The expression pattern of GhLhc proteins shows that Lhc family proteins are important for photosynthetic processes in leaves. Results of subcellular localization and qRT-PCR in different cotton varieties showed that GhLhcb2.3 is closely related to chloroplast chlorophyll. Y2H found extensive heteromeric interactions between the GhLhcb2.3 and GhLhcb1.4. Subcellular localization revealed that GhLhcb1.4 is located in chloroplasts. VIGS showed that GhLhcb2.3 influenced chlorophyll a synthesis. We comprehensively identified Lhc family genes in cotton, characterized these genes and reveal the influence of GhLhcb2.3 on chlorophyll a synthesis.