(Apr. 2011HZ - NARO

Major research outcomes are classified into 4 categories

� Intellectual contribution

� Development of technologies

� Agricultural production

� Bioindustry

(Apr. 2011 Mar. 2012; FY 2011)

　Discovery of rules and principals; construction of inno-vative theory; and prediction of unknown phenomena and discovery. These outcomes are published as papers, which contribute to intellectual creativity.

　Contribution to key researches for innovative techno-logical developments relevant to efficiency of production and improvement in the quality corresponding to consumption needs.

　Contribution to society through production activities for agriculture, forestry and fisheries industry.

　Contribution to development of technologies bringing about direct convenience using our outcomes.

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Summary of the Third Five-year Plan (FY 2011-2015)

1-1 Conservation of genetic resources for food and agriculture and intensification of their use

1-2 Development of a robust infrastructure for genomics-based approach in agricultural research

1-2-1 Enhancing the potential of the genome sequence and resources from agriculturally important organisms

1-2-2 Bioinformatics approach for advancement of agrobiological research 1-2-3 Genomics approach for advancement of research in crop improvement 1-2-4 Genomics approach for advancement of research in livestock production 1-2-5 Structural and functional analysis of biomolecules related to agriculture

2-1 Research platforms on biological functions associated with improved productivity of agricultural crops and livestock

2-1-1 Elucidation of the mechanisms involved in biomass production, growth, differentiation, and environmental response of agricultural crops

2-1-2 Elucidation of the regulatory mechanisms involved in insect growth, development and differentiation

2-1-3 Elucidation of the molecular mechanisms involved in the development and differentiation of germ cell and stem cell of livestock

2-1-4 Elucidation of the mechanisms involved in the control of the behavior and reproduction of livestock

2-2 Research platforms on biological interactions associated with improved productivity and development of novel technologies for their applications

2-2-1 Elucidation of the mechanisms involved in plant pathogenic microbe infections and development of innovative technologies for their applications

2-2-2 Elucidation of the mechanisms involved in crop response to microbial infection and development of crop strains with multiple pathogen resistance

2-2-3 Elucidation of the mechanisms involved in plant and soil microbe symbioses 2-2-4 Elucidation of the mechanisms involved in insect pest infestation and plant

resistance to insects 2-2-5 Elucidation of insect-insect, insect-plant and insect-microbe interactions and

their applications 2-2-6 Elucidation of molecular mechanisms in animal immune systems

3-0-1 Innovation of technologies for development of genetically modified crops and intensification of their use

3-0-2 Development of novel technologies for efficient use of genetically modified silkworm

3-0-3 Development of novel technologies for efficient use of genetically modified animals

3-0-4 Development of novel technologies using biomaterials based on silk proteins 3-0-5 Elucidation of insect-specific biological functions and development of novel

technologies for their applications

Enhancement of biological resources and development of research systems for improvement of innovative crops and livestock

1

2

3

Understanding life phenomena leading to enhancement of biological functions and developing novel technologies for their applications

Development of innovative technologies based on biological functions to create new bio-industries

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Contents

Summary of the Third Five-year Plan 1 Contents 2

Research Highlights for 2011 Plant and Microbial Sciences

� Cloning and functional analysis of Eibi1 essential for the retention of leaf water in barley 4

� Database construction and data release of 24,783 barley full-length cDNAs 6

� Metabolic engineering by site-directed mutagenesis via gene targeting in plants 8

� A new cryopreservation method for vegetatively propagated plant genetic resources using aluminum cryo-plates 10

� A database of plant diseases in Japan to provide unified linkage of information for plant and microorganism genetic resources 12

� Agricultural significance of the circadian clock of rice in paddy fields 14

� A common symbiosis gene CCaMK plays a central role in symbiosis between plants and soil microbes 16

� Identification of ARL8, a host protein that is required for tomato mosaic virus multiplication 18

Insect and Animal Sciences

� Structural mechanism of juvenile hormone delivery in hemolymph by juvenile hormone-binding protein of silkworm 20

� Development of a processing technique for antimicrobial fibers using beetle defensin-derived antimicrobial peptide-polymer conjugates 22

