2.4 DIVISION OF APPLIED BIOSCIENCES

The Division of Applied Biosciences was created in 1996 to consolidate the three divisions of Agricultural Biology (founded in 1953), Fisheries Science (1953), Animal Science (1976) and two research laboratories from Division of Tropical Agriculture (1981). The division is conducting the research and education on the subjects aiming at the efficient utilization, conservation and creation of species for a variety of living organisms including microorganisms, plants, animals and fishes living in land and sea from the molecular to population levels. The division consists of 17 laboratories in which 116 master students including 4 students from abroad and 75 Ph D. students including 7 students from abroad are enrolled.

Chair of Science of Plant Resources

2.4.1 Laboratory of Plant Genetics

Staff Professor : Endo, Takashi, D. Agric. Sci Associate Professor : Miyashita, Naohiko, Ph. D. Assistant Professor : Nasuda, Shuhei, Ph. D. Students and research fellows Doctor's program : (2) JSPS Postdoctoral Fellow : (1) Master's program: (7) JSPS Foreingn Research Fellow: (1) Undergraduate : (2)

A. Research Activities (2004.4-2005.3) A-1. Main Subjects a) Cytogenetic analysis of a genetic genome rearrangement system in wheat Chromosomal structural changes frequently occur in specific lines of common wheat carrying certain alien chromosomes from wild species related to wheat. These chromosomal aberrations can be identified by the chromosome banding and in situ bybridization techniques. Using this genome rearrangement system, we have established deletion and translocation lines of wheat and are conducting studies on chromosome mapping and the introduction of useful genes from alien species, such as barley and rye, into bread wheat. Also, we have initiated a basic study aimed at cloning the gene responsible for the unique genome rearrangement system. b) Molecular cytogenetic analysis of wheat and its relative species We have been trying to shed light on the structure of the three wheat genomes by employing various molecular cytogenetic techniques. In order to characterize specific regions of wheat chromosomes efficiently, we have developed a high-throughput marker system derived from the AFLP (amplified fragment length polymorphism) technique. We have also been conducting molecular cloning experiments aimed at isolating genes encoding for centromeric

222 proteins in wheat to elucidate the structure and function of the plant centromers. A major characteristic of wheat genomes is the abundance of repetitive elements. To reveal their structure and evolutionary consequence in the genomes of wheat and its relatives, we are trying to clone and analyse species-specific repetitive elements. c) Molecular population genetics study on DNA variation in the genera Arabidopsis and Arabis, and Oryza. To establish a model system for plant population genetics, we have chosen two genera Arabidopsis and Arabis. So far, several genic regions have been analyzed to quantify DNA variation at a specific genic region. In addition, microsatellite and AFLP analyses were conducted to investigate the pattern of DNA polymorphism over the entire genome. Currently, we are analyzing genes involved in herbicide resistance in A. thaliana. To examine a more general picture of molecular variation in plant species, we are planning to compare these two genera and Oryza species, which are monocots. We have already analyzed DNA variation in two Adh locus regions in Oryza species, and are analyzing Blast-related and flowering genes.

A-2. Publications and presentations a) Publications Original papers Matsuoka, Y., and S. Nasuda: Durum wheat as a candidate for the unknown female progenitor of

bread wheat: an empirical study with a highly fertile F1 hybrid with Aegilops tauschii Coss. Theor. Appl. Genet.109; 1710-1717, 2004. Katto, M. C., T. R. Endo, and S. Nasuda: A PCR-based marker for targeting small rye chromatin introgressions in wheat. Genes Genet. Syst. 79; 245-250, 2004. Kimbara, J., T. R. Endo, and S. Nasuda: Characterization of the genes encoding for MAD2 homologues in wheat. Chromosome Res. 12; 703-714, 2004. Ito, H., S. Nasuda, and T. R. Endo: A direct repeat sequence associated with the centromeric retrotransposons in wheat. Genome 47; 747-756, 2004. Sasanuma, T., K. Chabane , T.R. Endo, and J. Valkoun: Characterizaion of genetic variation in and phylogenetic relationships among diploid Aegilops species by AFLP: incongruity of chloroplast and nuclear data. Theor Appl Genet 108; 612-618, 2004. Yoshida, K., N. T. Miyashita and T. Ishii, 2004 Nucleotide polymorphism in the Adh1 region of the wild rice species Oryza rufipogon. Theor. Appl. Genet. 109: 1406-1416. b) Conference and seminar papers presented The 75th Annual meeting of the Genetic Society of Japan: 6 presentations Annual meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry 2005: 1 presentation

A-3. Off-campus activities Membership in academic societies Endo, T.: Genetic Society of Japan (Editor-in-Chief) Endo, T.: Genetic Resources Committee and Resource Center, National Institute of Genetics (Member) Miyashita, N.: Genetic Society of Japan (Council Board)

223 Research grants Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture (A) (Nasuda) Japan Science and Technology Corporation Project (Endo, Nasuda) National Bioresource Project (the Ministry of Education, Science, Sports and Culture) (Endo and Nasuda)

A-4. International cooperations and overseas activities Scholars from abroad Dr. Neil Jones (United Kingdom)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Basic Bioresource Science I (Endo), Outline of Bio-production Science I (Endo), Genetics I (Endo), Genetics II (Miyashita), laboratory of Bioresource Science I, II (Endo and Miyashita), Seminar in Plant Resource Science (Endo and Miyashita) Graduate level: Genetics (Advance Course) I (Endo), Genetics (Advance Course) II (Miyashita), Seminar in Plant Genetics (Endo and Miyashita), Research in Plant Genetics (Endo and Miyashita)

B-2. Off-campus teaching, etc Part-time lecturer Endo, T.: Faculty of Education, Kyoto University of Education (Genetics)

B-3. Overseas teaching Students and research fellows from abroad Doctor’s Program: one (Colombia) JSPS Postdoctoral Fellow, Ali Masoudi-Nejad (Iran)

224 2.4.2 Laboratory of Crop Evolution

Staff Professor : Ohnishi, Ohmi, Ph.D. Associate professor : Kawahara, Taihachi, Dr. Agr. Sci. Assistant Professor : Yasui, Yasuo, Dr. Agr. Sci. Research associate : Yamane, Kyoko, Dr. Agr. Sci. Students and research fellows: Doctor's program: (3) Master's program: (5)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Studies on the origin and evolution of cultivated plants Buckwheat: This year we surveyed cultivated and wild buckwheat species in the Nu river and its tributary valleys of southeastern China. There found no wild ancestor of cultivated buckwheat, Fagopyrum esculentum ssp. ancestrale in the Nu river valley. Hence we can exclude the Nu river valley from the candidate of the original birthplace of common buckwheat (Now, the Lancan river valley and the Jinsha river valley are the strong candidate of the original birthplace of common buckwheat.). Both common and Tartary buckwheat were cultivated in the Nu river valley. The cultivated populations of the Nu river valley were phylogenetically distinct from those in the Lancan river and Jinsha river valleys, rather they are close to the cultivated populations from eastern Sichuan province. This implies that common buckwheat in the Nu river valley did not come from the adjacent Lancan river valley beyond high mountains, they probably recently came from the lower part of the Nu river. As for F. homotropicum, both diploid and tetraploid cytotypes were growing as a mixed population in the Nu river valley. So the hypothesis that tetraploid F. homotropicum originated in northwestern corner of Yunnan province, and it diffused to cooler northward or to upper stream of big rivers must be reconsidered. For the detailed analyses of phylogenetic relationships among Fagopyrum species and for the study of gene flow between cultivated and natural populations of common buckwheat, about 50 SSR markers have been developed. The study on gene flow between cultivated and natural populations of common buckwheat is now in progress. Perilla crops: We traveled around Hubei, Hunan, Jiangxi, Guandong, Guanxi and Fujaing provinces of southern China to search wild tetraploid Perilla species and cultivated P. frutescence var. crispa and var frutescence. Wild tetraploid Perilla were found everywhere south of the Yangtse river. Previously we insisted the border area of Hunan, Guanxi and Guandong to be the candidate for original birthplace of Perilla crops. From the distribution of wild Perilla species, it may be slightly eastward of the area previously insisted. Radish: Chloroplast SSR markers have been investigated for their variation among 82 accessions of cultivated and wild radish. Those accessions included wild radish in East Asia, Raphanus raphanistrum and R. landra from Europe and cultivated radish from Europe, India and the Himalayan region and Far East (China Korea and Japan). The experimental results showed that Spanish black radish situated quite different phylogenetic position from other

225 cultivated radish, indicating distinct origin of Spanish black radish. European cultivated radish, R. sativus var. sativus and East Asian cultivated radish, R. sativus var. hortensis were also genetically distinct, indicating independent origin of these varieties. Studies on chrolophyl deficient and other detrimental genes concealed in natural populations of wild radish, R. raphanistrum, were conducted for Poland, and Italian populations this year. b) Studies on the evolution of wheat and its wild relatives Ae. tauschii: Variation of Aegilops tauschii, the D genome donor to hexaploid bread wheat was surveyed by using cpDNA SSR regions. We analysed 20 SSR regions in 210 accessions of Ae. tauschii covering the whole distribution range. Enough polymorphisn was obtained for the analysis of intraspecific variation. We found this method qite useful for the analysis of intraspecific variation of wheat and its relatives because this can be applied to other species easily. Genus Triticum and Aegilops: Phylogenetic relationships of polyploid species belonging to Triticum and Aegilops were estimated from cpDNA sequence. The following results were obtained: 1) All the species belonging to Section Aegilops, except for Ae. kotschyi, Ae. peregrina and one haplotype of Ae. triuncialis, formed a cluster and then this formed a cluster with diploid Ae. umbellulata. 2) Ae. kotschyi and Ae. peregrina formed a monophyletic clade and clustered with Subsection Emarginata Eig. 3) Ae. cylindrica, without intraspecific differentiation, formed a cluster with Ae. tauschii and this further clustered with Ae. ventricosa. 4) Ae. crassa, Ae. juvenalis, Ae. vavilovii formed a cluster and then clustered with the above branch including Ae. tauschii. 5) A haplotype of Ae. triuncialis formed a cluster with several haplotypes of Ae. caudata. Earlier findings of at least two independent origin with different maternal lineage were confirmed in this study. 6) T. araraticum and T. dicoccoides clustered together and formed a cluster with Ae. speltoides. c) Studies on the self-incompatibility gene of common buckwheat In common buckwheat, the self-incompatibility gene is closely linked with the genes controlling several morphologies related with heterostyly and they form the S supergene. So far, the linkage map and genomic library have been constructed as a first step for the cloning of the S gene by the positional cloning method. Screening of the genomic library is now under progress. Since we have already got a clone closely linked (0.07 cM, about 210kb) to the S gene, we are expecting to have clones closer than this in near future. We are also working towards the construction of transformants in common buckwheat.

A-2. Publications and presentations a) Publications Books Ohnishi, O: Chapt. 6 Population Genetics and Evolution. Introduction to Plant Genetics (ed. by Tetsuo Mikami). Asakura-shoten, Tokyo, 2004 Original papers Konishi, T., H. Iwata, Y. Yasui, O. Ohnishi, Y. Tsumura and R. Ohsawa: Development of microsatellite markers for common buckwheat (Fagopyrum esculentum Moench). Proc. 9th Intl. Symp. Buckwheat at Prague; 212-218, 2004 Matsuoka, Y., N. Mori and T. Kawahara: Genealogical use of chloroplast DNA variation for intraspecific studies of Aegilops tauschii Coss. Theor. Appl. Genet. 111; 265-271, 2005

226 Ohnishi, O.: Wild buckwheat species in the San Jiang (three rivers) area of southwestern China. Proc. 9th Intl. Symp. Buckwheat at Prague; 226-232, 2004 Ohnishi, O.: On the origin of cultivated buckwheat. Proc. 9th Intl. Symp. Buckwheat at Prague;16-21, 2004 Tomiyoshi, M. and O. Ohnishi: Morphological and genetic characteristics of Fagopyrum homotropicum plants with red-winged seeds discovered in Changbo village, Batang district of Sichuan province in China. Fagopyrum 21; 7-13, 2004 Yamane, K., Na Lu and O. Ohnishi: Chloroplast DNA variations of cultivated radish and its wild relatives. Plant Science 168; 627-634, 2004 Yamane, K., K. Tsuji and O. Ohnishi: Speciation of Fagopyrum tataricum inferred from molecular data. Proc. 9th Intl. Symp. Buckwheat at Prague; 317-322, 2004 Reports Kawahara, T.: Catalogue of Aegilops-Triticum Germ-plasm Preserved in Kyoto University, No. 3. Plant Germ-plasm Institute, Graduate School of Agriculture, Kyoto University. p. 313. 2005. b) Conference and seminar papers presented 105th annual meeting of the Japanese Society of Breeding: 4 presentations 106th annual meeting of the Japanese Sciety of Breeding: 2 presentations Annual meeting of the Japanese Society in 2004: 1 presentation

A-3. Off-campus activities Membership of academic societies (roles) Ohmi Ohnishi and Taihachi Kawahara: Kihara Memorial Yokohama foundation for the Advancement of Life Sciences (Visiting researchers) Ohmi Ohnishi: Japanese Society of Breeding (A member of Editorial Board) Research grants Grant-in-Aid for Scientific Research from the Japanese Society for Promotion of Science (B)(2): Is the San Jiang area the original birthplace of common and Tartary buckwheat? ─its genetic study─ (Ohnishi, Yasui) Grant-in-Aid from the Ministry of Education, Science, Sports and Culture for Exploratory Research: How to estimate plant phylogeny by using a novel plant SINE sequence, Au family (Kawahara, Yasui) Grant-in-Aid from the Japanese Society for Promotion of Science for Encouragement of Young Scientists (B): Evaluation of gene bank accessions of wild relatives of wheat as gene resources by using SINE sequence (Yasui) National Bioresource Project KOMUGI (the Ministry of Education, Science, Sports and Culture) (Kawahara, Yasui)

A-4. International cooperation and oversea activities International meeting (roles) Ohnishi, O.: the 9th International Symposium on buckwheat at Prague (International advisor, Editor-in-Chief of Journal, Invited lecturer) Membership in international academic societies Ohnishi, O.: Editor-in-Chief of FAGOPYRUM (International journal of IBRA), A member of the

227 editorial board of Genetic Resources and Crop Evolution (the Netherlands) Kawahara, T.: A member of the editorial board of Wheat Information Service (Japan) International joint Researches, oversea research surveys Ohnishi, O.: Field survey of cultivated and wild buckwheat species in the San Jiang area of southwestern China (China), Field survey of cultivated and wild Perilla in southern China (China) Kawahara, T.: Studies on the variation of wheat and its relatives (Russia) Yasui, Y.: Construction of AFLP linkage map in buckwheat (Germany)

B. Educational Activities (2004.4-2005.3) B-1. On-Campus teaching a) Courses given Undergraduate level: Plant Genetic resources (Ohnishi, Yasui) Graduaate level: Origin of Cultivated Plants (Ohnishi), Ethnobotany (Kawahara) Seminar in Origin of Cultivated Plants (Ohnishi, Kawahara, Yasui) Laboratory Course in Origin of Cultivated Plant (Ohnishi, Kawahara, Yasui)

B-2. Off-campus teaching, etc. Part-time lecturer Ohnishi, O.: Fac. Agr. Okayama Univ. (Origin of Cultivated Plants), Grad. School of Agr. Kyoto Prefectural Univ. (Plant Genetic Resources), Fac. Agr. Yamagata Univ. (Plant Genetic Resources), Fac. Agr. Kobe Univ. (Population genetics) Kawahara, T.: Fac. Agr. Kyoto pefectural Univ. (Genetics)

B-3. Oversea teaching Students and research fellows from abroad Graduate student (1) from China

228 Chair of Science of Plant Protection

2.4.3 Laboratory of Plant Pathology

Staff Professor : Okuno, Tetsuro, Dr. Agric. Sci. Associate Professor : Mise, Kazuyuki, Dr. Agric. Sci. Assistant Professor : Takano, Yoshitaka, Dr. Agric. Sci. Assistant Professor : Kaido, Masanori, Dr. Agric. Sci. Students and research fellows Doctor's Program: (4) Research fellow of JSPS: (1) Master's Program: (10) Research student : (1) Undergraduate : (4)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Studies on plant RNA virus infection Plant viruses cause serious diseases in many important plant species, but few effective antiviral strategies have been developed. This is likely due to our inadequate understanding of the basic biology of plant viruses. We are studying the mechanisms of infection and replication of plant viruses and their interaction with host plants. Viruses used for the studies are Diantho-, Bromo-, and Tobamoviruses. Plants used for the studies are Arabidopsis, barley, cowpea, tobacco and others. We have been analyzing the molecular structures and the functions of viral RNAs and proteins in the infection process of viruses in both plant and protoplast systems. Researches include investigations on the early events of infection to the late maturation stage such as protein synthesis, RNA replication, the cell-to-cell movement of virus, and packaging of viral RNA into virions. b) Identification of host factors involved in plant virus infection Viruses utilize host proteins to establish infection in plants. Plant virus-encoded proteins are proposed to have some physical interactions with putative host-derived factors. Surveys for plant genes involved in RNA virus infections have been progressing by developing a PTGS-based knock-out plant system and using Arabidopsis mutants. c) Studies on plant RNA virus genes in host specificity Plant viruses have host ranges different from each other. We analyze the molecular basis for such “host specificity” phenomena using tobamoviruses and bromoviruses. Reassortant and chimeric viruses have been constructed among those viruses to know the molecular determinants involved in the host range and to analyze host adaptation mechanisms of plant viruses. d) Studies on infection mechanism of phytopathogenic fungi Plant pathogenic fungi cause serious diseases on a wide range of crops and ornamental plants. For development of novel strategies for protecting plants from fungal infection, it is necessary to understand both of fungal infection and plant resistant mechanisms at molecular level. Researchers are studying infection mechanism of a plant pathogenic fungus Colletotrichum lagenarium that is the causal agent of cucumber anthracnose. Molecular genetic analysis has

229 identified many genes involved in pathogenicity of C. lagenarium. Based on information about identified pathogenicity-related genes, we are especially focusing on relation of cellular signaling pathways and peroxisomal metabolic function with fungal infection mechanism. We are also studying nonhost resistance mechanism of plants using Arabidopsis thaliana and several Colletotrichum species that have different host ranges.

