Poster: Emerging Technologies
Abs #
880: Extraction of total RNA from leaves of Eucalyptus and some other woody and herbaceous plants using sodium isoascorbate
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Presenter: |
suzuki, yuji , yujisuzu@cc.gifu-u.ac.jp |
Authors | suzuki, yuji (A) hibino, takashi (B) kawazu, tetsu (B) wada, tatsumi (C) kihara, tomonori (C) koyama, hiroyuki (C) | | Affiliations: |
(A): new energy and industrial technology development organization
(B): forestry research institute, oji paper co. ltd.
(C): laboratory of plant cell technology, faculty of agriculture, gifu university
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Eucalyptus is one of the attractive resources for the production of biomass and industrial materials, since it has good properties such as a fast growth rate, the ability to adapt to various environments and aptitudes as lumber, pulp and firewood. Further improvements in their properties would be related to breeding of transgenic Eucalyptus. To characterize such transformants, it is necessary to analyze changes in the patterns of transgene expression. However, RNA extraction from leaves of Eucalyptus is troublesome in many cases, possibly due to the high content of polyphenolics and polysaccharides. Here, a rapid and simple RNA extraction method using an extraction buffer containing sodium isoascorbate at a concentration of 500 mM was presented. This method consisted of one or two chloroform extractions, one acid guanidium-phenol-chloroform extraction and isopropanol precipitation alone. The yield of the RNA fraction were 1260 } 70 mg g-1 fresh weight and the contamination of the RNA fractions by proteins and polysaccharides was very limited as judged spectrophotometrically. When the RNA fraction was subjected to agarose gel electrophoresis, intact rRNA bands were detected. This RNA fraction could be used for reverse transcript-PCR. These results indicate that our new method achieves a simple and rapid preparation of high quality RNA from leaves of Eucalyptus. Furthermore, this method could be applied to leaves of some other woody and herbaceous plants such as Prunus mume, Thea sinensis, Osmanthus fragrans var. aurantiacus, Fagopyrum esculentum, Momordica charantia.