� Identification of the gene responsible for the dimolting mutant of the silkworm Bombyx mori 24

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Intellectual contribution

Takao Komatsuda1, Guoxiong Chen2, Jian Feng Ma3, Christiane Nawrath4, Mohammad Pourkheirandish1, Akemi Tagiri1, Yin-Gang Hu5, Mohammad Sameri1, Xinrong Li2, Xin Zhao2, Yubing Liu2, Chao Li2, Xiaoying Ma2, Aidong Wang2, Sudha Nair1, Ning Wang1, Akio Miyao1, Shun Sakuma1, Naoki Yamaji3, Xiuting Zheng5, Eviatar Nevo6

1Plant Genome Research Unit, NIAS, 2Cold and Arid Regions Environmental and Engineering Institute (China), 3Okayama University, 4University of Lausanne, 5Northwest Agriculture and Forestry University

6University of Haifa

A spontaneous mutation, eibi1.b, in wild barley has a low capacity to retain leaf water, a phenotype associated with reduced cutin deposition and a thin cuticle. Map-based cloning revealed that Eibi1 encodes an HvABCG31 full size transporter. Two transposon insertion lines of rice mutated in the ortholog of HvABCG31 also were unable to restrict water loss from detached leaves.

Keywords: ABC transporter, cuticular layer, drought resistance, eibi1

Background

Drought stress is one of most severe agricultural problems affecting plant growth and crop yield. The aerial surface of land plants is protected by the cuticle, a multilayered structure composed of polyester, cutin, and wax. The naturally occurring drought-hypersensitive wild barley (Hordeum spontaneum Koch) mutant eibi1.b suffers from a particularly severe level of water loss and displays a defective cuticle. The high transpiration rate of eibi1 is not related to an abnormal stomatal density. The defective eibi1 cuticle not only enhances the plant’s sensitivity to drought stress, but also is associated with the rapid loss of water from detached leaves. eibi1 was isolated and its biological function was analyzed in this study. Results and Discussion 1. eibi1.b was isolated by a map-based approach (Fig. 1A). Eibi1 encoded an HvABCG31 full

transporter, predicted to participate in the transport of secondary metabolites. eibi1.b revealed a point mutation that resulted in a loss of function of HvABCG31. A de novo wild barley mutant named “eibi1.c” mimicked the behavior of the eibi1.b mutant in its drought-hypersensitivity and dwarfism (Fig. 1B,C).

2. A reverse genetic approach was applied to rice by taking advantage of a library of established transposon insertions. Two independent transposon Oryza sativa 17 (Tos17) events within OsABCG31 (osabcg31.b and osabcg31.c) were identified. The two mutants were both highly sensitive to drought stress.

3. Transmission electron microscopy showed that the thickness of the mutant’s cuticle was only ∼ 25% of that of the wild type in all three zones of leaves. In addition, the quantity of the major cutin monomers of wild barley was reduced to ∼ 50% in the eibi1.b mutant, suggesting that the ABC transporter gene is specifically involved in the formation of cutin and retention of water.

4. The monocot rice and the dicot Arabidopsis ABCG31 shared 92% and 71% sequence identities with EIBI1, respectively, indicating the conservation of EIBI1 in monocot and dicot plants. The green alga Volvox carteri also contains an EIBI1 homolog, but the sequence identity was 35%. One may infer that V. carteri has a cutin-like layer, although experimental evidence is not available. It may be concluded that EIBI1 is highly conserved in the evolution of land plants.

Cloning and functional analysis of Eibi1 essential for the reten-tion of leaf water in barley

(China),

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Future prospects 1. This work underlines the importance of an intact cuticle and its structural polyester in

protecting leaves from uncontrolled water loss and the crucial evolution of HvABCG31 homologs in colonization of land by plants, a major milestone in terrestrial evolution of life.

