Yamamoto Kotaro T, Watahiki Masaaki K, Matsuzaki Jun, Satoh Soichirou, Shimizu Hisayo
Division of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, 060-0810, Japan.
Biosystems Science Course, Graduate School of Life Science, Hokkaido University, Sapporo, 060-0810, Japan.
J Plant Res. 2017 Jul;130(4):765-777. doi: 10.1007/s10265-017-0932-6. Epub 2017 Apr 10.
Imaging analysis was carried out during the gravitropic response of etiolated Arabidopsis hypocotyls, using an IAA19 promoter fusion of destabilized luciferase as a probe. From the bright-field images we obtained the local deflection angle to the vertical, A, local curvature, C, and the partial derivative of C with respect to time, [Formula: see text]. These were determined every 19.9 µm along the curvilinear length of the hypocotyl, at ~10 min intervals over a period of ~6 h after turning hypocotyls through 90° to the horizontal. Similarly from the luminescence images we measured the luminescence intensity of the convex and concave flanks of the hypocotyl as well as along the median of the hypocotyl, to determine differential expression of auxin-inducible IAA19. Comparison of these parameters as a function of time and curvilinear length shows that the gravitropic response is composed of three successive elements: the first and second curving responses and a decurving response (autostraightening). The maximum of the first curving response occurs when A is 76° along the entire length of the hypocotyl, suggesting that A is the sole determinant in this response; in contrast, the decurving response is a function of both A and C, as predicted by the newly-proposed graviproprioception model (Bastien et al., Proc Natl Acad Sci USA 110:755-760, 2013). Further, differential expression of IAA19, with higher expression in the convex flank, is observed at A = 44°, and follows the Sachs' sine law. This also suggests that IAA19 is not involved in the first curving response. In summary, the gravitropic response of Arabidopsis hypocotyls consists of multiple elements that are each determined by separate principles.
利用不稳定荧光素酶的IAA19启动子融合体作为探针,在黄化拟南芥下胚轴的向重力性反应过程中进行了成像分析。从明场图像中,我们获得了相对于垂直方向的局部偏转角A、局部曲率C以及C对时间的偏导数[公式:见正文]。在将下胚轴旋转90°至水平方向后的约6小时内,每隔19.9 µm沿着下胚轴的曲线长度,以约10分钟的间隔确定这些参数。同样,从发光图像中,我们测量了下胚轴凸侧和凹侧以及下胚轴中线的发光强度,以确定生长素诱导的IAA19的差异表达。这些参数作为时间和曲线长度的函数进行比较,结果表明向重力性反应由三个连续的要素组成:第一次和第二次弯曲反应以及一个反弯曲反应(自动伸直)。第一次弯曲反应的最大值出现在下胚轴全长A为76°时,这表明A是该反应的唯一决定因素;相比之下,反弯曲反应是A和C的函数,正如新提出的重力感知模型所预测的那样(巴斯蒂安等人,《美国国家科学院院刊》110:755 - 760,2013)。此外,在A = 44°时观察到IAA19的差异表达,凸侧表达较高,且遵循萨克斯正弦定律。这也表明IAA19不参与第一次弯曲反应。总之,拟南芥下胚轴的向重力性反应由多个要素组成,每个要素都由不同的原理决定。