A-2. Publications and presentations a) Publications Books Okuno, T.: Plant virus and bioscience. In New Strategies in Bioscience (Edited by Faculty of Agriculture, Kyoto University). pp. 102-115, Maruzen, Tokyo, 2004 (in Japanese) Mise, K. and I. Uyeda: Systemic movement and transmission of plant viruses. In Molecular Mechanisms of Disease Resistance in Plants (Edited by Shimamoto, K. et al.). pp. 172-181, Shujunsha, Tokyo, 2004 (in Japanese) Original papers Tatsuta, M., H. Mizumoto, M. Kaido, K. Mise and T. Okuno: The Red clover necrotic mosaic virus RNA2 trans-activator is also a cis-acting RNA2 replication element. J. Virol. 79(2); 978-986, 2005 Hamada, H., S. Takeuchi, A. Kiba, S. Tsuda, K. Suzuki, Y. Hikichi and T. Okuno: Timing and extent of hypersensitive response are critical to restrict local and systemic spread of Pepper mild mottle virus in pepper containing the L3 gene. J. Gen. Plant Pathol. 71(1); 90-94, 2005 Takeuchi, S., H. Hamada, A. Kiba, Y. Hikichi and T. Okuno: Discrimination of tobamoviruses and their pathotypes for L gene-mediated resistance in green pepper (Capsicum annuum L.) by reverse transcription polymerase chain reaction. J. Gen. Plant Pathol. 71(1); 60-67, 2005 Iwahashi, F., K. Fujisaki, M. Kaido, T. Okuno and K. Mise: Synthesis of infectious in vitro transcripts from Cassia yellow blotch bromovirus cDNA clones and a reassortment analysis with other bromoviruses in protoplasts. Arch. Virol. 150 (7); 1301-1314, 2005 Sasaki, N., M. Kaido, T. Okuno and K. Mise: Coat protein-independent cell-to-cell movement of bromoviruses expressing brome mosaic virus movement protein with an adaptation-related amino acid change in the central region. Arch. Virol. 150 (6); 1231-1240, 2005 Takeda, A., W. Nakamura, N. Sasaki, K. Goto, M. Kaido, T. Okuno and K. Mise: Natural isolates of Brome mosaic virus with the ability to move from cell to cell independently of coat protein. J. Gen. Virol. 86 (4); 1201-1211, 2005 Watanabe, T., A. Takeda, K. Mise, T. Okuno, T. Suzuki, N. Minami and H. Imai: Stage-specific expression of microRNAs during Xenopus development. FEBS Lett. 579(2); 318-324, 2005 Fujisaki, K., F. Hagihara, Y. Azukawa, M. Kaido, T. Okuno and K. Mise: Identification and characterization of the SSB1 locus involved in symptom development by Spring beauty latent virus infection in Arabidopsis thaliana. Mol. Plant-Microbe Interact. 17(9); 967-975, 2004 Takeda, A., M. Kaido, T. Okuno and K. Mise: The C terminus of the movement protein of Brome

230 mosaic virus controls the requirement for coat protein in cell-to-cell movement and plays a role in long-distance movement. J. Gen. Virol. 85(6); 1751-1761, 2004 Sasaki, N., M. Kaido, T. Okuno and K. Mise: Effects of artificial codon changes in the movement protein gene on adaptation of a hybrid bromovirus to cowpea. Microbiol. Immunol. 48(2); 131-135, 2004 Iwamoto, T., Y. Okinaka, K. Mise, K. Mori, M. Arimoto, T. Okuno and T. Nakai: Identification of host-specificity determinants in betanodaviruses by using reassortants between Striped jack nervous necrosis virus and Sevenband grouper nervous necrosis virus. J. Virol. 78(3); 1256-1262, 2004 Yamauchi, J., N. Takayanagi, K. Komeda, Y. Takano and T. Okuno: cAMP-PKA signaling regulates multiple steps of fungal infection cooperatively with Cmk1 MAP kinase in Colletotrichum lagenarium. Mol. Plant-Microbe Interact. 17; 1355-1365, 2004 Kojima, K., Y. Takano, A. Yoshimi, C. Tanaka, T. Kikuchi and T. Okuno: Fungicide activity through activation of fungal signaling pathway: Mol. Microbiol. 53(6); 1785-1796, 2004 Patents Patent pending Patent Application No. 2004-117384 “Poly-nucleotides capable of regulating translation in temperature dependent manner”, inventors: Okuno, T., H. Mizumoto and T. Ido, patentee: Kyoto University, application date: April 12, 2004 Reviews Okuno, T., H. Mizumoto and M. Tatsuta: Regulation mechanisms of gene expression in a plant RNA virus. Plant Disease Reserach (Edited by S. Takamatsu et al.). pp.94-100, Plant Disease Research Press, 2004 (in Japanese) Takano, Y.：Molecular genetic studies on infection mechanism in Colletotrichum lagenarium. Journal of General Plant Pathology 70; 390, 2004 Takano, Y.：Molecular genetic studies on infection mechanism in Colletotrichum lagenarium. Japanese Journal of Phytopathology 70; 161, 2004 (in Japanese) Takano, Y., M. Asakura, A. Kimura, S. Ninomiya and T. Okuno: Penetration structure formation and its functionality in phytopathogenic fungi: Bio-frontier of recognition and response in plant-microbe interactions. PSJ Plant-Microbe Interactions Symposium Report Vol. 40; 43-52. 2004 (in Japanese). b) Conference and seminar papers presented Annual Meeting of the Phytopathological Society of Japan: 10 presentations Kansai Meeting of the Phytopathological Society of Japan: 9 presentations Annual Meeting of the Japanese Society for Virology: 4 presentations Annual Meeting of the Molecular Biology Society of Japan: 3 presentations

A-3. Off-campus activities Membership in academic societies Okuno, T.: The Phytopathological Society of Japan (Councilor), The Japanese Society for Virology (Director), Iwate Biotechnology Research Center (Councilor), The Molecular Biology Society of Japan, The RNA Society of Japan, The Kansai Plant Protection Society (Editor in chief) Mise, K.: The Phytopathological Society of Japan, The Japanese Society for Virology, The

231 Molecular Biology Society of Japan Takano, Y.: The Phytopathological Society of Japan, The Molecular Biology Society of Japan The Kansai Plant Protection Society (Editorial Manager) Kaido, M.: The Phytopathological Society of Japan Research grants Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan: Grant-in-Aid for Scientific Research on Priority Area (A)(2) (Okuno, head), Grant-in-Aid for Scientific Research on Priority Area (A)(2) (Mise, member), Grant-in-Aid for Scientific Research on Priority Area (A)(1) (Takano, member) Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science: Grant-in-Aid for Scientific Research (B)(2) (Mise, head), Research for the Future (Mise, member)

A-4. International cooperations and overseas activities International meetings (roles) The 23rd Annual Meeting of American Society for Virology, 7.10-14 (2004) Montreal, Quebec, Canada, 3 poster presentations (Okuno, Mise, Kaido). The 23rd Fungal Genetics Conference 3.15-20 (2005) Pacific Grove, California, 1 oral presentation (Takano), 1 poster presentation (Takano), 2 poster presentations (Okuno, Takano)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Plant Pathology I (Okuno), Plant Pathology II (Mise), Seminar in Plant Protection (Okuno), Biotechnology-Novel Strategies for Agriculture (Okuno), Basic Bioresource Science II (Okuno), Microbiology (Okuno), Outline of Bioresource Science IV (Mise), Laboratory Course in Bioresource Science I, II (Okuno, Mise). Resource, Environment and Technology Factors and World Food Production (Okuno) Graduate level: Plant Pathology (Advanced Course) I (Okuno), Seminar in Plant Pathology (Okuno, Mise), Research in Plant Pathology (Okuno, Mise).

B-2. Off-campus teaching, etc. Part-time lecturer Okuno, T.: Kochi University (Plant Virology), Nagoya University (Plant Virology) Mise, K.: Kobe University (Plant Virology), Kyoto Institute of Technology (Plant Pathology) Takano, Y.: Iwate Biotechnology Research Center (Plant Pathology)

C. Other remarks Okuno, T.: The Safety Committee of Kyoto University for Recombinant DNA Experiments, The Steering Committee of Kyoto University Museum, A special research scientist in the Research Center for Science System in Japan Society for the Promotion of Science Mise, K.: Grand coordinator of Laboratory Course in Bioresource Science I, II Takano, Y.: The Young Scientist Award of the Phytopathological Society of Japan

232 2.4.4 Laboratory of Insect Ecology

Staff Professor : Fujisaki, Kenji, Dr. Agric. Sci. Associate Professor : Ohsaki, Naota, Dr. Agric. Sci. Assistant Professor : Nishida, Takayoshi, Dr. Agric. Sci. Students and research fellows Doctor's program : (12) Master's Program : (8) Undergraduate : (4) Post doctoral fellow : (4)

A. Research Activities (2004.4-2005.3) A-1. Main subjects The studies in this laboratory are concentrated on the ecology, dynamics, evolution and management of insect populations or communities with reference to agriculture and forestry. On-going main topics of research by the staff members are as follows. a) Analysis on life history strategy in insects - Insects have evolved various life history strategies in response to spatio-temporal heterogeneity of habitats. Among them, life history traits such as migration and diapause are especially important. These life history traits correlate with each other and constitute a syndrome. The evolutionary pathway of wing polymorphism as an example of such a syndrome is going to be clarified through the physiological, ecological and genetic analysis. b) Role of parasitic natural enemies in host plant choice by Pieris butterflies - Herbivorous insects do not always use nutritionally superior plants as food but use sometimes only inferior ones. This suggests the existence of other ecological factors influencing the food plant choice by herbivores. By studies of the host plant - herbivore - parasitoid interactions using Pieris butterflies, field and laboratory evidence is now being accumulated to infer that the difference in defense mechanisms of the insects against their specific parasitoids determines their host plant preference. c) In Batesian mimicry, mimetic forms of some species are female limited, other species are male limited, whereas the other species are not sex limited. To explain these phenomenon, trade-offs between predation by avian predators and sexual selection are analyzed. d) Indirect effect of predation on long-term population dynamics - Field observations and laboratory experiments have gradually revealed that prey often suffers more from indirect effects of predation (non-lethal injury, starvation, postponed reproduction and so on) than from direct predation events. This may explain why traditional analyses of population dynamics have long underestimated the effect of predation as the regulatory factor for prey-predator systems. e) Reproductive interference as a major underpinning mechanism of interspecific competition – Theoretical studies have revealed that reproductive interference between closely related species can explain a wide spectrum of ecological phenomena, from parapatry or allopatry to host shift of herbivorous insects.

233 A-2. Publications and presentations a) Publications Books Fujisaki, K. and S. Tanaka (eds). The mystery of flying insects and non-flying insects. pp.268, Tokai University Press, 2004 Original papers Ide, J.-Y.: Selection of age classes of Sasa leaves by caterpillars of the skipper butterfly Thoressa varia using albo-margination of overwintered leaves. J. Ethol. 22; 99-103, 2004 Ide, J.-Y.: Diurnal and seasonal changes in the mate-locating behavior of the satyrine butterfly Lethe diana. Ecol. Res. 19: 189-196, 2004 Ide, J.-Y.: Leaf trenching by Indian red admiral caterpillars for feeding and shelter construction. Popul. Ecol. 46; 275-280, 2004 Imai K. and N. Ohsaki: Internal structure of developing aucuba fruit as a defense increasing oviposition costs of its gall midges Asphondylia aucubae. Ecol. Entomol. 29 ; 420-428, 2004 Imai K. and N. Ohsaki: Oviposition site of and gall formation by the fruit gall midge Asphondylia aucubae (Diptera: Cecidomyiidae) in relation to internal fruit structure. Entomol. Sci. 7; 133-137, 2004 Izumi, Y., K. Anniwaer, H. Yoshida, S. Sonoda, K. Fujisaki and H. Tsumuki: Comparison of cold hardiness and sugar content between diapausing and nondiapausing pupae of the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae). Physiol. Entomol. 30; 36-41, 2005 Matsumoto, T., T. Itioka and T. Nishida: Why arrowhead scales, Unaspis yanonensis Kuwana (Homoptera: Diaspididae), which burrow and settle below conspecifics can successfully avoid attacks by its parasitoid, Coccobius fulvus Compere et Annecke (Hymenoptera: Aphelinidae)? Appl. Entomol. Zool. 39(1); 147-154, 2004 Matsumoto, T., T. Itioka, T. Nishida and T. Inoue: A test of temporal and spatial density dependence in the parasitism rates of introduced parasitoids on host, the arrowhead scale (Unaspis yanonensis) in stable host-parasitoids system. J. Appl. Entomol. 128(4); 267-272, 2004 Matsumoto, T., T. Itioka and T. Nishida: Is spatial density-dependent parasitism necessary for successful biological control? Testing a stable host-parasitoid system. Entomol. Exp. Appl. 110(3); 191-200, 2004 Miura K. and N. Ohsaki: Diet mixing and its effect on polyphagous grasshopper nymphs. Ecol. Res. 19; 269-274, 2004 Miura K. and N. Ohsaki: Relationship between physical leaf characteristics and growth and survival of polyphagous grasshopper nymphs, Parapodisma subastris (Orthoptera: Catantopidae). Popul. Ecol. 46; 179-184, 2004 Nishida, S., A. Naiki and T. Nishida: Morphological variation in leaf domatia enalbles coexitence of antagonistic mites in Cinnamomum camphora. Can. J. Bot. 83; 93-101, 2005 Ohsaki, N.: A common mechanism explaining the evolution of female-limited and both-sex Batesian mimicry in butterflies. J. Anim. Ecol. 74; 728-734, 2005 Sato, Y. and N. Ohsaki: Response of the wasp Cotesia glomerata (L.) to larvae of an invasive host, the large white butterfly (Pieris brassicae L.). Ecol. Res. 19; 445-449, 2004

234 b) Conference and seminar papers presented The 48th Annual Meeting of Japanese Society of Applied Entomology and Zoology: 18 presentations The 51st Annual Meeting of Ecological Society of Japan: 8 presentations The 64th Annual Meeting of Japanese Society of Entomology: 2 presentations The 22nd Symposium of Population Ecology:6 presentations XXII International Congress of Entomology, Brisbane, Australia：3 presentations 10th International Behavioural Ecology Congress, Jyväskylä, Finland：1 presentation

A-3. Off-campus activities Membership in academic societies (roles) Fujisaki, K.: The Japanese Society of Applied Entomology and Zoology (Executive Committee Member), The Society of Population Ecology (President, Editor) Ohsaki, N. : The Japanese Society of Ethology (Editor) Nishida, T.: Ecological Society of Japan (Editor) Research grants 21st century COE program: Innovative food and environmental studies pioneered by entomomimetic sciences (Fujisaki, K. project leader). Monbu-kagakusyo Research Grant: Scientific Research (C) (2) Effects of parasitoids on the evolution of host range of Pieris butterflies (Ohsaki, N.) Scientific Research: Nonlethal effects of predation on the community structure of grasshoppers in old fields (Nishida, T.)

A-4. International cooperations and overseas activities International joint research, overseas research surveys Fujisaki, K. Integrated biological control of Pueraria thunbergiana and Polygonum perfoliatum, invasive alien weeds in USA (USDA) Scholars from abroad JSPS research fellow (Argentina)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Insect ecology l (Fujisaki), Introduction to general bio-resource science II (Fujisaki), Biology of biosphere (Fujisaki), Insect ecology II (Ohsaki), Laboratory course in bio-resource science I and II (Ohsaki, Nishida), Seminar in plant protection (Fujisaki, Ohsaki) Graduate level: Insect evolutionary ecology (Ohsaki), Seminar in insect ecology (Fujisaki, Ohsaki), Research in insect ecology (Fujisaki, Ohsaki)

B-2. Off-campus teaching Part-time lecturer Fujisaki, K.: Kobe University, Kinki University, Kyusyu University Ohsaki, N.: Nagoya University

235 B-3. International education Fujisaki, K.: JICA (Hyogo)

2.4.5 Laboratory of Insect Physiology

Staff Professor : Sakuma, Masayuki, Dr. Agric. Sci. Assistant Professor : Fukui, Masao, Dr. Agric. Sci. Students and research fellows Doctor's program: (4) Master's program: (7) Undergraduate : (2)

A. Research Activities (2004.4-2005.3) A-1. Main subjects. In spite of their simple structure and tiny body size, the insects search for food, mates, shelter and other resources, with surprising precision and efficiency, by reference to cues of various modalities. We studied the unique mechanisms of insect behavior from the multidisciplinary aspects of insect physiology, organic chemistry, acoustics, robotics and neuroethology. a) Study on the spatial orientation of insects and other arthropods To investigate the causation of the searching behavior of insects and mites, we developed locomotion-compensator apparatus, i.e., the digitalservosphere and the micro locomotion compensator (MLC). In these apparatuses, an insect or mite walking freely on a moving floor can be retained at one point, because the floor continually returns the animal to the previous point. Since directional cues can be presented to a freely walking animal in a precisely controlled way, we can create a sensory field that is virtually the same as what the animal perceives spatially and temporally in the natural field. This year, we analyzed the chemo-orientation behavior of the cheese mite, Tyrophagus putrescentidae, induced by the odor of dry-yeast methanol extracts. Mites were individually allowed to walk freely on a glass plate in the MLC, where the exposure to the attractant odor was temporally controlled. The wavelet analysis of the turning velocity in the mites revealed that a characteristic oscillation at low frequency, which could be the principle for the attraction, was generated just after the cessation of the odor. Since the behavior was promoted without any directional cue, this principle would be the primordial element of the behavior leading animals to an attractant source. Turn alternation behavior in the pill bug, Armadillidium vulgaris, was re-investigated. The bug changes turn direction alternatively at every branching point in a T-maze array. A series of experiments in a servosphere demonstrated that this turn alternation response was owing to an intense thigmotactic or wall-following nature of the animal together with an artificial architecture of a T-maze array. The animal with gills may readily find hiding place by thigmotaxis to protect itself from desiccation.

236 b) Odor learning in insects The odor-associative learning of the German cockroaches has been demonstrated for the first time. They exhibited odor-conditioned upwind anemotaxis in a servosphere apparatus after being trained for general odor, whereas they always did for the odor of aggregation pheromone. c) Acoustic features of calling song in female mate choice in a field cricket. Male field crickets, Telleogryllus taiwanemma, produce calling songs to attract females. Variations in the calling song phrases consisting of chirp sequences were recognized in Taiwan, phrases were composed of single, double and seven chirps (Chow et al. 1990). Using a Japanese cricket population having phrase variations in calling songs, we tested what types of the phrase variations synthesized with computer contribute to female preference. However, females did not discriminate between variations in calling song phrases consisting of identical chirp rates. Females preferred higher calling song chirp rates; that is, the preference function shape was open-ended. The preference function shape by females toward two other acoustic features of song chirp was also determined. Open-ended preference function was found in the pulse number of chirps, and unimodal preference was shown toward carry frequency, but which features are correlated with male qualities remains unclear. On the other hand, female phonotaxis was investigated on two-choice preference test to determine the temporal parameter combination (pulse duration – interval duration in chirp) of the male advertisement call used for song recognition. The phonotactic field, which was determined on the base of the parameter combination in chirps preferred by females, revealed that female recognizes the nonspecific advertisement call by pulse rate.

