Soil alkalization is a global ecological problem that constrains food security and sustainable socio-economic development. As a wild relative of soybean, wild soybean (Glycine soja) exhibits strong salt and alkali stress resistance and its cotyledons play a key role during the emergence (VE) stage. This study aimed to compare variations in growth parameters, cotyledon ultrastructure, photosynthetic physiology, mineral ion and metabolite contents, and gene expression in two ecotypes of wild soybean to elucidate the regulatory mechanisms underlying alkali stress resistance in salt-tolerant wild soybean cotyledons during the VE stage. The results showed that salt-tolerant wild soybean cotyledons exhibited relatively stable growth parameters, dense and orderly chloroplast structure, high photosynthetic rates, as well as high K