Abstracts of Posters


P-01 : Molecular Diversity of diploid populations of Dendranthema zawadskii and its related species (Asteraceae)
Jung Sung KIM*1, Hiroshi AZUMA1 and Hiroshi TOBE1
(1Department of Botany, Graduate School of Science, Kyoto University, Kyoto 606-8502, JAPAN; 2Institute of Biotechnology, Yeungnam University, Gyeongsan 712-749, KOREA )
Dendranthema, comprising about 37 species mostly distributed in East Asia, is greatly diversified morphologically. Phylogenetic relationships within the genus based on molecular data have not been analyzed so far because of the lack of diploid individuals. Recently, however, we have reported that diploid individuals occur in four populations of D. zawadskii and D. indicum in Korea in which otherwise only polyploids have been known. In this paper, based on analyses of chloroplast DNA sequences and allozyme, we report the genetic diversity in the diploid individuals of D. zawadskii and related species from Korea as well as in part from Japan, mainland China and Taiwan. For the molecular phylogenetic analyses, 39 populations of 8 species of Dendranthema were used along with one species of Ajania, and also one species of Nipponanthemum was used as out-group. About 2,603 bp from 6 non-coding regions of chloroplast DNA were determined and used for constructing phylogenetic trees. For the allozyme analyses, the frequency of alleles was investigated on the basis of 14 loci of 9 isozymes using only diploid Korean taxa and constructed a cladogram. The cpDNA trees showed that Ajania was included in Dendranthema, and that very complicated relationships were formed among the species. In all the species examined, the species did not form a monophyletic clade because the populations of each species were positioned at different clades. Moreover, several populations of different species shared the same haplotype and thus were placed in the same clade. The cladogram obtained from the allozyme data was similar to the DNA tree. Three major clusters, each composed of different populations, were formed. Therefore, the morphological species concept was not supported by molecular data.The results suggest that the gene flow has occurred among D. zawadskii and its related species, so that each species has such a high genetic diversity. This agrees well with the suggestion given on the basis of karyotype analyses of metaphase chromosomes in our previous study. It seems likely that the gene flow has occurred not only among the species of Dendranthema but also even between Dendranthema and Ajania.

P-02 : Molecular Phylogeny of tribe Forsythieae (Oleaceae) based on Nuclear Ribosomal DNA Internal Transcribed Spacers and Plastid DNA matK gene Sequences
Dong-kap KIM1, Chansoo KIM2, Mark W. CHASE3 and Joo-hwan KIM*1
(1Division of Life Science, Daejeon University, Deajeon 300-716, Korea; 2Dept. Forest Genetic Resources, Korea Forest Research Institute, Suwon 441-350, Korea; 3Jodrell Lab., Royal Botanic Gardens, Kew, Surrey TW9 3DS, UK )
The tribe Forsythieae Taylor ex Johnson (Oleaceae) comprises two genera(Abeliophyllum Nakai and Forsythia Vahl), which are mostly distributed on Far Eastern Asia except only one species (Forsythia europaea Degen et Bald.) in southeast europe. The genus Forsythia comprises approximately 12-15 species, and it is distinguished from related genera by some distinct morphological characters such as capsule, membraneous or vacant pith and 4-merous yellow flowers. And Abeliophyllum distichum Nakai is a Korean monotype which is distributed on middle part of South Korea. Although taxonomists have suggested many results on the systematic relationships of the tribe Forsythieae and the taxonomic status of Abeliophyllum distichum, there is no confidential agreement up to now. But those ones can be summarized as three opinions: 1) Abeliophyllum shows closer relationships with Fontanesia than Forsythia based on the samara, anatomical characters and palynological characters, 2) Abeliophyllum is the most related taxon with Forsythia based on basic chromosome numbers and molecular systematic data, 3) there is no congruence by flavonoid and chemotaxonomic results. Recently, Kim et al (2001) conducted the morphological review of Korean Forsythieae and discuss the systematic relationships among the Forsythia, Abeliophyllum and Fontanesia based on the numerical and RAPD analyses. But unfortunately, most of previous molecular systematic data included very small, represented and limited taxa of tribe Forsythieae. We sequenced the nuclear ribosomal DNA ITS region including 5.8S coding gene and plastid DNA trnK sequences to discuss the phylogenetic relationships among the species of tribe Forsythieae and its related genera. Also, we tried to reevaluate the appropriate sister group and outgroup of tribe Forsythieae.

P-03 : Phylogenetic Relationship in the Genus Panax: inferred from Chloroplast trnK Gene and Nuclear 18S rRNA Gene Sequences
Shu ZHU*
(Institute of Natural Medicine, Toyama Medical and Pharmaceutical University)
Shu ZHU1,3, Hirotoshi FUSHIMI1, Shaoqing CAI3, Katsuko KOMATSU1,2(1Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Japan;221st Century COE Program, Toyama Medical and Pharmaceutical University, Japan;3Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, China.) Chloroplast trnK gene and nuclear 18S rRNA gene sequences of 13 Panax taxa, collected mainly from Sino-Japanese floristic region, were investigated in order to reconstruct phylogenetic relationship and to assist taxonomic delimitation within this genus. The length of trnK gene sequence varied from 2537 bp to 2573 bp according to the taxa, whereas matK gene sequences, embedded in the intron of trnK gene, were of 1512 bp in all taxa. Species-specific trnK/matK sequence provided much insight into phylogeny and taxonomy of this genus. 18S rRNA gene sequences were of 1808 or 1809 bps in length, only 10 types of 18S rRNA sequences were observed among 13 taxa. Parsimony and Neighbor-joining analyses of the combined data sets of trnK-18S rRNA gene sequences yielded a well-resolved phylogeny within genus Panax, where three main clades were indicated. P. pseudoginseng and P. stipuleanatus formed a sister group located at a basal position in the phylogenetic tree, which suggested the relatively primitive position of these two species. Monophyly of P. ginseng, P. japonicus (Japan) and P. quinquefolius, which are distributed in northern parts of Asia or America, was well supported (Northern Clade). The remaining taxa distributed in southern parts of Asia formed a relatively large clade (Southern Clade). The taxonomic debated taxa traditionally treated as subspecies or varieties of P. japonicus or P. pseudoginseng showed various nucleotide sequences, but all fell into one cluster. It might suggest these taxa are differentiated from a common ancestor and are in a period of high variation, which is revealed not only on morphological appearance, but also on molecular divergence. A new variety of Panax L. has been discovered during our field investigation in Yunnan province of China, and named as P. vietnamensis Ha et Grushv. var. fuscidiscus K. Komatsu, S. Zhu & S. Q. Cai. Furthermore, quantitative analysis of 11 ginsenosides with 4-type aglycones which are widely distributed in Panax taxa with relatively high content revealed that each taxon possessed its own characteristic pattern and similarities of constitution were observed among the phylogenetically-related taxa, except P. japonicus and P. zingiberensis.

P-04 : Molecular phylogeny in subtribe Aeridinae (Orchidaceae) based on matK and ITS sequence
Topik HIDAYAT*1, Tomohisa YUKAWA2 and Motomi ITO1
(1Department of Biological Science,Graduate School of Science,The University of Tokyo; 2Tsukuba Botanical Garden,National Science Museum )
Phylogenetic analysis using two regions matK and ITS has been performed to clarify the relationships among 88 taxa in subtribe Aeridinae, one of the most complicated groups in Orchidaceae. Since analyses of matK sequences provided results highly concordant with those via ITS sequences, combined data sets were used for further analysis. The result showed many unexpected relationship in the subtribe. The subtribe included the following two major clades: Clade 1, which is characterized by absence of distinct column foot, consists of Robiquetia, Gastrochilus, Luisia, Armodorum, Stereochilus, Dyardorchis, Renanthera and others; Clade 2 is represented by Vanda, Dyakia, Brachypeza, Arachnis, Phalaenopsis, Acampe, Pomatocalpa, Trichoglottis, Pelatantheria and Thrixspermum. Several monophyletic groups uncovered in this study have been discussed.

P-05 : Dating the Monocot-Dicot Divergence and the Origin of Core Eudicots Using Whole Chloroplast Genomes
Shu-miaw CHAW1, Chien-chang CHANG*1, Hsin-liang CHEN1 and Wen-hsiung LI2
(1Institute of Botany, Academia Sinica, Taipei, 115, TAIWAN; 2Department of Ecology and Evolution, University of Chicago, Chicago, IL 60637, USA )
We estimated the dates of the monocot-dicot split and the origin of core eudicots using a large chloroplast (cp) genomic dataset. Sixty-one protein-coding genes common to the twelve completely sequenced cp genomes of land plants were concatenated and analyzed. Three reliable split events were used as calibration points and for cross references. Both the method based on the assumption of a constant rate and the Li-Tanimura unequal-rate method were used to estimate divergence times. The phylogenetic analyses indicated that nonsynonymous substitution rates of cp genomes are unequal among tracheophyte lineages. For this reason, the constant-rate method gave overestimates of the monocot-dicot divergence and the age of core eudicots, especially when fast evolving monocots were included in the analysis. In contrast, the Li-Tanimura method gave estimates consistent with the known evolutionary sequence of seed plant lineages and with known fossil records. Combining estimates calibrated by two known fossil nodes and the Li-Tanimura method, we propose that monocots branched off from dicots 140–150 Myr ago (late Jurassic–early Cretaceous), at least 50 Myr younger than previous estimates based on the molecular clock hypothesis, and that the core eudicots diverged 100–115 Myr ago (Albian–Aptian of Cretaceous). These estimates indicate that both the monocot-dicot divergence and the core eudicot's age are older than their respective fossil records.

P-06 : Molecular phylogeny of Hypericum based on nuclear ribosomal ITS sequence data
Seon-joo PARK * and Ki-joong KIM
(Department Biology,Yeunganm University, South Korea)
Our sampling included 54 species of the genus Hypericum, either least one species from the 30 sections of Robson. Outgroup were chosen from the closely related genera Triandnum. Phylogenetic analysis of the ITS sequence data provide several insights into relationship in Hypericaceae. The phylogeneric position of the genus Hypericum as a monophyletic group. The present analysis of ITS sequence data corroborates the earlier hypothesis from morphological data. On the other hand, the Phylogenetis implications of the ITS analysis indicate a number of surprising incongruities. Section 2, 20, 21, 22, 23, 24, 25, 26, 27, 28 and 30 were evolved from section 1. Also, section 28 explained that is group that evolves more than section 27. But section 27 and section 28 are difficult to be forming one major clade as is different from data of Robson and reason their relation in data that see. Also, according to opinion of Robson, spoke that is grouping that section 22 evolves more than section 21. We could get such conclusion in ITS data. There is a strong correlationship between the ITS phylogeny and the geographic distribution of Hypericum. One of the four major clades is restricted to America, Africa, Cuba, Madagascar, Australia, whereas the other two Mexico, Turkey, Pakistan to Taiwan and Celebes, Philippines, Asia and S. Balkan Penn. Mediterranean, Madeira, Canary island.

P-07 : Phylogenetic analysis of nuclear ITS/5.8S regions in Calocedrus (Cupressaceae), an Eastern Asian and Western North American Disjunctive Genus
Chih-hui CHEN*1, Chi-chu TSAI2, Robert G. NICOLSON3 and Shu-miaw CHAW4
(1Endemic Species Research Institute, Taiwan; 2Kaohsiung District Agricultural Improvement Station, Taiwan; 3Botanic Garden of Smith College, USA; 4Institute of Botany, Academia Sinica, Taiwan )
Relationships among the three disjunctive taxa of Calocedrus are not fully known. Phylogenetic analyses were conducted on nuclear ITS and 5.8S rDNA regions in all 19 accessions of the genus from west North America (C. decurrens) and eastern Asia (Taiwan, C. formosana; China and Vietnam, C. macrolepis). Phylogenetic trees were inferred using distance based neighbor-joining (NJ) and maximum parsimony (MP) criteria with the tree nodes evaluated by bootstrapping method. The ITS/5.8S sequences range from 1067 to 1088 bp and the ITS1 sequences are most informative. Both NJ and MP tree topologies strongly indicate that the Asian accessions and the American ones are two separate and highly supported clades. Each clade has its own unique 34 bp and 11bp deletions. Within the Asian accessions, the six accessions of C. formosana as whole form a highly robust monophyletic grade nested within the Vietnamese and Chinese C. macrolepis. Since the K2 distances suggest that C. formosana is closer to C. macrolepis of China than Vietnam, C. formosana is likely descended from the Chinese C. macrolepis. Perhaps due to geographical isolation and little gene flow between the two taxa, they became genetically distinct. The distinctness of C. formosana is further reinforced by an unusual one bp deletion in the ITS1 regions, rendering their secondary structure quite different from those of the other Asian accessions. We estimated the substitution rate of ITS/5.8S sequences of Calocedrus to be 4.61 x 10 -10 – 5.30 x 10 -10 bp per site per year and the C. macrolepisand the C. formosana diverged during the period from 8.69 to 7.54 MYA (late Miocene).

P-08 : Phylogenetic Relationships of the Genera of Cycadales Inferred from the sequences of LEAFY orthologue
Rumi MORI*1, Takeshi A OHSAWA1, Akira YOSHIDA2, Tatsuo KONISHI3, Hiroshi ISHIKAWA1 and Yasuyuki WATANO1
(1Faculty of Science,Chiba University; 2Research Institute of Evolutionary Biology; 3Tsukuba Botanical Garden,The National Science Museum,Tokyo )
The Cycadales is one of the four major groups of gymnosperms. Recently the Cycadales is classified into 11 genera in three morphologically identical families.The phylogenetic relationships of the Cycadales inferred from rbdL gene sequences indicated the monophyly of Cycadaceae, but the relationships of the other genera were not solved. The analysis using a total of 17 chloroplast genes and associated non-coding regions sequences supported the monophyly of Cycadaceae ,consistent with the rbcL study. Dioon is part of the next most basal split. Two of the three commonly recognized families of cycads (Zamiaceae and Stangeriaceae) are not monophyletic ; Stangeria is embedded within Zamiaceae, close to Zamia and Ceratozamia, and not closely allied to the other genus of Stangeriaceae, Bowenia. However, the phylogenetic relationships of all genera of the Cycadales has not been clearly identified in these chloroplast studies. In this study, the relationships of the genera of the Cycadales were investigated from the sequences of nuclear single copy gene, LEAFY orthologue, determined from 9 genera of the Cycadales. The LEAFY orthologue was amplified by PCR method, and the heterozygosity was checked by SSCP analysis. The PCR products from the homozygous materials were directly sequenced. The PCR products from the hererozygous materials were cloned. The clones were typified by SSCP, and then both alleles were sequenced. Phylogenetic relationships were examined by MP,NJ, ML methods using Cycas as an outgroup. All phylogenetic trees showed that Dioon was the most basal split. The remaining genera were divided in two monophyletic groups and Bowenia. One monophyletic group was composed of the genera of Tribe Encephalarteae, and the other consisted of the genera of Subfamily Zamioideae and Stangeria. Although uncertainty remains over the precise placement of Bowenia, Bowenia and Stangeria (Stangeriaceae) were well separated on all trees with moderate bootstrap supports, and Stangeriaceae was nested within Zamiaceae ;suggesting that these families were not monophyletic. From these results, futher studies are necessary for taxonomic rearangement of Zamiaceae ,Stangeriaceae and Subfamily Encephalartoideae, as none of these taxa are monophyletic.

P-09 : Comparison among gene trees of paternal cpDNA, maternal mtDNA and biparental nrDNAs in the genus Pinus subgenus Strobus
Kiyomi TSUTSUI*, Kei-ichi SAWADA, Toshihide KATO, Takeshi A. OHSAWA and Yasuyuki WATANO
(Department of Biology, Faculty of Science, Chiba University)
Interspecific hybridization and introgression have been considered to be one of the main factors that lead to phylogenetic incongruence among different data sets at lower taxonomic levels. In the plants of Pinaceae, chloroplast and mitochondrial DNAs may have different evolutionary histories through introgression because cpDNA and mtDNA are inherited paternally and maternally, respectively. In fact, species of the genus Pinus have insufficient reproductive barriers, and many examples of introgressive hybridization have been reported. Pinus species, therefore, could be susceptible to phylogenetic bias by hybridization and introgression. In the present study, we reconstructed cpDNA tree, mtDNA tree, and two nuclear DNA trees in the genus Pinus subgenus Strobus, and compared them in order to detect cryptic and ancient introgression episodes which caused incongruence among gene trees. Two nrDNA trees were constructed based on partial sequences of the plastid NAD-specific glyceraldehyde-3-phosphate dehydrogenase (GapCp) and a sequence tagged site of Pinus taeda (PtIFG_9008). MtDNA tree were generated based on partial sequences of intron 2 of nad1 gene. CpDNA tree were based on the dataset of Wang et al. (1999) and our additional data. In two nrDNA trees, species of the section Strobus were monophyletic, and the species of the other section (Parrya) were paraphyletic. In the sec. Parrya, two species of the subsection Gerardianae formed a clade, which was a sister to the sec. Strobus. CpDNA tree was consistent with the two nrDNA trees in these relationship among sections and subsections. On the other hand, mtDNA tree showed three clades, and in the two of them, each clade consisted of the combined members both from sec. Strobus and from sec Parrya subsec. Gerardianae. Because phylogenetic relationships suggested by the two nrDNA and cpDNA trees are congruent and also reasonable in morphological respects, the cause of incongruence could be attributed to the bias in the mtDNA tree. Possibly, mtDNA introgression had occurred anciently between species of different sections.

