Os01g0201600

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Gibberellins (GAs) are a class of phytohormones that regulate many aspects of plant growth and development processes including stem elongation,flowering, and seed germination.
OsGAE1 is involved in various GAregulated developmental processes such as leaf sheath elongation, heading, and panicle development in rice.Elucidating the exact functions of OsGAE1 will contribute to understanding of GA action and the regulation of stem elongation and flower development in rice.

To study the hormonal and temporal expression patterns of OsGAE1 mRNA, expression was analyzed in response to different hormones during development.The effect of several plant hormones was examined on OsGAE1 mRNA abundance (Fig. 3A). The basal parts of rice seedlings were incubated with 5 lM each GA3,IAA, ABA, and 1 lM BL for 24 h. It was observed that GA3 up regulated the expression of OsGAE1 while there was little effect of other hormones such as BL, IAA, and ABA. GA dose-dependent expression patterns were determined in rice leaf sheaths. OsGAE1 expression was up-regulated with the increase in GA3 concentration and it continued to increase up to concentration of 50 lM GA3 (Fig. 3B). During temporal gene expression, OsGAE1 mRNA accumulation in leaf sheath segments increased with GA3 as its expression was up-regulated starting from 1 h and this effect was observed at 24 h as well (Fig. 3C). For more careful analysis, reverse transcription-PCR (RT-PCR) was used to examine GA time-dependent expression of OsGAE1. OsGAE1-specific oligonucleotides were used to amplify transcripts from total RNA isolated from leaf sheath segments treated with 5 lM GA3 for 0, 10, 30, 60, and 180 min. The expression of OsGAE1 increased starting from 10 min and continued to increase till 180 min (Fig. 3D).

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In order to examine the organ-specific expression of OsGAE1,total RNAs from rice callus, root, leaf blade, and leaf sheath were hybridized with OsGAE1-specific cDNA probe. A strong signal was detected in leaf sheath and callus but weak signal was observed in leaf blade, and root. Moreover, OsGAE1 expression in leaf sheath was increased when rice seedlings were treated with 5 lM GA3 for 24 h (Fig. 4A).In situ hybridization was performed using basal part of rice seedlings to learn more about the expression pattern. Hybridization of the longitudinal sections of rice basal part with gene-specific OsGAE1 antisense probe revealed the expression of OsGAE1 delimited to specific cell types, though significant hybridization was detected in shoot apex meristem region, young primary leaves, and vascular bundles (Fig. 4B, right). No significant signal was visible when sense probe of OsGAE1 was used (Fig. 4B, left).

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