Noriaki Murakami

Systematics and evolutionary biology of the fern genusHymenasplenium (Aspleniaceae)

In Aspleniaceae,Hymenasplenium is a well-defined group with dorsiventral creeping rhizomes. Members ofHymenasplenium are widely distributed in the tropic zones of the world and have great variation in morphology and ecolgy, making it a good model group for plant systematics and evolutionary biology. I have worked on this group using techniques such as comparative morphology, ecology, cytology (for examining chromosomes and reproductive modes), phytochemistry and molecular biology. I considered the evolution of various phenetic characters based on a molecular phylogenetic tree which I recently obtained from sequence comparisons ofrbcL. In this paper, I will summarize the results.

Chloroplast DNA variation and the phylogeny ofAsplenium sect.Hymenasplenium (Aspleniaceae) in the New World tropics

Asplenium sect.Hymenasplenium is a well-defined group in Aspleniaceae, distinguished by several morphological and cytological characters. However, interspecific relationships in the section were not clear. In this paper, we report the phylogenetic relationships of 9 Neotropical species of sect.Hymenasplenium determined by chloroplast DNA restriction site variation analyses. From the obtained phylogenetic tree, two major clades: one withA. obtusifolium, A. riparium, A. volubile andA. repandulum and another withA. delitescens, A. ortegae, A. purpurascens, A. laetum andA. hoffmannii were recognized.Asplenium delitescens was shown to have a polyphyletic origin. It was also shown that the epiphytic habit evolved only once in the New World species of sect.Hymenasplenium.

Stereochemistry of 2-aminopimelic acid and related amino acids in three species of asplenium

Abstract We previously reported two free D -amino acids, D-2-aminopimelic acid ( D -APA) and trans -3,4-dehydro- D -2-aminopimelic acid (D-Δ-APA), from Asplenium unilaterale . In the present work we isolated 4-hydroxy-2-aminopimelic acid (OH-APA) from the same plant and determined it to be the α- L -form. We also investigated the configurations of these amino acids isolated from A. prolongatum and A. wilfordii which are morphologically distinct from A. unilaterale . In A. prolongatum , APA was the D - and OH-APA was the L -isomer. In contrast, APA from A. wilfordii was partially racemized and the degree of racemization was significantly different in plant material collected in July and November, L:D = 3:2 and 3:7, respectively. In A. wilfordii OH-APA was almost pure L - and Δ-APA was mostly the D -isomer.

d-2-Aminopimelic acid and trans-3,4-dehydro-d-2-aminopimelic acid from Asplenium unilaterale

Abstract d -2-Aminopimelic acid and a novel amino acid, trans-3,4-dehydro- d -2-aminopimelic acid were isolated and characterized from Asplenium unilaterale. Their distribution in Aspleniaceae was also investigated.

Chemotaxonomic studies ofAsplenium sect.Hymenasplenium (Aspleniaceae)

We previously isolated and characterized a new free amino acid withd-configuration at the α-carbon,trans-3, 4-dehydro-d-2-aminopimelic acid and its related amino acids,d-2-aminopimelic acid and 4-hydroxy-l-2-aminopimelic acid fromAsplenium unilaterale. In this paper, we report that the biosynthetic relationshps among these three amino acids were studied using14C-and3H-labeled compounds as tracers. Glutamate and aspartate were shown to be good precursors and it was suggested that 4-hydroxy-l-2-aminopimelic acid is biosynthesized first and the twod-amino acids are derived from it. Furthermore, the distribution patterns of these non-protein amino acids inAsplenium sect.Hymenasplenium were examined in detail and they were evaluated by their biosynthetic pathway. Morphological characters especially on their rhizomes were also examined and their character phylogeny was determined by outgroup comparison. Taking all the characters available into account, the phylogenetic relationship among 7 species ofAsplenium sect.Hymenasplenium in Japan and Taiwan is discussed by the transformed cladistic method.

Stelar structure of Asplenium obtusifolium and its allied species in the new world tropics, with comparison to the Asian members of Asplenium sect. Hymenasplenium

The stelar structure ofAsplenium obtusifolium and its related species (A. repandulum, A. riparium, A. triquetrum, A. volubile, A. purpurascens, A. ortegae, A. delitescens, A. hoffmannii, andA. laetum) in the New World tropics was observed and compared to that of Asian species ofAsplenium sect.Hymenasplenium. Both of the groups were found to share peculiar stelar structures: steles with two meristeles, a broader ventral and a narrower dorsal, each providing one of the two leaf traces; leaf gaps arranged in two rows between the dorsal and ventral strands, which are connected by thin meristeles, delimiting the leaf gaps. These structures are distinct from the radial symmetrical ones general inAsplenium. Together with cytological evidence, this strongly indicates that the New and Old World groups are closely related. Thus, these Neotropical species should be included in sect.Hymenasplenium.

Structure of cDNA clones for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from a fern (Asplenium cataractarum rosenstock, aspleniaceae), and its comparison with those of various seed plants

We isolated the small subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO SSu) from a fern,Asplenium cataractarum and determined its 34 N-terminal amino acid sequence. We obtained a cDNA clone that contains the entire coding region of the SSu from the same fern species, using synthetic oligonucleotide probes derived from the above amino acid sequence. It contains a 525 bp open reading frame capable of coding for a polypeptide with 174 amino acids, 31 bp 5′-and 206 bp 3′-noncoding regions. It was also elucidated that the precursor to the SSu contains a transit peptide of 53 amino acid residues and a mature protein of 121 residues. We compared the deduced amino acid sequence of the fern SSu with those of 11 other vascular plant species (including gymnosperms, monocots and dicots). As low as 55% homology was observed between those of a fern and seed plants. Constancy of the amino acid substitution rate in RuBisCO SSu was supported by our relative rate test. Amino acid substitution rate per year per site for RuBisCO SSu was calculated to be 0.81×10−9 assuming that the separation between pteridophytes and seed plants arose 380 million years ago.

Phylogeny ofTyphonium (Araceae) inferred from restriction fragment analysis of chloroplast DNA

The interrelationship of the ten species of the genusTyphonium and related genera in subtribe Arinae of the Araceae was inferred by chloroplast DNA restriction fragment analysis. A total of 42 site mutations were observed and 26 site mutations were shared by two or more species. A majority rule consensus tree was made by performing 100 bootstrap replicates using Wagner Parsimony. Two groups ofTyphonium were recognized significantly as monophyletic groups, i.e. 1)Typhonium larsenii andT. kunmingense, and 2)T. trilobatum, T. blumei andT. flagelliforme.