P-10 : The evolution of epiphytes in higher leptosporangiate ferns, Davalliaceae and Polypodiaceae
Chie TSUTSUMI* and Masahiro KATO
(Department of Biological Sciences, Graduate School of Science, University of Tokyo)
The evolution of epiphytes in the higher leptosporangiate epiphytic ferns, Davalliaceae and Polypodiaceae, and non-epiphytic related ferns was examined by field observations and phylogenetic analyses. Field observations revealed that epiphytes in a broad sense are classified into climbers (C), secondary hemiepiphytes (SH), obligate epiphytes (E), characterized by the places of spore germination and sporophyte growth. In climbers, e.g. Arthropteris backleri and Lomariopsis spectabilis, the spores germinate on the ground and sporophytes climb on tree trunks, having the roots connected to the soil. Secondary hemiepiphytes, e.g. Oleandra pistillaris and Nephrolepis cordifolia, are similar to climbers in the germination and growth places, but can live even without the roots connection to the soil after separating from the old part of the rhizomes. Although most Davalliaceae and Polypodiaceae are obligate epiphytes, in which spores germinate and plants live exclusively on trees, some species, e.g. Araiostegia hymenophylloides and Microsorum nigrescens, are secondary hemiepiphytes. Results of phylogenetic analyses using rbcL and accD (and also atpB for Davalliaceae) sequences include that the davallioid-polygrammoid group is sister to Oleandra (SH), and next sister to Arthropteris (C), and then a clade of Nephrolepis (SH) and Lomariopsis (C). The primitive in Davalliaceae is Araiostegia with two life forms (SH&E). Analyses of life-form evolution based on the phylogenetic relationships infer that in the davallioid-polygrammoid group, obligate epiphytes evolved from secondary hemiepiphytes or less likely from climbers. We hypothesize a scenario for life-form evolution in the davallioid-polygrammoid ferns and sister groups. The first step from terrestrial plants to climbers involved climbing habit of rhizomes, the second step from climbers to secondary hemiepiphytes required development of absorbing roots on epiphytic rhizomes, and the last step to obligate epiphytes accompanied a change of a spore germination place from the ground to trees. Characters related to Life-form epiphyte include rhizome dorsiventrality, dense cover of the rhizome by peltate or pseudopeltate scales, leaf or pinna articulation, and large spores. A syndrome of these characters may be key innovations of the epiphytic davallioid-polygrammoid ferns.

P-11 : A phylogenetic classification of the Ophioglossales inferred from rbcL sequences and spore morphology
(1Faculty of Pharmaceutical Sciences, Toho University, Funabashi, 274-8510 Japan; 2Graduate School of Science, Kyoto University, Kyoto, 606-8502 Japan; 3Laboratory of Plant Morphology, Faculty of Horticulture, Chiba University, Chiba, 271-8510 Japan )
We determined 1,194bp nucleotide sequences of rbcL gene for 38 plant samples of Ophioglossales. From the obtained molecular tree, two clades were recognized. One of them consisted of Ophioglossaceae s. str. which was clearly divided into three existing genera, viz Ophioglossum s. str., Ophioderma and Cheiroglossa. The latter two were more closely related each other than to Ophioglossum s. str. Cheiroglossa was supported to be separable genus from Ophioderma based on their large number of nucleotide substitutions between the two genera.The other clade consisted of those of Botrychiaceae and Mankyua which was recently described from Korea. By the molecular data, Mankyua is genetically well differentiated from Helminthostachys and others existing genera of Botrychiaceae. It should belong to Botrychiaceae, and it status as a new genus was also well supported by the molecular tree. In Botrychiaceae, Sceptridium were recognized as a monophyly also its bootstrap support was not so high (57%). Above phylogenetic relationship was also almost supported from the detail spore morphology except a new genus Mankyua that was included into Helminthostachys at least from the SEM observations. According to the TEM observations, the perispore of Sceptridium ternatum consists of two layers (P1, P2), but only one layer (P1) recognized in Ophioglossum thermale. Further, at the tetrad stage on the outer surface of the former was surrounded by the lamella structures on the proximal face, while on the latter was surrounded all over the surface of the spore.

P-12 : Taxonomic study of Deparia petersenii (Woodsiaceae) based on cytological and molecular genetic information
Wataru SHINOHARA* and Noriaki MURAKAMI
(Department of Botany, Graduate School of Science, Kyoto University)
According to the recent monograph of genus Deparia by Kato (1984), D. petersenii is a species defined by single character of its special serrated indusia. This species is morphologically very variable both in size and leaf shape. However, its variation is completely continuous when we compare the plants from distant localities of its wide distribution ranges from East Asia, Southeast Asia, Oceania and Pacific Islands. The purpose of this study is to investigate whether these morphological variations are completely due to plasticity or because several biological distinct units are contained in D. petersenii. In this study, we collected about 135 plant samples from total of 29 different localities in Japan, Taiwan and China, covering its morphological variation. We used cytological (chromosome number) and molecular (nucleotide sequence data of rbcL and ndhF genes) information to reveal biological units in D. petersenii. As our result of cytological analysis, we newly found diploid (2x) and pentaploid (5x) plants in addition to the tetraploid (4x) and hexaploid (6x), which had been recorded from D. petersenii by Kurita (1960, 1967). Although 2x and 4x plants had large variation in all measured morphological characters, no significant differences were found in any characters between the 2x and 4x plants. In contrast, the 6x plants were smaller in size than 2x or 4x plants, and 5x plants were intermediate between 4x and 6x plants. Based on the molecular analysis of chloroplast DNA, we found 14 haplotypes. The 6x plants did not share any haplotypes with either 2x or 4x plants. Thus, the 6x were shown to be genetically differentiated. Moreover, it was strongly indicated that the 4x cytotypes were evolved from 2x at least twice because they share two high frequency haplotypes. To investigate whether several biological units were contained in the 2x D. petersenii or not, 41 2x individuals from two populations of Taiwan were analyzed by electrophoresis. The result indicated that the 2x plants belong to a single Mendel population even though they still have large morphological variation.


P-13 : Phylogeography of the component species of broad-leaved evergreen forests in Japan, based on mtDNA variation of phytophagous insects
Kyoko AOKI*1, Makoto KATO2 and Noriaki MURAKAMI1
(1Department of Botany, Graduate School of Science, Kyoto University; 2Graduate School of Human and Environmental Studies, Kyoto University )
In order to elucidate the past distribution and colonization routes of Castanopsis type forests, we have compared the intraspecific phylogeographical patterns of chloroplast DNA (cpDNA) among 6 plant species with a similar geographic distribution growing in Castanopsis-dominant forests. However, the molecular evolutionary rate of plant cpDNA is relatively slow at the nucleotide sequence level. In order to investigate recent phylogeographic processes, for example detailed postglacial colonizing routes from the refugia in the Quaternary, we need to use DNA markers with a faster evolutionary rate. In this study, we tried to use intraspecific variation in mitochondrial DNA (mtDNA) of phytophagous animals for the phylogeographic study of host plants. The substitution rate of animal mtDNA appears to be about 100 times the rate in plant cpDNA or mtDNA. We examined whether information about intraspecific mtDNA variation in the seed-parasitic insect Curculio hilgendorfi can be used for the phylogeographic study of Castanopsis type broad-leaved evergreen forests in Japan. The Curculio mtDNA actually proved to have a greater amount of intraspecific variation than that of the cpDNA of the plants distributed in the same geographical region. Furthermore, we observed the pattern suggesting that anciently originated haplotypes were geographically widespread, whereas recently originated haplotypes were restricted to one locality. Therefore, the phylogenetic relationship among the haplotypes can also be used for the phylogeographic study of C. hilgendorfi, which could not be used for the study of plant cpDNA variation. Consequently, we concluded that it is highly probable that studying the variation in the mtDNA of the phytophagous insect C. hilgendorfi will be also useful for the phylogeographic study of Castanopsis type broad-leaved evergreen forests in Japan. The geographical boundary between the two groups of mtDNA haplotypes found in C. hilgendorfi lies in the Chugoku-Shikoku region. This pattern could be consistent with the result based on the intraspecific phylogeographical patterns of cpDNA variation among 6 plant species growing in Castanopsis-dominant forests which suggested the existence of additional impotant refugia from Muroto to the Kii Peninsula along the Pacific coast of Japan during the glacial ages.

P-14 : Geographical pattern of genetic variation of Angelica japonica, Farfugium japonicum and Peucedanum japonicum from Japan and Taiwan
Akihiro SEO* and Noriaki MURAKAMI
(Department of Botany, Graduate School of Science, Kyoto University)
Molecular studies of plants from the Ryukyu Islands suggested a recurrent pattern of genetic differentiation and geographic structuring. A discussion on the historical process of any area flora should compare geographical patterns of genetic differentiation among several phylogenetically independent plant species. We at first 19 taxa screened to find plant species with large intraspecific variation using an enzyme electrophoresis method. Based on the results of this screening, Angelica japonica A. Gray, Farfugium japonicum (L. fil.) Kitamura and Peucedanum japonicum Thunb., which had the largest genetic variation, were selected for further analyses. In each of three plant species, allozyme data show three common geographical pattern of genetic differentiation. First, the southern Ryukyus (Yaeyama Islands) populations of each plant species were well differentiated from the other Ryukyu populations. Moreover, many rare alleles were found only in the southern Ryukyus populations of A. japonica, F. japonicum and P. japonicum. And Taiwan populations of those three plant species did not show high genetic similarity to those from the southern Ryukyus populations, although these populations are geographically not so distant from each other. Furthermore, Taiwan populations showed high genetic similarity to the populations of the other part of Japan. These results suggested that past and current gene flow between the plant populations of Yaeyama Islands and the adjacent area has been limited maybe due to some geographical barrier between these areas. Second, genetic diversity (heterozygosities) of each plant species decline from the central Ryukyus populations to northward. Few rare or area-unique alleles were found in the northern Ryukyus or northward populations of those plant species. These geographical patterns, which were common for these plant species, indicated that the northern Ryukyus and populations northward might have recolonized rapidly from the central Ryukyus after the last glaciation. Third, the central Ryukyus (Amami and Okinawa Islands) populations contained several genetically well differentiated populations. Heterogeneous environment and larger size of the central Ryukyus might have helped to maintain a higher genetic diversity in these area. The geographical patterns of chloroplast DNA in A. japonica, F. japonicum and P. japonicum will be also compared with allozyme patterns.

P-15 : Geographic structure as evidenced by ploidy levels and DNA variations of Aucuba japonica (Aucubaceae) in the Japanese Archipelago
Tetsuo OHI-TOMA*1, Tadashi KAJITA2 and Jin MURATA1
(1Botanical Gardens, Graduate school of Sciences, The University of Tokyo; 2Department of Biology, Faculty of Science, Chiba University )
In order to elucidate the evolutionary process of geographic diversification of Aucuba japonica in the Japanese archipelago, we investigated ploidy levels and cpDNA haplotypes. Based on the combination of nine haplotypes and two cytotypes, eleven "haplo-cytotypes" were defined, in which two haplotypes included both diploids and tetraploids, and seven others were tetraploids. Three diploid haplo-cytotypes are distributed in the southwestern part of the archipelago, the island of Korea and northernmost Taiwan, and eight tetraploid ones in the northeastern part of the archipelago. Diploid and tetraploid haplo-cytotypes with the identical haplotype are, in one case disjunctively distributed, and in another case, in contact. Their genealogy indicates that A. japonica is paraphyletic and includes two lineages: one consisting of diploid and tetraploid haplo-cytotypes and A. chinensis, and another consisting of four tetraploid haplo-cytotypes. Tetraploidizations have occurred independently at least three times: twice in the former lineage, and once at the base of the latter. The two lineages show a distinct geographic structure in the continuous range of the species. It is inferred the present geographic structure was caused by postglacial expansion from separate glacial refugia. Then, we examined on genetic variations of nuclear DNA between two cpDNA lineages and between diploid and tetraploid cytotypes, by utilizing the GapC gene of nuclear DNA. Within monophyletic two species, A. japonica and A. chinensis, a total 32 alleles were found, and A. chinensis has specific eight alleles. Their phylogenetic relationship was poorly resolved with several polytomies, but the topology was significantly discordant with that of the cpDNA phylogeny. Within A. japonica, six of the 24 observed alleles were shared between the cpDNA lineages, of which three alleles were shared among only tetraploids and the others among both diploid and tetraploid cytotypes. The sharing identical alleles indicated that gene flow between the chloroplast lineages have occurred rather than the retention of common ancestral polymorphisms. That is, the gene flow from diploid to tetraploid in the same lineage, by the repeated occurrences of tetraploids, and subsequent gene flow between the tetraploids of different lineages, might occur within a polyploid complex species.

P-16 : Molecular phylogeography of pan-tropic sea-drift seed dispersal plants, Hibiscus tiliaceus
Koji TAKAYAMA*1, Tadashi KAJITA2, Jin MURATA1 and Yoichi TATEISHI3
(1Botanical Gardens, Graduate School of Science, The University of Tokyo; 2Faculty of Science, Chiba University; 3Faculty of Education, University of the Ryukyus )
To understand how genetic differentiation leading to speciation proceeds in pan-tropic sea-drift seed dispersal plants, we need information on barriers to gene flow, their antiquity, and their efficacy. Hibiscus tiliaceus (Malvaceae) is one of sea-drift seed dispersal plants widely distributed in the tropics of the Old World. One of its allied species, H. pernambucensis, is only in the New World, and the sum of distribution ranges of the two species almost spread over the earth in tropics. Three other related species, H. elatus (in the West Indies), H. glaber (in the Bonin Islands), and H. hamabo (in East Asia) are distributed in rather restricted areas. To elucidate speciation process of H. tiliaceus and its allied species, we performed molecular phylogenetic and SSCP analyses using chloroplast DNA. Phylogenetic analysis based on 7,800 bp of nucleotide sequence produced the most parsimonious tree that consisted of two major clades. Samples of H. tiliaceus scattered over both clades and its allied species except H. pernambucensis are nested in either of the clades. Samples of H. pernambucensis appeared in both clades: Pacific samples possessed a unique haplotype in one of the clades, while Atlantic ones shared a haplotype with H. tiliaceus in the other clade. SSCP analysis using population samples also revealed that almost all Pacific populations of H. pernambucensis were fixed with a unique haplotype, but most Atlantic populations were fixed with the haplotype that was shared with H. tiliaceus. Several haplotypes of H. tiliaceus, including the shared haplotype with H. pernambucensis, were distributed throughout the Old World. These results indicated that 1) the extensive distribution of haplotypes would suggest frequent migration by sea-drift seeds among the populations of H. tiliaceus, 2) the Isthmus of Panama clearly prevented gene flow by seeds in the populations of H. pernambucensis, 3) genetic subdivisions between H. tiliaceus and H. pernambucensis occur over the east Pacific Ocean, and 4) gene exchange between the two species would have happened over the Atlantic Ocean.

P-17 : Is Entada koshunensis (Leguminosae; Mimosoideae) endemic to Taiwan and Japan? : Evidences from morphological and molecular data.
Norihisa WAKITA*1, Tadashi KAJITA2, Tetsuo OHI-TOMA3, Jin MURATA3 and Yoichi TATEISHI4
(1Graduate School of Science and Technology, University of the Chiba; 2Faculty of Science, University of the Chiba; 3Graduate School of Science, University of the Tokyo; 4Faculty of Education, University of the Ryukyus )
Entada koshunensis Hayata et Kanehira and E. phaseoloides (L.) Merr. are closely related species which have sea-drift seeds. Although the former is endemic to Taiwan and the Ryukyus, the latter is distributed over very wide range from the Indian to Western Pacific regions. As they are morphologically quite similar, systematic treatments of these species were sometimes confused in Japan. In order to reveal their relationships in detail, we performed molecular and morphological analyses using extensive samples obtained from 27 populations of the Ryukyus, Taiwan, Vietnam, Samoa and Vanuatu, including samples obtained from the type locality of E. koshunensis in Taiwan. Molecular phylogenetic tree constructed by nucleotide sequences of chloroplast trnK (including matK gene site) and trnL-F regions, ca. 3,700 bp in total, produced 38 most parsimonious trees. The strict consensus tree was cleary separated into two clades with high bootstrap values. One of the clades mostly consisted not only the samples of southern part of Taiwan (including the sample obtained from the type locality), Okinawa Isl. and Yeyama Isls., but also of Samoa and Vanuatu. All samples included in this clade were determined as E. koshunensis by seed characters, although samples of Samoa and Vanuatu have been recognized as E. phaseoloides based on gross morphology (mainly leaf character). The other clade consisted of samples obtained from northern part of Taiwan, Vietnam, Yaku and Amami Isl., that were determined as E. phaseoloides by seed morphology. These results indicates that 1) both E. koshunensis and E. phaseoloides are clear species, 2) seed morphology can be used as key character to distinguish the two species, and 3) E. koshunensis, which has been thought as an endemic species to Taiwan and the Ryukyus, can be distributed widely to the Pacific islands.