A-2. Publications and presentations a) Publications Books Sakuma, M.: Aggregation Pheromones. Encyclopedia of Agricultural Chemistry (edited by Suzuki & Arai). pp. 183-186, Asakura Shoten, Tokyo, 2004 (in Japanese) Original papers Gassa, A., M. Fukui, M. Sakuma, T. Nishioka and S. Takahashi: Rotenoids in the Yam bean Pachyrrhizus erosus: Possible defense principles against herbivores. Japanese Journal of Enviromnetal Entomology and Zoology 15(4); 251-259, 2004 Reports Fukui, M.: Advertisement call recognition by temporal cues in a field cricket, Teleogryllus taiwanemma. Trans. Tech. Common. Psychol. Physiol. Acoustc., The Acoustic Society of Japan 34(3); 165-169, 2004 (in Japanese) b) Conference and seminar papers presented The 64th Annual Meeting of Japanese Society of Entomology, Sapporo, August 2004: 1 presentation XXII International Congress of Entomology, Brisbane, Australia, August, 2004: 3 presentations The 1st International Symposium of the 21st Century Program (Entomomimetic Sciences), Kyoyo, March 2005: 4 presentations The 49th Annual Meeting of Japanese Society of Applied Entomology and Zoology, Tokyo, March 2005: 2 presentations

237 A-3. Off-campus activities Resarch grants 21th Century COE program: COE for Innovative Food and Environmental Studies Pioneered by Entomomimetic Sciences (Sakuma).

A-4. International cooperation and overseas activities International meetings Sakuma, M.: XXII International Congress of Entomology, Brisbane, Australia, August 2004. (2 presentations) Fukui, M. : XXII International Congress of Entomology, Brisbane, Australia, August 2004. (1 presentation)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Animal Physiology (Sakuma et al.), Outline of Bioresource Science IV (Sakuma et al.), Insect Physiology (Sakuma), Laboratory in Bioscience I, II (Sakuma and Fukui), Seminar in Plant Protection (Sakuma) Graduate level: Current Topics in Insect Physiology (Sakuma), Seminar in Insect Physiology (Sakuma), Research in Insect Physiology (Sakuma)

B-2. Off-campus teaching, etc. Part-time lecturer Sakuma, M. Faculty of Agriculture, Kobe University, (Integrated Pest Management Course II for JICA students) Fukui, M. Faculty of Engineering, Kyoto Sangyo University, (Fundamentals of Biological Experiments)

238 Chair of Animal Genetics and Reproduction

2.4.6 Laboratory of Animal Breeding and Genetics

Staff Professor : Sasaki, Yoshiyuki, Dr. Agric. Sci. Associate Professor : Yamada, Takahisa, Dr. Medic. Sci. Assistant Professor : Taniguchi, Yukio Assistant Professor : Miyake, Takeshi Invited professor from abroad : (1) Students and research fellows Master's program : (4) Research fellow : (1) Undergraduate : (2)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Studies on Genetic Evaluation Procedure and Breeding Scheme for Beef Cattle The genetic evaluation method was developled which accounts for the heterogeniety of variances across subclasses for fixed effects. With the method, bias for the predicted breeding values of sires and dams was decreased by adjusting heterogeneity of variances. Furthermore, By using the field records collected from the two Japanese Black populations (Oita and Hyogo prefectures) and one Japanese Brown population (Kumamoto) during 1988 to 2003, the genetic trends for the carcass traits of Wagyu were clarified. Based on the comparisons of the genetic gains among different breeding programs, the effectiveness of the sire evaluation method using field records was clarified. b) Studies on Analysis of Quantitative Trait Loci in livestock animals In order to detect the quantitative trait loci (QTL) for the economically important traits in beef cattle, MQEM-HS method which is the multiple QTL mapping method with epistatic interactions for the half-sib families in beef cattle was developed and applied to the multiple QTL mapping using Japanese Black half-sib family in Oita prefecture. Furthermore, the estimation method of the degree of linkage disequilibrium between genetic markers in Japanese Brown populations was investigated, suggesting the linkage disequilibrium mapping would be useful for the fine mapping of QTLs. In addition, the multiple QTL mapping for outbred populations of general pedigree was investigated by using descent graph sampling. c) Exploration of the Genes Responsible for Beef Marbling The genes responsible for beef marbling have been explored using differential display approach with somatic nuclear-derived cloned steers, which were genetically destined for high marbling. A total of 77 genes, including 35 annotated genes and 42 novel sequences, have been detected as the genes which showed the expression pattern associated with intramuscular fat deposition. The expression patterns in all of the 35 annotated genes and in 29 of the 42 novel genes were validated using real-time PCR. Furthermore, 32 annotated genes and 34 novel genes were mapped to bovine chromosomes using bovine radiation hybrid panel or bovine-human comparative map, and their mapped positions were compared with chromosomal locations for

239 marbling QTLs. As a result, 5 annotated genes and 7 novel genes were regarded as possible condidates for marbling genes, taken together with functional annotation in case of the annotated genes. d) Study on the Molecular Mechanism Underlying Marbling in Beef To study the molecular mechanism of intramuscular-adipogenesis in beef cattle, the temporal and spatial expression patterns in bovine longissimus muscle tissue of transcription factors, membrane proteins and extracellular matrix proteins regulating the adipogenesis, were investigated using molecular biological and immunohistochemical methods. The promoter analysis of ADAM12 gene that is assumed to control the intramuscular fat deposition, was carried out. To establish the mouse model showing intramuscular fat deposition, transgenic mice expressing the bovine ADAM12 gene specifically in muscle tissue are now producing. e) Study on Genetic Dissection of Marbling Traits in Mouse or Rat Model To identify a positional candidate gene, Pnlip, as the gene responsible for intramuscular fat deposition, genetic analysis has been performed using OLETF rat model. The VNTR polymorphism in Pnlip promoter completely cosegregated with intramuscular fat content in backcross progenies of F344. OLETF-marbling QTL congenic and F344 parental recipient strains, providing circumstantial evidence that Pnlip is causal gene.

A-2. Publications and presentations a) Publications Original papers Ito, M., K. Moriya and Y. Sasaki: Relationship between age of sires and production ability of their progenies. Bull. Beef Cattle Sci. 77; 3-10, 2004 Sakoyama, R., Y. Taniguchi, M. Kitagawa, T. Yamada and Y. Sasaki: Comparison of age-dependent expression patterns of C/EBP family and PPARγ in musculus longissimus between Japanese black and holstein breeds. Anim. Sci. J. 75; 527-533, 2004 Fujita, T., M. Ito, W. Sato, T. Kurahara, T. Miyake, K. Shiga and Y. Sasaki: Association between the deletion mutant allele of MCSU deficiency and carcass traits in Japanese Black cattle. J. Anim. Genet. 32; 11-16, 2004 Ibi, T., T. Miyake, S. Hobo, H. Oki, N. Ishida and Y. Sasaki: Estimation of heritability of laryngeal hemiplegia in the thoroughbred horse by Gibbs sampling. J. Equine Sci.(J) 41; 155-162, 2004 Narita, A. and Y. Sasaki: Detection of multiple QTL with epistatic effects under a mixed inheritance model in an outbred population. Genet. Sel. Evol. 36; 415-433, 2004 Oishi, M., H. Gohma, H.Y. Lejukole, Y. Taniguchi, T. Yamada, K. Suzuki, H. Shinkai, H. Uenishi, H. Yasue and Y. Sasaki: Generation of a Total of 6483 expressed sequence tags from 60 day-old bovine whole fetus and fetal placenta. Anim. Biotechnol. 15:1-8, 2004 Odani, M., H. Nakaoka, A. Narita, T. Ibi, Y. Sasae and Y. Sasaki: A national genetic evaluation of carcass traits for the Japanese Black cattle is possible. Anim. Sci. J. 75; 353-361, 2004 Reviews Sasaki, Y., H. Hirooka and M. Tsuiki: Systems analysis applied to animal husbandry. J. JASS, 20; 125-137, 2004 Miyake, T., T. Yamada and Y. Sasaki: Current and future subjects for QTL analysis methods in livestock. J. Anim. Genet., 32; 43-52, 2004

240 b) Conference and seminar papers presented The 29th International Conference on Animal Genetics : 8 presentations Workshop in the 29th International Conference on Animal Genetics : 1 presentation JICA seminar in the 29th International Conference on Animal Genetics : 1 presentation The 104th Annual Meeting of Japanese Society of Animal Science : 8 presentations The 42th Annual Meeting of the Society of Beef Cattle Science : 2 presentations The 27th Meeting of the Molecular Biology Society of Japan: 1 presentation

A-3. Off-campus activities Membership in academic societies Sasaki, Y: Japanese Society of Animal Breeding and Genetics (President), Japanese Society of Animal Science (Representative), Animal Breeding and Genetics Symposium (Head of Organizing Committee), Society of Beef Cattle Science (President), Animal Science Systems Society (Director), Kinki Committee of New Technology for Bioscience in Agriculture, Forestry, Fisheries and Food (Sub-chairman, Technological participation) Membership in Science Council of Japan, etc Sasaki, Y.: Communication Committee for Research of Genetic Resources (Member), Technical Committee for Reformation of Agricultural Production (Member) Research grants Monbusho research grant; Grant-in-Aid for Scientific Research on Priority Areas(C)(2) Positional candidate cloning of NIDDM causative gene in OLETF rat (Head: Yamada), Grant-in-Aid for Scientific Research (B)(2) Exploration of genes responsible for marbling using somatic nuclear cloning technology (Head: Sasaki, Sharer: Yamada, Taniguchi), Promotion Research Establishment of genome-shift strategies for identification of genes involved in QTLs using livestock animal models (Head: Sasaki, Sharer: Yamada, Taniguchi) Grant-in-Aid-(Bio Design Program) from National Agricultural Research Organization; Spatial regulation of intra-muscular adipocyte differentiation in beef marbling (Head: Sasaki) Grant-in-Aid-(Bio Design Program) from National Agricultural Research Organization; Exploration of genes involved in developmental abnormality in somatic nuclear cloned animals (Head: Sasaki) Grant-in-Aid from Kumamoto Agricultural Research Station; Research of the re-construction of sire evaluation systems adapting personal computers (Head: Sasaki)

A-4. International cooperations and overseas activities International meetings (roles) Sasaki, Y.: The 29th International Conference on Animal Genetics, Tokyo (Chairman of the Organizing Committee) Yamada, T.: The 29th International Conference on Animal Genetics, Tokyo (Staff), The 2nd Seminar for New Animal Breeding and Genetics, Tokyo (Staff) Taniguchi, Y.: The 29th International Conference on Animal Genetics, Tokyo (Staff), The 2nd Seminar for New Animal Breeding and Genetics, Tokyo (Staff) Miyake, T. : The 29th International Conference on Animal Genetics, Tokyo (Staff), The 2nd Seminar for New Animal Breeding and Genetics, Tokyo (Staff)

241 Membership in international academic societies Sasaki, Y.: World Congress on Genetics Applied to Livestock Production (Permanent Committeeman), The 29th International Conference on Animal Genetics (Chairman of the Organizing Committee), Journal of Animal Breeding and Genetics (Editor), Asian-Australasian Journal of Animal Science (Section Editor), Bioimaging Society (Editor)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Outline of Agricultural Science II (Sasaki et al.), Outline of Bioresource Science II (Sasaki et al.), Animal Breeding and Genetics (Sasaki), Animal Breeding (Sasaki), Introduction to Foreign Literature in Bioresource Science IV (Sasaki and Yamada et al.), Basic Laboratory Course in Bioresource Science I, II (Sasaki, Yamada, Taniguchi, Miyake et al.), Technology theory of Animal Husbandry and Practice II (Sasaki, Yamada, Taniguchi, Miyake et al.), Laboratory Course in Bioresource Science I, II (Sasaki, Yamada, Taniguchi, Miyake et al.), Molecular Biology (Yamada), Seminar in Applied Animal Science I, II (Yamada et al.), Domestic Animal Genome Science Biotechnology (Yamada et al.) Graduate level: Animal Breeding and Genetics (Sasaki), Animal Genomics (Yamada), Seminar in Animal Breeding and Genetics (Sasaki, Yamada), Laboratory Course in Animal Breeding and Genetics (Sasaki, Yamada)

B-2. Off-campus teaching, etc. Part-time lecturer Sasaki, Y.: Kinki University (Animal Science), Kyoto Prefectural University (Animal Breeding and Genetics), Ishikawa Agricultural College (Animal Breeding)

B-3. Overseas teaching Students and research fellows from abroad Master’s program: 1 (Kyrgyz Republic)

C. Other remarks Sasaki, Y.: Faculty of Agriculture, Public Announcements Committee for Kyoto University (Member), Division of Public Announcements Technology on the Public Announcements Committee for Kyoto University (Sub-director), Promotion Conference on Research Project, “Development of Efficient Basic Technology in Fisheries Breeding” in the Ministry of Agriculture, Forestry and Fisheries (External Assessment Committeeman), Committee for Beef Cattle Genomic Research, Development and Promotion in Livestock Technology Association (Member), Agricultural Establishment Research Committee of Kinki, Chugoku, Shikoku Region in National Agriculture and Bio-oriented Research Organization (Member)

242 2.4.7 Laboratory of Reproductive Physiology

Staff Professor : Imai, Hiroshi, Dr. Agric. Sci. Associate Professor : Yamada, Masayasu, Dr. Medic. Sci. Assistant Professor : Minami, Naojiro, Dr. Agric. Sci. Assistant Professor : Nagao, Yasumitsu, Ph. D. Students and research fellows Doctor's program: (4) Master's program : (13) Undergraduate : (3) Research student: (1)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Development and Differentiation in Mammalian Embryos The regulatory factors required for in vitro maturation, fertilization and development of eggs are studied. We found that IVM/IVF embryos require some unknown factors from follicle cells during maturation for their further development, so now we are focusing on elucidation of the molecular mechanisms of this intracellular communication between oocytes and follicle cells. In addition, the genes and the proteins that are involved in the preimplantation development and differentiation are also studied using mouse embryo culture system. b) Production of clone animals derived from somatic cells It has become possible to produce clone animals derived from differentiated somatic cells using nuclear transfer technique, however, it is still unknown how can the differentiated cells acquire the totipotency during development. In addition, the great majority of reconstituted embryos die after nuclear transfer without involvement of ontogenesis. In our laboratory, reprogramming mechanisms of differentiated cells and embryonic anomaly during embryogenesis are examined using cell and molecular biological techniques. c) Establishment of embryonic stem cells and production of transgenic animals Development of techniques to establish the embryonic stem cell having totipotency from mammalian embryos are undertaken. Reconstitution of embryos and production of transgenic animals using the stem cells are studied. We are intending to utilize the embryonic stem cells to apply the transgenic technique directly to chromosome for the improvement of livestock animals. d) Relationship between mitochondria and phenotype of the mouse Effects of mitochondria derived from different species on the phenotype of the mouse are examined using individuals produced by microinjection or nuclear transfer. In addition, the interaction between mitochondria of different species and cell nuclear is examined using cell and molecular biological techniques.

A-2. Publications and presentations a) Publications Books and reviews Imai, H.: Basic Molecular Biology for Biotechnology. Transgenic animals (Ohshima, T., K. Kitamoto, S. Harashima and M. Miyagawa eds). pp.181-183, Kagaku-Dojin, Kyoto, 2004 (in Japanese)

243 Imai, H.: Gakumon Kamifusen - Advanced Technology in Developmental Biology and Reproductive Biology (edition), Guideline 11, pp.56-57, Kawaijuku, 2004 (in Japanese) Yamada, M.: Control of the preimplantation development of mammalian embryos, pp.1-104, Asakura shoten, Tokyo, 2004 (in Japanese) Original papers Watanabe, T., A. Takeda, K. Mise, T. Okuno, T. Suzuki, N. Minami and H. Imai: Stage-specific expression of microRNAs during Xenopus development. FEBS Lett. 579; 318-324, 2005 Shimatsu Y, M. Uchida, R. Niki, H. Imai: Effects of a synthetic progestogen, altrenogest, on oestrus synchronisation and fertility in miniature pigs. Veterinary Record 155; 633-635, 2004 Review Ikeda, S., M. Kitagawa, H. Imai and M. Yamada: The roles of vitamin A for cytoplasmic maturation of bovine oocytes. J. Reprod. Dev. 51; 23-35, 2005 Reports Yamada, M.: Effects of midkine and retinoic acid on apoptosis in cumulus cells during in vitro maturation of bovine cumulus-enclosed oocytes. Final Reports for Research Grants for Meat and Meat Products Vol. 22; 10-16, 2004 Patents Imai, H., Y. Hoshino, M. Miyake, M. Uchida and M. Shimazu: Methods of efficient production of nuclear transfer animals from somatic cells. No. 2004-16115, 2004 b) Conference and seminar papers presented The 104th Annual meeting of Japanese Society of Zootechnical Science: 3 presentations The 11th Annual meeting of Japan Embryo Transfer Society: 2 presentations The 97h Annual meeting of Japanese Society of Animal Reproduction: 4presentations The 127th Annual meeting of Japanese Society of Fertility and Sterility –Kansai Branch-: 1 presentation The 27h Annual meeting of Japanese Society of Molecular Biology: 1 presentation

A-3. Off-campus activities Roles in academic societies Imai, H.: Japanese Society of Animal Reproduction (Director), Japan Embryo Transfer Society (Vice President, Director), Japan Society of Fertility and Sterility (Director), Japan Society of Fertilization and Implantation (Director), Japanese Society of Zootechnical Science (Councilor), Kansai Society of Zootechnical Science (Councilor), Japanese Society of Reproductive Endocrinology (Councilor) Yamada, M.: Japan Society of Fertility and Sterility (Councilor), Japan Embryo Transfer Society (Officer), Japanese Society of Animal Reproduction (Editorial Board) Minami, N.: Japan Embryo Transfer Society (Officer), Japan Society of Fertility and Sterility (Officer) Research grants Monbusho Research Grants, Grant-in-Aid for Scientific Research (B)(1): Search for abnormal production of cloned animals by exhaustive gene analysis (Sharer: Imai), Grant-in-Aid for Scientific Research (C)(2): Studies on the promotion of cytoplasmic maturation of bovine immature oocytes by cumulus cells (Head: Yamada), Grant-in-Aid for Scientific Research (B)(2): Functional analysis of a novel gene, Oogenesin, which localizes in

