ABSTRACT Ruffled vegetables, having wrinkled leaves with a crispy and tender texture, are highly favored by consumers. However, the genetic resources of crinkled leaf vegetables in the Brassica genus are limited. In this study, distant hybridization was performed between ‘Mottle-leaf Tai-cai’ (Brassica campestris L. ssp. chinensis (L.) Makino var. tai-tsai Hort. AA, 2n = 20) and ‘Big Yellow Flower Chinese Kale’ (B. oleracea var. alboglabra Bailey. CC, 2n = 18) by hand emasculation followed by tissue culture. As a result, all obtained three embryos were identified as true hybrids (AC, 2n = 19) through morphology, cytology, and molecular markers. After chromosome doubling, an allotetraploid hybrid (AACC, 2n = 38) was obtained with a giant leaf, serrated leaf margin, and wrinkled leaf surface; therefore, its most agronomical characters were biased toward the female parent of ‘Mottle-leaf Tai-cai’. In comparison with diploid hybrid, the pollen stainability of allotetraploids were significantly improved and similar to their parents. Additionally, all 5 pairs of SSR primers showed the authenticity of the hybrids. Furthermore, the contents of soluble sugar, vitamin C, and cellulose in the hybrids increased when compared with their parents. Briefly, the obtained hybrid is interspecific euhybrid, and has wrinkle-leaf characteristics, improved nutritional quality, so it is a novel Brassica wrinkle-leaf vegetable germplasm.