P-18 : Chloroplast DNA variation and geographic structure of Aristolochia kaempferi group
Kana WATANABE*1, Tadashi KAJITA2 and Jin MURATA1
(1Botanical Gardens, Graduate School of Science, The University of Tokyo, Tokyo 112-0001, Japan; 2Departmet Biology, Faculty of Science, Chiba University, Chiba 263-8522, Japan )
The Aristolochia kaempferi group (Aristolochiaceae), that consists of five species of Aristolochia subgen. Siphisia, i.e. (A. kaempferi, A. liukiuensis, A. shimadai, A. cucurbitifolia, and A. mollissima), forms a monophyletic group according to the molecular phylogenetic study of Murata et al. (2001). Although samples and molecular data used in their study were insufficient for the phylogeny of the A. kaempferi group, their tree revealed that three species shared one haplotype and that two species had intraspecific haplotype variation. Their results imply that geographic structure of chloroplast haplotypes will be observed in the whole range of distribution of the A. kaempferi group. To examine the geographic distribution of chloroplast haplotypes in detail, we performed molecular phylogenetic study using 120 samples of 61 populations from the entire geographic range of the group in Japan and Taiwan, as well as 2 samples of A. mollissima from eastern China. Combined sequences of the matK gene (ca.1200bp), atpB-rbcL intergenic spacer region (ca.700bp) and trnS-trnG intergenic spacer region (ca.900bp) produced the 16 most parsimonious trees. The strict consensus tree showed geographically well-structured clades but that was not consistent with recent taxonomic classifications. Two major clades including 34 haplotypes were recognized in the tree. One of the clades consisted of 21 samples of A. kaempferi distributed over southern Kanto District, and the other one was further divided into five sub-clades: The first one consisted of 24 samples of A. kaempferi and 4 samples of A. shimadai distributed in Kanto District to Kinki District; the second of 14 samples of A. kaempferi in Seto-uchi region and northern Kyushu District; the third of 15 samples of A. kaempferi in southern Kyushu District; and the fourth of 4 samples of A. kaempferi in southwest Shikoku District; the fifth of 40 samples of four species (19 samples of A. liukiuensis, 17 samples of A. shimadai, 2 samples of A. cucurbitifolia and 2 samples of A. mollissima), which were distributed in Nansei Islands, Taiwan, and eastern China. The incongruence of molecular phylogeny and taxonomic species may suggest the possibility of introgression and / or parallel evolution etc.

P-19 : Genetic differentiation of Fagopyrum cymosum supports recent uplift hypothesis of Himalayan-Tibet region
Kyoko YAMANE*, Yasuo YASUI and Ohmi OHNISHI
(Graduate School of Agriculture, Kyoto University)
We investigated the phylogenetic and biogeographic relationships of twelve natural populations of Fagopyrum cymosum. Intraspecific variation of chloroplast DNA sequences was detected in three regions approximately 5 kb long in total: the end of rbcL, accD and associated intergenic spacer region, the trnC (GCA)-rpoB spacer region, the trnK (UUU) intron, and the matK region. The accessions of F. cymosum were divided into two major groups, a Tibet-Himalayan clade and a Yunnan- Sichuan clade, with a high bootstrap probability. These two groups were geographically separated by the Hengduanshan mountains. What was the reason for intraspecific differentiation in the genus F. cymosum? We estimated the time of divergence of these two clades as about 0.7 million years ago. The Himalayan range and the adjacent Tibetan Plateau, including the Hengduanshan mountains, constitute an exceptional topographic anomaly as a result of the collision of the Indian and Asian plates. Hsü (1978) and Li et al. (1979) suggested a hypothesis of the recent uplift (Late Pliocene and Pleistocene) of this region. On the other hand, Fort (1996) and other authors have claimed that the uplift of the Tibet-Himalayan group was probably ended by the Late Tertiary. Our estimated divergence period is in agreement with the recent uplift (Late Pliocene and Pleistocene) theory and the geographical and climatic interruption by the Hengduanshan mountains caused the genetic divergence in F. cymosum.

P-20 : Origin of polyploidy and biogeography of the Ludwigia sections Dantia (Onagraceae ) based on organelle and nuclear rITS DNAs
Kuo-hsiang HUNG1, Barbara A. SCHAAL2, Ching-i PENG3, Tzen-yuh CHIANG1 and Tsai-wen HSU*4
(1Department of Life Science, Cheng-Kung University, Tainan, Taiwan 701; 2Department of Biology, Washington University, St. Louis, USA; 3Institute of Botany, Academia Sinica, Taipei, Taiwan 115; 4Institute of Endemic Species Research, Ch-Chi, Nantou, Taiwan 501 )
Dantia, an indigenous section of Ludwigia in North America consisting of five species, including one diploid (L. palustris), two tetraploids (L. spathulata and L. arcuata), and two hexaploids (L. brevipes and L. repens), is phylogenetically related to Miquelia, a monotypic section exclusively occurring in East Asia. In this study, phylogeny of the section Dantia was reconstructed to recover a history of reticulation based on nucleotide variation of cpDNA atpB-rbcL intergenic spacer and nuclear ribosomal ITS region. Reciprocal monophyly of the two sections was supported, a pattern consistent with long division based on geological evidence. Within section Dantia, most species are paraphyletic at both genes, indicating recurrent hybridization or recent speciation. In the cpDNA network, rooted at L. ovalis (sect. Miquelia), haplotypes of tetraploid L. spathulata are nested as the most interior nodes, suggesting a likely autopolyploidy origin of this tetraploid from an extinct diploid, which was phylogenetically affined to L. palustris and L. arcuata. Ludwigia arcuata, another autopolyploid, contributed its organelle DNAs to hexaploid L. repens. Ludwigia brevipes may have arisen via bi-directional hybridization between L. plaustis and L. arcuata. nrITS data suggests that a putative, diploid ancestor of L. repends (6X) may have been extinct. In total, two extinct and one extant diploid species were involved in polyploidization in the sect. Dantia. Based on a calibrated molecular clock, divergence between sections Dantia and Miquelia was dated to 4.04 MYA, a time consistent with fragmentation of a continuous forest that occurred throughout Northern Hemisphere due to globally climatic changes. Polyploidy events of the formation of tetraploid L. spathulata and hexaploid L. brevipes and L. repens occurred 1.14 and 0.42-0.51 MYA, respectively.

P-21 : Phylogeography of Pinus luchuensis species complex based on genetic variation of cp- and mtDNAs
Tzen-yuh CHIANG1, Xue-jun GE2, Kuo-hsiang HUNG1, Tsai-wen HSU1, Yu-chung CHIANG3 and Shann-jye MOORE*4
(1Department of Life Science, Cheng-Kung University, Tainan, Taiwan 701; 2South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, PRC ; 3Department of Biology, Washington University, St. Louis, MO; 4Department of Life Science, Taiwan Normal University, Taipei, Taiwan )
Plants distributed across mainland and adjacent islands provide ideal materials for examining species and genetic divergence. Within a chain of islands of an archipelago, a stepping-stone migratory model is expected. Compared to mainland relatives, island species/ populations would display lower levels of genetic variability and significant, genetic differentiation given small population number and size due to limited, available habitats and geographical isolation. In this study, paternally transmitted chloroplast DNA and maternally inherited mitochondrial DNA were used to estimate the population structure and phylogeographical patterns of Pinus luchuensis, a species distributed across Ryukyu Archipelago (ssp. luchuensis), Taiwan (ssp. taiwanensis), and eastern China (ssp. hwangshanensis). In total, two major clades X and Y of cpDNA, consisting of subclades A-D and E-G, respectively, and two major clades α and β of mtDNA, consisting of subclades I-IV and V-VII, respectively, were identified. High levels of genetic diversities of both organelle DNAs were detected in these three ecologically dominant subspecies, except for that of cpDNA in ssp. hwangshanensis. Unexpectedly, the two island subspecies possess greater genetic polymorphisms than the mainland taxon. Given low possibilities of dispersal of winged seeds of these hard pines across straits, great Nm values deduced from mtDNA are likely to be attributable to ancestral polymorphisms that are shared between subspecies, a result consistent with recent divisions between mainland and islands populations based on geological evidence. In contrast to a general assumption of long-distance pollen dispersal in pines, lacking of the major clade X and subclade E of cpDNA in ssp. hwangshanensis indicates low possibilities of across-strait dispersal. A pattern of spatial partitioning of the organelle DNAs showing higher genetic diversity in islands than in the mainland, and in south than in north indicates that the P. luchuensis taxa experienced postglacial expansion. Subspecies. hwangshanensis may have been colonized via founders from islands, followed by expansion to its modern range.

Masaya SATOH*1 and Terumitsu HORI2
(1Faculty of Integrated Arts and Sciences, The University of Tokushima; 2Laboratory for Ginkgo Sciences )
Ginkgo biloba is a very popular tree in Eastern Asia. The old records on G. biloba of the Chinese historical literature were written in the 11th century. G. biloba is considered to be introduced to Japan from China (via Korea?) at the period between the late 13th and the early 14th centuries, according to the Japanese literatures and illustrations. However, the details of the incidents have not been traced yet in any field of sciences. In order to know details of transplantation and propagation of G. biloba we analyzed DNA polymorphism of giant trees (more than 2 m in trunk diameter at breast height, which are believed to be several hundred years old) in Japan, using their leaves, and compared with some results of the preliminary research made in Korea and China. Polymorphism is detected when the intron of the mitochondrial nad2 gene is analyzed. Up to the present 18 DNA types have been identified in the three countries, Japan (14), Korea (4) and China (11). All the four DNA types got in Korea and seven in China are common to the ones of Japanese, respectively. Judging from geological distribution of the DNA polymorphism in Japan, it is suggested that G. biloba was independently brought into Japan from different regions in Eastern Asia.

3.Species Biology and Reproduction

P-23 : Intraspecific differentiation of Nepnthes vieillardii Hook.f. in New Caledonia
Kaoruko KURATA*1, Tanguy JAFFRE2 and Hiroaki SETOGUCHI1
(1Graduate school of Human and Environmental Studies, Kyoto University; 2Reseach Institute for Development, New Caledonia )
Marked speciation of plants has been reported in the soils that develop on serpentine rocks and limestone. The present study was conducted to study the infraspecific differentiation of pitcher plants associated with serpentine in New Caledonia. Molecular and morphological studies of Nepenthes vieillardii in the extensive serpentine area in New Caledonia were conducted, and cpDNA haplotypes, pitcher morphology, floral and inflorescence morphologies and soil components were examined. The morphologies of pitchers, flowers and inflorescence varied widely in all examined species, however, morphology was relatively stable in each localized small population. The number of digestive glands in pitchers, density of flowers in inflorescence, and length of pedicles were particularly differentiated among populations. DNA sequences of five spacers on cpDNA were showed 11 haplotypes from 11 populations. Ten of the 11 populations possessed single or two haplotypes while four haplotypes were sympatrically distributed in one population. Soil analyses were also performed for each population, three types were recognized based on Mg, Ca, Ni and Fe content as well as five other factors. Correlations between the morphological characteristics, cpDNA haplotypes and soil types were examined, however, no relationship was recognized between the three factors. Furthermore, edaphic conditions within serpentine sites were not associated with the morphologies or haplotypes. We speculated that the morphological differentiation between populations was related to the differences of prey-fauna and pollinators in their microhabitats. Fixation into single or two cpDNA haplotypes in most populations may be attributed to founder effect and/or gentic drift in populations. We consider that very littele gene flow occurred between populations associated with seed dispersal and/or pollen movement based on the results of this study.

P-24 : Genetic variation of Syzygium (Myrtaceae) in the Bonin Islands, and fine-scale spatial genetic structure within a population
Taku FUJITA*1, Saeko KATO2 and Hidetoshi KATO2
(1Japan Wildlife Research Center; 2Tokyo Metropolitan University )
Three groups of Syzygium (Myrtaceae) in the Bonin Islands occurs their own way of living to adapt to the specific environment of their respective habitats (xeric rocky place, xeric shrubby forest and mesic forest; Fujita et al. 2002). These groups occasionally grow adjacent to each other. The flowering periods among groups are slightly different, but overlap considerably each other. Therefore, the reproductive isolation among these groups may be imperfect. In order to understand the population differentiation of Syzygium in the Bonin Islands corresponded closely to their respective habitats, it is important to reveal what is the mechanism that the morphological differentiation among the groups persists.The purpose of this study is to reveal the genetic variation of Syzygium in the Bonin Islands, and to assess the gene flow within a population (6 m x 20 m) adjacent to two groups, and to clarify the effects of environmental condition and geographical distance for the gene flow.We examined 155 individuals from seven localities in the Bonin Islands using amplified fragment length polymorphism (AFLP) with three primer pairs. Genetic variation significantly resided among groups (7.15%,p<0.0001; AMOVA). Genetic relationships among populations determined by neighbor-joining dendrogram showed assemblages that nearly corresponded to three groups. Assignment tests proposed that estimated random mating assemblages did not equal the populations given by localities and groups, suggesting that each group might not be a simple monophyletic group.In the population (6 m x 20 m) adjacent to two groups that grow xeric rocky place and xeric shrubby forest, assignment tests detected genetic structure corresponded with nearly each group, although there was slightly high gene flow between two groups (Nm=3.39). Genetic distance was affected significantly geographical one within each group and in all individuals, and also by vegetation height, which indicated soil moisture, only within the group that occur in xeric rocky place (Partial Mantel tests; p<0.05), suggesting that geographical distance and soil moisture persist genetic structure in the population. We are now adding primer pairs of AFLP and sampling localities to further confirm these results.

P-25 : Genetic structure of an endemic genus Boninia (Rutaceae) on the Bonin (Ogasawara) Islands.
Saeko KATOH*1 and Hidetoshi KATO2
(1Graduate School of Science, Tokyo Metropolitan University; 2Makino Herbarium, Tokyo Metropolitan University )
Many species of oceanic islands show the low genetic diversities in spite of the high morphological variation, which can be explained by restricted gene flow, small population size and rapid adaptive radiation. The genus Boninia (Rutaceae), endemic to the Bonin Islands, shows considerable morphological and ecological diversities at the inter- and intrapopulational level. Based on the allozyme analyses, however, high levels of genetic diversities (Hs: 0.233-0.354) were observed within local populations of Boninia. In order to elucidate the complexity of genetical, morphological and ecological diversities of Boninia, we used amplified fragment length polymorphism (AFLP) loci to examine the distribution of genetic diversity within and among seven local populations all over the islands and to detect genetic population structure without external information. The assignment analysis indicated that the assigned subpopulations did not consist with the nominal populations given by localities and morphological features. AFLP data also provides considerable resolution for the genetic structure within population. The association between genetic distance and geographic distance, or other factors (e.g., morphological differentiation and phenological variation) will be discussed among the assigned subpopulations.

P-26 : Agamospermous triploids (Taraxacum officinale) meet sexual diploids in Japan
Hiroyuki SHIBAIKE*1, Misuzu YAMANO2, Tetsuichi HAMAGUCHI3, Tatsuyoshi MORITA4 and Makoto IDE1
(1National Institute for Agro-Environmental Sciences, Japan; 2Graduate School of Agriculture & Life Sciences, University of Tokyo, Japan; 3Hiratsuka City Museum, Japan; 4Faculty of Education, Niigata University, Japan )
Recently, hybrid dandelions between introduced agamospermous triploids (3n=24) and native sexual diploids (2n=16) have been discovered in Japan. Since floral morphology of these hybrids is quite similar to those of the introduced dandelions, it is difficult to discriminate the hybrids in the wild by eye. This difficulty in species recognition makes it hard to treat with the dandelions already identified as the introduced ones in descriptions of vegetation. This study developed new molecular markers to detect the hybrid dandelions. Out of 844 plants morphologically identified as introduced dandelions, namely Taraxacum officinale, 713 plants (85%) were found to be the hybrids. These hybrids were further classified into three types according to the combinations of nuclear and chloroplast genomes, and chromosome numbers. To understand their geographical distributions, localities of the sampling sites were plotted on the map. Several floral traits including pollen fertility were also analyzed. The results suggested that constitutions of chromosomes derived from each of the parental species in nuclear genomes were significantly associated with the differentiation patterns in the distributions and floral morphology of the hybrids. Finally, hybridization between introduced and native dandelions in East Asian countries was discussed from the point of view of conservation of rare species.

