Rice is a major staple food worldwide. Making hybrid rice has proved to be an effective strategy to significantly increase grain yield. Current hybrid rice technologies rely on male sterile lines and have been used predominantly in indica cultivars. However, intrinsic problems exist in the implementation of these technologies, such as limited germplasms and unpredictable conversions from sterility to fertility in the field. Here, we describe a photoperiod-controlled male sterile line, carbon starved anther (csa), which contains a mutation in an R2R3 MYB transcription regulator of pollen development. This mutation was introduced into indica and japonica rice, and it rendered male sterility under short-day conditions and male fertility under long-day conditions in both lines. Furthermore, F 1 plants of csa and a restorer line JP69 exhibited heterosis (hybrid vigor), suggesting the feasibility of using this mutation to create hybrid rice. The csabased photoperiod-sensitive male sterile line allows the establishment of a stable two-line hybrid system, which promises to have a significant impact on agriculture.PGMS line | hybrid breeding | sugar partitioning R ice (Oryza sativa L.) is a major staple food that feeds more than one-half of the human population (1). The three-and two-line hybrid rice breeding technologies take advantage of heterosis (hybrid vigor) and have been successfully applied in many countries, leading to a more than 20% yield increase over inbred varieties (2). The three-line hybrid rice breeding system was developed in the 1970s after the revolutionary discovery of the wild abortive cytoplasmic male sterile cytoplasm (CMS-WA) for the breeding of CMS lines, which solved the problem of selfpollination in the varieties used for hybrid rice production (3). In this technology, a restorer line that contains the restorer gene(s) is used as the pollen donor to recover male fertility in F 1 plants (hybrid rice) after its cross with the CMS line (Fig. S1A). To maintain CMS, a maintainer line, which has viable pollen grains and normal seed sets, is used as the pollinator to cross with the CMS line (Fig. S1A). Although this system has been widely used for hybrid rice seed production, the requirement of a maintainer line with normal cytoplasm and a restorer line with nuclearencoded fertility restorer genes restricts breeders from exploiting more rice germplasms and complicates the procedures for generating hybrid seeds (4).As a replacement of the three-line technology, a two-line system has been developed and applied for hybrid rice seed production after the discovery of the environmentally sensitive genic male sterile lines controlled by nuclear recessive gene(s). A japonica cultivar, Nongken 58S (NK58S), was the first spontaneous photoperiod-sensitive genic male sterile (PGMS) mutant identified (5). Because indica varieties are predominant in most rice planting regions, the PGMS loci/locus in NK58S were/was introduced into indica to generate breeding cultivars, such as Peiai 64S (PA64S) and Xin'an S. These ...