244 nucleus at the time of zygotic gene activation (Head: Minami) Research project for utilizing advanced technologies in agriculture, forestry and fisheries: Establishment of stable and efficient technology of the production of bovine somatic clones (Head: Imai, Sharer: Minami)

A-4. International cooperation and overseas activities International meetings (roles) 37th Society for the Study of Reproduction, Vancouver (presentation: Imai, Minami) 15th International Congress of Animal Reproduction, Port Segro (presentation: Imai, Minami) The 5th Royan International Congress Tehran (Award lecture) 31st International Embryo Transfer Society Copenhagen (presentation: Imai, Yamada, Minami, Nagao)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Reproductive Physiology (Imai), Outline of Bioresource Science III (Imai et al), Outline of Japanese Agriculture (Imai), Program of International Education (Imai), Developmental and Reproductive Technology (Yamada), Introduction to Animal Science Literature I (Yamada and Moriya), Methods and Techniques in Animal Reproduction Experimentation (Imai, Yamada, Minami, Nagao), Biotechnology (Yamada et al) Graduate level: Advance Course of Reproductive Physiology (Imai), Reproductive Physiology-Seminar (Imai, Yamada), Laboratory Course in Reproductive Physiology (Imai, Yamada), Genetic Engineering in Developmental Biology (Yamada)

B-2. Off-campus teaching, etc. Part-time lecture Osaka City University Medical School (Yamada) Open seminar, etc Imai, H.: A traceability system for Wagyu production. Workshop for Artificial Insemination for Domestic Species (Lecturer), 2004. Imai, H.: Present status of somatic cell cloning technology. Symposium for the Japanese Society for Bioscience, Biotechnology, and Agrochemistry (Kansai branch) (lecturer), 2004. Imai, H.: On-site study and training for high-school students (Sagae High school in Yamagata Prefecture) (guidance and suggestion), 2004. Imai, H.: Clone Technology, Training course of animal production in National Livestock Breeding Center in 2004. (lecturer), 2004. Minami: Oog1, an oocyte-specific gene and preimplantation development in the mouse. 2nd Reproductive Medicine and Biology symposium (lecturer), 2004]

B-3. Overseas teaching Lecture and seminars Students and research fellows from abroad Graduate Student: 2 (Korea), Researcher (Russia)

245 C-1. Other remarks Imai, H.: Member of Conference Committee of Very-low Temperature Research Laboratory at Kyoto University, Member of Domestic Animal Improvement Committee of Kyoto Prefecture, Research Adviser of Technology Conference of Agriculture, Forestry and Fishery of Ishikawa Prefecture, Committee of Research Investigation of, Agriculture, Forestry and Fisheries Technical Information Society, Committee of Kyoto University Livestock Farm, Assessment Committee of Project Research of Ministry of Agriculture, Forestry and Fisheries, Assessment Committee of Research Grant of Ministry of Agriculture, Forestry and Fisheries, Member of Administration Committee of Human Genome and Gene Analysis of Kyoto University Yamada, M.: Guest Researcher of Gifu Prefectural Livestock Institute, Guest Researcher of Toyama Prefectural Agriculture Research Center

Chair of Animal Anatomy, Nutrition and Resources

2.4.8 Laboratory of Nutritional Science

Staff Professor : Yano, Hideo, Dr. Agric. Sci. Associate Professor : Matsui, Tohru, Dr. Agric. Sci. Assistant Professor : Kawachi, Hiroyuki, Dr. Eng. Students and research fellows Doctor's program : （3） Research student: （1） Master's program: （6） Undergraduate : （4）

A. Research Activities（2004.4-2005.3） A-1. Main subjects a) Mineral nutrition and metabolism We have studied mineral metabolisms using stable isotopes as tracers in rats. We found that magnesium concentration in drinking water affected its absorption and that coexisted calcium with magnesium in drinking water did not affect magnesium absorption but increased urinary excretion of magnesium. Rats showed iron and magnesium deficiencies induced by excess calcium in the form of carbonate. Excess calcium in the form of citrate induced iron deficiency but not magnesium deficiency. Thus, we indicate that the adverse effects of excess calcium depends its form. Exercise induced hemolysis and reduced plasma iron in racehorses but iron absorption was stimulated by exercise, which suggested the exercise did not induce iron deficiency when horses were given common diets. b) Adipocyte differentiation and function We cloned bovine leptin receptor gene and determine the nucleotide sequences of its 3 isoforms. These genes were expressed in many tissues and were highly expressed in liver and

246 C-1. Other remarks Imai, H.: Member of Conference Committee of Very-low Temperature Research Laboratory at Kyoto University, Member of Domestic Animal Improvement Committee of Kyoto Prefecture, Research Adviser of Technology Conference of Agriculture, Forestry and Fishery of Ishikawa Prefecture, Committee of Research Investigation of, Agriculture, Forestry and Fisheries Technical Information Society, Committee of Kyoto University Livestock Farm, Assessment Committee of Project Research of Ministry of Agriculture, Forestry and Fisheries, Assessment Committee of Research Grant of Ministry of Agriculture, Forestry and Fisheries, Member of Administration Committee of Human Genome and Gene Analysis of Kyoto University Yamada, M.: Guest Researcher of Gifu Prefectural Livestock Institute, Guest Researcher of Toyama Prefectural Agriculture Research Center

Chair of Animal Anatomy, Nutrition and Resources

2.4.8 Laboratory of Nutritional Science

Staff Professor : Yano, Hideo, Dr. Agric. Sci. Associate Professor : Matsui, Tohru, Dr. Agric. Sci. Assistant Professor : Kawachi, Hiroyuki, Dr. Eng. Students and research fellows Doctor's program : （3） Research student: （1） Master's program: （6） Undergraduate : （4）

A. Research Activities（2004.4-2005.3） A-1. Main subjects a) Mineral nutrition and metabolism We have studied mineral metabolisms using stable isotopes as tracers in rats. We found that magnesium concentration in drinking water affected its absorption and that coexisted calcium with magnesium in drinking water did not affect magnesium absorption but increased urinary excretion of magnesium. Rats showed iron and magnesium deficiencies induced by excess calcium in the form of carbonate. Excess calcium in the form of citrate induced iron deficiency but not magnesium deficiency. Thus, we indicate that the adverse effects of excess calcium depends its form. Exercise induced hemolysis and reduced plasma iron in racehorses but iron absorption was stimulated by exercise, which suggested the exercise did not induce iron deficiency when horses were given common diets. b) Adipocyte differentiation and function We cloned bovine leptin receptor gene and determine the nucleotide sequences of its 3 isoforms. These genes were expressed in many tissues and were highly expressed in liver and

246 testis. We found that activin, a member of TGF- β superfamily, suppressed adipocyte differentiation in the early stage and this suppressive effect was reduced by follistatin. Additionally, we found that activin suppressed the expressions of PPARγ and C/EBPαthrough inhibiting binding capacity of C/EBPβ. c) Others We found that heat stress decreased plasma vitamin C level in lactating cows, which suggested that the supplementation with vitamin C is preferable for lactating cows in summer. Additionally, we studied the availability of vitamin C supplements in cattle. We developed the PCR-based method for detection of bovine bone meal. This method was sensitive and highly selective for bovine bone meal. Thus this method resolved the problem of conventional PCR-based methods that cannot distinguish between bovine bone meal and other bovine-derived materials such as skim milk in feeds.

A-2. Publications and presentations a) Publications Books Matsui, T. and Y. Inoue: Minerals, Japanese Feeding Standard for horses-2004 (edited by Equine Research Institute, Japan Racing Association), pp.26-39, Animal Media, Tokyo, 2004 (in Japanese) Matsui, T. and Y. Inoue: Vitamins, Japanese Feeding Standard for horses-2004 (edited by Equine Research Institute, Japan Racing Association), pp.40-48, Animal Media, Tokyo, 2004 (in Japanese) Original papers Shimada, T., T. Saitoh and T. Matsui: Does acclimation reduce the negative effects of acorn tannins in the wood mouse Apodemus speciosus? Acta Theriologica 49; 203-214, 2004 Hirai, S., H. Kawachi, T. Matsui and H. Yano: Effects of protein deficiency on the mRNA levels of insulin-like growth factors and myostatin in skeletal muscle of weaned lambs. Animal Science Journal 75; 207-212, 2004 Toyoda, A., M. Nakajo, H. Kawachi, T. Matsui and H Yano: PCR detection of bovine mitochondrial DNA derived from meat and bone meal in feed. Journal of Food Protection 67; 2829–2832, 2004 Jeon, T., S.G. Hwang, S. Hirai, T. Matsui, H. Yano, T. Kawada, B.O. Lim and D.K. Park: Red yeast rice extracts suppress adipogenesis by down-regulating adipogenic transcription factors and gene expression in 3T3-L1 cells. Life Sciences 75; 3195-3203, 2004 Hirai, S., M. Yamanaka, H. Kawachi, T. Matsui and H. Yano: Activin A inhibits differentiation of 3T3-L1 preadipocyte. Molecular and Cellular Endocrinology 232; 21-26, 2005 Fukada, M., H. Kawachi, A. Fujikawa and M. Noda: Yeast substrate-trapping system for isolating substrates of protein tyrosine phosphatases: Isolation of substrates for protein tyrosine phosphatase receptor type z. Methods 35; 54-63, 2005 Review Yano, H., S. Hirai and M. Kitagawa: Recent advances in research for nutritional physiology of beef cattle. Proc. Soc. Anim. Nutr. Metab. 48; 79-140, 2004 b) Conference and seminar papers presented The 104th Meeting of Japanese Society of Animal Science: 3 presentations

247 The 54th Meeting of Kansai Society of Animal Science: 2 presentations The 21st Symposium on Trace Nutrition Research: 1 presentation The 24th Meeting of the Japanese Society for Magnesium Research: 1 presentation

A-3. Off-campus activities Membership in academic societies Yano, H.: Japanese Society of Animal Science (Director), Japanese Association of Pet Animal Nutrition (Vice President), Japanese Trace Nutrients Research Society (President), Society of Beef Cattle Science (Councilor), Japanese Society of Environmental and Animal Production (Director, Trustee), Japanese Society of Nutrition and Food Science (Councilor), Japanese Society of Nutrition and Food Science-Kinki Section (Councilor), Kansai Society of Animal Science (Councilor), Japanese Society for Magnesium Research (Director), Japanese Society of Animal Nutrition and Metabolism (Councilor), Japanese Society of Veterinary Science (Councilor) Matsui, T.: Japanese Trace Nutrients Research Society (Councilor), Japanese Society for Magnesium Research (Councilor, Editor), Japanese Society of Animal Nutrition and Metabolism (Editor), Japanese Association of Pet Animal Nutrition (Editor), Society of Beef Cattle Science (Secretary) Membership in Science Council of Japan, etc. Yano, H.: Science Council of Japan (Member), Council of Animal Science (Chairman) Research grants Yano, H.: Research Project for Utilizing Advanced Technologies in Agriculture, Forestry and Fisheries: Evaluation for safety of feeds in relation to BSE and the development of techniques for inactivation and utilization of bone and meat meal (Head: Yano, Sharer: Kawachi), Research Project for Utilizing Advanced Technologies in Agriculture, Forestry and Fisheries: Development of palatable beef by integration of breeding information (Head: Yano, Sharer: Kawachi), Japan Livestock Technology Association: The improvement of beef quality by zinc supplementation (Head: Yano, Sharer: Matsui and Kawachi) Kawachi, H.: Monbusho Research Grants, Grant-in-Aid for Encouragement of Young Scientists (A): Studies on the physiological roles of leptin soluble receptor in fattening cattle (Head: Kawachi).

A-4. International cooperations and overseas activities International meetings (roles) Yano, H.: The 39th International Congress of the International Society for Applied Ethology, International Organizing Committee, Tokyo (Chairman), The 9th World Conference on Animal Production, Brazil (Presentation), The 11th Asian-Australasian Animal Science Congress, Malaysia (Presentation) Matsui, T.: The 11th Asian-Australasian Animal Science Congress, Malaysia (Presentation). Membership in international academic societies Yano, H.: World Association for Animal Production (Director)

248 International joint researches, overseas research surveys Yano, H., Matsui, T., and Kawachi, H.: Effect of conjugated linoleic acids on bovine adipogenesis (USA)

B. Educational Activities（2004.4-2005.3） B-1. On-campus teaching Courses given Undergraduate level: Basic Bioresource Science Ⅱ (Yano et al), Outline of Bioresource Science Ⅱ (Yano et al), Applied Animal Sciences (Yano et al), Physiological Nutrition of Animals (Matsui and Yano), Animal Physiology (Matsui et al), Animal Nutrition (Matsui and Yano), Fundamentals for the Experiments for Bioresource Science (Matsui and Kawachi et al), Laboratory Course in Bioresource Science Ⅰ and Ⅱ (Matsui and Kawachi et al), Introduction to Foreign Literature in Bioresource Science Ⅳ (Matsui et al), Seminar for Applied Animal Science Ⅰ and Ⅱ(Matsui et al) Graduate level: Metabolism and its Regulation of Animals (Matsui), Seminar in Nutritional Science of Animals (Yano and Matsui), Laboratory Course in Nutritional Science of Animals (Yano and Matsui)

B-2. Off-campus teaching, etc. Part-time lecturer Yano, H.: School of Environmental Science, The University of Shiga Prefecture (Environmentology for Animal Production) Matsui, T.: School of Environmental Science, The University of Shiga Prefecture (Environmentology for Animal Production), Graduate School of Agriculture, Kyushu University (Advanced Animal Science)

B-3. Overseas teaching Yano, H.: National Livestock Research Institute of Korea (Lecture) Students and research fellows from abroad Doctoral course: 2 students (Korea and Honduras) Research student: 1 student (Korea)

C. Other remarks Yano, H.: Subcommittee of Agricultural Technology, Evaluation Committee of Incorporated Administrative Agency, The Ministry of Agriculture, Forestry and Fishery (Specialist). Committee on Preparation of Tables of Feed Composition for Animals, The Ministry of Agriculture, Forestry and Fishery (Member). Policymaking Advisory Board of Food, Agriculture and Agricultural Community, The Ministry of Agriculture, Forestry and Fishery (Temporal Member). Committee of Grant-in-Aid for Scientific Research, Japan Society for the Promotion of Science (Member). Advising Committee of Science and Technology of Gifu Prefecture (Member). Evaluation Committee of Research Institutes of Gifu Prefecture (Chairman)

249 Matsui, T.: Committee on Standard Methods for Feed Analyses, Fertilizer and Feed Inspection Services (Member). Subcommittee of Agricultural Industry, Evaluation Committee of Incorporated Administrative Agency, The Ministry of Agriculture, Forestry and Fishery (Specialist). Japanese Committee on ISO TC34/SC10, The Ministry of Agriculture, Forestry and Fishery (Specialist).

2.4.9 Laboratory of Animal Physiology and Functional Anatomy

Staff Professor : Kume, Shin-ichi, Dr. Agric. Sci. (2004.6～) Associate Professor : Manabe, Noboru, Dr. Agric. Sci. (～2004.6) Assistant Professor : Sugimoto, Miki, Dr. Agric. Sci. Students and research fellows Research fellows of the Japan Society for the Promotion of Science: (2) Doctor's Program: (5) (Including 2 of Research fellows of the Japan Society for the Promotion of Science) Master's Program: (6) Undergraduate : (4)

A. Research Activities（2004.4-2005.3） A-1. Main subjects a) Clarification of the effect of environmental pollution on physiology and reproductive functions of mammals. Endocrine disrupters can bind to nuclear receptors including estrogen receptors and interfere with reproductive functions in animals. We evaluate effects of endocrine disrupters and other environment load substances on biological functions in mammals to develop a method that reduces their undesired effects. Effects of endocrine disrupters on nuclear receptors in bones, kidneys and small intestine are examined in pregnant mice and domestic animals. b) Assessments of the effect of global warming on physiology and productivity of mammals. Animals convert ingested feed into heat energy and maintain their body temperature with homeostasis, while they yield livestock products for human food. Global warming, however, impairs physiology and productivity of mammals. Effects of heat stresses on metabolisms at organ- to whole body levels are examined using biochemical and pathological techniques. In addition, methods for effective utilization of energy and materials upon environmental changes are developed. c) Functional morphology of mammalian reproduction and its regulation mechanism Domestic animals with high productivity often have low reproductive ability. We analyze the factors involved in different processes in mammalian reproduction and develop techniques for efficient utilization of germ cells and for avoidance of reproductive disorders.