P-27 : Polyploid speciation through hybridization in East-Asian Taraxacum
Tatsuyoshi MORITA*1, Motomi ITOH2, Hiroyuki SHIBAIKE3, Ching-i PENG4 and Jae-hong PAK5
(1Faculty of Education, Niigata University, Japan; 2Graduate School of Arts and Sciences, University of Tokyo, Japan; 3National Institute for Agro-Environmental Sciences, Japan; 4Institute of Botany, Academia Sinica, Taiwan; 5Kyongpook University, Korea )
The genus Taraxacum (Asteraceae) includes polyploids of 3X to 10X (X=8), as well as diploids. While the diploids are sexuals, the polyploids are asexual reproducers through apomixis. However, the apomictic polyploids have potential to make hybridization with the diploid sexuals, and as the results, new clonal species with the same or higher ploidy level can be occurred. We found two different cases of hybridization in Taraxacum . In one case the polyploid performs as a father (pollen parent) and in another one, it performs as a mother (seed parent). The former example is hybridization event progressing at the present time between the Japanese diploid species (mother) and the European triploids (father). The latter examples are past events when the Korean and Japanese white-dandelions, T. coreanum, T. shinanense and T. albidum originated. By means of chloroplast DNA sequencing (non-coding region of trnF-trnT), rflps of ITS (nuclear DNA) and isozyme analysis, it was clarified that Korean tetraploid species, T. coreanum is a hybrid derivative between a triploid species T. mongolicum as a mother and a diploid species T. hallaisanense (tentative name) as a father. Similarly, Japanese tetraploid species, T.shinanense is of hybrid origin between T. mongolicum (3X, mother) and T. japonicum (2X, father). Pentaploid species T. albidum is also supposed to originate through hybridization between T. coreanum (mother) and T. japonicum (father). T. mongolicum is regarded as an autotriploid of Chinese diploid, T. formosanum and facultative sexual reproduction in the apomictic polyploids of T. formosanum lineage is an extremely rare case in Taraxacum.

P-28 : The occurence of Aster ageratoides var. ovalifolius in Korea and its cytogeographycal feature
Akiko SOEJIMA*1, Jae-hong PAK2 and Motomi ITO3
(1Osaka Pref. Univ.; 2Kyungpook National Univ.; 3Tokyo Univ. )
Aster ageratoides complex is one of the Sino-Japanese elements distributed widely from southwestern China, Korea, Taiwan to Japan, and has many intraspecific taxa and correlated species. Among them, A. ageratoides var. ovalifolius is a variety known to be Japanese endemic, distributed along Japan Sea side region. This variety has larger, ovate to oval leaves in contrast to the generally smaller, lanceolate leaves of A. ageratoides var. ageratoides.In Japan, the variety ovalifolius occurs westward from Yamagata prefecture to Tsushima island of Nagasaki prefecture. Its distribution is overlapped with the distribution range of the variety ageratoides, and sometimes they grow in the populations closely located. In Korea, only variety ageratoideshas been recorded, but we found the plants with ovate, large leaves quite similar to the variety ovalifolius in the south part of the Korean Peninsula. Because of the geographic continuity and the morphological resemblance, we can consider them as the variety ovalifolius.In this study, we made researches in the field, and counted the chromosome numbers of the varieties ovalifolius and ageratoides. It elucidated that the variety ovalifolius is tetraploid in Japan, but in Korea, both diploids and tetraploids were found within the variety; tetraploids at the southeastern part of the Peninsula, and diploids around them. Meanwhile, the variety ageratoides is totally diploid in Korea, but tetraploid in Japan (as far as in the overlapped region of these two varieties). These results suggest that the variety ovalifolius was originated in the southern part of Korea, and spread eastward to Japan.

P-29 : Floral scents in Akebia (Lardizabalaceae)
Hiroshi AZUMA* and Hiroshi TOBE
(Department of Botany, Graduate School of Science, Kyoto University)
Floral scent is an important factor to maintain the plant-insect interaction that has prompted the divergence of angiosperms. However, our knowledge on chemical profiles of the floral scent in wild species are extremely limited. In this study, we conducted chemical analysis of floral scent emitted from flowers of three Akebia species (A. quinata, A. trifoliata, and A. x pentaphylla native to Japan to reveal inter- and intraspecific difference and to discuss the significance of the scent in the reporductive biology of the species. The floral scents were collected from a total of 26 (A. quinata), 4 (A. trifoliata), and 5 (A. x pentaphylla) individuals distributed in the Kinki district of Japan, by means of the head-space method using Tenax GR (adsorbent). Chemical analysis by GC-MS (gas chromatography-mass spectrometory) suggests that A. quinata is entomophilous (scented), while A. trifoliata is anemophilous (scentless), and that A. x pentaphylla is intermediate. There is no difference between chemical profiles of male and female flower of scented A. quinata except one compound, benzenacetaldehyde, which is detected in male flowers only. The scent of A. quinata is mainly composed of ß-myrcene, limonene, ocimene, etc. Also, analysis based on a limited number of samples available show that A. quinata has three types of floral scent chemistry. Among the 26 samples, nine were composed of ß-myrcene (47-92%) as a main scent compound with a little amount of limonene (ca. 3%) (type 1); 11 were composed of ß-myrcene (36-53%) and limonene (33-50%) with almost equal amount (ratio 0.85-1.39) (type 2); the remaining six samples were composed of ß-myrcene (20-26%) and limonene (57-76%) with ratio 0.25-0.45 (type 3). Such chemical profiles in the samples were consistent throughout 2003-1004, indicating that they are genetically fixed. A brief discussion is provided concerning interrelationships among the three species based on the floral scent chemistry.

P-30 : Genetic comparison of three Japanese Magnolia species: M. stellata, M. kobus and M. salicifolia
Miyuki NAKASHIMA* and Yoshimichi SAKAI
(Gifu Prefecgtural Forest Science Research Insititute)
Five Magnolia species are distributed in Japan: Magnolia stellata, M. salicifolia, M. kobus, M. sieboldii, and M. obovata. The first three species belong to section Buergeria and share a common feature, in that their flowers appear before the leaves, whereas M. sieboldii and M. obovata belong to sections Oyama and Rytidospermum, respectively. Recent phylogenetic studies have revealed the close relationship of the three species (Azuma et al. 1999, Kim et al. 2001), although they are distinguished by morphological traits and their geographic distributions. However, it is not clear how their genetic features differ at the interspecific level. Therefore, we carried out allozyme electrophoresis for 22 populations (eight M. stellata populations, five M. kobus populations, and nine M. salicifolia populations) to examine the genetic relationships of the three species.Twenty-seven alleles were detected at nine loci encoding five enzymes in the three species: Adh-1, Adh-2, Dia-1, Dia-2, Est, Got-1, Got-2, Got-3, and Lap. All nine loci were polymorphic in the three species. Six alleles were common to all populations at high frequency. The mean genetic identity and genetic distance between any pair of populations from different species was 0.72 and 0.33, respectively. A dendrogram of the 22 populations generated using UPGMA clustering of the genetic distances produced three distinct groups at the species level. The populations of M. salicifolia were further divided from the populations of the other two species. In addition, principal component analysis was performed on the frequencies of the 27 alleles for the 22 populations. Our results suggest that these three species in sect. Buergeria are highly differentiated at four loci. The first principal component suggested that M. salicifolia was featured by allele frequency at two loci: Est and Dia-1. By contrast, the second principal component suggested that the allele frequency at Got-2 and Got-1 characterized M. kobus and M. stellata, respectively.Azuma et al. 1999. Journal of Plant Research 112: 291-306.Kim et al. 2001. American Journal of Botany 88: 717-728.

P-31 : Molecular identifications of endomycorrhizal fungi inhabiting in Cephalanthera (Orchidaceae)
(Graduate School of Life Sciences, Tohoku Univeristy)
The genus Cephalanthera L.C.Richard (Orchidaceae) comprises about 15 species distributed widely in northern hemisphere. Cephalanthera exhibits distinctive diversification of nutritional forms. Most species are photosynthetic as normal plants but some species of the genus reduce or lack foliage leaves and have strongly mycotrohiphic nutritional existences. In this study, we conducted molecular identifications of endomycorrhizal fungi in Cephalanthera to reveal its relationship to nutritional forms. Two types of fungi were found in Cephalanthera roots; one is original orchid mycorrhizal fungi (Rhizoctonia spp.) and the other is ectomycorrhizal fungi of trees. The semi-mycotrohiphic species, C. subaphylla, had ectomycorrhizal fungi as a previously reported case of holo-mycotrohiphic species, C. austinae in North America. Moreover, photosynthetic species, C. erecta and C. longibracteata also had ectomycorrhizal fungi in their roots. The change of fungal associates from orchid mycorrhizal fungi to tree ectomycorrhizal fungi in photosynthetic species may be the precursory states of complete mycotrophism in Cephalanthera.

P-32 : Habitat shifts lead to morphological and nutritional innovations in Orchidaceae: a case study in Cymbidium
Tomohisa YUKAWA*1, Kazumitsu MIYOSHI2, Jun YOKOYAMA3 and William L. STERN4
(1Tsukuba Botanical Garden, National Science Museum; 2Department of Biological Production, Akita Prefectural University; 3Department of Ecology & Evolutionary Biology, School of Life Sciences, Tohoku University; 4Department of Botany, University of Florida )
Cymbidium Sw., an orchid genus distributed from East and Southeast Asia to Australia, comprises about 50 species. The genus exhibits distinctive ecological diversification and occurs in terrestrial, epiphytic, and lithophytic life forms. Moreover, Cymbidium section Pachirhizanthe lacks foliage leaves and has an obligate mycotrophic nutritional existence. In this study correlation between various morphological and anatomical characters and ecological characters was tested for most species of Cymbidium and its sister groups. Furthermore, a molecular phylogenetic tree of Cymbidium (Yukawa et al. 2002) was used to examine phylogenetic constraint of these characters. The following insights were provided: (1) In Cymbidium, lithophytic and terrestrial life forms were likely to be derived from epiphytic habitat at least three times. (2) The obligate mycotrophic lineage, Cymbidium section Pachyrhizanthe diverged from a terrestrial progenitor. (3) The seed of epiphytic species showed higher hydrophilicity and larger dimensions than that of terrestrial species. (4) The seed size of obligate mycotrophic species decreased greatly in comparison with its sister photosynthetic species. (5) Development of ramified rhizomes was found only in terrestrial species. We found that this type of the rhizome is the principal dwelling for mycorrhizal fungi. (6) In this genus, development of sclerenchymatous fibers in leaves is correlated to the degree of epiphytism. (7) Differentiation of palisade tissue and amphistomaty are likely adapted to high light stress because the species possessing these characters inhabit trees at exposed positions. (8) Orchid-type endomycorrhizal fungi and tree ectomycorrhizal fungi were found in Cymbidium roots and/or rhizomes. Although terrestrial Cymbidium species symbiose with both types of fungi, epiphytic species show symbiosis only with Orchid-type endomycorrhizal fungi. On the other hand, obligate mycotrophic species symbiose exclusively with tree ectomycorrhizal fungi.

P-33 : Sex ratio and floral dimorphism of a gynodioecious pan tropical shrub, Arugusia argenta (Boraginaceae) in Japan
Junko OKAZAKI*1, Emi AZEGAMI1 and Shunichi MATSUMURA2
(1Osaka Kyoiku University; 2Tohoku University )
We investigated the floral sex expression of a pan tropical gynodioecious shrub, Argusia argentea (Boraginaceae), which is distributed in the coastal regions of Nasei Islands and Bonin Islands, Japan. The sex ratios of hermaphrodites to females are not significantly deviated from 1:1 in 11 natural populations which we examined. The percentages of fruit-set in females are significantly higher than those in hermaphrodites in each population. Hermaphrodite plants have larger flowers, and more deeply serrated corolla. These characters might play a role in promoting the floral display for pollinator attractions in hermaphrodites.

P-34 : Observation on floral behavior and pollinators of flexistylous ginger, Alpinia nieuwenhuizii Val. (Zingiberaceae) from Borneo
Atsuko TAKANO*1, Johnny GISIL2, Mashitah YUSOFF2 and Takuji TACHI3
(1Museum of Nature and Human Activities, Hyogo; 2Institute for Tropical Biology and Conservation, University Malaysia Sabah; 3Biosystematics Laboratory, Kyushu University )
Flexistyly is a unique floral mechanism involving extreme stigmatic movement, first described in Chinese ginger (Alpinia, Zingiberaceae). This is the first report on flexistylous gingers in Malesia with the highest species diversity of the family. We observed floral behavior and flower visitors in Alpinia nieuwenhuizii Val., a Bornean endemic. Although floral behavior and effective pollinator (carpenter bee, Xylocopa) were similar to that of the previously reported flexistylous Alpinia, behavior of the pollinators were strikingly different in that visit frequency of the pollinators showed bimodal pattern during the day. It was better match for the behavior of flexistylous ginger in Borneo. Besides, some gender differentiation between the two morphs is also discussed.

P-35 : Structure based large scale co-evolution analysis Structure based large scale co-evolution analysis
Yoobok CHO*1, Wan Kyu KIM2, Dan M BOLSER3, Donghoon OH1 and Jong Hwa PARK4
(1Object Interaction Technologies, Inc., 201 Jueun Building, 29-4 Jamwon-dong, Seocho-gu, Seoul 137-904 Korea; 2MRC Rosalind Franklin Centre for Genome Research, Hinxton, Cambridge, CB10 1SB, UK; 3MRC-DUNN Human Nutrition Unit, Hills Road, Cambridge, CB2 2XY, UK; 4Biomatics Lab, BioSystems Dept. KAIST, DaeJeon, Korea )
Previously the propensity of co-evolution has been observed for various ligand-receptor pairs. However, it has not been studied systemically in the global scope. We were able to investigate the degree of co-evolution for more than 900 family pairs using a global protein structural interactome map (PSIMAP - a map of all the structural domain-domain interactions whose information is extracted from Protein Data Bank). If interacting pairs of proteins are co-evolve, such a pair of protein families will show similarity between their phylogenetic trees. We observed that there was significant correlation in 45% of the total SCOPs Family level pairs, rising to 78% in 454 reliable family interactions. Expectedly, the intra-molecular interactions between protein families showed stronger co-evolution than inter-molecular interactions. However, both types of interaction have a fundamentally similar pattern of co-evolution except for cases where different interfaces are involved. These results validate the use of co-evolution analysis with predictive methods such as PSIMAP to improve the accuracy of prediction based on homologous interaction. The tendency of co-evolution enabled a nearly 5-fold enrichment in the identification of true interactions among the potential interlogues in PSIMAP. The estimated sensitivity was 79.2%, and the specificity was 78.6%.

Chan-ho PARK* and Masahiro KATO
(Department of Biological Sciences, Graduate School of Sciences, The University of Tokyo)
Apomixis is an asexual reproduction via unreduced spores and the subsequent apogamous reproduction of a sporophyte from vegetative cells of a gametophyte. In pteridophytes, about 10% of the world's species and about 15% of Japanese species are apomictic. It is often argued that apomixis, like polyploidy, plays a significant role in speciation and as an escape from sterility caused by hybridization. Our understanding of the origin of apomictic reproduction in sterile hybrids is poor, partly due to our lack of the understanding of incipient apomixis. Cornopteris christenseniana is a sterile interspecific triploid hybrid of the diploid C. crenulatoserrulata and tetraploid C. decurrenti-alata, but a preliminary observation suggested that it may propagate by incipient apomixis in a semiartificial environment in the Fern Garden of Botanical Gardens, University of Tokyo. Cytogenetic observations and culture experiments show that C. christenseniana produces normal spores in various proportions in some sporogenetic pathways that are aberrant from ordinary ones of sexual and apomictic ferns. Under culture conditions the normal spores germinate in rough proportion to the frequency of normal spores, and sporophytes are apogamously produced in rough proportion to the frequency of spore germination. As a whole, the rates of spore germination and apogamous sporophyte development are various according to plants. Taken together, it is suggested that C. christenseniana is an incipient apomict.Furthermore, we analyzed allozyme electrophoretic polymorphism of 10 isozymes and sequence variation of the trnL-trnF intergenic spacer region in C. christenseniana and its two closely related species. From the data obtained, the parents of most offspring plants produced in the Fern Garden were identified. The results strongly support the idea that C. christenseniana often propagates by apomixis in the artificial environment and not by in situ hybridization. C. christenseniana appears here to be an incipient apomictic species of remarkably multiple and polytopic origin, and its apomixis is not so strongly regulated as obligate apomixis. The multiple origins of incipient apomicts implies that the evolution of apomixis in C. christenseniana began with unreduced sporogenesis, and it may be that pleiotropic effect led to the subsequent apogamy.

