Department of Botany, University of Wisconsin, 53706, Madison, WI, USA.
Planta. 1987 Nov;172(3):371-7. doi: 10.1007/BF00398666.
The abundance and molecular mass of phytochrome in germinating embryos of A. sativa (oat) grown in light or darkness have been monitored using immunoblot and spectrophotometric assays. Immunoblot analysis shows that imbibed but quiescent embryos have two immunochemically distinct species of phytochrome with monomeric molecular masses of 124 and 118 kDa (kdalton). The 118-kDa species has the properties of the 118-kDa phytochrome extracted from fully green oat tissue (J.G. Tokuhisa, S.M. Daniels, P.H. Quail, 1985, Planta 164, 321-332), whereas the 124-kDa polypeptide appears similar to the well-characterized photoreceptor of etiolated tissue. The capacity of antibodies directed against etiolated-oat phytochrome to immunoprecipitate the 124-kDa species but not the 118-kDa species has been exploited to quantitate the levels of each separately over a 72-h time course of germination and seedling development. The abundance of the 124-kDa molecule increases at least 200-fold in etiolated seedlings over 72 h whereas in light-grown seedlings the level of this molecule is relatively constant. In contrast, the amount of the 118-kDa species increases only twofold in both dark- and light-grown seedlings over the same period of time. These data indicate that whereas the abundance of 124-kDa phytochrome is regulated at the protein level by the well-documented, differential stability of the red- and far-red-absorbing forms in vivo, the 118-kDa molecule is present at a low constitutive level, presumably reflecting no such difference in the stability of the two spectral forms.
用免疫印迹和分光光度法监测了在光或黑暗中生长的 A. sativa(燕麦)萌发胚胎中光敏素的丰度和分子量。免疫印迹分析表明,吸胀但静止的胚胎有两种免疫化学上不同的光敏素,单体分子量为 124 和 118 kDa(千道尔顿)。118 kDa 种具有从完全绿色燕麦组织中提取的 118 kDa 光敏素的特性(J.G. Tokuhisa、S.M. Daniels、P.H. Quail,1985,Planta 164,321-332),而 124 kDa 多肽似乎与发育良好的组织的典型光受体相似。针对黄化燕麦光敏素的抗体能够免疫沉淀 124 kDa 种但不能沉淀 118 kDa 种的特性被用来定量分析在萌发和幼苗发育的 72 小时时间过程中每种物质的水平。在黄化幼苗中,124 kDa 分子的丰度在 72 小时内至少增加了 200 倍,而在光照下生长的幼苗中,这种分子的水平相对稳定。相比之下,在黑暗和光照下生长的幼苗中,118 kDa 种的量在同一时间段内仅增加了两倍。这些数据表明,虽然 124 kDa 光敏素的丰度在蛋白质水平上受到体内红光和远红光吸收形式的已有充分记录的差异稳定性的调节,但 118 kDa 分子以低组成性水平存在,可能反映了两种光谱形式的稳定性没有差异。