Harrak H, Chamberland H, Plante M, Bellemare G, Lafontaine J G, Tabaeizadeh Z
Department of Biological Sciences, University of Quebec, P.O. Box 8888, Station Centre Ville, Montreal, Quebec, Canada H3C 3P8.
Plant Physiol. 1999 Oct;121(2):557-64. doi: 10.1104/pp.121.2.557.
A cDNA clone encoding a proline-, threonine-, and glycine-rich protein (PTGRP) was isolated from a wild tomato species (Lycopersicon chilense) (L.X. Yu, H. Chamberland, J.G. Lafontain, Z. Tabaeizadeh [1996] Genome 39: 1185-1193). Northern-blot analysis and in situ hybridization studies revealed that PTGRP is down-regulated by drought stress. The level of the mRNA in leaves and stems of 8-d drought-stressed plants decreased 5- to 10-fold compared with that in regularly watered plants. The mRNA re-accumulated when drought-stressed plants were rewatered. Antibodies raised against a glutathione S-transferase/PTGRP fusion protein were used to elucidate the subcellular localization of the protein by immunogold labeling. In regularly watered L. chilense plants, PTGRP protein was found to be localized in xylem pit membranes and disintegrated primary walls. Examination of sections from drought-stressed plants revealed a significant decrease in the levels of labeling. In these samples, only a few scattered gold particles were detected in the same areas. In the leaf tissues of plants that had been rewatered for 3 d following an 8-d drought stress, the labeling pattern was similar to that of the regularly watered plants. To our knowledge, PTGRP is the first drought-regulated protein that has been precisely localized in the cell wall.
从野生番茄物种(智利番茄,Lycopersicon chilense)中分离出一个编码富含脯氨酸、苏氨酸和甘氨酸的蛋白质(PTGRP)的cDNA克隆(L.X. Yu、H. Chamberland、J.G. Lafontain、Z. Tabaeizadeh [1996] Genome 39: 1185 - 1193)。Northern杂交分析和原位杂交研究表明,PTGRP在干旱胁迫下表达下调。与正常浇水的植株相比,干旱胁迫8天的植株叶片和茎中的mRNA水平下降了5至10倍。当干旱胁迫的植株重新浇水后,mRNA重新积累。利用针对谷胱甘肽S-转移酶/PTGRP融合蛋白产生的抗体,通过免疫金标记来阐明该蛋白的亚细胞定位。在正常浇水的智利番茄植株中,发现PTGRP蛋白定位于木质部纹孔膜和解体的初生壁中。对干旱胁迫植株切片的检查显示标记水平显著降低。在这些样品中,在相同区域仅检测到少数分散的金颗粒。在经历8天干旱胁迫后再浇水3天的植株的叶片组织中,标记模式与正常浇水的植株相似。据我们所知,PTGRP是第一个在细胞壁中精确定位的干旱调节蛋白。