Method of DNA Extraction from Pinus rigida Wood Pretreated with Sandpaper

Abstract

Species identification of wood provides important information for archaeology, restoration of cultural assets, preventing illegal logging, and more. Wood species are usually identified based on their anatomical features with the use of a microscope. However, this method may not be able to distinguish between anatomically similar species or subspecies. To overcome this problem, wood species need to be identified at the molecular level using DNA sequencing. However, unlike living plant cells, wood is difficult to pulverize using a mortar, and DNA extraction from dried wood is challenging.To solve these problems, we propose a pretreatment method in which wood is pulverized using 60-grit sandpaper and hydrated with water for 2 days. Using this method, we were able to stably amplify the rpoB gene from the extracted DNA of Pinus rigida. In addition, sequence analysis of the rpoB gene revealed six single nucleotide polymorphisms (SNPs), which classified the rpoB sequences in the genus Pinus into five groups. Our data indicate that although these SNPs were not suitable for species identification, they can potentially be used to determine the origin of different wood subspecies or individual samples of wood.

Genetic Species Identification Using ycf1b, rbcL, and trnH-psbA in the Genus Pinus as a Complementary Method for Anatomical Wood Species Identification

“…In previous studies, matK, rbcL, rpoB, and ycf1 were suggested as genes for DNA barcoding [31][32][33]. matK and rbcL were suggested as DNA barcodes for land plants by the Consortium for the Barcode of Life-Plant Working Group [34]. rpoB was suggested to be more efficient in family level identification than genus level identification [35].…”