DNA Phylogeny of Boreal Conifers from the Last Glacial Age in Japan

在最后一次冰河时代，本州岛的大部分地区被北方针叶林覆盖。从当时的沉积物中，有大量保存完好的圆锥体化石。但是，由于它们在形态上非常相似，仅通过形态特征在物种水平上进行鉴定是非常困难的。因此，我们试图从这些化石中提取毛泡体DNA，并通过DNA测序的方法对其进行鉴定。为此，我们收集了现存的云杉、热带云杉、油松、小叶云杉、大叶云杉、白云杉和石泽松等针叶树种，建立了基于DNA序列的系统发育树。将标本放入液氮中保存，提取总DNA，用聚合酶链式反应方法进行扩增。我们成功地从几个化石中读取了DNA序列。从青森县Dekijima(约2.4万年前)埋藏的森林中获得的化石DNA与青海云杉相同，而从青森县贡诺县(约1.36万年前)埋藏的森林中获得的DNA与青海云杉相同。在这两个地方都没有这些云杉物种的分布。从另外两个地方，我们还不能成功读取DNA。

The most parts of Honshu Island were covered by boreal conifer forests during the Last Glacial Age. From sedimentations of those age, there are much amounts of fossil cones with beautiful preservation. But, to identify them in species level only bymorphological characters is very difficult because of close similarity in morphology. Therefore, we tried to extract chioroplast DNA from those fossils and to identify there by mean of DNA sequence. For this purpose1 we collected extant conifers such as Picea bicolor r, P jezoensis, P glehnii, P koyamae, P.maximowiczii, P.palita , and P shirasawae and established phylogenetic tree basing on DNA sequence.Fossil cones were collected from four localities in northern Honshu. Those samples were kept in liquid nitrogen and extracted total DNA, then multiplied by mean of PCR method. We succeed to read DNA sequence from several fossils. DNA of fossils obtained from buried forest in Dekijima, Aomori Prefecture (about 24,000 years ago) was identical with Picea glehnii , while that from buried forest in Gonohe county, Aomori Prefecture (about 13,600 years ago) was identical with Picea koyamae. In both localities there is no distribution of those Picea species. From other two localities, we can not yet success to read DNA.