P-37 : Hybridization and Reticulation in Japanese /Polystichum/ (Dryopteridaceae)
Su-juan LIN*1, Atsushi EBIHARA2, Misuzu NAGAMOTO1 and Kunio IWATSUKI3
(1Faculty of Life and Environmental Science, Shimane University; 2Department of System Sciences (Biology), Graduate School of Arts and Sciences, University of Tokyo; 3University of the Air )
Polystichum is a large genus of Dryopteridaceae, estimated to have more than 200 species, best represented in the mountains of warm-temperature areas, especially in east to south-east Asia (Kramer & Green 1990). Hybridization and polyploidization play important roles in species diversification and reticulation in Polystichum (Reichstein, 1972; Daigobo, 1972; W. Wagner, 1973; D. Wagner, 1979; Barrington, 1985, 1986, 1990). Systematic researches have been made in resolving evolutionary and taxonomic problems in European and American regions (Manton, 1950; Reichstein, 1972; W. Wagner, 1973; D. Wagner, 1979; Barrington, 1986, 1990, 1992). Thirty-two species were recorded in Japan, and about 40 interspecific hybrids among 13 putative parentage species were presumed based on morphological characters (Iwatsuki 1992). The taxonomy and discrimination of closely allied species are difficult because of overlapping in variations of various characters. It is necessary to have a combination of morphological, cytological and electrophoretic analyses to trace evolutionary relationships among Japanese species. In this study, the putative parentage species, i.e. five diploid species ( P. fibrilloso-paleaceum, P. retroso-paleaceum, P. igaense, P. otomasui, P. ohmurae) and seven tatraploid species ( P. longifrons, P. makinoi, P. ovato-paleaceum, P. polyblepharum, P. pseudo-makinoi, P. tagawanum) , were collected from several regions (Central and NW Honshu, Kyusyu) in Japan. The polyploidy and genetic patterns of these species were determined by cytology, flow cytometry and allozyme data. The results are: 1) the species-specific marker alleles were detected in three loci of PGM, PGI, SkDH enzymes in each of four diploid species P. fibrilloso-paleaceum, P. igaense, P. otomasui, P. ohmurae. 2) electrophoretic variants were detected in tetraploid species, P. longifrons (two types) and P. polyblepharum (four types), and both of them are allopolyploid. 3) Based on these data, six triploid hybrids and five tetraploid hybrids were examined erectrophoreticlly, and the putative parents of these hybrids were confirmed. This is a preliminary study to document the evolutionary relationships of Japanese Polystichum, and provides insights into hybridization and speciation among them. Further synthetic analysis is expected. (This study was supported by Showa Seitoku Memorial Foundation)

4.Conservation and Informatics

P-38 : Conservation genetic study of Magnolia tomentosa using nuclear and chloroplast microsatellite markers
Saneyoshi UENO*1, Suzuki SETSUKO2, Takayuki KAWAHARA3 and Hiroshi YOSHIMARU4
(1Genome Analysis Laboratory, Department of Forest Genetics, Forestry and Forest Products Research Institute; 2Laboratory of Forest Ecology and Physiology, Graduate School of Bioagricultural Sciences, Nagoya University; 3Forest Dynamics and Diversity Group, Hokkaido Research Center, Forestry and Forest Products Research Institute; 4Ecological Genetics Laboratory, Department of Forest Genetics, Forestry and Forest Products Research Institute )
Genetic diversity and differentiation were investigated for 11 populations of Magnolia tomentosa Thunb. (Magnoliaceae), endangered deciduous tree endemic to Tokai district, Japan. Four nuclear microsatellite (nSSR) and three chloroplast microsatellite (cpSSR) loci were simultaneously analyzed for the same 11 populations, which represented the whole distributional range of the species. The populations were classified into 3 groups (A, B and C) for population genetic analysis. The two populations in group A are located in the southern part of Aichi prefecture; the five populations in group B in the northern part of Aichi prefecture and the southern and western parts of Gifu prefecture; and the four populations in group C in the northern part of Mie prefecture. Combination of the three cpSSR markers generated 13 haplotypes. Group B harbored the highest gene diversity (H) and allelic richness (Ar) for nSSRs (H = 0.74 and Ar = 8.0). In contrast, group C was the richest in diversity for chloroplast haplotypes (H = 0.79 and Ar = 6.8). Different contribution to total diversity of each population was observed for nSSRs and chloroplast haplotypes. Populations with higher contribution for nSSRs were not always ranked high in terms of cpSSRs. Significant correlation between population pairwise distance and FST was detected for nSSRs (r = 0.614; p < 0.001) but not for chloroplast haplotypes. Population dendrograms were constructed using the standard genetic distance of Nei for nSSR loci by UPGMA and neighbor-joining method, suggesting that populations in group C were diverged from other populations and those in group B were similar to each other. For group B, fragmentation between populations should be avoided to maintain gene flow, while for group C, uniqueness of each population should have the highest priority for genetic conservation of the species. The present study used both nSSRs and cpSSRs and confirmed that their simultaneous use highlighted the importance of each population and suggested the possible future management plans.

P-39 : Genetic diversity of Miyamasukashi lily (Lilium maculatum var 'Bukosanense'), an endemic and endangered species in Buko Mountain, Saitama, Japan
Amaury ARZATE*, Makoto MIWA, Tomohide SHIMADA, Tetsushi YONEKURA and Kazuo OGAWA
(Center for Environmental Science in Saitama, Environmental Biology Division, Kamitanadare 914,)
Lilium maculatum var 'Bukosanense', called Miyamasukashi lily, grows endemically in Saitama and Ibaraki Prefectures, Japan. The natural population size of this species have been reduced drastically due to human activities, especially land use change, but overcoat by damages caused by monkey (Macaca fuscata), and deer (Cervus nippon) in Buko Mountain, Saitama Prefecture. Before they are lost, population genetic studies can provide information for efficient in-situ conservation as well as for possible future use in breeding programs. Although L. maculatum var 'Bukosanense' is listed in the Saitama Prefecture Red Data Book as a critically endangered plant, articles about its genetic diversity and population genetic structure have not been reported. Therefore, we carried out a population genetic study of natural Miyamasukashi lily using nuclear and chloroplast DNA markers. In this study, fresh leaves of Miyamasukashi lily were collected from 46 plants in Buko Mountain. By PCR-RFLP analysis of chloroplast DNA (cpDNA), we estimated the maternal origin of individuals sampled. Sixteen cpDNA primer pairs were tested, and only four of them (SHO20+SHO21, SHO19+SHO22, psbC+trnS, and trnC+trnD) produced clear bands, then combined with six restriction enzymes, thus 24 combinations were assayed. PCR-amplified products could be digested with 75% primer-enzyme combinations, producing 2 to 4 fragments. All individuals of Miyamasukashi lily analyzed were virtually identical for all primer-enzyme combinations. These findings suggest that the 46 individuals analyzed have the same haplotype, and its maternal origin is close similar. In addition, we also used ASSR (Anchored-Simple Sequence Repeat) markers to provide mainly biparentally inheritance. PCR reactions using five out of six ASSR primers showed polymorphism, indicating that individuals sampled are not a clone, and then it appears that all of them could be reproduced by seeds. The primers yielded a total of 30 polymorphic bands, i.e., an average of six polymorphic bands per primer. Cluster analysis showed that the individual plants were rather dispersed in the dendogram. From these results some strategies for its conservation might be considered.

P-40 : Recent Progress in the Flora of the Philippines Project
Domingo Allado MADULID*
(Philippine National Herbarium)
The Flora of the Philippines project was originally an undertaking of the Philippine National Museum with the Bishop Museum, Hawaii and Botanical Research Institute of Texas with financial support from the US-National Science Foundation and the John D. and Catherine T. MacArthur Foundation from 1990-2000. Through the project over 70,000 specimens were collected from various parts of the country and distributed to different major herbaria in the US, Europe and Southeast Asia. The collection records were databased at the National Museum and BRIT. It has also curated, databased and processed over 100,000 back-log specimens. The remarkable achievements of the project staff slowed down with the termination of the financial grants. Despite the present limited financial support from the Philippine government the Flora of the Philippines project continues to accumulate specimens and contribute to taxonomic knowledge through different cross-cutting approaches, i.e. ecosystem and protected area research, threatened flora research, ethnobotany and sustainable tourism. Collaboration with individual scientists in the writing of the flora continue. Data management has improved through the Southeast Asian Botanical Collections Information Network (SEABCIN) with other herbaria in the Malesian region. Taxonomic capacity building and infrastructure development in the museum also continue though slowly. The Project hopes to explore new collaboration and linkages with local and foreign institutions to continue the collection phase and eventually publish the first volume of the Flora. It also plans to circulate taxonomic data in more accessible media through the internet.


P-41 : Taxonomy and genetic diversity of Korean Suaeda
Hyun-bo SHIM1 and Byoung-hee CHOI*1
(1Department of Biological Science, Inha University, Korea; 2 )
The genus Suaeda is one of the most popular plants on salt marsh areas in Korean Peninsular. The entities that comprise taxa in Korea exhibit widely overlapping ranges in all morphological attributes. In this study the taxonomic delimitation and genetic diversity on Korean species of Suaeda were studied by gross morphology, ramdom amplified polymorphic DNA (RAPD) makers and ITS sequences of nrDNA. From the morphological and molecular data S. glauca of sect. Schanginia is the most distinctive species, compared with others of sect. Heterosperma. A newly recorded species from Korea of S. malacosperma, which has been treated as a varity of S. maritima, is clearly separated from the latter in having the stellate shape of fruits and molecular evidences. In RAPD analysis six decamer primers amplified a total of 65 scorable bands, of which 61 were polymorphic. S. japonica, S. maritima, and S. malacosperma, which have been hardly distinguished each other by external morphology, were readily recognized by its specific DNA bands. The characteristic RAPD markers were identified between local populations of S. japonica, but this feature was not revealed within population. The phylogenetic relationships among Korean species of Suaeda and its allied genera are discussed based on ITS sequences and the molecular data was compared with the morphological evidences.

P-42 : Checking taxonomic relationships in Umbelliferae-Apioideae of Ligusticum alliance by ITS rDNA sequencing and immunochemical comparison of storage seed proteins
Michael Georgievich PIMENOV*, Carmen Manuelvna VALIEJO-ROMAN, Victoria Semenovna SHNEYER, Tagir Halafovich SAMIGULLIN and Elena Igorevna TERENTIEVA
(Botanical Garden Moscow State University)
Relationships within the taxa of the East Asian Umbelliferae, presumably close to Ligusticum and Selinum ("Foenicilinae") were investigated by two independent molecular taxonomic methods. 133 ITS 1-2 sequences were analyzed (31 new species) and immunochemical comparison of storage seed proteins for 38 species of Apioideae was performed, eight reference systems (antisera) were used. Both approaches showed extremely polyphyletic nature of this group in the Umbelliferae. Similarly, a lot of traditional genera of "Foeniculinae", i.e. Ligusticum, Selinum, Cnidium, Pachypleurum and some others are polyphyletic, and must be revised. Molecular data confirm the result of partial revisions in these taxa, that yielded separation of Sphaenolobium (from Selinum), Dimorphosciadium and Lomatocarpa (from Pachypleurum), Paraligusticum and Arafoe (from Ligusticum), Cnidiocarpa (from Cnidium). The description of a separate genus Magadania, which was composed from two NE Asian species, previously treated in unrelated genera Cnidium and Conioselinum, was completely confirmed too. The revision of Selinum /Cnidium was confirmed only partly: The type of the genus, annual C. monnieri, is distant from C. davuricum. Early described genera, Mutellina and Oreocome, were shown to be distant from Ligusticum and Selinum respectively. The situation in the group of East Asian "Foeniculinae" is the most complicated. A big clade, comprising the species of Oreocome, Ligusticopsis, Cortia and Cortiella formed in tree with rather high support of the whole group and unstable inner structure. Oreocome, recently restored from Selinum, and Ligusticopsis, are undoubtedly related taxa. The difference between them is mainly carpological Some species of Ligusticopsis and Oreocome are very similar (as L. wallichiana and O. candollei). So, congenerity in these two genera is possible. The sequencing data also provide information on structural biodiversity in plants, but at a new (molecular) level. What comes without any doubt, however, that correlation of molecular data with geographical origin is stronger than with many morphological characters. This work was supported by the Russian Foundation of Basic Research (project 03-04-48831a) and Scientific Schools Program (project 1712.2003.4)

P-43 : Taxonomic diversity in Eastern Asian Umbelliferae in context of the whole continent data
Michael G. PIMENOV*
(Moscow State University, Botanical Garden)
In Moscow University Botanical Garden the multidisciplinary investigations in systematics and botanical geography of Umbelliferae have a long history, and Asia was a main field of analysis. This continent has the greatest diversity in genera (279) and species (2108) of the family, covering more than a half of the World richness. In computer databases GNOM (Generis NOMenclator) and ASIUM (ASIatic UMbelliferae) nomenclatural data, typification, synonyms, and essential citations are retrieved, together with distribution information according to countries and provinces. The highest numbers of genera and species are known for China (106 genera; 671 species), Turkey (109; 456), Iran (114; 364), Russia (105; 279), India (78; 227), and Afghanistan (73; 199). China, Turkey, Iran and Afghanistan, together with countries of mountainous Middle Asia, are the regions of concentration of endemics. One of reputed regions of the high family diversity is the Mediterranean area (s.l.) - up to Middle Asia and Afghanistan, but Eastern Asia, especially SW Chinese provinces and adjacent Himalaya, are of no less importance. The maximal number of taxonomic enigmas remains also in this area. Numerous critical revisions were made during last decades to elucidate taxonomy and phylogeny of Asian, especially East Asian, Umbelliferae. The main objects of these revisions were genera Pleurospermum, Hymenolaena, Hymenidium, Keraymonia, Trachydium, Pterocyclus, Angelica, Ostericum, Tordyliopsis, Semenovia, Notopterygium, Seseli, Sinocarum, Conioselinum, Ligusticopsis, Oreocome, Arcuatopterus, Hansenia, Rupiphila and some other genera. In taxonomic revisions morphological, karyological and molecular data were used side by side. Although exact and detailed floristic regioning of Asia is not elaborated, botanical Eastern Asia (or Sino-Japanese region) can be limited using Hara (1966), Takhtadjan (1986), and Wu ZhengYi & Wu SuGong (1996) proposals - from Amur basin, Sakhalin and Japan to Western Himalaya (partly). 717 species, belonging to 92 genera of the Umbelliferae, are distributed at this area. The numbers of genera and species are calculated for each country of Eastern Asia, including Russian Far East, without the northern part (35 genera; 67 species), Korea (36; 83), Japan (37; 104), N Burma (20; 43), Bhutan (42; 91), Nepal (49; 141), Indian E Himalaya (34; 88), Indian E Himalaya (61; 124).

P-44 : Hydrastis and Glaucidium, the phytochemical relationship using flavonoid markers
Tsukasa IWASHINA*1 and Hiroshi TOBE2
(1Tsukuba Botanical Garden, National Science Museum; 2Department of Botany, Graduate School of Science, Kyoto University )
The genera Hydrastis and Glaucidium are endemic to North America and Japan, respectively, and consist of each one species, H. canadensis L. and G. palmatum Sieb. & Zucc. It has been shown that they are phylogenetically correlated each other. A few chemical components in H. canadensis and G. palmatum have been reported. The alkaloids, berberastine, canadine, canadaline, hydrastidine and hydrastine, were isolated from the roots of the former species. On the other hand, a coumarin, glaupalol was reported from the roots of the latter species. The flavonoids of G. palmatum were studied by Iwashina and Ootani (1990), and three new or rare flavonol allosides, i.e., quercetin 3-O-alloside, kaempferol 3-O-alloside and rhamnocitrin (kaempferol 7-methyl ether) 3-O-alloside were isolated from the leaves. However, those of H. canadensis are not surveyed until now. In this study, the flavonoids of Hydrastis canadensis were isolated and identified, and the phytochemical relationship between Glaucidium and Hydrastis using the flavonoids as chemotaxonomic markers was discussed. The leaves of H. canadensis were extracted with MeOH. After concentration, the extracts were applied to preparative paper chromatography using solvent systems: BAW, 15%HOAc and then BEW, and finally purified by Sephedex LH-20 column chromatography. Two flavonoids were isolated and characterized as quercetin 3-O-glycosides by UV spectral, TLC and HPLC properties. The flavonoids of G. palmatum were flavonol 3-O-allosides based on quercetin, kaempferol and rhamnocitrin which are very rare glycosides in nature. However, though those of H. canadensis were flavonol 3-O-glycosides, these glycosidic sugars which attached to 3-position were not allose. thus, it was shown that the flavonoid characters of H. canadensis and G. palmatum were essentially similar to each other (the presence of flavonol 3-O-glycosides), but they were slightly different by quality of the glycosidic sugars.

