The quality and weight of grain, which stores sugars and proteins, are controlled by nutrient import during the grain-filling period. Understanding the mechanism of nutrient import is therefore important for the improvement of agricultural productivity.We previously identified a 23 kDa protein, P23k, highly expressed in the scutellum of germinating barley (Hordeum vulgare L. cv Minorimugi) seeds and analyzed its characteristics in young seedlings (Kidou et al. 2006). The results showed that P23k is a paralogous protein of JIP-23, a jasmonic acid (JA) and methyl ester (MeJA)-induced protein identified in barley leaf (Andresen et al. 1992). Moreover, the expression of P23k was shown to be dependent on sugars and its localization was observed around membranes where sugar transport is active. Additionally, regardless of the high homology between P23k and JIP-23, P23k was not induced by JA and MeJA, and JIP-23 was not expressed at the young seedling stage. These results indicate that these two paralogous proteins have different roles in barley and suggest that P23k plays a role in sugar translocation. If this is correct, P23k should also be expressed in other organs such as the filling grain of developing seeds, which are active in sugar import. To test this hypothesis, we examined the expression and localization of P23k in barley filling grains. Moreover, to further clarify the functional difference between these two paralogous proteins, we also examined the expression of JIP-23 with P23k.The filling grain was divided into three stages ( Figure 1A): stage I (0ϳ3 days after fertilization), stage II (4ϳ6 days after fertilization) and stage III (7ϳ12 days after fertilization). In the scutellum of germinating barley seed, P23k was expressed higher than JIP-23 (Kidou et al. 2006). Based on this result, we expected the same expression pattern observed in filling grains. To confirm this, RT-PCR analysis was performed. Gene-specific reverse primers were used to differentiate expression of the two genes encoding P23k and JIP-23. Surprisingly, P23k mRNA was not detected in the PCR condition described in the figure legend, and only JIP-23 mRNA was observed in the filling grains ( Figure 1B). Additional reaction cycles were required to detect P23k mRNA (data not shown). In contrast, JIP-23 mRNA was not detected, and only P23k mRNA was observed in the scutellum. This result indicates that these two paralogous proteins have different roles in barley. It is possible that P23k acts on sugar transport in the scutellum of Abstract We previously showed the high expression of a 23 kD protein (P23k) in germinating barley seeds. In this study, we examined whether P23k is involved in supplying sugar to developing barley grains. RT-PCR analysis revealed that, although P23k expression was not detectable in developing grain, the 23 kD jasmonate-induced protein (JIP-23) exhibited a high level of RNA expression. JIP-23 belongs to the same protein family as P23k. Its remarkable expression was specifically confirmed at early development stage w...