Clark K R, Sims T L
Department of Molecular Genetics, Ohio State University, Columbus 43210.
Plant Physiol. 1994 Sep;106(1):25-36. doi: 10.1104/pp.106.1.25.
To determine the ability of isolated S-locus promoter sequences to direct organ-specific gene expression, we used microprojectile bombardment to introduce chimeric S-allele/beta-glucuronidase genes into different tissues of Petunia hybrida for transient expression. Histochemical staining showed that S-locus/beta-glucuronidase fusions were expressed in pistil, ovary, and petal tissue. No expression of the chimeric genes was detected in leaves or in mature pollen, either by histochemical staining or by fluorescence assays. RNA blot hybridization confirmed that low levels of S-locus mRNA accumulate in petals and ovaries in vivo. Analysis of the expression pattern of S-locus promoter deletions showed that sequences in the immediate vicinity of the TATA box were sufficient to confer qualitatively correct organ-specific expression of beta-glucuronidase. To further investigate the potential for S-ribonuclease expression in pollen, we used the polymerase chain reaction to amplify RNA accumulated in developing anthers. These assays demonstrated that mRNA for the S-ribonuclease accumulates to low levels in developing anthers several days prior to corolla opening and pollen anthesis. We discuss these results in light of current models of self-incompatibility.
为了确定分离的S-位点启动子序列指导器官特异性基因表达的能力,我们使用微粒轰击法将嵌合S-等位基因/β-葡萄糖醛酸酶基因导入矮牵牛的不同组织中进行瞬时表达。组织化学染色显示,S-位点/β-葡萄糖醛酸酶融合体在雌蕊、子房和花瓣组织中表达。通过组织化学染色或荧光测定法,在叶片或成熟花粉中均未检测到嵌合基因的表达。RNA印迹杂交证实,体内花瓣和子房中积累有低水平的S-位点mRNA。对S-位点启动子缺失表达模式的分析表明,TATA框紧邻区域的序列足以赋予β-葡萄糖醛酸酶定性正确的器官特异性表达。为了进一步研究花粉中S-核糖核酸酶表达的可能性,我们使用聚合酶链反应扩增发育中的花药中积累的RNA。这些测定表明,在花冠开放和花粉散粉前几天,发育中的花药中S-核糖核酸酶的mRNA积累到低水平。我们根据当前的自交不亲和模型讨论了这些结果。