P-45 : Calcium sulphate crystals in the Iranian species Sophora and Ammodendron (Leguminosae, Sophoreae)
(1Biology Department, University of Arak, P. O. Box: 38156-879, IRIB; 2Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond Sorrey TW9 3AB, UK; 3Department of Biology, Birkbeck College, University of London, Mallet Street, WC1E 7HX, UK )
The Leguminosae are one of the many families of higher plants that include many species in which crystals of calcium oxalate are a prominent structural feature. The crystal forms or combination of forms are characteristic of species or higher taxonomic grouping, so that they are useful for classification and identification and are of value (for example) in pharmacognosy. The Sophora group from Leguminosae: Papilionoideae is represented in Iran by three genera: 1. Sophora L., 2. Ammodendron Fisch and 3. Ammothamnus Bunge. Sophora consists of three species: S. alopecuroides L., S. mollis (Royle) Baker and S. pachycarpa C. A. Mey. Both S. alopecuroides and S. mollis have two subspecies: S. alopecuroides ssp. alopecuroides and S. alopecuroides ssp. tomentosa (Boiss.) Yakovlev and S. mollis ssp. griffithii (Stocks) Ali and S. mollis ssp. mollis. Ammothamnus and Ammodendron each have one species in Iran. Ammothamnus lehmanni Bunge and Ammodendron conollyi Bunge respectively. For the study of crystals both fresh and herbarium material was used. We developed two methods for testing crystal based on the information in Paulsen (1884) using anatomy and microchemistry. Preparations were viewed under light and polarizing microscope, and then photomicrographs were taken for each sample. Results showed all taxa studied, except Ammothamnus lehmanni, contained calcium sulphate (gypsum) crystals. Crystals occurred inside the vacuole of epidermal cells at the base of thricomes and around the stomata. They were 7.4-24 μ in size but there was variation in the expression of this character in different conditions. All of gypsum crystals have an amber colour. Gypsum crystal may have a protective defensive role against absorbency of UV and/ or defensive against?

P-46 : Consideration for the origin of Kinugasa japonica
Ichiro FUKUDA*
(The Asian Ecology-Evolution Bot. Inst)
Kinugasa japonica grows the restricted area in the central mountain region in Japan. The species has characteristics as large variations of morphological characters and high number of chromosomes (2n=40). By means of morphological and chromosomal analyses, it is considered that Kinugasa japonica is hybrid species between Trillium tschonoskii and Daiswa polyphylla.

P-47 : Mode of Hybrid species in Asian Trillium (Trilliaceae).
Ichiro FUKUDA*
(The Asian Ecology-Evolution Bot. Inst)
Trillium species have a disjunct pan Asian-North American distribution accross the Pacific Ocean. In Asia the 10 species of Trillium have developed a polyploid system of speciation, whereas in North America the 39 species have all thrived as diploids. Why have such differences evoluved in the speciation between the Asian and the American species? A comparison in the approaches that have taken place in their genomic chromosome arrangements and in their breeding systems will be discussed. Special attention will be given to the Asian Trillium species that have undergone speciation through hybridization.

P-48 : Berberidaceae in Thailand
(Faculty of Science, Chiangmai University)
Mahonia siamensis Takeda ex Craib is a species of Berberidaceae in Thailand.It grows naturally in hilly limestone areas, especially on Doi Phuka in the north of Thailand.The studies shows the detail in botanical description and anatomical characteristics. The botanical tradiyional knowledge is also given.

P-49 : Study of scanning on seed coat of species groups of linaria sec. Linaria (Scrophulariaceae) to linear leaves in Iran .
Seyyed Mohammad Mahdi HAMDI*1, Fatolah FALAHIAN 2 and Mostafa ASSADI3
(1Islamic Azad University , Garmsar branch ,Garmsar, Iran; 2Islamic Azad University, Science &Recearch branch, Tehran, Iran; 3Institute Forestes and Ranglands P. O. Box: 13185-116, Tehran, Iran )
Recently, Drew attention to the importance and impact of SEM in the study of systematic problem , and the most valid information the used to this teqnic . Heywood stydy systematic in 1971 , afterwards , Davise , study pay to taxonomy of Antirhineae and Sutton study to revision of the Tribe Antirhineae in 1988 .In resereach and study microscopy of seed coat surface linariain taxonomy of species . This group of Linaria sec. Linaria are complex , separation between of species are difficult . Genus of Linaria is 150 species in world ,that distribution in South Europe , W-South Asia , North africa and E-asia .In the base , form and seed coat divid to section six that about 30 species in Iran . 15 species of linaria are section lLinaria in iran . In this section ,species are linear leaves and discform seed . These species have complex and separation between of species are difficult that between species for management problem of scanning microscopy seed coat picture to SEM -LEO i-440, provid . Species that investigated are as fllow : Linaria lineolata(BOSS.), Linaria kurdica (BOISS.), Linaria leptoceras KUPP., Linaria steriatella KUPP., Linaria elymatica(BOISS.)KUPP., Linaria odora(BIEB.)FICHER, Linaria bamianica PATEZK, Linaria azerbaijanensis HAMDI&ASSADI , Linaria shahroudensis HAMDI&ASSADI.Seed coat microsculpturing shows diagonestic characters, inclosed species these characters are almost similar as awole but these are some difference in flower and leaf detials that to scanning separation of species .Reference:1)David A. Sutton 1988 :A Revision Of The Antirhineae , Oxford University Press-London&Oxford 2)Heywood V. H. (ed)1971 : Scanning electron microscopy , Systematic and Evolutionary applications -London .

P-50 : Inter-Specific Relationship by Leaf Character of Prunus from Jeju Island, Korea
Chan Soo KIM1, Myung Ok MOON2, Sim Hee HAN*1 and Joo Hwan KIM3
(1Department of Forest Genetic Resources, Korea Forest Research Institute, Suwon, 441-350, Korea; 2Warm-Temperate Forest Research Center, Korea Forest Research Institute, Seogwipo, 697-050, Korea; 3Department of Biology, College of Science, Taejon University, Taejon, 300-716, Korea )
This study is an approach to clarify the taxonomic identity of Prunus yedoensis, which is native in Jeju Island, Korea. Twelve Prunus species growing in Hallasan (Mt. Halla), Jeju were selected for investigation to determine inter-specific relationship. P. buergeriana, P. donarium, P. jamasakura, P. jamasakura var. quelpaertensis, P. japonica, P. maximowiczii, P. mume, P. padus, P. pendula var. ascendens, P. persica, P. sargentii and P. yedoensis were included. We analyzed leaf characters such as leaf blade length and width, petiole length, width of upper 1/8 and 1/4 leaf blade, the width of 1/2 leaf blade and lower 3/4 leaf blade, number of veins, distance of left and right gland from leaf base, and the angle of left and right leaf base. Leaf shapes of examined species were mainly divided two groups, eliptic and oblong. P. donarium, P. jamasakura, P. jamasakura var. quelpaertensis, P. maximowiczii, P. mume, P. padus, P. sargentii, and P. yedoensis were involved in elliptic shape group. The ratio of leaf blade length to width, number of veins, gland position and leaf base angle are distinct characters among species. These characters can be used to determine inter-specific relationship of Prunus species in Jeju Island. We created standard diagrams of leaf shapes of each taxa and similarity diagram based on examined characters.

P-51 : Taxonomy and distribution of Symplocos fasciculata s. lat. (Symplocaceae) in Malesia
Hidetoshi NAGAMASU*
(The Kyoto University Museum, Kyoto University, Kyoto 606-8501, Japan)
Symplocos fasiculata Zoll. s. lat. is an evergreen tree characterized by axillary inflorescenses of fasciculate racemes and is widely distributed from the Nicobars to the Philippines and Sulawesi. This species has been treated as a single species without recognizing infraspecific taxa (Nooteboom 1975, 1978). However, morphological studies revealed that two taxa can be recognized in this 'species': one with slender brown twigs, smaller leaves with minute serration and smaller fruits; and another with thick greenish twigs, larger leaves with distinct serration, and larger fruits. These taxa are referable to S. phanerophlebia Merr. originally described from the Philippines and S. fasciculata s. str., respectively. The distributions of these two species are clearly different. Symplocos fasciculata s. str. covers the western part: the Nicobals, Peninsular Thailand, Malay Peninsula, Sumatra, Java and Borneo. While that of S. phanerophlebia does the eastern part: Borneo, the Philippines and Sulawesi. In Borneo, these two taxa are overlapping, but the former is not found in Sabah and the latter is quite rare and restricted to the central mountainous area in Sarawak.The distribution pattern of these two taxa partly overlapping in Borneo suggests that these two taxa should be distinguished at specific rank, and shows the floristic complexity of Borneo with a contrast between the eastern coast and the western coast.

P-52 : Taxonomical analysis of Skimmia japonica (Rutaceae) in Japan and Taiwan
Tomoko FUKUDA*1 and Hidetoshi NAGAMASU2
(1Department of Botany, Graduate School of Science, Kyoto University; 2Kyoto University Museum )
The genus Skimmia comprises 4 species (Taylor 1987), occurring from the Himalaya to Far East Asia. Skimmia japonica is distributed from Taiwan to Sakhalin, and is found in various habitats. In the southern part of Japan, and in Taiwan, the plants grow in evergreen Quercus forests, and in the northern part of Japan and in Sakhalin, in deciduous forests of Fagus crenata, or forests of Abies sachalinensis.Skimmia japonica has unisexual, 4-merous flowers. Three varieties have been recognized; var. intermedia, var. japonica and var. lutchuensis. The first one has creeping stems, the latter two have erect stems, and the last one is distinguished by its tree-like habitat and large leaves (Yamazaki 1993). These varieties, however, have not been universally accepted because of the lack of distinctive characteristics and geographic distributuions. In our study we found new, useful characteristics of Skimmia japonica; the density of the hairs on the midrib on the adaxial surface of the leaves, and leaf size. Skimmia japonica var. intermedia has dense hairs (40-60pcs/4mm2), and relatively small leaves. Var. japonica and var. lutchuensis have highly variable hair density (glabrous in the Ryukyus and in southwestern Japan, but 5-50pcs/4mm2 in other localities), and leaf size. S. japonica var. japonica in Taiwan has small leaves with some hairs (20-30pcs/4mm2 and does not appear to be identical with Japanese var. japonica. On the bases of these observations, we consider that S.japonica with erect stems may not be delimitated as two varieties, but plants with creeping stems may be delimitated fairly well. All chromosome counts of S. japonica from Taiwan, Japan, and Sakhalin from 40 localities were 2n=30. The karyotype of S. japonica var. japonica in Taiwan, however, appears different from that of Japanese plants. In Taiwan, besides these plants, there are at least two types of plants. One is dioecious and has 5-merous flowers, and another is hermaphroditic and has 5-merous flowers. The chromosome counts for the latter was 2n=60, as reported (Johnson & Taylor 1989). Taylor (1987) recognized these two types as one species, and treated as subspecies of S.japonica, but needed further study, including chromosome analysis.

P-53 : Taxonomy and Biodiversity of Aegilops native to Iran
(1Biology Dept. Azzahra University, Vanak, Tehran, Iran; 2Biology Dept. Isfahan University, Hezar jarib, Isfahan )
The genus Aegilops is one of the wheat relatives that comprise main part of its gene pool. Identification of Aegilops species is difficult due to their vast morphological similarities and gene flow among different species of Aegilops and cultivated wheat. Aegilops has a wide distribution in different parts of Iran, and is capable of making different complexes with each other and with genus Triticum . This genus is a western Asia - Mediterranean element found around Mediterranean Sea and the western central part of Asia. Iran is one of the centers of the distribution and variation of this genus in the world. Due to the importance of cultivated wheat, having a better knowledge about new genetic resources is really necessary to improve wheat races and different biotechnological process with a correct and fundamental classification of the genus Aegilops. A lot of characters have been used to identify this taxon. In this study, 246 accessions of 13 Aegilops species were collected in the form of seed accessions and herbarium specimens from different part of Iran. Results show that there are 13 species of Aegilops in Iran as below: Diploids (Ae. tauschii, Ae. speltoides, Ae. caudata & Ae. umbellulata), Tetraploids (Ae. triuncialis, Ae. neglecta, Ae. geniculata, Ae. biuncialis, A. kotschyi, Ae. cylindrica, Ae. columnaris & Ae. crassa) and Hexaploids (Ae. juvenalis). Diploid species have a limited distribution and tetraploid ones are the most common species in Iran. Distribution of species in different height is discussed and some possible pattern for species distribution is given.

Kartini Binti SAIBEH*
Selected species from the genus Scindapsus, Epipremnum and six other genera of the tribe Monstereae and Anadendrum, were treated numerically using 11 micromorphological and anatomical characters. The computer software programme used for the analyses was MVSP Plus Verson 2.2 concentrating on Nearest Neighbour Cluster Analysis and Furthest Neighbour Cluster Analysis. The analyses constantly clustered the genera Scindapsus and Epipremnum. Both genera might have been congenerics and it is suggested the established reliance on gynoecial characters that define these genera be reassessed.


P-55 : Taxonomic revision of the genus Trismegistia (Musci, Sematophyllaceae)
Hiroyuki AKIYAMA*
(Museum of Nature and Human Acitivities, Hyogo)
The genus Trismegistia (Sematophyllaceae; Musci) can be distinguished from allied genera (Wijkia, Brothrella, Mastopoma and Pseudotrismegistia) by the the procumbent or upright stems with more or less developed stipes from prostorate primary shoots, differentiation of shapes among stipe, stem, and branch leaves, well-bordered and deeply serrate or dentate (rarely only serrulate) leaf margins, and segmented alar cells at least in the ourter one or two tiers. The genus is distributed mainly in southeast Asia and south Pacifics, and disjunctly reported from Eastern Nepal. Total nine species and one variety are recognized in this study; they are T. brachyphylla Fleisch., T. calderensis (Sull.) Broth., T. complanatula (Muell.Hal.) Muell.Hal., T. lancifolia (Harv.) Broth, T. malayana H.Akiyama, sp. nov., T. maliauensis H.Akiyama & M.Suleiman, T. panduriformis (C.H.Wright) Broth. var. panduriformis, T. panduriformis var. prionodontella (Broth.) H.Akiyama, comb. nov., T. plicata H.Akiyama sp. nov., and T. spinosodentata (Zant.) H.Akiyama, comb. nov., Although species with wide distribution ranges, that is, T. brachyphylla, T. calderensis, T. lancifolia, and T. panduriformis have a number of local morphological variations, we considered them as mere morphs and recognized no infraspecific taxon except for T. panduriformis var. prionodontella that is endemic to the higher portion of Mt. Kinabalu, Borneo. Inter-relationship between the genus Mastopoma and Trismegisita is also discussed, and Mastopoma deningeri (Herz.) H.Akiyama comb. nov., M. pulchellum (Herz.) H.Akiyama comb. nov., and M. delicatulum (Broth.) H.Akiyama comb. nov. are proposed. The only species previously reporte from Africa, Trismegistia monodii (P. de la Varde) Ando is not recognized as a member of Trismegisita.

P-56 : The monospecific genera Giraldiella and Pylaisiopsis should be synonymized with Pylaisia (Hypnaceae, Musci)
Tomotsugu ARIKAWA*
(Department of Biology, Keio University)
The genus Pylaisia (Hypnaceae, Musci) is distributed mainly in temperate regions of the Northern Hemisphere, and many species occur in East Asia. Arikawa (2004) revised the genus and recognized fifteen species conclusively. The monospecific genus Giraldiella, which had been treated as an endemic genus in China, was synonymized with Pylaisia (Arikawa 2004), based on the molecular phylogenetic analysis (Arikawa & Higuchi 2003) and the following morphological characters: (1) foliose pseudoparaphyllia, (2) seta which is sinistrorsely twisted below and dextrosely twisted above, and (3) smooth outer surface of lower part of exostome teeth (Arikawa 2004). The new combination Pylaisia levieri (M?ll. Hal.) Arikawa was published with description, illustlation, representative specimens examined, habitat, distribution, and diagnostic characters. The genus Pylaisiopsis is distributed over the eastern Hymalayas and southwesten China, and it is also treated as a monotypic genus. Detailed morphological examination had proved that the following features are characteristc of the genus: (1) strongly projecting perichaetial leaves, (2) spores growing into bullet-shaped endosporic protonemata within the capsule. However, the characters mentioned as the basis of the synonymy of Giraldiella to Pylaisia are also shared with the genus Pylaisiopsis. The phylogenetic analysis using the nucleotide sequence of chloroplast rbcL gene also suggests that Pylaisiopsis speciosa should be placed in the genus Pylaisia. Arikawa et al. presented those results in the 67th Annual Meeting of the Botanical Society of Japan (September 2003). In Arikawa (2004), however, I continued conventional treatment about the genus Pylaisiopsis and could not published those results which had been presented in the meeting because of insufficent examinations. I have not examined enough specimens, including the type of the genus. In this symposium, I will show that the genera Giraldiella and Pylaisiopsis should be treated as the synonym of Pylaisia because of the morphological characters and morecular phylogenetic analysis again. New figures and data will be presented.

