1 Gene duplication has played an important role in the evolution and domestication of flowering plants. 2 Yet little is known about how plant duplicate genes evolve and are retained over long timescales, 3 particularly those arising from small--scale duplication (SSD) rather than whole--genome duplication 4 (WGD) events. Here we address this question in the Poaceae (grass) family by analyzing gene 5 expression data from nine tissues of Brachypodium distachyon, Oryza sativa japonica (rice), and 6 Sorghum bicolor (sorghum). Consistent with theoretical predictions, expression profiles of most grass 7 genes are conserved after SSD, suggesting that functional conservation is the primary outcome of SSD 8 in grasses. However, we also uncover support for widespread functional divergence, much of which 9 occurs asymmetrically via the process of neofunctionalization. Moreover, neofunctionalization 10 preferentially targets younger (child) duplicate gene copies, is associated with RNA--mediated 11 duplication, and occurs quickly after duplication. Further analysis reveals that functional divergence of 12 SSD--derived genes is positively correlated with both sequence divergence and tissue specificity in all 13 three grass species, and particularly with anther expression in B. distachyon. Therefore, as found in 14 many animal species, SSD--derived grass genes often undergo rapid functional divergence that may be 15 driven by natural selection on male--specific phenotypes. 16 17 12 60 MYA (Bowers, et al. 2005; Bennetzen 2007; Paterson, et al. 2009; International Brachypodium 13 Initiative 2010). Grasses represent an interesting evolutionary system because they are agriculturally 14 important (Davidson, et al. 2012) and, thus, have undergone domestication events in their recent 15 evolutionary histories. Further, these three grass species are ideal for comparison due to the availability 16 of RNA--seq data from the same nine tissues (leaf, anther, endosperm, early inflorescence, emerging 17 inflorescence, pistil, embryo, seed 5 days after pollination, and seed 10 days after pollination) that were 18 obtained in a single lab under similar experimental conditions (Davidson, et al. 2012). Hence, we have a 19 powerful toolkit with which to assess expression divergence after SSD in grasses. 20 21 RESULTS 22 Retention mechanisms of SSD--derived duplicates in grasses Assis R, Bachtrog D. 2015. Rapid divergence and diversification of mammalian duplicate gene functions. 5 metabolic network gives rise to relative and absolute dosage constraints. The Plant Cell 23:1719--6 1728. 7 Bennetzen JL, Ma J, Devos KM. 2005. Mechanisms of recent genome size variation in flowering plants.