Artificial selection on storage protein 1 possibly contributes to increase of hatchability during silkworm domestication

摘要

Like other domesticates, the efficient utilization of nitrogen resources is also important for the only fully domesticated insect, the silkworm. Deciphering the way in which artificial selection acts on the silkworm genome to improve the utilization of nitrogen resources and to advance human-favored domestication traits, will provide clues from a unique insect model for understanding the general rules of Darwin's evolutionary theory on domestication. Storage proteins (SPs), which belong to a hemocyanin superfamily, basically serve as a source of amino acids and nitrogen during metamorphosis and reproduction in insects. In this study, through blast searching on the silkworm genome and further screening of the artificial selection signature on silkworm SPs, we discovered a candidate domestication gene, i.e., the methionine-rich storage protein 1 (SP1), which is clearly divergent from other storage proteins and exhibits increased expression in the ova of domestic silkworms. Knockout of SP1 via the CRISPR/Cas9 technique resulted in a dramatic decrease in egg hatchability, without obvious impact on egg production, which was similar to the effect in the wild silkworm compared with the domestic type. Larval development and metamorphosis were not affected by SP1 knockout. Comprehensive ova comparative transcriptomes indicated significant higher expression of genes encoding vitellogenin, chorions, and structural components in the extracellular matrix (ECM)-interaction pathway, enzymes in folate biosynthesis, and notably hormone synthesis in the domestic silkworm, compared to both the SP1 mutant and the wild silkworm. Moreover, compared with the wild silkworms, the domestic one also showed generally up-regulated expression of genes enriched in the structural constituent of ribosome and amide, as well as peptide biosynthesis. This study exemplified a novel case in which artificial selection could act directly on nitrogen resource proteins, further affecting egg nutrients and eggshell formation possibly through a hormone signaling mediated regulatory network and the activation of ribosomes, resulting in improved biosynthesis and increased hatchability during domestication. These findings shed new light on both the understanding of artificial selection and silkworm breeding from the perspective of nitrogen and amino acid resources. Author summary Like other domesticates, nitrogen resources are also important for the only fully domesticated insect, the silkworm. Deciphering the way in which artificial selection acts on the silkworm genome to improve the utilization of nitrogen resources, thereby advancing human-favored domestication traits, will provide clues from a unique insect model for understanding the general rules of Darwin's theory on artificial selection. However, the mechanisms of domestication in the silkworm remain largely unknown. In this study, we focused on one important nitrogen resource, the storage protein (SP). We discovered that the methionine-rich storage protein 1 (SP1), which is divergent from other SPs, is the only target of artificial selection. Based on functional evidence, together with key findings from the comprehensive comparative transcriptome, we propose that artificial selection favored higher expression of SP1 in the domestic silkworm, which would influence the genes or pathways vital for egg development and eggshell formation. Artificial selection also consistently favored activated ribosome activities and improved amide and peptide biosynthesis in the ova, like what they may act in the silk gland to increase silk-cocoon yield. We highlighted a novel case in which artificial selection could directly act on a nitrogen resource protein associated with a human-desired domestication trait.