P-57 : Taxonomic studies of Riccardia elata (Steph.) Schiffn. and its related species in Southeast Asia
Tatsuwo FURUKI*
(Nat. Hist. Mus. & Inst., Chiba)
The genus Riccardia is a one of the dominant in mossy forest of Southeast Asia, and it is very difficult to determine because of its simple thalli. Riccardia elata (Steph.) Schiffn. was described from Java, and it was reported from Southeast Asia and ist adjacent regions. This is very characteristic of large thalli and erect main thalli; however, it may be confused with R. plumosa (Mitt.) E.Campb. and some species, and it was treated as a synonym of R. plumose. In this study, this species and its related species, species with erect thalli, are taxonomically revised, and this species is distinguished from other species by large thalli, wide wings of ultimate thalli and indistinct trigones of wings. R. plumosa, R. diminuta Schiffn., R. anguste-alata (Steph.) Hewon, R. robinsii Hewson et Grolle and some species are recognized.

P-58 : Taxonomy of Himalayan species of Cryptoleptodon (Musci)
Masanobu HIGUCHI*
(Department of Botany, National Science Museum, Tokyo)
Among the four species of Cryptoleptodon, three species, C. pluvinii (Brid.) Broth. (= C. flexuosus (Harv.) Renauld et Cardot), C. rigidulus (Wilson ex Mitt.) Broth. and C. acuminatus M.Fleisch., have been known from the Himalayas, except African C. longisetus Enroth. Taxonomic position of the genus is controversial. That is, the genus was placed by the different researchers in Neckeraceae, Leptodontaceae or Pterobryaceae. These attempts were based on mainly gametophytic characters and sporophytic characters of C. pluvinii. Because no sporophytes have been known from the remaining three species. My examination of the specimens with sporophytes of Cryptoleptodon collected from Pakistan adds some taxonomic data of the genus. They show that the species has dioicous sexual condition, shortly exserted capsules and reduced peristomes. The sporophytic characters of the plants are different from those of C. pluvinii. The gametophytic characters are similar to those of C. rigidulus, but it differs from C. rigidulus by having asexual reproductive organs. It is also similar to Leptodon smithii (Hedw.) F.Weber et D.Mohr in having shortly exserted capsules, but is different from that in having reduced peristome. The peristome structure is significant at the generic level. Although the identity of C. acuminatus is uncertain, the Pakistan plants are considered to be a new species.


P-59 : On an endemic species Agarum Oharaense Yamada (Laminariaceae, Phaeophyta) in Japan
Masahiko MIYATA*
(Natural History Museum & Institute, Chiba)
In orderto taxonomically reexamine the position of Agarum oharaense Yamada in Genus Agarum Dumortier (Laminariaceae, Phaeophyta) , which is an ebdemic species in Japan, morphological study of characters in detail was done. The results of this study of characters, twisty and flattened stipes with rhzoidal hapters, insist that A. oharaense is distinct species in Genus Agarum. Then molecular analysis of the nuclear ribosomal DNA(18SrDNA)internal transcribed spacer indicate that A. oharaense is distinct species (Miyata & Yotukura, 2002).

P-60 : Taxonomic study of two species of Chlorogonium (Volvocales, Chlorophyceae) in Japan
Takashi NAKADA* and Hisayoshi NOZAKI
(Department of Biological Sciences, Graduate School of Science, University of Tokyo)
Chlorogonium is an unicellular genus characterized by having spindle-shaped vegetative cells. Recently, Nozaki et al. (1998, J. Phycol.) studied taxonomy of six species of Chlorogonium in culture. They demonstrated that physiological examination, transmission electron microscopy and molecular phylogenetic analysis are essential for correct species identifications. However, only three species (C. neglectum; C. fusiforme and C. capillatum) were identified based on Japanese materials. Therefore, further taxonomic studies of Japanese strains of Chlorogonium are needed to evaluate correct species composition of Chlorogonium in Japan.Here, we report two species of Chlorogonium newly identified based on cultured materials. These were isolated from Chiba Prefecture, Japan. They were studied with a light microscope equipped with Nomarski interference optics, a transmission electron microscope and DNA sequencing. They were identified as C. elongatum (from soil of a paddy field) and C. kasakii (from bottom mud of a pond).C. elongatum has two spherical pyrenoids with penetrating thylakoid lamellae. Their pyrenoids were stable (distinct) under photoheterotrophic conditions. The stigma of C. elongatum consists of a single layer of globules. Light and transmission electron microscopy of this species is very similar to those of C. euchlorum, and both species were briefly reported from Japan (Hirose and Yamagishi, 1977). However, the pyrenoids of C. euchlorum are unstable (reduced) under photoheterotrophic conditions.C. kasakii has two polygonal pyrenoids without penetrating thylakoid lamellae, stable under photoheterotrophic conditions. The stigma of C. kasakii consists of three layers of globules. This is the first record for C. kasakii from Japan. Formerly, no strain of C. kasakii other than the authentic strain has been reported and no comparative morphological study has been performed. Therefore, this is the first such study of C. kasakii.Phylogenetic analysis of the rbcL genes was also performed. It revealed that the Japanese strain of C. elongatum forms a robust clade with the other C. elongatum strains from England and U. S. A., and the English strain is the closest relative of the Japanese strain. The Japanese strain and the authentic strain of C. kasakii are sister to each other within the Chlorogonium and related species.

P-61 : Taxonomic significance of internal morphology of the oospore wall in the genus Nitella (Charales, Charophyceae)
Hidetoshi SAKAYAMA*1, Kazuyuki MIYAJI2, Tamotsu NAGUMO3, Masahiro KATO4, Yoshiaki HARA5 and Hisayoshi NOZAKI4
(1Biodiversity and Phylogenetic Study Section, Environmental Biology Division, National Institute for Environmental Studies; 2Department of Biology, Faculty of Science, Toho University; 3The Nippon Dental University; 4Department of Biological Sciences, Graduate School of Science, University of Tokyo; 5Department of Biology, Faculty of Science, Yamagata University )
In order to construct natural taxonomic system of Nitella, 18 species of Nitella subgenus Tieffallenia were re-examined based on SEM observations of internal morphology of the oospore wall (IMOW), as well as atpB-rbcL-psaB-ITS-5.8S rRNA phylogeny. Our SEM observations demonstrated three types of IMOW; homogeneous (HG), weakly spongy (W-SG) and strongly spongy (S-SG) types. Based on differences in IMOW, the reticulate and tuberculate oospore walls previously recognized in external morphology of the oospore wall (EMOW) were classified into two distinct types corresponding to the HG and S-SG types, respectively, which are separated from one another in the present molecular phylogenetic analyses. Our molecular phylogeny demonstrated that the subgenus Tieffallenia consists of at least four robust monophyletic groups - three HG type clades (one of them is further subdivided into two sister monophyletic groups) and a spongy (S-SG and W-SG) type clade - that were characterized by differences in IMOW and EMOW. Moreover, our SEM observations of IMOW also demonstrated that N. megaspora identified by LM includes at least two distinct species characterized by the differences in IMOW. Therefore, detailed IMOW and EMOW seen with SEM is efficient for characterizing phylogenetic groups within the subgenus Tieffallenia and should be used to construct a natural taxonomic system at the sectional level, after examinations of a large number of taxa.

P-62 : Rediscovery of the wild-extinct species Nitellopsis obtusa from Lake Kawaguchi, Japan (Charales, Charophyceae)
Syou KATO*1, Hisayoshi NOZAKI1 and Jiro TANAKA2
(1Department of Biological Sciences, Graduate School of Science, University of Tokyo; 2Department of Ocean Science, Tokyo University of Marine Science and Technology )
Charalean algae constitute one of the green algal groups. They occur in mainly fresh water, such as lakes, ponds, swamps, paddy fields. Especially in the lake, they thrive below the vegetation zones of aquatic vascular plants and form a "Chara zone". Nowadays, biodiversity of freshwater habitats of aquatic macrophytes in Japan is becoming reduced because of the recent pollution of the Japanese inland waters. The Charales seem to be especially endangered in such a habitat. Seventy-four taxa of Charalean have been known from Japan (Imahori and kasaki 1977), but about half of the taxa are listed in Red Data Book (Environmental Agency of Japan 2000). Although Nitellopsis obtusa Groves had once grown in four Japanese lakes, Lake Ashinoko, Lake Kawaguchi, Lake Nojiri, Lake Yamanaka (Kasaki 1964), they were thought to be extinct, based on the recent field sueveys in 1992-1993 (Nozaki et al. 1994). During our studies of the Charalean flora of Japan in 2003, N. obtusa was collected from natural populations of Lake Kawaguchi. The thalli formed huge bract-cells and stars, asexual reproductive organs. These are the diagnostic characters of N. obtusa (Wood 1965). Furthermore, the rbcL gene phylogeny demonstrated that the present alga and N. obtusa from Germany (McCourt et al. 1996) formed a robust small clade.

P-63 : Sexual Reproduction of Dilophus okamurae (Dictyotales, Phaeophyceae)
Zhongmin SUN* and Jiro TANAKA
(Tokyo University of Marine Science and Technology, Phycology Laboratory)
Dilophus okamurae distributes in Pacific coast of Asia. Basing on a multilayered medulla in marginal area of a branch, it can be easily distinguished from Dictyota species which have a unilayered medulla. Sporophyte of Dilophus okamarae was described by Okamura(1931), but gametophytes have never been reported. Sporophytes, male and female gametophytes were collected at Tateyama, the Boso Peninsula , Japan. Frond attached on a red alga is 15-20 cm high and much branched. Reproductive organs are spread on both surfaces of a branch. Sporangial sori are scattered over all the surface irregularly. Sporangia is spherical or ellipsoid. When becoming mature, tetraspores(63.6±3.5µm in diameter) and a few big spores are released. Each ooganial sorus is seen as a dot with naked eyes, which contains about 10 to 15 oogania. Released eggs(68.5±2.1µm in diameter) are almost of the same size. Antheridial sori are blister-like, and mature antheridia with 8 layers of loculi are 43.3±3.8µm in height. Sperm(5.1±0.7µm in diameter) is rounded with only one flagellum. The unilayered medulla and the reproductive structure of Dilophus okamurae which occurs in early spring are similar to those of Dictyota, that supports the closer relation between the two genera.


P-64 : Endogenous Leaf Initiation in Apical-meristemless Shoot of Cladopus queenslandicus (Podostemaceae) and implications for evolution of the shoot morphology
Satoshi KOI*1, Ryoko IMAICHI2 and Masahiro KATO1
(1Department of Biological Sciences, Graduate School of Science, University of Tokyo; 2Department of Chemical and Biological Sciences, Faculty of Science, Japan Women )
Podostemaceae, comprising three subfamilies with about 270 species assigned to about 47 genera, are an aquatic angiosperm family that live in rapids and waterfalls in the tropics and subtropics of the world. For adapting to such an extreme habitat, their morphology is specialized and the plant organization has been described as exhibit a misfit or fuzzy morphology. From light and transmission electron microscopic observations, we discovered a peculiar organogenetic pattern of endogenous leaf initiation at the shoot tip in the absence of an apical meristem in Cladopus queenslandicus, subfamily Podostemoideae. The pattern, like the one described previously for some other species, is in a marked contrast to the general pattern seen in most angiosperms that the shoot apical meristem is essential to leaf formation on the flank. Our noteworthy finding is that leaf formation involves two unique processes, i) cell separation that occurs on the outside of the incipient leaf primordium, and ii) vertical tissue splitting in a region where a shoot apical meristem exists in other angiosperms. Comparison in the presence or absence of a shoot apical meristem, cell separation, and tissue splitting in Podostemaceae shows that this organogenetic pattern may be a defining evolutionary novelty of the Asian-Australian lineage of the subfamily Podostemoideae.

P-65 : Developmental morphology of inflorescences of Monophyllaea glabra (Gesneriaceae) .
Madoka AYANO*1, Ryoko IMAICHI1 and Masahiro KATO2
(1Department of Chemical and Biological Sciences Japan Women's University; 2Department of Biological Science, Graduate School of Science, University of Tokyo )
The Monophyllaea or one-leaf plants are distributed in limestone regions of Southeast Asia. The body is constructed of only an enlarged cotyledon (macrocotyledon) and a long stalk with no vegetative shoots. Inflorescences generally arise from the macrocotyledon base, and show so complex structure that they remain to be interpreted. We examined the developmental morphology of inflorescences of M. glabra, a species occupying the base position in the molecular phylogenetic tree of the Monophyllaea (H. Setoguchi, unpublished data). At the beginning of the reproductive stage, the groove meristem formed at the site of the embryonic shoot apex between the two cotyledons enlarges to give rise to the first inflorescence axis. Concomitantly, a small meristem detaches from the groove meristem toward the macrocotyledon. Four or five inflorescence axes branch successively from the detached meristem in a zigzag line. They are subtended by a bract at every branching point, and have a bracteole at their base. As a result, a primary inflorescence is constructed of the first axis with no bract and bracteole along with five or six axes with pairs of bracts and bracteoles. After formation of the primary inflorescence, additional inflorescences arise from the axillary meristems of both the microcotyledon and of bracts and bracteoles of the primary inflorescence. Every additional inflorescence also branches several times forming bracts and bracteoles is equal in structure to the primary inflorescence with the first axis removed. In other words, the first axis of the primary inflorescence is specific in lacking bracts and bracteoles as well as in occupying the central position between two cotyledons. The first axis with no bract and bracteole might be derived from the groove meristem itself (embryonic shoot apex) the other axes derived from the axillary bud of the macrocotyledon. The evolution of inflorescence structure in the Monophyllaea, is discussed based on these data.

P-66 : Basal meristem of Isoetes asiatica rhizomorph
Sunyoung YI* and Masahiro KATO
(Department of Sciences, Graduate school of Science, University of Tokyo)
The rhizomorph is root-bearing organ seen only in Isoetes (Lycopsida, Microphyllophyta) among living plants and in the Carboniferous Lepidodendrales. The Isoetes rhizomorph occupies the basal half of the corm and bears roots in orthostichies. The Isoetes rhizomorph is assumed to have evolved by a series of reduction from that of lepidodendrids via those of the Pennsylvanian Paurodendron and Triassic Pleuromeia. The apex of the Paurodendron rhizomorph, like that of Stigmaria, is covered by an apical plug circumscribed by a circular groove, and a parenchymatous rim. The circular groove of Paurodendron above the basal meristem in the rhizomorph is hypothesized to be a homologue of the linear groove of Pleuromeia, Isoetes, and others. The basal meristem of Isoetes is linear, lies in parallel to and under the furrow of the rhizomorph, and produces root intermittently. We examined the development of roots from the basal meristem in Isoetes asiatica. Prior to root initiation, an area distal to the basal meristem is small in cross section, and then expands by cell increment. Subsequently a new root forms in the area between developing roots and occupies it. The new root is shifted to the flank of the basal meristem, due to expansion of the area, and the flank of the basal meristem becomes a lateral meristem, which contributes to the secondary thickening of the rhizomorph. Therefore, there is a considerable difference between the rhizomorphs of Isoetes and extinct lepidodendrids.

