Abstract
Many metabolites in leaf tissue disturbed plant genomic DNA isolation and always varied when leaves was harvested from different environments. Objective of this study was to investigate whether season, environment stress and refrigerated storage affect genomic DNA isolation of tung tree leaves. Five types of young leaves and two DNA isolation protocols, the recycling CTAB protocol I and II, were adopted to carry out the experiment. Our results showed that both leaf type and protocol affected DNA isolation of tung tree. Using the recycling CTAB protocol II, though little DNA were obtained from three types of young leaves, the other two have satisfying results. Whereas the recycling CTAB protocol I could produce high yield genomic DNA from all the five types of young leaves. All the detectable DNA samples in agarose gel electrophoresis were good templates for PCR reaction. Season, environment stress and refrigerated storage had a big effect on genomic DNA isolation of tung tree. The recycling CTAB protocol I was proved to be an effective and universal protocol for DNA isolation of tung tree. Five types of young leaves could all act as the tissue for isolation of genomic DNA, but the summer healthy young leaves without long-time refrigerated storage are the best. The optimal leaf tissue will benefit DNA isolation of plant species.
Highlights
Plant often produces many metabolites, such as polysaccharides, polyphenols, protein and tannin
Our results showed that both leaf type and protocol affected DNA isolation of tung tree
Environment stress and refrigerated storage had a big effect on genomic DNA isolation of tung tree
Summary
Plant often produces many metabolites, such as polysaccharides, polyphenols, protein and tannin These compounds always interfere with DNA isolation and downstream DNA-based experiments [1,2,3,4,5,6]. Leaf age was reported to affect the properties of extracted DNA, which was inferred to be related with the accumulation of defense compounds during the leaf development [19]. Environmental factors such as light intensity, temperature and seasonal variance were reported to affect the production of polysaccharide [25], phenolic compounds [28,29] and other metabolites in leaves. The recycling CTAB protocol I was proved to be a universal method for DNA isolation of tung tree
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