250 A-2. Publications and presentations a) Publications Books Kume,S.: Dairy Cattle. Livestocks (A.Abe ed.). p.122-159, Nobunkyo, Tokyo, 2005 Original papers Kume,S., T.Toharmat, M.Ridla, K.Nonaka, T.Oshita, M.Nakamura, Y.Yamada and J.H.Ternouth : Effect of high potassium intake from alfalfa silage on mineral status in sheep and periparturient cows. Res. Bull. Natl.Agric. Res. Cent.for Hokkaido Reg. 181; 1-14, 2004 Kume,S., K.Nonaka, T.Oshita, T.Kozakai and H.Kojima: Potassium excretion of dry, pregnant and lactating cows fed forage. Nihon Chikusan Gakkaiho 75; 179-184. 2004 Oshita,T, K.Nonaka, S.Kume and T.Nakui: Effects of forage type on particle size distribution of ruminal digesta and faeces ob non-lactating cows fed high quality forage. Livestock Production Science 91; 107-115, 2004 Yamaguchi-Yamada, M., N. Manabe, K. Uchio-Yamada, N. Akashi, Y. Goto, Y. Miyamoto, M. Nagao, Y. Yamamoto, A. Ogura and H. Miyamoto: Anemia with chronic renal disorder and disrupted metabolism of erythropoietin in ICR-derived glomerulonephritis (ICGN) mice. J. Vet. Med. Sci. 66 (4); 423-431, 2004 Wollenhaupt, K., H. Welter, R. Einspanier, N. Manabe and K.P. Brussow: Expression of epidermal growth factor receptor (EGF-R), vascular endothelial growth factor receptor (VEGF-R) and fibroblast growth factor receptor (FGF-R) systems in porcine oviduct and endometrium during the time of implantation. J. Reprod. Dev. 50 (3); 269-278, 2004 Yamaguchi-Yamada, M., N. Manabe, Y. Goto, S. Anan, K. Miyamoto, Y. Miyamoto, M. Nagao, Y. Yamamoto and A. Ogura: Improvement of anemia associated with chronic renal failure by recombinant human erythropoietin treatment in ICR-derived glomerulonephritis (ICGN) mice. J. Vet. Med. Sci. 66 (7); 883-886; 2004 Goto, Y., F. Matsuda-Minehata, N. Inoue, T. Matsui, A. Maeda and N. Manabe: Porcine (Sus scrofa) cellular FLICE-like inhibitory protein (cFLIP): Molecular cloning and comparison with the human and murine cFLIP. J. Reprod. Dev. 50 (5); 549-555, 2004 Uchio-Yamada, K., N. Manabe, Y. Goto, S. Anann, Y. Yamamoto, K. Takano, A. Ogura and J. Matsuda: Decreased expression of matrix metalloproteinases and tissue inhibitors of metalloproteinase in the kidneys of hereditary nephrotic (ICGN) mice. J. Vet. Med. Sci. 67 (1); 35-41, 2005 Kagawa, N., Y. Sakurai, T. Miyano and N. Manabe: Effects of long-term grafting on follicular growth in porcine ovarian cortical grafts xenoplanted to severe combined immunodeficient (SCID) mice. J. Reprod. Dev. 51 (1); 77-85, 2005 Reviews Kume, S.: Improvement of reproduction and nutrient in high producing cows (1). Kachiku Jinkojyusei 226; 24-39, 2005 Kume, S.: Improvement of reproduction and nutrient in high producing cows (1). Kachiku Jinkojyusei 227; 40-55, 2005 b) Conference and seminar papers presented The 137th Annual Meeting of Japanese Society of Veterinary Science : 4 presentations The 97th Annual Meeting of Japanese Society of Animal Reproduction: 3 presentations

251 The 104th Annual Meeting of Japanese Society of Animal Science: 3 presentations

A-3. Off-campus activities Membership in academic societies Kume, S.: Japanese Society of Equine Science (Editor), Hokkaido Animal Science and Agriculture Society (Editor), Hokkaido Society of Grassland Science (Editor) Manabe, N.: Japanese Society of Animal Reproduction (Senior Director), Japanese Society of Veterinary Science (Councilor), Kansai Society of Animal Science (Councilor), Japanese Society of Veterinary Anatomists (Councilor), Society for the Study of the SAM (Councilor), Journal of Reproduction and Development (Associate Editor-in Chief) Membership in Science Council of Japan etc. Manabe, N.: Evaluator for Research fellows of the Japan Society for the Promotion of Science Research grants Manabe, N.: Monbusho Research Grant: Creative Scientific Research; Regulation mechanism of follicular atresia in mammalian ovaries and artificial control for availableness of potential oocytes in the ovaries (head) Sugimoto, M.: Monbusho Research Grant: Creative Scientific Research; Regulation mechanism of follicular atresia in mammalian ovaries and artificial control for availableness of potential oocytes in the ovaries (Sharer)

A-4. International cooperation and overseas activities International meetings Manabe, N.: XIII International Workshop on the Development and Function of the Reproductive Organs, Copenhagen (Presentation) Sugimoto, M.: The 37th Annual Meeting of the Society for the Study of Reproduction, Vancouver (presentation), 16th International Congress of the International Federation of Associations of Anatomists, Kyoto (presentation), XIII International Workshop on the Development and Function of the Reproductive Organs, Copenhagen (Presentation), 3th Copenhagen Workshop on Environment, Reproductive Health and Fertility, Copenhagen (Presentation) Editorial work for international journals (roles) Manabe, N.: Journal of Reproduction and Development (vice chief editor), Reproductive Biology and Endocrinology (editor), European Journal of Cellular Pathology (editor), FEBS letters (reviewer), Biochemica et Biophysica Acta (reviewer), Molecular Reproduction and Development (reviewer), American Journal of Obstetrics and Gynecology (reviewer), Life Science Journal (reviewer). Overseas researcher receiving Inviting researchers: (Dr. Abraham Amsterdam, Weismann Institute of Science, Professor; Dr. Lynn R. Fraser, Kings College London, Professor)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Outline of Bioresource Science III (Kume et al.), Physiology (Manabe et al.),

252 Fundamentals for the Experiments of Bioresource Sciences (Sugimoto et al.), Anatomy and Physiology (Kume, Manabe), Introduction to Foreign Literature in Bioresource Science II (Manabe, Kume et al.), Laboratory Course in Bioresource Science I and II (Kume, Manabe, Sugimoto et al.), Applied Animal Sciences (Kume et al.), Molecular Cell Biology (Kume), Livestock Production Techniques and Practice II (Kume et al.), Seminar in Applied Animal Science I and II (Manabe et al.), Graduate level: Anatomy and Cell Biology (Kume), Molecular Cell Biology (Kume), Seminar in Anatomy and Cell Biology (Kume), Laboratory Course in Anatomy and Cell Biology (Kume)

C. Other remarks Kume,S. : Subcommitee of Japanese Standard for Dairy Cattle (member), Assessment Committee of Research Grant of Ministry of Agriculture, Forestry and Fisheries (member)

2.4.10 Laboratory of Animal Husbandry Resources

Staff Professor : Hirooka, Hiroyuki, Dr. Agric. Sci. Lecturer : Ishida, Joken, Dr. Agric. Sci. Students and research fellows Doctor's program: (4) Master's program: (5) Undergraduate : (3)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Total evaluation for animal production systems. Data and information on genetic performance, nutrition, management and economic situations for beef, dairy, pigs, sheep and goats were collected from various research fields concerning animal sciences in order to evaluate total animal production systems. Modeling and simulation of such systems were carried out. b) Studies on livestock production systems in tropical areas Surveys have been conducted to investigate various performances of native livestock and feeding systems in tropical areas. Such information will be used to search better strategies for improving the existing systems. Sustainable farming systems integrated between livestock and crops in smallholders in the developing countries were evaluated through modeling works. c) Studies on regional properties of vegetation and nomadic livestock in arid areas. For the purpose of defining and settling the problems found in traditional animal production systems in arid areas, the evaluations of regional woody and herbaceous plants as the feed resources with reference to the physiological properties of nomadic animals have been investigated. The studies performed in 1996 were as follows. 1) Strategy of reclaiming the grassland devastated by over-grazing in Syria. 2) Feeding and management techniques of

253 nomadic animals in Gobi district, Mongolia. 3) Feeding and management techniques of farm animals in northern step, southern desert and eastern mountains of Kazakhstan. d) Studies on environmental problems in animal production. In recent, because animal industries have been specialized and scaled up, environmental pollution caused by such animal industries led to serious problems in society in Japan. Models for predicting excretion of nitrogen and phosphorus from animal production systems were developed and a country-wide fact-finding studies of the connection between crop and animal industries were performed in order to define the role of animal industry in establishing a sustainable agriculture.

A-2. Publications and presentations a) Publications Books Hirooka, H.: (Chapter 9 Animal science) Environmental problems in animal production-with special reference of cattle production-. Theory and politics for environmental issues (Edited by H. Terada). pp.181-201, Koyo Shobo, 2005 (in Japanese) Original papers Xiong, Y., M. Muramatsu, M. Hirata, K. Oishi, I. Kaihotsu, T. Takamura, S. Furumi and N. Fujiwara: Approximation method for time-integral of photosynthesis for NPP estimation using remote sensing data : case study in Mongolia. Journal of The Remote Sensing Society of Japan 25; 79-190, 2005 Kojima, H., S. Kume, K. Nonaka, T. Oshita, T. Kozakai and H. Hirooka: Effects of feeding and animal performance on nitrogen, phosphorus and potassium excretion by Holstein cows. Animal Science Journal.76; 139-146, 2005 Choumei, Y., Y. Nakagawa, H. Kojima and H. Hirooka: An analysis for impression of residents in livestock waste management system. J. JASS 21(1); 15-24, 2005 (in Japanese) Sasaki, Y., H. Hirooka and M. Tsuiki: Systems analysis applied to animal husbandry. J. JASS 20(2); 125-137, 2004 (in Japanese) Reviews Hirooka, H., F. Terada and J.B. Liang: Development and application of simulation models for sustainable cattle production systems. In: Wong, H. H. eds. New Dimensions and Challenges for Sustainable Livestock Farming. Vol. 2, p.107-112, Proceedings of the 11th Animal Science Congress, The Asian Australasian Association of Animal Production Societies, Kuala Lumpur, Malaysia, 2004 Kojima, H., S. Kume, H. Hirooka, K. Nonaka, T. Oshita and T. Kozakai: Effects of intakes and animal performance on nitrogen, phosphorus, and potassium excretions by Holstein cows. In: Wong, H. H. eds. New Dimensions and Challenges for Sustainable Livestock Farming. Vol.3, p.6-9, Proceedings of the 11th Animal Science Congress, The Asian Australasian Association of Animal Production Societies, Kuala Lumpur, Malaysia, 2004 Hirooka, H. and J. B. Liang: Comparison of protein and fat deposition for Malaysia indigenous cattle and buffaloes with temperate cattle breeds. In: Wong, H. H. eds. New Dimensions and Challenges for Sustainable Livestock Farming. Vol. 3, p.463-465, Proceedings of the 11th Animal Science Congress, The Asian Australasian Association of Animal Production Societies, Kuala Lumpur, Malaysia, 2004

254 b) Conference and seminar papers presented The 104th Annual Meeting of Japanese Society of Animal Science: 3 presentations Proceedings of the 11th Animal Science Congress, The Asian Australasian Association of Animal Production Societies: 3 presentations The Autumn Symposium of Japanese Agricultural System Society in 2004: 1 presentation The 42th Annual Meeting of Beef Cattle Science: 1 presentation

A-3. Off-campus activities Membership in academic societies Hirooka, H.: Japanese Agricultural System Society (Director, Editor), Society of Beef Cattle Science (Secretary) Research grants Monbushou Research Grants, Promotion Research: Theoretical Studies on Animal Genetic Conservation and Construction of a Database system (Head: Hirooka), Evaluation of Advanced Japanese Beef Cattle Breeding Schemes Incorporating Performance Testing for Blood Serum Insulin-Like Growth Factor I and Carcass Traits (Head: Hirooka), Grant-in-Aid for Scientific Research (C): Comparative research of methodology and practice about agricultural field science and agriculture (Head: Suehara, Sharer: Hirooka)

A-4. International cooperation and overseas activities Overseas researcher receiving Inviting researcher: Doctor Kahi Alexander Kigunzu (Assistant professor of Animal Husbandry at Egerton University)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Outline of Bioscience Ⅲ (Hirooka et al.)，International Animal Agriculture (Ishida) Graduate level: Seminar of Animal Husbandry Resources (Hirooka, Ishida), Laboratory Course of Animal Husbandry Resources (Hirooka, Ishida)

B-2. Off-campus teaching, etc. Open seminar, etc. Hirooka, H.: Professional education I at Kyoto Prefecture (Lecture: Field researches and Data analysis) Kumamoto Prefectural Agricultural Research Center (Lecture: Effects of cow’s age on performance)

B-3. Overseas teaching Students and research fellows from abroad Doctor course student: 1 (Honduras)

255 Chair of Marine Biological Resources

2.4.11 Laboratory of Fisheries and Environmental Oceanography

Staff Professor : Fujiwara, Tateki, Dr. Agric. Sci Associate Professor : Kasai, Akihide, Dr. Agric. Sci. Students and research fellows Doctor's program : (3) Master's program : (８) Undergraduate : (４)

A. Research activities (2004.4-2005.3) A-1. Main subjects a) Analysis of the mechanism maintaining high productivity of coastal seas Coastal seas are highly productive areas. To reveal mechanisms maintaining this high productivity and to find effective measures to sustain this productivity, we are studying nutrient dynamics in the Seto Inland Sea, Ohmura Bay and Ise Bay. Seto Inland Sea (SIS): Nutrient pools of the oceanic origin are located in the lower layer at the openings of SIS and that of terrestrial origin lies on the bottom of Harima-nada. These nutrients are transported to the central SIS by the estuarine circulations and then upwell into the upper layer at Bisan Strait (central SIS). b) Study on eutrophication and hypoxia in semi-enclosed coastal seas Excess amount of loads of nitrogen and phosphorus flowing into Ise Bay, Tokyo Bay and Seto Inland Sea causes eutrophication and hypoxia in these seas. Mechanisms generating hypoxic water mass in Ise bay and Hiuchi-nada were studied in this year. A cold dome is created at the bottom in the eastern part of Hiuchi-nada. Temperature is well correlated with oxygen concentration in the dome. This indicates that the currents are weak in the dome, so that the water exchange is insufficient and then oxygen is depleted. c) Analysis of long term variability in coastal water qualities To restore water qualities in semi-enclosed coastal seas (Ise Bay, Tokyo Bay and Seto Inland Sea), reduction of anthropogenic loads of organic matter (COD), phosphorus and nitrogen have been conducted for more than 30 years. Owing to these measures, near-shore water quality have been significantly improved. However, water qualities in basin-wide scale (WQBS) are not so. We gathered data of water qualities, hydrographic parameters and meteorological parameters over 20 ~ 30 years to make data-base. It was revealed that WQBS varies with low correlation with the amount of pollutant loads, but varies in relation to hydrographic and meteorological conditions. In Seto Inland Sea, variations in WQBS are governed mainly by the through flow of SIS. A similar study was conducted for an estuary area of Bangpakong River, Thailand. d) Analysis of short-term variability in coastal water qualities The composition and behavior of allochtonous particulate organic matter (POM) in Ise Bay were investigated to elucidate the short-term variation in POM accompanying changes in river discharge. The behavior of POM was significantly regulated by hydrographic conditions, but behavior was different in the upper layer versus the middle & lower layers. The former showed

256 simple dynamics controlled by the river plume, while the latter showed complex dynamics because of the changes in river discharge and subsequent variation in estuarine circulation. During normal discharge, the contribution of riverine materials to POC in the surface water within the bay was negligible because most riverine organic matter is deposited before flowing into the bay. During high discharge, on the other hand, the contribution of riverine organic matter to total POM increased to 50% at ~10 km from the river mouth. Though riverine organic matter loads increased, the total amount of POC decreased around the river mouth due to flushing of phytoplankton. After river discharge, the contribution decreased rapidly. The behavior of POM in the middle and lower layers differed from that in the surface layer. At normal discharge, the influence of riverine organic matter was weak. During high discharge, high flooding temporarily weakened the bottom inflow, resulting in heavier riverine organic matter distributed from the river mouth to mid regions within the bay in the lower layer. The maximum contribution of riverine organic matter to total POM was estimated to be ~60% around 25 km from the river mouth. After high discharge, riverine POM in the lower layer was pushed to the bay head by enhanced estuarine circulation and was uplifted to the middle layer. The behavior of riverine POM dynamically changed in relation to river discharge, and exerted a significant influence on bottom water conditions in the bay. e) Analysis of food chain by using stable isotope ratio Food chain is analyzed by stable isotope ratio of nitrogen and carbon in muscular tissue of marine animals and particulate organic matter. The bivalve corbicula (Corbicula japonica) in Lake Ogawara and Lake Shinji showed enriched isotope composition, while those in Lake Jusan were depleted. In addition, the difference in the isotope ratios between the sampling sites was remarkable in Lake Jusan. Chlorophyll concentrations were significantly higher in Lake Ogawara and Lake Shinji than those in the inflow rivers, although that in Lake Jusan was equivalent to that in the river. The bivalves in Lake Ogawara and Lake Shinji essentially assimilate autochthonous phytoplankton, while those in Lake Jusan have to assimilate terrestrial matter in the upper reaches and marine phytoplankton in the lower reaches due to low production in the lake.

