Arend Matthias, Stinzing Andrea, Wind Christa, Langer Katharina, Latz Andreas, Ache Peter, Fromm Jörg, Hedrich Rainer
FG Holzbiologie der TU München, Winzererstrasse 45, 80797 Münich, Germany.
Planta. 2005 Dec;223(1):140-8. doi: 10.1007/s00425-005-0122-y. Epub 2005 Oct 29.
In previous studies, we have shown that annual expression profiles of cambial and wood tissue with respect to the Shaker K+ channel PTORK correlate with cambial activity. To follow PTORK-gene activity on the cellular level, we isolated the respective promoter regions and generated transgenic Arabidopsis plants expressing the GUS gene under the control of the PTORK promoter. Cross-sections of petioles showed PTORK-driven signals predominantly in the xylem parenchyma surrounding the vessels and in the phloem. Antibodies raised against a unique N-terminal region of PTORK in histo-immunochemical analyses recognised this K+-release channel in growth-active poplar plants only. PTORK labelling was found in differentiating xylem cells (young fibres) and mature xylem (vessel-associated cells of the ray parenchyma). Patch-clamp measurements on fibre cell protoplasts, derived from young poplar twigs, identified outward-rectifying K+ channels as the major K+ conductance of this cell type, which resembled the biophysical properties of PTORK when expressed in Xenopus oocytes.
在之前的研究中,我们已经表明,形成层和木质组织中与摇床型钾离子通道PTORK相关的年度表达谱与形成层活性相关。为了在细胞水平上追踪PTORK基因的活性,我们分离了各自的启动子区域,并培育出了在PTORK启动子控制下表达GUS基因的转基因拟南芥植株。叶柄的横截面显示,PTORK驱动的信号主要出现在围绕导管的木质部薄壁细胞和韧皮部中。在组织免疫化学分析中,针对PTORK独特的N端区域产生的抗体仅在生长活跃的杨树植株中识别出这种钾离子释放通道。在正在分化的木质部细胞(幼嫩纤维)和成熟木质部(射线薄壁组织的与导管相关的细胞)中发现了PTORK标记。对来自杨树嫩枝的纤维细胞原生质体进行膜片钳测量,确定外向整流钾离子通道是这种细胞类型的主要钾离子传导途径,其类似于在非洲爪蟾卵母细胞中表达时PTORK的生物物理特性。