Reference 1. Chen G, Komatsuda T, Ma J.F, Nawrath C, Pourkheirandish M, Tagiri A, Hu Y-G, Sameri M, Li X,

Zhao X, Liu Y, Li C, Ma X, Wang A, Nair S, Wang N, Miyao A, Sakuma S, Yamaji N, Zheng X, Nevo E (2011) An ATPbinding cassette subfamily G full transporter is essential for the retention of leaf water in both wild barley and rice Proceedings of the National Academy of Sciences of the United States of America 108(30):12354-12359

Fig. 1. Barley drought-hypersensitive eibi1 mutant and its functional assignment

(A) Exon/intron structure of barley Eibi1. The single-nucleotide difference between the wild-type and the eibi1.b mutant sequence is indicated. The red dashed line indicates the 9-bp deletion in eibi1.c exon 10. (B) The γirradiation-induced eibi1.c mutant and its wild type at the flowering stage. (C) Water-loss test of detached leaves from eibi1.c and OUH602.

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Takashi Matsumoto1, Tsuyoshi Tanaka2, Hiroaki Sakai2, Hiroyuki Kanamori1, Kanako Kurita3, Kiyosumi Hori4, Takeshi Itoh2, Kazuhiro Sato5

1Plant Genome Research Unit, NIAS, 2Bioinformatics Research Unit, NIAS, 3Advanced Genomics Laboratory, NIAS, 4Rice Applied Genomics Research Unit, NIAS, 5Okayama University

Full-length cDNA libraries of barley, whose complete genome is yet undetermined, were created and 24,783 representative cDNAs were fully sequenced. The information about these cDNAs and their functions was released from our database.

Keywords: barley, full-length cDNA, database Background

The amount of barley production in the world is fourth largest. Barley is widely used as a food crop, a feed grain and raw material for beer. Several breeding projects to improve useful traits, such as disease resistance and vivipary, are currently underway. However, since the complete barley genome sequence is yet undetermined, large-scale studies have not been expedited so far. In this study, we aimed to collect from a barley cultivar a large amount of full-length cDNAs that reflect the functional forms of genes. Sequencing of the cDNAs will help provide comprehensive understanding of the barley genes and should ultimately lead to elucidation of the function of individual genes and agronomically important phenomena. Results and Disucussion 1. RNAs were extracted from a barley cultivar, Haruna Nijo, which was grown under

several conditions, such as cold and dry stresses and exposure to hormones. We obtained ~172,000 clones of full-length cDNAs (FLcDNAs). After removal of redundant sequences, 24,783 were selected as representatives and were sequenced (Fig. 1). Of these FLcDNAs, 85% were inferred to contain complete protein-coding regions. Comparisons with the sequences in the RefSeq and UniProtKB databases resulted in predictions of 85% of the gene functions.

2. The barley sequences obtained in this study were compared to publicly available sequences of three cultivated species (rice, maize, sorghum) and one wild relative (Brachypodium). As a result, it was found that the four cultivated species share 87% of the genes. This observation indicates that the grass species sequences are highly conserved. Nonetheless, 1,699 barley genes did not show any homology to the known genes of the other grasses, so these genes might be related to characteristics specific to barley.

3. The FLcDNAs deciphered in this study are available in the public DNA database, DDBJ. In addition, the annotation and expression data were released from our barley full-length cDNA database (http://barleyflc.dna.affrc.go.jp/hvdb/index.html) on March 17th, 2011.

Future prospects1. This study presented a comprehensive set of expressed functional genes of barley. It

is expected that comparisons with genomes and genes of other species will accelerate studies of individual gene functions and development of molecular markers, so that

Database construction and data release of 24,783 barley full-length cDNAs

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the sequence information will be used for breeding of barley in the future. 2. In general, FLcDNAs contain the complete protein-coding regions. Transformation by

introgression of FLcDNAs will help investigations of individual gene functions. 3. International collaborations of barley and wheat genome-wide sequencing efforts are

currently underway. The barley FLcDNAs presented in this study will be vital for genome annotations of barley and wheat.