A-2. Publications and presentations a) Publications Books Fujiwara, T.: Tide and Tidal Current. A handbook on Fisheries Science (edited by Takeuchi et al.). pp. 4-9, Seibutsukenkyusha, Tokyo, 2004 (in Japanese) Kasai, A.: Physical properties of ocean water, A handbook on Fisheries Science (edited by Takeuchi et al.). pp. 11-16, Seibutsukenkyusha, Tokyo, 2004 (in Japanese) Original papers Fujiwara, T.: Long-term variations in water quality and ecosystem in the Seto Inland Sea. Sea and Sky 80 (2); 91-97，2004. Sasakura, S., T. Noda, A. Ishino, and T. Fujiwara: Long-term changes of phosphorous loads and water qualities in the Seto Inland Sea. Sea and Sky 80; 99-107，2004 Kobayashi, S., T. Takashi, and T. Fujiwara: Long-term variation of nutrient transport and its effect on the ecosystem of the Seto Inland Sea. Sea and Sky 80 (2); 61-67，2004 Kakehi, S., T. Fujiwara, and Y. Sugiyama: Upwelling induced by intermittent bottom intrusion of

257 oceanic water into Ise Bay. Oceanography in Japan 13; 537-551, 2004 Fujiwara, T., A. Kishimoto, and M. Nakajima: Long and short term variation in hypoxic water in Osaka Bay. Proceedings of Coastal Engineering 51; 931-935, 2004 Fujiwara, T., M. Kunii, and N. Uno: Coastal sea level decided by the temperature of the shlf-slope water,. Proceedings of Coastal Engineering 51; 376-380, 2004 Kasai, A. and A. Nakata: Utilization of terrestrial organic matter by the bivalve Corbicula japonica estimated from stable isotope analysis. Fisheries Science 71; 151-158, 2005 Kasai, A., T. Fujiwara, T. Kimura and H. Yamada: Fortnightly shifts of intrusion depth of oceanic water into Ise Bay. Journal of Oceanography 60; 817-824, 2004 Sugimoto, R., A. Kasai, S. Yamao, T. Fujiwara and T. Kimura: Variation in particulate organic matteraccompanying changes of river discahrge in Ise Bay. Bulletin of the Japanese Society of Fisheries Oceanography 68；142-150, 2004 Reports Fujiwara, T.: Impact of a flooding on circulation and mass-transport in Ise Bay. Kaiyo Monthly 36 (3); 196-199，2004 Fujiwara, T.: Tranport of nitrogen and phosphorus from the Ocean into the Seto Inland sea. Scientific Forum of the Seto Inland Sea 40; 10-16, 2004 Fujiwara, T: Long term variation in water quality of Osaka Bay. Scientific Forum of the Seto Inland Sea 39; 31-33，2004 Ohsawa, T., H. Mizutani and T. Fujiwara: Development and verification of the coupled atmosphere-ocean model for Ise Bay, Japan. Proceedings of the 29th International Conference on Coastal Engineering, ASCE; 690-700, 2004 Kasai, A.: Numerical model of transport of fish eggs and larvae. Elucidation of variability in fish stocks and development of advanced prediction of variability; 64-65, 2004 b) Conference and seminar papers presented 2004 Spring meeting of the Oceanographic Soc. Japan: 2 presentations 2004 Autumn meeting of the Oceanographic Soc. Japan: 9 presentations 2004 Annual meeting of the Japan. Soc. Fish. Sci.: 1 presentation 2004 Annual meeting of the Japan. Soc. Fish. Oceanogr.: 1 presentation Seto Inland Sea Forum in Kyoto: 3 presentations 51st meeting of Japanese Coastal Engineering Soc.: 2 presentations The 34 Workshop on Marine Research and Technology: 3 presentations Challenger Conference for Marine Science, Liverpool (U. K.)：1 presentation International symposium on long-term variations in the coastal environments and ecosystems: 1 presentation 2004 Autumn meeting of Kinki Branch of the Fisheries Soc. Japan: 1 presentation Hokkaido Univ.- Kyoto Univ.- Ryukyu Univ. Collaborative field science symposium: 1 presentation 13th Hiuchinada symposium: 1 presentation

A-3. Off-campus activities Membership in academic societies Fujiwara, T.: The Oceanographic Society of Japan (Counselor), Coastal Oceanography Division of the Oceanographic Society of Japan (Committeeman), The Marine Meteorological Society (Director, Editor), The Japanese Society of Fisheries Oceanography (Committeeman),

258 Japanese Society of Civil Engineers (Committeeman) Kasai, A.: Coastal Oceanography Division of the Oceanographic Society of Japan (Editor, Committeeman), The Japanese Society of Fisheries Oceanography (Committeeman) Research grants Grant-in-aid for Scientific Research (A); Removal of terrestrial organic matters by bivalves (Kasai) Fisheries Agency; Variability of fisheries resources and its prediction (Kasai)

A-4. International cooperation and overseas activities International meetings (roles) Kasai, A.: International symposium on long-term variations in the coastal environments and ecosystems (Matsuyama, Japan, Poster presentation)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Marine Environment (Fujiwara), Marine Ecosystem (Kasai), Practical Course in Marine Bioscience and Technology (Fujiwara), Fundamentals for the experiments of bioresource science (Fujiwara, Kasai), Laboratory Course in Bioresource Science (Fujiwara, Kasai), Outline of Bioresource Science (Fujiwara), Seminar in Marine Bioresources Science (Fujiwara, Kasai), Introduction to Foreign Literature in Bioresource Science 3 (Fujiwara) Graduate level: Physical Oceanography (Fujiwara), Fisheries and Environmental Oceanography (Kasai), Seminar in Fisheries Physics (Fujiwara, Kasai), Laboratory Course in Fisheries Physics (Fujiwara, Kasai)

B-2. Off-campus teaching, etc Part-time lecturer Fujiwara: Nagoya University, Graduate School of Engineering Fujiwara: Kobe University, Faculty of Human Development Kasai: Kinki University, Faculty of Agriculture (Statistics)

C. Other Remarks Fujiwara: Researcher of the Maritime Safety Agency, Researcher of the Disaster Prevention Research Institute, Researcher of the Disaster Science Research Institute, Kyoto University

259 2.4.12 Laboratory of Marine Stock-Enhancement Biology

Staff Professor : Tanaka, Masaru, Dr. Agric. Sci. (Nakabo, Tetsuji, Dr. Agric. Sci. Ichthyology, Kyoto University Museum) Associate Professor : Tagawa, Masatomo, Dr. Sci. Assistant Professor : Nakayama, Kouji, Dr. Agric. Sci. COE Post doctor :Shoji, Jun, Dr. Agric. Sci Students and Research Fellows Doctor's program : (8) Master's program : (7) Undergraduate : (5) JSPS post-doctor : (1)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Early life history of fishes Occurrence, distribution, food habits and growth of larvae and juveniles in coastal marine fishes as flounder, sea bass, sea breams, tunas, and mackerels have been observed in the sea, being combined with physiological laboratory experiments concerning endocrine, osmoregulatory, and digestive functions. Recent particular focuses have been placed on the stock structure and impacts of mass-release of juvenile flounder on coastal ecosystem, and origin of the Ariake sea bass population related to its amphidromous life history. b) Studies of systematic ichthyology We are proceeding the systematic study of marine fishes based on the specimens from the Indo-Pacific region. We have an another project to study the taxonomy of some common coastal fishes cooperated with the study of early life histories of them (supervised by Prof. T. Nakabo, Kyoto University Museum). c) Endocrinological study of fish development Controlling mechanisms of fish metamorphosis, neoteny and migration are investigated, mainly during early life history using endocrinological methods. Involvements of maternal hormones (thyroid hormones and cortisol contained in unfertilized eggs) in early survival and osmoregulation before the onset of larval endocrine organ are examined in flounders, ice goby, and tilapia.

A-2. Publications and Presentations a) Publications Books Tanaka, M., M. Tagawa and K. Nakayama: Fishes in Ariake estuary. Connections among forest, sato and sea―Challenge by the Field Science Education and Research Center, Kyoto University. pp64-68, Ei Press, 2004 Original papers Shoji J. and M. Tanaka: Feeding of piscivorous Japanese Spanish mackerel juveniles on larvae with an elongate and laterally compressed body. Journal of Fish Biology 65; 282-286,

260 2004 Shoji J. and M. Tanaka: Effects of prey concentration on growth of piscivorous Japanese Spanish mackerel Scomberomorus niphonius larvae in the Seto Inland Sea, Japan. Journal of Applied Ichthyology 20; 271-275, 2004 Shoji J., R. Masuda, Y. Yamashita and M. Tanaka: Moon jellyfish or juvenile piscivore? A possible change in the relative importance of trophic pathways from fish larvae to their predators caused by hypoxia in the Seto Inland Sea. Proceedings of International Symposium on Long-term variations in the Coastal Environments and Ecosystems; 173-178. Tagawa, M., T. Kaji, M. Kinoshita and M. Tanaka: Effect of stocking density and addition of proteins on larval survival in Japanese flounder, Paralichthys olivaceus. Aquaculture 230; 517-525, 2004 Dou, SZ, R. Masuda, M. Tanaka and K. Tsukamoto: Size hierarchies affecting the social interactions and growth of juvenile flounder, Paralichthys olivaceus. Aquaculture 233; 237-249, 2004 Islam, Md. S., Md. J. Saker, T. Yamamoto, Md. A. Wahab and M. Tanaka: Water and sediment quality, partial mass budget and effluent N loading in coastal brackishwater shrimp farms in Bangladesh. Mar. Pol. Bull. 48; 471-485, 2004 Islam, Md. S., Md. A. Wahab and M. Tanaka: Seed supply for coastal brackishwater shrimp farming: environmental impacts and sustainability. Mar. Pol. Bull. 48; 7-11, 2004 Islam, Md. S. and M. Tanaka: Impacts of pollution on coastal and marine ecosystems including coastal marine fisheries approach for management: a review and synthesis. Mar. Pol. Bull. 48; 624-649, 2004 Ueda, H., A. Terao, M. Tanaka, M.Hibino and Md. S. Islam: How can river-estuarine planktonic copepods survive river floods? Ecological Research, 19; 625-632, 2004 Wada, T., M. Aritaki, M. Tanaka: Effects of low-salinity on the growth and development of spotted halibut Verasper variegates in the larva-juvenile transformation period with reference to pituitary prolactin and gill chloride cells responses. J. Exp. Mar. Biol. Ecol. 308; 113-126, 2004 Aritaki, M., K. Ohta, Y. Hotta, M. Tagawa and M. Tanaka: Effects of different water temperatures on occurrence of morphological abnormalities relating to development and metamorphosis in spotted halibut larvae. Nippon Suisan Gakkaishi 70; 8-15, 2004 (in Japanese) Reports Tanaka, M.: Ariake bay and its environmental problems- restoration from a new aspect of connection among forest, river and sea. Hodaoki 36: 12-21, 2004 (in Japanese) Tanaka, M.: Forest grows sea, and sea grows forest. Kankyo-kaigi(fall issue): 222-225, 2004 (in Japanese) Yamashita, Y., Y. Tanaka, M. Tanaka: Early ecology of Japanese flounder and south-north differences in its fluctuation pattern. Gekkan-Kaiyo 412: 766-770, 2004 (in Japanese) Tagawa, M., K. Nakayama, M. Tanaka: Metamorphosis and early mortality in fishes. Report of Monbu-kagaku-sho Research Grant (Creative-basic Research), 249-256, 2004 (in Japanese) Tanaka, M., K. Nakayama: Genetic stock structure and regional characteristics of ecophysiology in Japanese flounder. H13-15 Research report on the development of seed production and

261 release techniques for establishing ecosystem-conservation culture system, 55-58, 2004 (in Japanese) Tanaka, M., K. Nakayama: Genetic stock structure and regional characteristics of ecophysiology in Japanese flounder H16 Research report on the development of seed production and release techniques for establishing ecosystem-conservation culture system. Plan for the 2nd Phase (H16-18). 50-51, 2004 (in Japanese) b) Conference and seminar papers presented 2004 Annual Meeting of the Japan Society of Fisheries Science (Kagoshima, 2004. 4.1-5): 15 (Tanaka, Tagawa, Nakayama). 11th International Symposium on nutrition and feeding in fish (Puket, 2004.5.2-7): 1 (Tanaka, Tagawa) 5th International symposium on fish endocrinology (Castellon, 2004.9.5-9):1 (Tagawa) Symposium on Present status and future perspective of fisheries sciences (Hiroshima, 2004.8.28):1 (Tanaka) 2004 Kinki-Chushikoku Divisions Meeting of the Japan Society of Fisheries Science (Okayama, 2004.9.13-14):1 (Tanaka) Seminar by Regional zero-emission research group (Shibecha, 2004.10.25):1 (Tanaka) Symposium on Scientific progress in Ariake sea and its possible restoration (Nagasaki, 2004.11.20):1 (Tanaka) Joint field science symposium by Hokkaido, Kyoto and Ryukyu Universities (Naha, 2004.12.4-5):3 (Tanaka) 2004 Symposium on Dynamics of the Ocean Biosystem (Tokyo, 2004.12.8-10):1 (Tagawa)

A-3. Off-campus activities Membership in academic societies (roles) Tanaka: The Japanese Society of Fisheries Science (Vice president, Council member of Kinki Branch), Japan Aquaculture Society (Council member), The Japanese Society of Fisheries Oceanography (council member), The Ichthyological Society of Japan (council member). Research grants Monbusho Research Grant: Scientific Research(A2), Fisheries resource biology and conservation ecology on the Chinese temperate bass and the hybrid population of Ariake temperate bass(Tanaka, Tagawa, Nakayama), Scientific Research(C2), Eye translocation in flatfish metamorphosis-mechanism of asymmetrical morphogenesis by thyroid hormone (Tagawa), Creative-basic Research, Dynamics of the Ocean Biosystem (Tagawa), Encouragement of young scientist (B), Genetic diversities of temperate bass around Japan (Nakayama), Scientific Research(S), Biological and biochemical researches on enhancement and culture for the bluefin tuna (Tanaka) Entrust Research Grant:「 Research on development of seed production and release techniques for establishing ecosystem-conservation culture system」 (Tanaka, Nakayama)

A-4. International cooperations and overseas activities International meetings (roles) Tanaka, M.: 6th International Flatfish Ecology Symposium (Chairman)

262 Tagawa, M.: 6th International Flatfish Ecology Symposium (Organizing committee) Nakayama, K.: 6th International Flatfish Ecology Symposium (Organizing committee) Membership in international academic society Tanaka, M.: Journal of Applied Ichthyology (editorial board), Aquatic Living Resources (Scientific Advisary Committee) International Joint Researches, Overseas Research Surveys Tanaka, M.: Cooperative research on the early development and rearing techniques for marine fish larvae (Malaysia). Tanaka, M., Nakayama, K.: Cooperative research on the sympatric distribution of spotted and non-spotted temperate bass in south-western Korea(Korea). Scholars from abroad Visiting scientists 3 (Associate Professor, University of Kyongsang; Professor, University of Toronto; Professor, University of Rhode Island)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Marine Ecology (Tanaka), Marine Physiology (Tagawa), Laboratory Course in Marine Resource Biology (Tagawa, Nakayama), Seminar in Fisheries Science (Tanaka, Tagawa, Nakayama), Course in Marine Bioscience and Technology I (Tagawa, Nakayama), Pocket Seminar (Tanaka, Tagawa, Nakayama) Graduate level: Seminar in Marine Resource Biology (Tanaka, Tagawa, Nakayama), Laboratory Course in Marine Resource Biology (Tanaka, Tagawa, Nakayama), Biology of Marine Resources (Tagawa)

B-2. Off-campus teaching, etc. Part-time Lecture Tanaka, M.: Faculty of Environmental Science, Shiga Prefectural University (Fisheries Biology)

B-3. Overseas teaching Students: Doctoral course student 1 (Bangladesh)

C. Other remarks Committees in the Faculty Tanaka, M.: Director of Field Education and Research Center, Council member, Curriculum committee, Pocket seminar committee (chairman), Council member of Graduate School of Global Environmental Studies, Council member of University Museum, Council member of Ecological Research center Committees out of the University Tanaka, M.: Shiga Prefectural committee for adjustment of Lake-Biwa Fisheries, Evaluating committee member of National Research Institute of Aquaculture

263 Chair of Marine Microbiology

2.4.13 Laboratory of Physiology and Genetics of Marine Microbes

Staff Professor : Sako, Yoshihiko, Dr. Agric. Sci.. Assistant Professor : Yoshinaga, Ikuo, Dr. Agric. Sci. Assistant Professor : Nomura, Norimichi, Dr. Agric.Sci. Students and research fellows Fellow of Cooperative Research among Industry, Academia and Government: (1) JSPS fellow (PD) : (1) Post-doctoral fellow : (1) Doctor's program : (2) (JSPS Fellow: 1) Master's program : (6) Research student : (1) Undergraduate : (4)

A. Research Activities (2004.4-2005.3) A-1. Main subjects In the sea, there are many different environmental conditions such as open ocean, deep sea, hydrothermal vents, polar circles, red tide sea, coastal area, where many kinds of marine microbes (bacteria, archaea, and microalgae) live. We study their diversity and strategies how to live and survive and their mutual relationships in such conditions. We, further, aim to develop the genetic resources from these marine microbes. Our main themes are as follows: a) Studies on novel marine hyperthermophilic microorganisms and their development of genetic resources (1) Pysiological and ecological studies on thermophilic microorganisms thriving at marine hydrothermal environments (2) Isolation and characterization of novel hyperthermophilic microbes from marine hydrothermal vents (3) Search for the useful genes and enzymes from hyperthermophilic micorobes b) Ecology of microbes in the marine environments (1) Bioremediation for marine environments by using coastal marine bacteria (2) Molecular ecological studies on novel denitrifying bacteria and denitifying genes in coastal area c) Ecological and molecular biological studies on the harmful algae (1) Molecular ecological studies of toxic dinoflagellates (2) Genomics of organelle DNA in toxic dinoflagellates

264 A-2. Publications and presentations a) Publications Books Sako, Y.: Deep-sea hydrothermal vents and thermophilic biotechnology. In “New Strategy of Bioscience”, pp.45-57, Maruzen, Tokyo, 2004(in Japanese). Original papers Nakagawa, S., K. Takai, K. Horikoshi and Y. Sako: Aeropyrum camini sp. nov., a strictly aerobic, hyperthermophilic archaeon from a deep-sea hydrothermal vent chimney. Int. J. Syst. Evol. Microbiol. 54(2); 329-335, 2004 Imagawa, T., H. Nakayama, N. Katsunuma, H. Sakuraba, T. Ohshima, T. Itoh, Y. Sako, N. Nomura and H. Tsuge: Crystallization and preliminary X-ray diffraction analysis of homing endonuclease I-Tsp061I. Acta. Crystallogr. D Biol. Crystallogr. 60 (11); 2006-2008, 2004 Kim C.-J., C.-H., Kim and Y. Sako: Molecular phylogenetic relationships within the genus Alexandrium (Dinophyceae) based on the nuclear-encoded SSU and LSU rDNA D1-D2 sequences. J. Kor. Soc. Oceanog. 39 (3); 172-185, 2004 Kim C.-J., C.-H., Kim and Y. Sako: Population analysis of Korean and Japanese toxic Alexandrium catenella using PCR targeting the area downstream of the chloroplast psbA gene. J. Fish. Sci. Tech. 7(3); 130-135, 2004 Kim C.-J., S. Yshimatsu and Y. Sako: Molecular identification of the toxic Alexandrium tamiyavanichii (Dinophyceae) by the whole-cell FISH method. J. Fish. Sci. Tech. 7(4); 175-183, 2004 Sako, Y., S. Hosoi-Tanabe and A. Uchida: Fluorescence in situ hybridization using rRNA-targeted probes for simple and rapid identification of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella. J. Phycol. 40; 598-605, 2004 Nakagawa, S., S. Nakamura, F. Inagaki, K. Takai, N. Shirai and Y. Sako: Hydrogenivirga caldolitus gen. nov., sp. nov., a novel extremely thermophilic, hydrogen/sulfur oxidizing bacterium from a coastal hydrothermal field. Int. J. Syst. Evol. Microbiol. 54; 2079-2084, 2004 Hosoi-Tanabe,S., S. Nagai and Y. Sako: Species-specific detection and quantification of the toxic dinoflagellate Alexandrium tamarense by Taq Man PCR using 5’-3’exonuclease activity. Marin. Biotechnol. 6; S30-S34, 2004 Maki, T., I. Yoshinaga, N. Katanozaka and I. Imai: Phylogenetic analysis of intracellular bacteria of a harmful marine microalga, Heterocapsa circularisquama (Dinophyceae). Aqua. Microb. Ecol. 36; 123-135, 2004 Kim C.-J., C.-H., Kim and Y. Sako: Paralytic shellfish poisoning toxin analysis of the genus Alexandrium (Dinophyceae) occurring in Korea coasral waters. Fish. Sci. 71(1); 1-11, 2005 Nakagawa, S., F. Inagaki, K, Takai, K. Horikoshi and Y. Sako: Thioreductor micantisoli gen. nov., sp. nov., a novel mesophilic, sulfur-reducing chemolithoautotroph within the epsilon-Proteobacteria isolated from the hydrothermal sediments in the Mid-Okinawa Trough. Int. J. Syst. Evol. Microbiol. 55; 599-605, 2005 Reports Sako,Y. : Development of molecular detection method of toxic planktons and cysts (in Japanese).