P-67 : Expression of STM and YABBY homologous genes in phylloclades of Ruscus aculeatus (Asparagaceae)
Yumiko HIRAYAMA*1, Yukiko OOYA1, Toshihiro YAMADA2, Motomi ITO3, Masahiro KATO4 and Ryoko IMAICHI1
(1Department of Chemical and Biological Sciences, Faculty of Science, Japan Women; 2Department of Geology and Paleontology, National Science Museum; 3Department of Systems Science, Graduate School of Arts and Sciences, University of Tokyo; 4Department of Biological Sciences, Graduate School of Science, University of Tokyo )
The phylloclades of Ruscus are flattened leaf-like organs occupying the position of an axillary bud and forming inflorescences on either their adaxial or abaxial surfaces. The inflorescence is subtended by a bract formed from the phylloclade itself. Organographically, the phylloclade has been interpreted in four ways as: a flattened stem; a leaf; a congenital fusion product of a stem and a leaf; or an intermediate organ between a stem and a leaf. To provide new insight into the phylloclade, we analyzed the expression pattern of the STM ( SHOOTMERISTEMLESS ) homologue expressed specifically in the shoot apical meristem (SAM), and of the YABBY2 ( YAB2 ) homologue expressed specifically in lateral organs. In young terminal shoots of R. aculeatus , expression of the STM homologue was detected in the SAM and in apices of young phylloclade primordia. Expression of the YAB2 homologue was detected in abaxial tissue of a slightly older phylloclade primordium with an inflorescence bract. The YAB2 homologue was also detected in the inflorescence bract itself. These data suggest that the phylloclade is developed under the same genetic programs as the SAM as well as the lateral organs of ordinary plants. Therefore, it is unlikely that the phylloclade is an additional leaf subtended by a scale leaf. It might be a flattened branch or intermediate organ with genetic programs of SAM and lateral organs. For complete clarification, we need more data on the exact gene expression patterns of the STM and YAB2 homologues during phylloclade development.

P-68 : Function of FLO/LFY genes in the moss Physcomitrella patens and the gene evolution regulating flower development in land plants
Takako TANAHASHI*1, Naomi SUMIKAWA2, Mitsuyasu HASEBE2 and Masahiro KATO1
(1Department of Biological Sciences, Graduate school of Science, The Univ. of Tokyo; 2National Institute for Basic Biology )
To understand plant diversity and evolution from the viewpoint of the regulation of flower development, we characterized FLORICAULA/LEAFY (FLO/LFY) genes in the moss Physcomitrella patens. FLO/LFY genes encode plant specific transcription factors that positively regulate floral homeotic genes. In angiosperms, FLO/LFY genes have largely conserved function in flower development among distantly related species. The transcriptional activation of floral homeotic genes by FLO/LFY genes is conserved even in gymnosperms, conifer and Gnetum; however this is not likely the case with ferns. The function of FLO/LFY genes in bryophytes has not been identified so far. We isolated two FLO/LFY genes from P. patens and designated them as PpLFY1 and PpLFY2. They were so similar to each other as to have nearly 90% identical nucleotide sequence in their coding regions. Their expressions were detected mainly in gametophore shoot apices, young leaves, archegonia and an egg cell in them and developing sporophytes. Functional analyses by gene disruption showed that double disruption of PpLFY1 and PpLFY2 caused severe defect in the sporophyte formation. The detailed observation of archegonia and crossing experiments with wild type strains showed that the gene disruptants were arrested at a round small zygote stage just after fertilization. Because the wild type zygote expands and fills the cavity of the archegonium, the zygote of the gene disruptants was interpreted to have defect in the expansion process. Moreover, a very little number of gametophores of the gene disruptants produced sporophytes. These sporophytes exhibited abnormal morphology and spores yielded by them seldom germinated. This indicated that PpLFY1 and PpLFY2 also regulate the sporophyte development and spore formation after the zygote stage.Our results demonstrate that FLO/LFY genes in the moss function in the zygotic maturation and sporophyte development afterwards. We suggest that FLO/LFY genes, which regulate the sporophyte development at various stages in bryophytes, evolved to specifically regulate the flower development in sporophytic generations in angiosperms during land plant evolution.

P-69 : Vegitative Anatomy of BEGONIA species
Adewale Taofiq ARIWOOLA*
Anatomical studies of vegetative structure of BEGONIA species were carried out. Leaf samples of different taxa of BEGONIA found in Nigeria from different geographical locations (habitat) were examined using light microscopy method. A microscopic observation shows that different species of BEGONIA possess similarities and as well as differences in their anatomical features. The epidermal cell shape of the species could be regular,polygonal or irregular. Some are curve.Trichomes are present in some and absent in others. Description statistics of mean ( X)standard deviation and standard errors ( S . E) Calculate for all quantitative characters each taxa also varies in the species of BEGONIA.

P-70 : Diversity of pollen-tube growth pattern in pistils of Fagales
Akiko SOGO* and Hiroshi TOBE
(Department of Botany, Graduate School of Science, Kyoto University)
In flowering plants, pollen tubes grow from the stigma to the embryo sac through female tissues, delivering a sperm cell to the egg cell. In a majority of them the fertilization occurs within 24 to 48 hours after pollination. In some fagalean species, however, it occurs over one month after pollination because ovules are not yet ripe at the time of pollination. We pursued the pollen-tube growth in pistils of Casuarina, Alnus and Fagus (Fagales), the former two of which are known to undergo chalazogamy instead of porogamy as in most angiosperms including Fagus. All the three genera agreed in that the pollen tube(s) grows intermittently in several steps in close association with the development of an ovary and ovules, but differed in the following aspects. In Casuarina and Alnus the ovules are not differentiated when the stigma received pollen grains, so that their pollen tubes stop growing in the style. Only one pollen tube resumed growing toward the ovary after the ovule underwent meiosis in a nucellus, and, after two- or three-time repetition of stop and growth, it reached the chalaza to wait for maturity of the embryo sac. In Fagus, on the other hand, young ovules are already differentiated at the time of pollination, and many pollen tubes grew downward in a pistil without stopping in the style but stopped growing around ovules. The pollen tubes resume growing when the ovules developed sporogenous cells in their nucellus, and several of them entered the micropyle and stopped growing in the exostome again to wait for maturity of the embryo sac. Therefore, a single pollen tube is selected from many in the style in Casuarina and Alnus and in the micropyle in Fagus. No matter how the pollen tube reaches the embryo sac, the pollen tube stops at the chalaza in Casuarina and Alnus and at the micropyle in Fagus, and waits there until the embryo sac matures. Thus the chalaza in Casuarina and Alnus seems to have the same function as the micropyle in Fagus. We also discuss the evolution of the chalazogamy in Fagales.

P-71 : Dimorphism of gall type induced by Pseudasphondylia neolitseae on Neolitsea sericea , representing geographic variations in Kyushu.
Misako MISHIMA*, Christiane Yuri YOSHIDA and Junichi YUKAWA
(Kyushu University Museum)
Neolitsea sericea (Bl.) Koidz. is an evergreen tree species of Lauraceae and distributed in Honshu, Shikoku, Kyushu, Okinawa, and southern part of Korean Peninsula (Momiyama, 1989). Pseudasphondylia neolitseae Yukawa (1974), a dipteran insect of Cecidomiidae, forms the gall on the leaves of N. sericea. The gall form shows two types, upper side is more projected (upper type) or the lower side is more (lower type). The geographic distribution of upper and lower types is different, southern part and northern part of Kyushu, respectively (Yukawa & Masuda, 1996). Yukawa observed the distribution of each type in western Kyushu, and which was changed alternatively in Yatushiro City (unpublished). Yatushiro City is the western end of tectonic line in Kyushu Island. We hypothesized that distribution border of two gall types is the tectonic line, and tested more than 20 sites including the Usuki-Yatushiro tectonic line, the predicted median tectonic line (MTL) in Kyushu. The distribution border of two gall types is MTL in Kyushu main land, while upper type was dominant beyond MTL in islands of western Kyushu.

P-72 : Usefulness of cuticular characters as a grouping system of the Asian Lauraceae species
Sachiko NISHIDA*
(The Nagoya University Museum)
Lauraceae contain many taxa that are important ecological and economical components of tropical Asia. Still, classification of the family in this region has not been sufficiently clarified. Although there are a few systematic studies for some genera based on the molecular phylogeny in progress, the large size of the genera and the difficulty of collecting samples have slowed their process. It is inevitable to recognize some subgeneric groups and use the representative species for each group for the phylogenetic analysis of the genera. Practical methodology to recognize the subgeneric groups has been needed for the progress of the systematics. In this study, the author proposes a grouping system by leaf cuticular characters as one of the practical methodologies to recognize the subgeneric groups with a test case of the Bornean Beilschmiedia species.Beilschmiedia is one of the largest genera of Lauracea, and contains about 30 members in Borneo. While revising the Bornean Beilschmiedia species, the author has recognized several subgeneric groups by the floral characters. On the other hand, in the former study with Christphel, the author proved that cuticular characters were informative to subdivide the neotropical Beilschmiedia species that had few differences in the flowers (Nishida & Christophel 1999, Nature and Human Activities). In this study, the author carries out the cuticular study on the Bornean species and compares the result with the subdivision by the floral characters, and discusses the usefulness of the grouping system by the cuticular characters. Cuticles are easily studied with herbarium materials, and the characters would provide valuable information to understand the systematics of the Asian Lauraceae.

9.Ecology and Vegetations

P-73 : Evaluation of New Elms from China for Cold hardiness in Northern Latitudes
(The Morton Arboretum, 4100 Illinois Route 53, Lisle, IL 60532, USA)
The high resistance of elms from China to Dutch elm disease and other problems makes the Chinese elm an excellent choice for the urban landscape. Many new Chinese elms are being developed through the tree breeding program at The Morton Arboretum. The stem cold hardiness of nine new elms from different provenances of China and several cultivars were evaluated using an artificial freezing test. The LT50 (the temperature at which 50% of the tissues were killed) of the most to least hardy genera for February 1998 were Ulmus macrocarpa ( -36°C), U. wilsoniana # 673 (-34°C), U. wilsoniana # 669 (-34°C), U. parvifolia (-34°C ), U wilsoniana # 997 ( -33.8°C ), U. szechuanica (-30°C), U. gaussenii (-30.7°C), U. bergmanniana, var. lasiophylla (-27.7°C), U. castaneifolia (-26°C). New elm cultivars cold hardiness were: U. Morton Plainsman Vangurd TM Elm, -40°C) , U. Morton Glossy - Triumph TM Elm (-40°C), U. Stalwart Commendation TM Elm ( -40°C), U. Patriot (-38°C), U. Morton Accolade TM Elm (-35.5°C), U. Morton Red Tip- Danada Charm TM Elm (-31°C), U. parvifolia AlleeR (-26.5°C) and U.parvifolia AthenaR Elm (-23.5°C).

P-74 : Searching for cold hardy "Lace-bark" elm (Ulmus parvifolia Jacq.)
(The Morton Arboretum, 4100 Illuinois Route 53, Lisle, IL 60532 USA)
"Lace-bark" elm (Ulmus parvifolia Jacq.) has extensive distribution in China. Mottled reddish "lace-bark" trunk is a distinctive feature of this tree. The high resistance of this elm to Dutch elm disease and other elm problems makes it an excellent tree for urban landscapes. We evaluated eight different U. parvifolia Jacq. groups from different provenances in China at The Morton Arboretum, Lisle, Illinois, USA. for stem cold hardiness and dormancy in October 1999, January and March 2000. There were significant differences between these groups for acclimation and deacclimation associated with their provenance origin. Chilling requirements for seed germination were determined. Seedeling from 6 different trees were planted and screening for potential new cultivars. Response of seeds to freezing temperature during winter and subsequent germination for potential invasiveness in warmer climate will be reported.

P-75 : Vegetation history in the vicinity of Kagaminuma in Niseko,southwestern Hokkaido,Japan
(Sapporo Daiichi High School)
The study area is located from 40km northeast Kuromatsunai lowland,the present northen limit of Fagus. It is a high land mire,called Tekagaminuma by Fujita et.al(1997), located 1km from Kagaminuma,and 580m a.s.l..There are dominant Betula ermanii and mixed needle leaved trees and decidous broad leaved trees. In the mire,Sphagnum,Drosera rotundifolia,Hosta recifolia,Ledum palustre var diversipilosum,Vaccinium oxycoccus,Carex sp.are distributed.Its base rock is a part of the Niseko volcanic mountains which had been active in the first ~middle Pliestoscene.(Ohba 1960;Ishida & Mimura 1991).The samples were taken 4.8m in thicknes,by a Thomas type handorger. The sediments were bluegrey fine sand,volcanic ash,peaty clay,land peat. The volcanic ashes are being analized now.5 pollen assemblage zones are recognized through pollen anaiysis according to the dominant pollen taxa.They are,from bottom to upper,Ni I:Picea-Betula zone,Ni II:Betula zone,Ni III:Quercus zone,Ni IV:Betula-Quercus zone,Ni V:Betula zone.Fagus pollen appears from sample No.170 which belongs to Ni IV,the Hypsithermal time, and countinously appears from No.120 which belongs to Ni III and its peak is No.60,11%.This fact indicates that Fagus was distributed around this mire before forming in the present northern limit.The change of Ni I~Ni II shows that it warmed suddenly from a cold climate.Thus,the origin of this mire seems to be around the last-term of the Late Plistosene.

P-76 : North-western Himalayas - A hidden treasure of diverse gene-pools of invaluable PGRs vis-a vis Opportunities to establish linkages with agri -horticultural based industry
(1Cytogenetics & Tissue Culture Lab,Dept of Plant Breeding & Genetics,HP Agricultural University,Palampur-176062 India; 2DSS,Nagri-Chachiyan ,Palampur-176061 India; 3Lab of Plant Molecular Genetics,Osaka Kyoiku University,4-698-1 Asahigaoka , Kashiwara, Osaka 582-8582 Japan )
North-western Himalayas are comprised of the dry & wet temperate regions of HP, Uttranchal and J&K states of India. Almost all the regions are endowed with rich and highly diverse plant genetic resources. As the Green revolution could not register much impact of inducing mono cropping culture in almost all the tribal valleys of Himalayan ranges due to certain communication, ethnic and social barriers, growing of mixture of various landraces is still in vogue in these areas and diversity of various crop species is being maintained. To provide food security and future posterity, there is an urgent need to conserve this precious wealth. An endeavour has been exercised to conserve various CGRs comprising cereals(wheat, rye, barley and maize), pseudo cereals (amaranth, buckwheat and chenopod), pulses (adzukibean, common bean, chickpea and peas), minor millets(foxtail and kodo millet),medicinal herbs(different species of Bacoppa, Centella, Viola, Ocimum, Asparagus, Mentha, Spilanthus, Withania, Bunium, Crocus and Saussurea) and certain alpine grasses from the different parts of the north-western Himalayas. Various landraces and wildly growing lines have been conserved as active collections in the seed and field gene banks. Sustainably utilizing these bio resources, a number of improved crop varieties were released for general cultivation in the valleys of their habitat. Most of these varieties are being grown as the cash crops and fetching excellent premium in the market. Besides, concerted efforts are being made by this group of scientists in the genetic amelioration of certain important crops of the Himalayan regions following various ultra modern approaches. Out of the locally available medicinal herbs, certain plant species of Bacoppa , Ocimum and Spilanthus have been brought under large scale cultivation and their products are floated in the adjoining markets by an associated volunteer Organisation, DSS, Nagri HP(India). This meagre effort has actually established an active linkage of the bio resources, their conservation, sustainable utilization , commercial cultivation and the industrial level exploitation. Following certain recommendations like, establishment of the gene banks in the vicinity of the PGRs, raising gene sancturies in the high variability regions for certain endangered plant species, creating perma-frost facility to conserve the PGRs as a back up mechanism(security from the explosives) in the permanent glaciers of the Himalayas, activate farmers' participatory approach and identification and documentation of the unexplored novel genes can actually open new vistas for providing food security, raising agri-horti based industry and managing the bio resources for future posterity

P-77 : Plant fossil assemblage of the Middle Jurassic (Callovian) Kaizara Formation
Toshihiro YAMADA*
(Division of Paleobotany, Department of Geology and Paleontology, National Science Museum)
Late Jurassic to Early Cretaceous florae of Japan are divided into three types, Tetori- (Siberian-) type flora, Ryoseki- (Gondowanian-) type flora and Mixed type flora of these two florae. Tetori-type flora flourished under the warm-temperate and humid climate is distributed in the Inner Zone of Japan and characterized by the abundance of the Ginkgopsids and Dicksoniaceous ferns. Ryoseki-type flora grown in the tropical-subtropical and arid climate is distributed in the Outer Zone of Japan and characterized by the abundance of the bennettitalean genus such as Zamites and Ptilophyllum. In the Tetori Group (distributed mainly in Fukui, Gifu, and Toyama Prefectures), the Tetori-type flora has been thought to back to the Middle Jurassic time based on the plant fossils from the non-marine strata, although the exact chronostratigraphic correlation is difficult in such strata with non-marine origin because of the lack of index fossils. Here I report some plant fossils from the Middle Jurassic (Callovian) Kaizara Formation of the Tetori Group, which is marine deposit with many ammonoid fossils, distributed in Izumi, Fukui Prefecture. Collected plant fossils included some species of Otozamites, Ctenozamites and conifers in addition to Zamites which is characteristic genus of Ryoseki-type flora. No characteristic genera of Tetori-type flora were obtained and floristic composition in species revel is completely different from those reported so far from the Tetori Group. Furthermore they have very thick lamina. These suggest that the area was under the arid climate in the Middle Jurassic time as in the Euro-Sinian Region.