GA3 and GA4 (gibberellins) play an important role in controlling gravitropism and tension wood formation in woody angiosperms. In order to improve our understanding of the role of GA3 and GA4 on xylem cell formation and the G-layer, we studied the effect of GA3 and GA4 and uniconazole-P, which is an inhibitor of GA biosynthesis, on tension wood formation by gravity in Fraxinus mandshurica Rupr. var. japonica Maxim. seedlings. Forty seedlings were divided into two groups;one group was placed upright and the other tilted. Each group was further divided into four sub-groups subjected to the following treatments: 3.43 × 10-9 μmol acetone as control, 5.78 × 10-8 μmol gibberellic acid (GA3), 6.21 × 10-8 μmol GA4, and 6.86 × 10-8 μmol uniconazole-P. During the experimental period, GAs-treated seedlings exhibited negative gravitropism,whereas application of uniconazole-P inhibited negative gravitropic stem bending. GA3 and GA4 promoted wood fibers that possessed a gelatinous layer on the upper side, whereas uniconazole-P inhibited wood formation but did not inhibit the differentiation of the gelatinous layer in wood fibers on the upper side. These results suggest that: (i) both the formation of gelatinous fibers and the quantity of xylem production are important for the negative gravitropism in horizontally-positioned seedlings; (ii) GA3 and GA4 affect wood production more than differentiation of the gelatinous layer in wood fibers;G-layer development may be regulated by other hormones via the indirect-role of GA3 and GA4 in horizontally-positioned F. mandshurica seedlings rather than the direct effect of GAs; and (iii) the mechanism for upward wood stem bending is different to the newly developed shoot bending in reaction to gravity in this species.