The application provides a domesticationgene THERMO TOLERANCE 5 (TT5) for regulating heat tolerance of plants and application thereof. In the application, a temperature-dependent Oryza minuta chromosome substitution line material for regulating rice quality and seed setting is identified, a QTL capable of polyfunctionally protecting yield and quality under high temperature is located and cloned by a method of map-based cloning, and the QTL is TT5. TT5 is a positive regulation transcription factor for heat tolerance, polyfunctionally regulates starch synthesis and storage protein accumulation, and endows high temperature tolerance. Cloning of TT5 has important significance for guiding de novo domestication of wild rice, and rapid domestication of wild rice can be realized by adding or erasing methylation of a genome.