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Identificaton and characterization of stamen- and tapetum-specific genes from tomato
Authors:Alan G Smith  Charles S Gasser  Kim A Budelier and Robert T Fraley
Institution:(1) Department of Horticultural Science, University of Minnesota, 55108 St. Paul, MN, USA;(2) Monsanto Co., 700 Chesterfield Village Parkway, 63198 St. Louis, MO, USA;(3) Present address: Department of Biochemistry and Biophysics, University of California, 95616 Davis, CA, USA
Abstract:Summary 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.
Keywords:cDNA cloning  Stamen development  Tapeturn  In situ hybridization  Differential screening
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