RNA In Situ Hybridization To Study Resveratrol Synthase (RS) Gene Expression In Vegetative Organs Of Peanut (Arachis Hypogaea L.)

Resveratrol (as a phytoalexin) is an antimicrobial substance, whichaccumulates in response to a pathogen and enhances the resistance within a plant. It is a kind of stilbenes that was found in a limited number of unrelated plant species, such as peanuts and grapevines. New roles of resveratrol as an antioxidant and antimutagenic agent of human health benefits, was first shown to act as its association with reduced the risk of coronary heart disease and has demonstrated therapeutic activities in a variety of tumors. This thesis details the research progress of the Resveratrol Synthase (RS) gene, and development and application of in situ hybridization (ISH). This research not only can be a method to identified and breed the transgenic farm crop, but also have the important leading meaning to the development of the resveratrol medicine.Using Resveratrol Synthase gene of Arachis hypogaea as a templet, synthesize Digoxigenin-labeled antisense RNA probe in vitro transcription with RNA polymerases. Utilizing the probe to detect specific RS gene expression within tissue sections of peanut vegetative orga'ns and regulation of the outsider inducement factor to the gene expression. In situ hybridization analysis revealed that the expression of RS gene was associated with phloem of vascular tissu and mesophyll cells. RS transcripts were predominant in phloem, but restricted to other tissues. Treated with yeast extract, the gene transcription level was increased. So we believed that the expression of RS gene has tissue specific expression pattern and inducible expression pattern.