Smith A G, Gasser C S, Budelier K A, Fraley R T
Department of Horticultural Science, University of Minnesota, St. Paul 55108.
Mol Gen Genet. 1990 Jun;222(1):9-16. doi: 10.1007/BF00283016.
Differential screening of a tomato cDNA library produced from pre-anthesis stamens resulted in the isolation of 25 cDNA clones that hybridized to probes made from stamen RNA and showed no hybridization to probes made from RNA of vegetative organs. The 25 clones were found to represent 11 noncross-hybridizing classes. The majority of these clones were derived from genes that were single or low copy in the tomato genome. Northern RNA blotting experiments of vegetative and floral organs at several stages of development demonstrated that expression in all 11 classes was confined to floral organs. Of the 11 classes 9 were found to be expressed exclusively in stamens prior to anthesis. Two classes showed expression in immature stamens and in petals, with one of these two additionally being expressed in mature stamens at anthesis. Clones from three of the classes that were expressed exclusively in stamens were used as probes for in situ localization of RNA in floral organs. These experiments demonstrated that expression of the genes corresponding to these clones was confined to the tapetal cells of the anthers. Expression of one of the three genes was found to be limited to a single cell type during the 5-6 day period from late meiosis to immature pollen formation.
对花期前雄蕊构建的番茄cDNA文库进行差异筛选，分离出25个cDNA克隆，这些克隆与由雄蕊RNA制备的探针杂交，但与营养器官RNA制备的探针无杂交信号。发现这25个克隆代表11个非交叉杂交类别。这些克隆中的大多数来自番茄基因组中单一或低拷贝的基因。对营养器官和花器官在几个发育阶段进行的RNA印迹实验表明，所有11个类别的基因表达都局限于花器官。在这11个类别中，发现9个类别仅在花期前的雄蕊中表达。两个类别在未成熟雄蕊和花瓣中表达，其中一个类别在花期的成熟雄蕊中也有表达。从仅在雄蕊中表达的三个类别中选取的克隆用作花器官中RNA原位定位的探针。这些实验表明，与这些克隆对应的基因表达局限于花药的绒毡层细胞。在从减数分裂后期到未成熟花粉形成的5 - 6天期间，发现三个基因之一的表达仅限于单一细胞类型。