References 1. Matsumoto T, Tanaka T, Sakai H, Amano N, Kanamori H, Kurita K, Kikuta A, Kamiya K,

Yamamoto M, Ikawa H, Fujii N, Hori K, Itoh T, Sato K (2011) Comprehensive sequence analysis of 24,783 barley full-length cDNAs derived from 12 clone libraries Plant Physiology 156(1):20-28

2. Mayer K.F.X, Martis M, Hedley P.E, Šimková H, Liu H, Morris J.A, Steuernagel B, Taudien S, Roessner S, Gundlach H, Kubaláková M, Suchánková P, Murat F, Felder M, Nussbaumer T, Graner A, Salse J, Endo T, Sakai H, Tanaka T, Itoh T, Sato K, Platzer M, Matsumoto T, Scholz U, Doležel J, Waugh R, Stein N (2011) Unlocking the barley genome by chromosomal and comparative genomics The Plant Cell 23(4):1249-1263

Fig. 1. Sequencing and data release of barley FLcDNAs RNAs were extracted from barley specimens, cDNAs were synthesized and full-length cDNAs were selected and cloned. Redundant sequences were discarded and 24,783 representatives were selected and sequenced. The sequences were released from our database.

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Hiroaki Saika1, Akira Oikawa2, Fumio Matsuda2, Haruko Onodera1, Kazuki Saito2,3, Seiichi Toki1,4 1Plant Genome Engineering Research Unit, NIAS, 2RIKEN Plant Science Center, 3Chiba

University, 4Yokohama City University

Using gene targeting (GT), we successfully introduced mutations that improve

enzymatic properties into an endogenous rice gene encoding a key enzyme of tryptophan biosynthesis. In mature seeds of GT plants, free tryptophan accumulated to levels up to 230-fold higher than in the original cultivar. This is the first demonstration of the application of GT-mediated targeted mutagenesis to metabolic engineering in crops.

Keywords: metabolic engineering, gene targeting, mutagenesis, tryptophan, rice

Background

Results and Discussion 1.

2.

3.

Metabolic engineering by site-directed mutagenesis via gene targeting in plants

　Structure-based protein engineering of metabolic enzymes can reveal point mutations that relieve feedback inhibition and enhance enzymatic activity. This information enables the design of modified enzymes with improved enzymatic properties. To date, however, it has not been possible to mutagenize endogenous enzymes as expected in plants. In this study, we demonstrate site-directed mutagenesis of an endogenous plant gene encoding a metabolic enzyme, thus showing that it is indeed possible to perform metabolic engineering in plants. In our gene targeting (GT) test system, we examined whether tryptophan-fortified rice can be produced by site-directed mutagenesis of the OASA2 gene that encodes a key enzyme in tryptophan biosynthesis of rice.

A GT vector containing a modified OASA2 gene encoding a key enzyme of tryptophan biosynthesis (Fig. 1) was transformed into rice (cv. Nipponbare). Cells in which GT occurred successfully were selected with a tryptophan analogue. Thus, we succeeded in the production of novel rice plants in which mutations that relieve feedback inhibition and enhance enzymatic activity were introduced simultaneously into the endogenous OASA2gene. In mature seeds of GT plants, free tryptophan accumulated to levels up to 230-fold higher than in the original cultivar (Fig. 2). Moreover, free amino acids other than tryptophan accumulated to levels up to 2- to 10-fold higher than in the original cultivar, suggesting that the whole metabolic pathway of amino acids is modified in GT plants (Fig. 3). Furthermore, in young leaves of GT plants, free tryptophan accumulated to levels up to 7-fold higher than in the original cultivar (Fig. 2), suggesting the possibility that additional tryptophan synthesized and accumulated in the leaves also translocates to seeds in GT plants. This translocation system may be a useful tool with which to accumulate specific metabolites to high levels in seeds.

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Future prospects 1.

2.

3.

Fig. 1. Modification of OASA2 gene via GT

GT vectors: a 7.0-kb fragment containing a partial OASA2 fragment with mutations (stars) that confer tryptophan insensitivity and enhance enzymatic activity was constructed. This truncated OASA2 was expected to be non-functional, thus recovery of cells showing tolerance to a tryptophan analogue is expected only after homologous recombination (HR) between the GT vector and the chromosomal.

Fig. 3. The ratio of free amino acids in seeds of GTplants to that in the original cultivar

References

Fig. 2. Free tryptophan contents in leaves and seeds of GT plants The free tryptophan content in leaves and mature seeds of GT plants is higher than in the original cultivar.

