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Light-harvesting chlorophyll a/b-binding (LHC) protein is a superfamily that plays a vital role in photosynthesis. However, the reported knowledge of LHCs in kiwifruit is inadequate and poorly understood. In this study, we identified 42 and 45 <i>LHC</i> genes in <i>Actinidia chinensis</i> (Ac) and <i>A. eriantha</i> (Ae) genomes. Phylogenetic analysis showed that the kiwifruit LHCs of both species were grouped into four subfamilies (Lhc, Lil, PsbS, and FCII). Expression profiles and qRT-PCR results revealed expression levels of <i>LHC</i> genes closely related to the light, temperature fluctuations, color changes during fruit ripening, and kiwifruit responses to <i>Pseudomonas syringae</i> pv. <i>actinidiae</i> (Psa). Subcellular localization analysis showed that AcLhcb1.5/3.1/3.2 were localized in the chloroplast while transient overexpression of AcLhcb3.1/3.2 in tobacco leaves confirmed a significantly increased content of chlorophyll a. Our findings provide evidence of the characters and evolution patterns of kiwifruit <i>LHCs</i> genes in kiwifruit and verify the <i>AcLhcb3.1/3.2</i> genes controlling the chlorophyll a content.