265 Reports of counterplanning for protection of shellfish poisoning damage (Ministry of Agriculture), 2005. Development of molecular identification method of red tide planktons (in Japanese). Reports of counterplanning for protection of red tide damage (Ministy of Agriculture), 2005 b) Conference and seminar papers presented The 2004 Annual Meeting of the Japanese Society of Fisheries Science: 5 presentations The 2004 Joint Meeting of the Chugoku-Shikoku and Kinki Branches of the Japanese Society of Fisheries Science: 5 presentations The 2004 Annual Meeting of the Kinki Branches of the Japanese Society of Fisheries Science: 1 presentation The 7th Meeting of the Japanese Society of Marine Biotechnology: 1 presentation The 20th Annual Meeting of the Japanese Society of Microbial Ecology: 6 presentations The 77th Annual Meeting of the Japanese Biochemistry Society: 1 presentation The 27th Annual Meeting of the Molecular Biology Society of Japan: 1 presentation

A-3. Off-campus activities Membership in academic societies (roles) Sako, Yoshihiko: Japanese Society of Fisheries Science (Referee, Trustee in Kinki Branch, Publication committee Member, Member of Symposium Organizing Committee, Award Committee Member), Japanese Society of Microbial Ecology (Trustee, Editor, Award Committee Member), Japan Society for Archaea (Executive Committee Member), Japanese Society of Marine Biotechnology(Trustee) Yoshinaga, Ikuo: Japanese Society of Microbial Ecology (Auditor) Nomura, Norimichi: Japanese Society of Fisheries Science (Member of Organizing Committee of the 2004 Joint Meeting of the Chugoku-Shikoku and Kinki Branches) Sako, Yoshihiko: Monbukagakusho Grants-in-Aid for Scientific Research (A)(2), Massive collection of metagenomes occurring at deep-sea hydrothermal environments and their application to industrial processes (Head: Sako, Collaborator: Nomura); Monbukagakusho Grants-in-Aid for Exploratory Research, Development of new genes encoding restriction enzymes in marine thermophilic archaeal genomes (Head: Sako, Collaborator: Nomura), Monbukagakusho Grants-in-Aid for Scientific Research (B)(2), Structural biological studies of target DNA recognitions by endonucleases (Head: Keiko Kita [Div. of Applied Life Sciences], Collaborator: Sako), Entrust Research Grant by Ministry of Agriculture, Forestry and Fisheries: Technical development in counterplanning for protection of shellfish poisoning damage (Head: Sako), Technical development in counterplanning for protection of red tide damage (Head: Sako), National Project on Protein Structural and Functional Analyses, Metabolic Enzymes (Head: Toshihisa Ohshima [Univ. Tokushima], Collaborator: Sako); COE for Microbial-Process Development Pioneering Future Production Systems (Head: Sakayu Shimizu [Div. of Applied Life Sciences], Collaborator: Sako) Nomura, Norimichi: Monbukagakusho Grants-in-Aid for Scientific Research on Priority Areas (“Genome Biology”), Hrizontal transfer and evolution of mobile introns spersely distributed in archaeal genomes (Head: Nomura); Monbukagakusho Grants-in-Aid for Encouragement of Young Scientist (B), Molecular design of artificial rare-cutting DNA

266 endonucleases (Head: Nomura); Monbukagakusho Grants-in-Aid for Scientific Research (A)(2), Massive collection of metagenomes occurring at deep-sea hydrothermal environments and their application to industrial processes (Head: Sako, Collaborator: Nomura); Monbukagakusho Grants-in-Aid for Exploratory Research, Development of new genes encoding restriction enzymes in marine thermophilic archaeal genomes (Head: Sako, Collaborator: Nomura), Monbukagakusho Grants-in-Aid for Scientific Research (B)(2), Structural biological studies of target DNA recognitions by endonucleases (Head: Keiko Kita [Div. of Applied Life Sciences], Collaborator: Nomura), National Project on Protein Structural and Functional Anelyses, Metabolic Enzymes (Head: Toshihisa Ohshima [Univ. Tokushima], Collaborator: Nomura) Yoshinaga, Ikuo: Monbukagakusho Grants-in-Aid for Scientific Research (B)(2) Protection against red tide by mixed culturing of fish and macroalgae attached with algicidal bacteria (Head: Ichiro Imai [Div. of Applied Biosciences], Collaborator: Yoshinaga), Research Grant by Steel Industry Foundation for the Advancement of Environmental Protection

Technology: Analytical research for physical and chemical factors effect on N2O

concentration in seawater –Quantification of N2O gas exchange between marine and atmosphere- (Head: Hiroshi Ishida [Kobe Univ.], Collaborator: Yoshinaga) Tanaka, Reiji: Monbusho Grants-in-Aid for for JSPS Fellows, Development of genetic manipulation technologies of marine hyperthermophilic archaea toward the genome-wide functional analyses (Head: Tanaka) Nakagawa, Satoshi: Monbukagakusho Grants-in-Aid for JSPS Fellows, Use of specific metabolism of new thermophilic microbes to environmental repair and geochemical studies on specific environmental ecology (Head: Nakagawa)

A-4. International cooperations and overseas activities International meetings (role) Nomura, Norimichi: The 5th New England Biolabs Meeting on Restriction/Modification, Bristol, UK (presentation), The 1st Pacific-Rim International Conference of Protein Science, Yokohama (presentation) Overseas research surveys Sako, Yoshihiko: Development of genetic diagnosis for toxic microalgae (USA, Korea), Search for extremophilic microorganisms at sub-seafloor biotopes in cooperation with IODP (USA), Research surveys of extremophilic microorganisms in hot springs at Yellowstone National Park (USA) Editorial work for international jounals (role) Nomura, Norimichi: Archaea, Heron Publishing, British Columbia, Canada (Editorial Review Board)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Marine Microbiology I (Sako), Marine Microbiology II (Sako), Seminar of Marine Microbiology (Sako), Fundamentals for the experiments of Bioresource Science (Sako, Yoshinaga, Nomura), Biotechnology - New Strategies in Agriculture- (Sako), Basic

267 Bioresource Science I (Sako, sharer) Graduate level: Current Topics in Marine Microbiology (Sako), Seminar in Marine Microbiology (Sako), Laboratory Course in Marine Microbiology (Sako)

B-2. Off-campus teaching, etc. Part-time lecturer Sako, Yoshihiko: Mie University Faculty of Bioresources (Marine microbial physiology), Nishinomiya High School (Specially Science Class) Yoshinaga, Ikuo: Kawaijuku Educational Institution (Special lecture from Kyoto University), Nishiyamato-High School (Super Science High School Program, Special lecture) Open classes Sako, Yoshihiko: Open Class of Nishinomiya City

C. Other Remarks Committees Committees in the Faculty Sako, Yoshihiko: Member of Research Promotion Committee of Research activities Committees out of the University Sako, Yoshihiko: Special Committee Member of Project on the Deep-sea Research in Japan Marine Science and Technology Center

2.4.14 Laboratory of Marine Environmental Microbiology

Staff Professor : Nakahara, Hiroyuki, Dr. Sci. Associate Professor : Imai, Ichiro, Dr. Agric. Sci Assistant Professor : Ajisaka, Tetsuro, Dr. Agric. Sci. Students and research fellows Doctor's program : (7) Master's program : (6) Undergraduate : (3) Research fellow : (1) Research student : (1)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Effects of eutrophication and loading of pollutants on fishery resources. The exponential expansion of human development, industrialization and exploitation of various resources has resulted in an undesirable nutrient enrichment and contamination of aquatic environment. The loadings from municipal, industrial urban and agricultural pollutants are a serious threat to the conservation of the global aquatic ecosystem and fisheries resources. So, we are studying the effects of nutrient enrichment, heavy metals and agricultural chemicals

268 on the aquatic primary producers, phytoplankton and benthic macroalgae, in laboratory and field conditions and in single species and community levels. b) Ecophysiological studies on red tides and toxic algal blooms in coastal sea . Red tide incidents with fishery damage and toxic algal blooms causing poisonings of bivalves have recently increased in frequency in the coastal seas of temperate and tropical areas. To understand the mechanisms of red tide and toxic algal bloom occurrences, the life history and physiological ecology of causative organisms are studied especially on toxic and fish-killing species. And further, algicidal bacteria are investigated for the trial for exterminating the red tides of harmful flagellates such as Chattonella spp. and Heterosigma akashiwo (Raphidophyceae). Diatom blooms occasionally cause fisheries damage to Nori culture in coastal sea such as Ariake Sea. The mechanisms for diatom bloom occurrences are investigated with reference to resting stage cell formation, vegetative growth and turbidity in the sea. c) Taxonomical study of Sargassum (marine algae) Populations variations of Sargassum plants from East Asia area (Japan, China and Korea) are studying from DNA analysis (co-operation with other scientists) and taxonomical/statistical analysis. Main target plants are Sargassum patens, S. piluliferum, S. horneri and S. filicinum, which are very common in the coasts. On the other hand, subgenus Sargassum those are growing in southern part of Japan, Southeastern Asia and Pacific countries, are studying by the taxonomical analysis. d) Multilateral studies of freshwater green algae in the Mekong Freshwater green algae (Cladophora glomerata and Spirogyra spp.) are used for food in the vicinity of the Mekong, South-east Asia. Multilateral studies on these algae is now studying in Laos.

A-2. Publications and presentations a) Publications Books Nakahara, H.: Handbook for marine environmental learning, NPO Institute of Environmental Restoration/Creation on Osaka Bay Coastal Zone, Osaka, 2004 (in Japanese) Nakahara, H.: Restration techniques using marine organisms. In “Studies of Environmental Symbiosis between marine and land” (Yamaguchi K. et al. eds). pp.155-178, Osaka University Press, Osaka, 2004 (in Japanese) Murata, B., H. Ueshina and H. Nakahara: Technology for the Regeneration of coastal Environment. In “Marine environment and human life” (Maeda H, and Kada R. eds). pp.173-191, Taga-shuppan, Tokyo, 2004(in Japanese) Imai, I.: Red tides. In “A Handbook on Fisheries Science” (Takeuchi, T., H. Nakata, T. Wada, H. Ueda, T. Arimoto, S. Watabe, A. Nakamae eds.). pp.53-58, Seibutu-Kenkyusha, Tokyo, 2004 (in Japanese) Ajisaka, T. : Seaweed bed, A Handbook on Fisheries Science(edited by Takeuchi et al. ). pp.76-78, Seibutsu Kenkyu-sha, Tokyo, 2004(in Japanese) Original papers Yamamoto, Y. and H. Nakahara: The formation and degradation of cyanobacterium Aphanizomenon flos-aquae blooms: the importance of pH, water temperature, and day length. Limnology 6; 1-6, 2005

269 Maki, T., I. Yoshinaga, N. Katanozaka and I. Imai: Phylogenetic analysis of the intracellular bacteria of a harmful marine microalga, Heterocapsa circularisquama (Dinophyceae). Aquatic Microbial Ecology 36; 123-135, 2004 Naito, K., M. Suzuki, M. Matsui and I. Imai: Secretion of iron-complexing ligands from Closterium aciculare (Charophyceae, Chlorophyta) under iron-deficient conditions. Phycologia 43; 632-634, 2004 Imai, I., M. Hatano and K. Naito: Development of a chemically defined artificial medium for marine red tide-causing raphidophycean flagellates. Plankton Biology & Ecology 51; 95-102, 2004 Naito, K., M. Matsui and I. Imai: Effects of organic iron complexes on the growth of red tide causative phytoplankton. Oceans’04 MTS/IEEE TECHNO-OCEAN’04 Conference Proceedings; 1774-1780, 2004 Naito, K., M. Matsui and I. Imai: Influence of iron chelation with organic ligands on the growth of red tide phytoplankton. Plankton Biology & Ecology 52; 14-26, 2005 Ajisaka, T. : Nutrient analysis of Cladophora glomerata from the Yahagi River and Cladophora spp. from the Mekong River. Rep. Yahagi River Inst. 8; 75-84, 2004 (in Japanese) Reviews Komatsu, T., K. Tatsukawa and T. Ajisaka: Distribution and ecology of drifting seaweeds. Kaiyo Monthly 36; 429-430, 2004(in Japanese) Ajisaka, T., S. Arai, Y. Ishihi, S. Uwai and K. Kogame: Study on the morphological variations in Sargassum (Brown algae) - especially on the vesicles - . Kaiyo Monthly 36; 779-783, 2004 (in Japanese) Reports Imai, I., G. Nishitani and Y. Matsuyama : Search for microorganisms responsible for diarrhetic shellfish poisoning and development of feasible monitoring. Report of Counterplanning for Protection of Shellfish Poisoning Damage (Ministry of Agriculture, Forest, and Fisheries); p.1-18, 2004 (in Japanese). Imai, I. and T. Shiraishi : Development of new techniques for monitorings of the harmful dinoflagellate Heterocapsa circularisquama by using of monoclonal antibodies. Report of Counterplanning for Fisheries Damages by Heterocapsa Red Tides (Fisheries Agency); p.41-59, 2004 (in Japanese). Imai, I., T. Watanabe and T. Ishida : Diatom red tides and the distribution of diatom resting stage cells in sediments of the Ariake Sea. Report of Counterplanning for Fisheries Damages by Diatom Red Tides (Fisheries Agency); p.8-18, 2004 (in Japanese). Imai, I. and K. Fukami : Report on the joint meeting 2004 of two branches of Chugokushikoku and Kinki in Okayama. Nippon Suisan Gakkaishi 70 (1); 86-90, 2005 (in Japanese). Imai, I., S. Itakura, Y. Watanabe, A. Ishikawa and Y. Fukuyo : Activity of Japanese delegates in the Harmful Algal Bloom section of PICES (North Pacific Marine Science Organization). Bulletin of the Plankton Society of Japan 52 (1); 50-53, 2005 (in Japanese). Ajisaka, T. and I. Wakana: Research of Cladophora sp. from the Mekong River, Laos A Transdisciplinary study on the regional eco-history in tropical Monsoon Asia: 1945-2005. Res. Inst. Humanity and Nature, pp.231-237, 2004 (in Japanese). Ajisaka, T. : Problems of the taxonomical study of Sargassum in the Southeast Asian regions, and a proposal of procedure for the identification. Proc. JSPS Coordinator Conference in

270 Chiang Mai, Thailand. JSPS. pp. 173-177, 2004 b) Conference and seminar papers presented 2004 Meeting of Oceanographic Society of Japan: 1 presentation The 29th Annual Meeting of Phycological Society of Japan: 5 presentations 2004 Meeting of the Japanese Society of Scientific Fisheries: 1 presentation 2004 Joint Meeting of Two Branches of Chugokushikoku and Kinki of Japanese Society of Scientific Fisheries: 4 presentations 2004 Joint Meeting of the Plankton Society and the Benthos Society: 4 presentations North Pacific Marine Science Organization 13th Annual Meeting: 2 presentations 11th International Conference on Harmful Algal Blooms (Cape Town 2004): 3 presentations First Korea-Japan Joint Symposium on Limnology (Pusan): 1 presentation The 29th Congress of the International Association of Theoretical and Applied Limnology SIL (Lahti, Finland): 2 presentations

A-3. Off-campus activities Membership in academic societies Nakahara, H.: The Japanese Society of Scientific Fisheries (Council Member), The Japanese Society of Phycology (Council Member), Imai, I.: The Japanese Society of Scientific Fisheries (Vice-Chairman of Committee of Protecting Fisheries Environment, Council Member of Kinki Branch), The Plankton Society of Japan (Editor in Chief of English Journal, Council Member), The Japanese Society of Phycology (Council Member, Member of Editorial Board), The Japanese Society of Fisheries Oceanography (Council Member) Ajisaka, T. : Japanese Society of Phycology (Council Member) Research grants Monbusho Research Grant: Scientific Research (B) (2), Comparative analysis of algicidal bacteria in red tide areas and seaweed beds, and application of algicidal bacteria in seaweed beds to prevention of harmful red tides (Imai, I. and H. Nakahara). Overseas Research (A): Distribution and ecological study of Sargassum beds in China and its contribution for drifting seaweeds (Ajisaka, T.) Entrust Research Fund by Ministry of Agriculture, Forestry and Fisheries: Technical development in counterplanning for protection of shellfish poisoning damage (Imai, I.). Urgent counterplanning for diatom red tides (Imai, I.). Urgent counterplanning for Cochlodinium polykrikoides red tides (Imai, I.) Fund by Salt Science: Mechanisms of iron uptake in the growth of red tide organisms (Naito, K. and I. Imai)

A-4. International cooperation and overseas activities International meetings Nakahara, H: First Korea-Japan Joint Symposium on Limnology, Pusan, Korea (presentation); The 29th Congress of the International Association of Theoretical and Applied Limnology, Lahti, Finland (presentaton) Imai, I.: North Pacific Marine Science Organization 13th Annual Meeting, Honolulu, U.S.A. (Delegate of Japanese committee members in Harmful Algal Bloom section,

271 Presentation); 11th International Conference on Harmful Algal Blooms (Cape Town 2004), Cape Town, South Africa (presentation) Ajisaka, T.: JSPS Multilateral Cooperative Research Program: Coastal Oceanography Regional Coordinators’ Meeting, Palawan, Philippines, 2-3 December 2004.(presentation) International Joint Researches, Overseas Research Surveys Ajisaka, T.: Biodiversity in Southeast Asian coastal area (Philippines), Distribution and ecological study of Sargassum beds in China and its contribution for drifting seaweeds (China), A Trans-Disciplinary Study on the Regional Eco-History in Tropical Monsoon Asia (Laos, Thailand), Utilization of Seaweed (Korea)

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Aquatic Microbial Ecology I (Nakahara), Aquatic Microbial Ecology (Imai), Importance of Global Environmental Study (Imai, Partaker), Practical Course in Marine Bioscience and Technology (Nakahara, Imai, partaker), Seminar in Marine Microbiology (Nakahara, Imai, partaker), Laboratory Course in Bioresource Science I, II (Imai, partaker), Marine Practice I, II (Imai, Ajisaka, partaker), Marine Practice III (Faculty of Science) (Ajisaka, partaker) Graduate level: Aquatic Environmental Microbiology (Nakahara), Aquatic Microbial Ecology (Imai), Seminar of Aquatic Environmental Microbiology (Nakahara, Imai and Ajisaka), Laboratory Course of Aquatic Environmental Microbiology (Nakahara, Imai), Seminar in Environmental Management (Summer Practice) (Nakahara, Imai,, Ajisaka, partaker)

