Abstract
Abstract In situ hybridization techniques provide an exquisitely sensitive method to study tissue-specific or organ-specific patterns of gene expression. The techniques described in this chapter and based on refs 1--4, have been developed to study the expression of several genes involved in the biosynthesis of pigment (anthocyanin) in different floral organs (petal, style, stamen) of Antirrhinum majus. The methods involve hybridization of tritium-labelled single stranded RNA probes to mRNA in sections of fixed and wax embedded plant material. Hybridization is visualized using photographic emulsion, where radioactive emissions produce latent images in the emulsion which form silver grains when developed. The silver grains are visualized by dark-field microscopy. The techniques are applicable to a wide range of plant tissue and to different probes, including those involved in plant-pathogen interactions. For example, these techniques have been used at the John Innes Institute to study: (a) storage protein gene expression in developing pea embryos (A. Hauxwell, personal communication); (b) tobacco rattle virus RNA in and around infection sites on tobacco leaves (V.
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