Reeves C D, Krishnan H B, Okita T W
Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340.
Plant Physiol. 1986 Sep;82(1):34-40. doi: 10.1104/pp.82.1.34.
The developmental accumulation pattern of messenger RNA transcripts and polypeptides for wheat gliadins and ADPglucose pyrophosphorylase was determined using cDNA and antibody probes. Gliadin mRNA was detected on Northern and RNA dot blots at 3 days after flowering, it increased 100-fold by 10 days and decreased subsequent to 14 days. The abundant mRNAs encoding alpha/beta- and gamma-type gliadins and mRNA for ADPglucose pyrophosphorylase, a key regulatory enzyme of starch biosynthesis, accumulated coordinately. Despite the coordinate accumulation of their mRNA transcripts, the accumulation of gliadin and ADPglucose pyrophosphorylase polypeptides, as determined by Western blot, differed significantly. The time at which gliadin and ADPglucose pyrophosphorylase mRNAs began accumulating was also the time when the overall pattern of gene expression, as seen by two-dimensional gel electrophoresis of in vitro translation products, changed most significantly. However, the accumulation of a number of other mRNAs or polypeptides having unknown function occurred at other times during endosperm development. The pattern of expression in the earliest stages of development was strikingly similar to that of coleoptile, another rapidly growing, nonphotosynthetic tissue. Thus, the pattern of gene expression reflects the program of development observed cytologically.
利用cDNA和抗体探针确定了小麦醇溶蛋白和ADP葡萄糖焦磷酸化酶信使核糖核酸转录本和多肽的发育积累模式。在开花后3天的Northern印迹和RNA斑点印迹上检测到醇溶蛋白mRNA,到10天时增加了100倍,14天后下降。编码α/β-和γ-型醇溶蛋白的丰富mRNA以及淀粉生物合成的关键调节酶ADP葡萄糖焦磷酸化酶的mRNA协同积累。尽管它们的mRNA转录本协同积累,但通过蛋白质印迹法测定,醇溶蛋白和ADP葡萄糖焦磷酸化酶多肽的积累差异显著。醇溶蛋白和ADP葡萄糖焦磷酸化酶mRNA开始积累的时间,也是体外翻译产物二维凝胶电泳所显示的基因表达总体模式变化最显著的时间。然而,许多其他功能未知的mRNA或多肽的积累发生在胚乳发育的其他时期。发育最早阶段的表达模式与另一个快速生长的非光合组织胚芽鞘的表达模式惊人地相似。因此,基因表达模式反映了细胞学上观察到的发育程序。