Male reproductive development in rice is very sensitive to various forms of environmental stresses including low temperature. A few days of cold treatment (<20°C) at the young microspore stage induce severe pollen sterility and thus large grain yield reductions. To investigate this phenomenon, anther proteins at the early stages of microspore development, with or without cold treatment at 12°C, were extracted, separated by two-dimensional gel electrophoresis, and compared. The cold-sensitive cultivar Doongara and the relatively cold-tolerant cultivar HSC55 were used. The abundance of 37 anther proteins was changed more than 2-fold after 1, 2, and 4 days of cold treatment in cv. Doongara. Among them, one protein was newly induced, 32 protein spots were up-regulated, and four protein spots were down-regulated. Of these 37 protein spots, we identified two anther-specific proteins (putative lipid transfer protein and Osg6B) and a calreticulin that were down-regulated and a cystine synthase, a -6 subunit of the 20 S proteasome, an H protein of the glycine cleavage system, cytochrome c oxidase subunit VB, an osmotin protein homologue, a putative 6-phosphogluconolactonase, a putative adenylate kinase, a putative cysteine proteinase inhibitor, ribosomal protein S12E, a caffeoyl-CoA O-methyltransferase, and a monodehydroascorbate reductase that were up-regulated. Identification of these proteins is available upon request. Accumulation of these proteins did not vary greatly after cold treatment in panicles of cv. Doongara or in the anthers of the cv. Rice (Oryza sativa L.) evolved in tropical and subtropical areas, and as a result it has the characteristic of being vulnerable to cold weather. In particular, the combination of the low temperature sensitivity and a maximal sensitivity to these temperatures at precise stages of pollen microspore development (1) makes cool midseason temperatures the major environmental limitation (other than soil and water) for growing rice in temperate regions and at high elevations in the tropics (2). During the life cycle of rice, pollen microspore development at the booting stage is known to be the most susceptible to cool weather damage (3). Cold damage at this reproductive stage of rice causes limitation to the yields of temperate-grown rice around the world and can reduce yields to 40% (3, 4). The main cause of this damage is pollen sterility resulting from low temperature damage at the early stage of microspore development (1). The developmental stage most sensitive to the various forms of environmental stress, including cold damage, is just after meiosis. That is tetrad to early microspore phase, or the young microspore stage (5). In cold-treated rice plants, both cytological and histological abnormalities are greater in the anthers than in the pistil or other organs of the flower, and the cold damage can be rescued by artificial pollination with healthy pollen (5). In this process of cold damage, anthers become smaller, and cells in the tapetal layer in the anthers that surrounds a...