Using the traditional pedigree method and marker assistance selection （MAS） technology, we successfully bred a TGMS rice line——Longke 638S, with ideal plant type, low sterile conversion temperature （< 23.5℃）, stable fertility, good outcrossing and high combining ability. In 2014, Longke 638S was approved by Hunan Provincial Crop Variety Approval Committee. The whole-genome resequencing analysis showed that Longke 638S carried about 56.0% of the early-season indica and about 8.7% of the japonica genetic background, which laid a foundation for making full exploit of the double-heterosis of “early-season indica ecotype with late-season indica ecotype, indica with japonica subspecies”. Based on the theory of molecular design and heterosis groups, and the principle of “parental resistance genes complementation” and “parental quality genes uniformity”, a series of hybrid rice varieties from Longke 638S with disease resistance, high quality, wide adaptability and high yield were efficiently developed. This series of varieties quickly became the main hybrid rice varieties in southern China, and promoting the updating of middle-season indica hybrid varieties in the Yangtze River Basin.