The study of molecular phylogeny and pedigree preservation in marine macroalgae

在本项目中，我们展示了爱胜门(Laminariales，菲藻门)和Enierornorplia(Ulvales，Chiorophyta)的克隆化保存和紫菜的分子系统发育。这些结果总结如下：1)我们尝试在液氮中超低温保存赤子爱胜子和爱默子的配子体。这一结果表明，紫菜的存活率与以前建立的紫菜属红藻的冷冻保存技术没有太大差异。2)对烟草赤霉菌(TU-3、T-1、T-8)和条斑病菌(TU-1)的SSU rDNA序列进行了分析。这一结果表明条斑病菌TU-1与条斑病菌TU-3关系密切，而条斑病菌T-1和T-8是条斑病菌TU-1和条斑病菌TU-3.3的姊妹群。分支分析的表型数据分解了两个不同的群体，在突触形态的基础上，“生殖细胞的排列”，其中一个进一步分为两个群体。我们在紫菜中新测定了9个SSU rDNA全序列。基于SSU rDNA的系统发育分析表明，紫菜分为两个类群，其中一个类群进一步分为三个类群，这些研究取得了很好的结果。近年来，多态序列(GT)n和(CAC)n广泛分布于凤眼莲中。我们还通过斑点杂交证实了这些核苷酸重复序列。目前，我们正在构建条斑病菌Tu-L基因组文库，并用(GT)10和(CAC)10寡核苷酸探针筛选这些核苷酸重复序列。

In the present project, we have demonstrated clyopreservation of Eisenia (Laminariales, Pheophyta) and Enierornorplia (Ulvales, Chiorophyta) and molecular phylogeny of Porphyra. These results are summarized as follows : 1) We attempted to cryopreserve the gametophyte of Eisenia and Emeroinorpha in liquid nitrogen. This result indicated that survival rates were not extremely different from red algal genus Porphyra in which the cryopreserved techniques were established previously. 2) We analyzed SSU rDNA sequences of P.tenera (TU-3, T-1 and T-8) and P.yezoensis (TU-1). This result suggested that P.yezocnsis TU-1 was closely related to P.tenera TU-3 and P.tenera T-1 and T-8 were sister group to P.yezoensis TU-1 and P.frnera TU-3.3) We investigated phylogenetic relationship on the basis both for cladistic analysis of the phenotypic data and molecular phylogeny of SSU rDNA data. Cladistic analysis of the phenotypic data resolved two distinct groups, on the basis of synapomorphy, "arrangement of reproductive cells" and one of them was further divided into two groups. We newly determined nine complete sequences of SSU rDNA in Porphyra. Phylogenetic analysis based on the SSU rDNA indicated that Porphyra divided into two groups and one of them was further divided into three groups.These studies obtained excellent results. Recently, it was reported that polymorphic sequences (GT)n and (CAC)n are widely interspersed in Poiphyra. We also confirmed these nucleotide repeats by using the dot blot hybridization. At present, we are constructing a genomic plasmid library from P.yezoensis TU-l and al-c screening these nucleotide repeats with (GT)10 and (CAC)10 oligonucleotide probes.