B-2. Off-campus teaching, etc. Part-time lecturer Ajisaka, T.: Mariculture of seaweeds, Faculty of Agriculture, Kinki University; Marine Practice, Faculty of Science, Nara Women’s University

B-3. Overseas teaching Ajisaka, T.: Ocean Research Institute of University of Philippines (Lecture), NAFRI Laos (Lecture) Students and research fellows from abroad Research Student 1 (Korea)

C. Other remarks Nakahara, H.: Kyoto Prefectural commitee for adjustment of inland fisheries Imai, I.: Fisheries Agency of Japan (Committee member), Ministry of Environments (Committee member), North Pacific Marine Science Organization (Delegate of Japanese Committee Member of Harmful Algal Bloom section)

272 Chair of Marine Bio-production

2.4.15 Laboratory of Technology of Marine Bio-products

Staff Professor : Hirata,Takashi, Dr.Agric.Sci. Associate Professor : Sugawara, Tatsuya, Dr.Agric.Sci. Assistant Professor : Kinoshita,Masato, Dr.Agric.Sci. Number of Students and research fellows Doctor's program : (2) Master's program : (6) Undergraduate : (4) Research fellow : (1)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Studies on functional components extracted from oceanic lives Oceanic life has adapted itself to the specific environment different from land. For this adaptation, various bio-active substance are incorporated into or biosynthesized in their bodies. The objectives of this work are to screen these components and to elucidate their functions for human use. The progress hitherto made is summarized as follows:(1) Heat treatment of pigment-proteins from Porphyra sp. provided a disappearance of their anti-oxidative activities. (2) Carotenoids and glycolipids from algae were studied from the perspective of their absorption, metabolism through intestinal tract, and anti-proliferative activities of the metabolites against various cell lines. (3) Pridinin from Dinophyta and sphingoid bases from Sea cucumber induced apoptosis in cancer cells. b) Studies on reguration of lipid metabolism via nuclear receptors and nuclear transcription factor. Modulation of lipid metabolism is important to decrease the risk of life-style related diseases. Recently, it has been known the mechanism of lipid metabolic gene expression through nuclear receptors. In this study, we examined the effect of structural modified EPA (trans-isomers and oxidaized forms) on lipogenetic gene expression via liver-X receptor (LXR) and sterol regulatory element binding protein-1c (SREBP-1c) in HepG2 human hepatoma cells. Our results show that trans-isomers of EPA decrease the expression of lipogenic gene as well as its intact from. Noteworthy, oxidized EPA decreased more potently the lipogenic gene expression than EPA. c) Studies on post-harvest development of melanogenesis cascade in crustaceans. Invertebrates including crustaceans lack acquired immunity system. And a phenoloxidase system, which is non-inducibly present, is one candidate for recognition of foreign materials. However, this cascade system is often triggered during post-mortem storage, resulting in black spot development and seriously reducing the market quality of crustaceans. So far we confirmed that hemocyanin-derived phenoloxidase activity plays the major role in the black spot development, especially in fronzen-thawed prawns. We also showed that dopachrome induced by the action of hemocyanin-derived phenoloxidase is further converted into melanin precursors by a

273 newly prepared converting-related enzyme. We also found that hemocyanin is present in the exoskeleton of kuruma prawn. d) Production of transgenic fish and regulation of expression of transgene Transgenic technique is one of the most powerful tools for the investigation of gene function. In this study, we used medaka as a model. I optimized the gene transfer conditions using microinjection method and particle delivery method with gene gun. And I also developed some vector cassettes, which regulate tissue specific gene expression (in muscle, nerve system, and liver). I also adopt Cre-LoxP system and IRES (Internal Ribosome Entry Site) system in medaka transgenesis. I also tried to develop gene transfer method into red sea bream, which is commercially important seawater fish in Japan. e) Development of transgenic medaka for the detection of endocrine disrupting substances. Recently, endocrine-disrupting substances in environmental water became big problem. In this study, we tried to produce transgenic medaka strains that told us the existence of estrogen-like substances in water. Using the regulatory region of choriogenin H gene, we successfully produced transgenic medaka strain that could detect estrogen at ppb order. We also established a unique transgenic medaka line which has GFP labeled oocyte using oocyte specific gene promoter.

A-2. Publications and presentations a) Publications Books and Review Song, X., T. Hirata, T. Kawai, N. Niizeki and M. Sakaguchi:Volatile compounds in the hepatic muscular tissue of common carp, Japanese flounder, Spanish mackerel and skipjak. In More Efficient Utilization of Fish and Fisheries Products (M. Sakaguchi, eds.). pp209-222, 2004 Adachi, K., T. Hirata, K. Nagai, A. Fujio and M. Sakaguchi: Hemocyanin-related reactions induce blackening of freeze-thawed prawns during storage. In More Efficient Utilization of Fish and Fisheries Products (M. Sakaguchi, eds.).pp317-330, 2004 Wang, X., T. Hirata, Y. Fukuda, M. Kinoshita and M. Sakaguchi: Aceptability and its improvement of kamaboko gel derived from silvercarp surimi. In More Efficient Utilization of Fish and Fisheries Products (M. Sakaguchi, eds.). pp387-394, 2004 Original papers Adachi, K., H. Endo, T. Watanabe, T. Nishioka and T. Hirata: Hemocyanin in the exoskeleton of crustaceans:enzymatic properties and immunolocalization. Pigment Cell Research 18; 136-143, 2005 Aida, K., M. Kinoshita, T. Sugawara, J. Ono, T. Miyazawa and M. Ohnishi: Apoptosis inducement by plant and fungus sphingoid bases in human colon cancer cells. Journal of Oleo Science 53; 503-510, 2004 Sugawara, T., M. Kinoshita, M. Ohnishi, T. Tsuzuki, T. Miyazawa, J. Nagata, T. Hirata and M. Saito: Efflux of sphingoid bases by P-glycoprotein in human intestinal Caco-2 cells. Bioscience Biotechnolgy and Biochemistry 68; 2541-2546, 2004 Aida, K., M. Kinoshita, M. Tanji, T. Sugawara, M. Tamura, J. Ono, N. Ueno and M. Ohnishi: Prevention of aberrant crypt foci formation by dietary maize and yeast cerebrosides in 1,2-dimethylhydradine-treated mice. Journal of Oleo Science 54; 45-49, 2005

274 Matsubara, K., C. Xue, X. Zhao, M. Mori, T. Sugawara and T. Hirata: Effects of middle molecular weight fucoidans on in vitro and ex vivo angiogenesis of endothelial cells. International Journal of Molecular Medicine 15; 695-699, 2005 Kinoshita M.: Transgenic medaka with brilliant fluorescence in skeletal muscle under normal light. Fisheries Sci. 70; 645-649, 2004 Ohtsuka M, N. Kikuchi, H. Yokoi, M. Kinoshita, Y. Wakamatsu, K. Ozato, H. Takeda, H. Inoko and M. Kimura: Possible roles of zic1 and zic4, identified within the medaka Double anal fin (Da) locus, in dorsoventral patterning of the trunk-tail region (related to phenotypes of the Da mutant). Mech. Dev. 121; 873-882, 2004 Tagawa M, T. Kaji, M. Kinoshita and M. Tanaka: Effect of stocking density and addition of proteins on larval survival in Japanese flounder. Paralichthys olivaceus. Aquaculture 230; 517-525, 2004 Reports Sugawara, T.: Study of Chemical Structures and Biological Functions on Glycolipids in Algae. Touwa Shokuhin Kenkyu Shinkoukai, p.43-49, 2004 b) Conference and seminar presented Annual meeting of the Society of Scientific Fisheries: 2 presentations Annual Meeting of the Japan Society for Bioscience, Biotechnology, and Agrochemistry, 2005:4 presentations The 43rd Annual Meeting of the Japan Oil Chemists’ Society: 1 presentation The 38th Annual Scientific Meeting and Symposium of the Tohoku Local Section of Japanese Society of Nutrition and Food Science: 1 presentation The 77th Annual meeting of the Japanese Biochemical Society: 1 presentation The 7th Annual meeting of Japan Society of Endocrine Disrupters Research: 1 presentation

A-3. Off-campus activities Membership in academic societies (role) Hirata,Takashi: The Japanese Society of Fisheries Science (Symposium planning committee, Fisheries science education committee), World fisheries congress (Program committee), The Society of Packaging Science and Technology (vice-President, Editor), The Japanese Society for Food Science and Technology (Councilor, Auditor) Research grants Monbusho Research Grant: Scientific Research; Studies on the novel pathway of melanogenesis in crustaceans and its postharvest control (Hirata, Sugawara, Kinoshita), Scientific Research; Visualization of water pollutants using transgenic medaka (Kinoshita), Creative Scientific Research; Establishment of Neuroglycobiology (Glycobiological approach for Neuroscience) (Kinoshita), Iwate Prefecture: Assessment and conservation of environmental water in Sanriku area (Kinoshita)

A-4. International cooperations and overseas activities International meetings (roles) Hirata, T.：The Ninth International Symposium on the Effcient Application and Preservation of Marine Biological Resources, Qingdao, China (Organizer, Chairperson, Speaker) Sugawara, T. : The Ninth International Symposium on the Effcient Application and Preservation

275 of Marine Biological Resources at Qingdao, China （Speaker） International Activity Kinoshita, M.: External examiner of a thesis (Ph.D) of National University of Singapore

B. Educational Activities (2004.4-2005.3) B-1. On-campus teaching a) Courses given Undergraduate level: Outline of Bioresource Science II (Hirata), Fundamentals for the experiments of Bioresource Science (Sugawara), Technology of Marine Bioproducts I (Hirata), Technology of Marine Bioproducts II (Hirata), Introduction to Foreign Literature in Bioresource Science III (Hirata), Seminar in Marine Bio-production (Hirata, Sugawara), Laboratory in Bioresource Science I, II (Hirata, Sugawara), Practical Course in Marine Bioscience and Technology III (Hirata), Technology of Marine Biological System (Sugawara) Graduate level: Laboratory Course in Technology of Aquatic Biological Products (Hirata, Sugawara), Seminar in Technology of Aquatic Biological Products (Hirata, Sugawara), Technology of Aquatic Biological Products (Hirata), Biochemistry of Aquatic Animals (Sugawara)

B-2. Off-campus teaching, etc. Part-time lecture Hirata,Takashi: Fukui Prefectural University Hirata,Takashi: Kochi University Kinoshita, Masato: Kyoto Prefectural University of Medicine Open seminar Hirata,Takashi: Packaging manager course (Lecturer) Kinoshtia, Masato: Lecture for high-school students (Lecturer)

C. Other Remarks Hirata, Takashi: Guest Researcher (Kagawa prefecture) Kinoshita, Masato: Guest Lecturer (Nagoya City Science Museum)

276 2.4.16 Laboratory of Marine Biological Function

Staff Professor : Hayashi, Isao, Dr. Agric. Sci. Associate Professor : Toyohara, Haruhiko, Dr. Agric. Sci. Students and research fellows Doctor's program: (4) Master's program: (6) Undergraduate : (4)

A. Research Activities (2004.4-2005.3) A-1. Main subjects a) Taxonomical and ecological studies on benthic organisms A new modified molecular biological method which has been newly established for identification of planktonic bivalve larvae has been published and got the 2004 Award of Excellence for Scientific Papers for the Japanese Society of Fisheries Science. Using this method some new information has been accumulated on the early life stage of one of hard clams from Kashima-nada collaborated with the National Research Institute of Fisheries Engineering. Taxonomic and zoogeographical studies have been carried on buccinid gastropods in the offshore deep bottom in the Sea of Japan collaborated with various fisheries research stations located along the coast of the Sea of Japan. DNA analysis has given some indications for solving serious taxonomical problems of these taxa argued for a couple of decades. It has been newly indicated that one of deep-sea whelks Buccinum tsubai in the Sea of Japan includes 4 local genetic populations separated genetically each other. This conclusion has been published. To obtain some basic information on the conservation of coral reef populations, an experimental ecological study has been taken, mainly focusing on recruiting process of coral larvae at Ishigaki Experimental Station of Seikai National Fisheries Research Institute, Ishigaki, Okinawa Prefecture. Some information has been obtained on the ecological responses of larval stage of Acropora sp. coral to various environmental factors. A part of the reslts was presented at the Annual Meeting of the Japanese Association of the Coral Reef Scientists held in Tokyo. b) Taxonomic and ecological studies on the polychaetes from Japan and adjacent seas. Polychaete samples obtained from Wakasa Bay and adjacent seas from 1971 through 1991 have been reexamined to make a precise faunal list of this bay area for these several years. The species belonging to the families Paraonidae, Maldanidae, Glyceridae and Goniadidae have been reexamined in this academic year. The result on Paraonidae was presented at the Annual Meeting of the Japanese Association of Behtology, Matsuyama. c) Studies on physiological function of marine invertebrates Molecular mechanism of heat tolerance, hypoxic tolerance, osmotic tolerance and heavy metal tolerance of marine invertebrates has been studied in our laboratory. Function of HSP 70 implicating in heat tolerance, transporters for amino acids and taurine implicating in osmotic tolerance, transporters for divalent metal ions implicating in heavy metal ions were investigated by using marine bivalves. Biomineralization mechanism including in shell formation was clarified especially focusing on calcium ion transporter to utilize these information to develop novel

277 architectural materials and adhering cement reagents. G-protein related genes implicating in the signal transduction of chemical sensor of marine organisms were also identified by using spiny lobster.

A-2. Publications and presentations a) Publications books Hayashi, I. and S. Nakao: Editor. Benthos and Fisheries, Koseisha-Koseikaku, 2005 Original papers Takagi, M., T. Kikko, M. Hosoi, I. Hayashi and H. Toyohara: cDNA cloning of oyster matrix metalloproteinase and its possible involvement in hypoxic adaptation. Fish. Sci. 70; 682-687, 2004 Hosoi, M., S. Hosoi-Tanabe, H. Toyohara, M. Ueno and I. Hayashi: Sequence and RFLP analysis of the large subunit rRNA gene for identification of multiple species of bivalve larva. Fish. Sci. 70; 629-637, 2004 Toyohara, H. and M. Hosoi: The role of taurine in the osmotic adaptation in the marine mussel Mytilus galloprovincialis. Marine Biotechnol. 6; S511-S516, 2004 Toyohara, H., M. Takagi, M. Hosoi, M. Kinoshita, I. Hirono, T. Aoki and S. Kubota: Establishment of a transgenic medaka line expressing Japanese flounder tissue inhibitor of metalloproteinase for the suppression of post-mortem meat tenderization. Mar. Biotechnol. 6; S413-417, 2004 Toyohara, H., S. Yamamoto, M. Hosoi, M. Takagi, I. Hayashi, K. Nakao and S. Kaneko: Scallop DMT functions as a Ca2+ transporter. FEBS Lett. 579; 2727-2730, 2005 Iguchi, A., M. Ueno, T. Maeda and I. Hayashi: Genetic population of the deep-sea whelk Buccinum tsubai in the Japan sea. Fish. Sci. 70; 569-572, 2004 Reports Toyohara, H: 2004 Annual report on the Bio-Design Project by the Ministry of Agriculture, Forestry and Fisheries, Japan. 2004 Annual report to Nippon Suisan Fund b) Conference and seminar papers presented The 2004 Annual Meeting of the Japanese Society of Fisheries Sci: 2 presentations The 2004 Annual Meeting of the Japanese Association of Benthology: 2 presentations The 2004 Annual Meeting of Marine Biotechnology Conference: 2 presentations The 2004 Annual Meeting of Asia-Pacific Marine Biotechnology Conference: 2 presentations 4th Enzymatic Chemistry on Food Conference: 1 presentation 7th Coral Conference: 1 presentation

A-3. Off-campus activities Membership in academic societies (roles) Hayashi, Isao: Japanese Society of Fisheries Science (Trustee, Trustee in Kinki Branch, Member of Symposium Organizing Committee) Toyohara, Haruhiko: Japanese Society of Fisheries Science (Editor), Japanese Society for Traditional Food Science (Secretary) Research grants Toyohara, Haruhiko: Grant-in-Aid for Scientific Research (B)(2), Studies on fish muscle

278 tenderization-analysis of transgenic red sea bream (Head: Toyohara); Grant-in-Aid for Scientific Research (promotion Research), Studies on biomineralization mechanism focusing on bivalve mantle (Head: Toyohara); The Bio-Design project from the ministry of Agriculture, Forestry and Fisheries, Japan; Research aid from Nippon Suisan Kaisha (Head: Toyohara); .

A-4. International cooperations and overseas activities Toyohara, Haruhiko: Asia Pacific Biotechnology Conference invited speaker.

B. Education activities (2004.4-2005.3） B-1. On-campus teaching a) Courses given Undergraduate level: Science of Biosphere-Life, Food and Environment (Hayashi), Biotechnology-New Strategy in Agriculture- (Toyohara), Basic Bioresource Science II (Hayashi), Outline of Bioresource Science III (Hayashi, Toyohara), Marine Zoology (Hayashi), Molecular Biology (Toyohara), Practical Course in Marine Bioscience and Technology I (Toyohara), Cell Technology of Marine Organisms (Toyohara), Laboratory Course in Bioresource Science I and II (Toyohara), Introduction to Foreign Literature in Bioresource Science III (Toyohara), Seminar in Marine Bio-production (Hayashi, Toyohara) Graduate level: Ecological Function of Marine organisms (Hayashi), Seminar in Function of Marine Organisms (Hayashi, Toyohara), Laboratory Course in Function of Marine Organisms (Hayashi, Toyohara)

B-2. Off-campus teaching, etc. Part-time lecturer Hayashi, Isao: Fukui Prefectural University (Planktology and Periphytology)

B-3. Overseas teaching Toyohara, Haruhiko: Shanghai Fisheries University.

C. Other remarks Committees in the Faculty Hayashi, Isao: A member of Fishery Adjustment Commission of Kyoto Prefectural sea area Toyohara, Haruhiko: Award of Fisheries Science of 2004, Committee on Self-Evaluation in Kyoto University, Committee on Biological Education in Kyoto University. Committee on Evaluation of Experimental Animals in Kyoto University.

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