Ohno Ryoko, Takumi Shigeo, Nakamura Chiharu
Laboratory of Plant Genetics, Department of Biological and Environmental Science, Faculty of Agriculture, Division of Life Science, Graduate School of Science and Technology, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.
J Plant Physiol. 2003 Feb;160(2):193-200. doi: 10.1078/0176-1617-00925.
We studied the kinetics of induction of the wheat (Triticum aestivum L.) gene Wdhn 13, which encodes a predominantly cold-responsive protein belonging to the LEA D-11 dehydrin family. The deduced polypeptide WDHN 13 (MW = 12.8 kDa) represented the smallest dehydrin member in cereals with three lysine-rich K-segments. Purified WDHN 13 was boiling-stable due to its high hydrophilicity. Western blot analysis using the polyclonal anti-WDHN 13-antibody revealed a number of cross-reacting proteins in mature embryos and endosperms and in seedling leaves under normal temperature (25 degrees C), while a single major protein and a transcript were induced in response to low temperature (4 degrees C) in the leaves. An increase in the amount of mRNA was temporary with a peak occurring at day 3 to 5 of the low temperature treatment, while the protein accumulation proceeded with a significant time lag and continued until the end of the experiment (day 10). Steady-state levels of the transcript and protein were much higher in the leaves than in the roots of low temperature-treated seedlings and were apparently modulated by light/dark conditions. The light/dark modulation of the transcript and protein levels suggested stabilization of the mRNA under the low temperature condition. Genomic Southern blot analysis showed that Wdhn 13 is located on the homoeologous group 7 chromosomes, unlike all other wheat Dhn genes that are located on the group 6 chromosomes.
我们研究了小麦(Triticum aestivum L.)基因Wdhn 13的诱导动力学,该基因编码一种主要对低温响应的蛋白质,属于LEA D - 11脱水素家族。推导的多肽WDHN 13(分子量 = 12.8 kDa)是谷物中最小的脱水素成员,具有三个富含赖氨酸的K片段。纯化的WDHN 13因其高亲水性而具有热稳定性。使用多克隆抗WDHN 13抗体进行的蛋白质印迹分析显示,在正常温度(25℃)下,成熟胚、胚乳和幼苗叶片中有许多交叉反应蛋白,而在叶片中,低温(4℃)诱导产生单一主要蛋白和转录本。mRNA量的增加是暂时的,在低温处理第3至5天出现峰值,而蛋白质积累则有明显的时间滞后,并持续到实验结束(第10天)。低温处理幼苗的叶片中转录本和蛋白质的稳态水平远高于根,且明显受光/暗条件调节。转录本和蛋白质水平的光/暗调节表明在低温条件下mRNA得到稳定。基因组Southern印迹分析表明,Wdhn 13位于第7同源群染色体上,这与所有其他位于第6群染色体上的小麦Dhn基因不同。