The study of molecular phylogeny and pedigree preservation in marine macroalgae

在本项目中，我们已经证明了clyopreservation的海带（Laminariales，Pheophyta）和Enierornorplia（Ulvales，Chiorophyta）和分子遗传学的紫菜。主要研究结果如下：1）尝试了用液氮超低温保存野牡丹属和野牡丹属的配子体。这一结果表明，存活率没有非常不同的红藻属紫菜，其中超低温保存技术建立以前。2)本研究分析了细叶对虾（TU-3、T-1和T-8）和条斑对虾（TU-1）的SSU rDNA序列。结果表明，条斑紫菜TU-1与细条紫菜TU-3亲缘关系较近，细条紫菜T-1、T-8与条斑紫菜TU-1、细条紫菜TU-3为姐妹群。分支分析的表型数据解决了两个不同的群体，在突触形态学的基础上，“生殖细胞的排列”，其中一个进一步分为两组。我们新测定了紫菜属9个SSU rDNA的全序列。基于SSU rDNA的系统发育分析表明，紫菜属可分为两个类群，其中一个类群又可进一步分为三个类群。近年来，有报道表明（GT）n和（CAC）n在紫菜中广泛分布。我们还通过斑点杂交证实了这些核苷酸重复序列。目前，我们正在构建条斑紫菜基因组质粒文库，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.