Telomere Length Detection of Ancient Tree Based on a Modified Southern Hybridization Assay Method

doi:10.11924/j.issn.1000-6850.casb18010104

Abstract

Abstract: Southern hybridization assay is reliable for detecting telomere length to predict plant age. However, secondary metabolites such as flavonoids, phenols, polysaccharides and terpenoids are rich in woody plants, which seriously affect the yield and quality of the extracted genomic DNA as well as subsequent results of enzyme digestion and hybridization. In this research, we modified the method of telomere length measuring by using 5 kinds of ancient trees collected from the Summer Palace in Beijing, on the basis of the conventional CTAB method, 2%-6% PVP (polyvinylpyrrolidone) was added to the DNA extracts of different tree species, which effectively removed the interference of secondary metabolites, therefore DNA could be digested completely by restriction enzyme HinfⅠ. At the same time, we also adjusted DNA content and the exposure time for the Southern hybridization assay, and detected telomere lengths of 3 kinds of trees. The results show that the average telomere length of ancient trees is longer than that of young trees. The improved Southern blotting assay is constructed and lays a foundation for predicting tree age related to telomere length.

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