The contents of free amino acids other than tryptophan in mature seeds of GT plants are higher compared to the original cultivar. Overall, the total amount of free amino acids is >8-fold higher than in the original cultivar.

The GT plants produced in this study could be used as a breeding line to produce tryptophan-fortified rice cultivars. Metabolic enzymes involved in amino acid biosynthesis are conserved in various organ-isms and much basic information on the design of native enzymes has been accumu-lated. Thus, besides tryptophan, crops fortified with other amino acids could be produced by modification of enzymes involved in amino acid biosynthesis as designed via GT and with subsequent selection on amino acid analogues. The site-directed mutagenesis of endogenous targeted genes based on information from comparative genomics and protein engineering will be a powerful tool in the field of functional genomics and molecular breeding in plants.

1.

2.

3.

Saika H, Oikawa A, Matsuda F, Onodera H, Saito K, Toki S (2011) Application of gene targeting to designed mutation breeding of high-tryptophan rice Plant Physiology 156(3):1269-1277 Saika H, Toki S (2011) Gene targeting applied to designed-mutation breeding of high-tryptophan rice ISB News Report (12):3-6 Patent application# JP-2009-177959 (Japan)

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Development of technologies

Shin-ichi Yamamoto, Kuniaki Fukui, Takao Niino Genetic Resources Conservation Research Unit, NIAS

A new cryopreservation method using aluminum cryo-plates was developed for clonally propagated crops. In this method, shoot tips are attached to the cryo-plates and can easily be manipulated for preservation treatments, such as dehydration. The ease of handling, minimal injury to shoot tips, and rapid rates of cooling and warming resulted in high regrowth rates. More than 80% regrowth was obtained for mint and tropical mulberry germplasms. It will be possible to utilize this highly effective method for long term preservation in genebanks. Keywords: vegetatively propagated, long term preservation, vitrification, cryo-plate,

liquid nitrogen, genetic resources Background

Results and Discussion

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To achieve easier manipulation of in–vitro-produced shoot tips, we designed and ordered aluminum plates that fit inside 2-ml cryotubes (Fig. 1). We standardized the storage procedure combined with the vitrification and droplet methods (Fig. 2) and monitored regrowth after liquid nitrogen storage using mint, tropical mulberry, carnation, and Dalmatian chrysanthemum as materials. It was necessary to optimize each treatment, such as pretreatment before excision, osmoprotection, and dehydration to improve regrowth after cryopreservation. We determined the optimal conditions for the crop species listed above (see references). With the V-cryo-plate method, high regrowth rates were observed under the optimal conditions for each crop tested: 88%, 87%, 81%, 95% and 77% for mint, tropical mulberry, strawberry, carnation and Dalmatian chrysanthemum, respectively. The V-cryo-plate method has many advantages as listed below and is suitable for large scale storage in genebanks. a) Shoot tips adhere to the plates, so only the plates need to be moved thus increasing the 　efficiency of treatment. b) The injury and loss of shoot tips are dramatically reduced. c) Regrowth rates are high because of rapid cooling and warming d) Technical staff can implement this method after brief training. Until now, a total of 100 accessions of germplasm such as mint, tropical mulberry, straw-berry, mat rush and others have been successfully cryopreserved (100 buds on 10 cryo-plates for each) in the NIAS Genebank.

　Although the cryopreservation methods such as slow freezing, vitrification, and encapsulation desiccation have been developed extensively, their application has not advanced to the same extent for storage of large scale samples in the genebanks of various countries. One reason for lack of widespread use is that these methods require a high level of technical skill. In this study, we designed a new aluminum plate (cryo-plate) and developed a new cryopreservation method (V-Cryo-plate method) that allows easy manipulation of plant material for improvement of vitrification and droplet vitrification methods. As a result, we have established a cryopreservation procedure suitable for large scale storage in genebanks.

A new cryopreservation method for vegetatively propagated plant genetic resources using aluminum cryo-plates

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Future prospects 1. The cryo-plate method can be expected to facilitate and increase the use of cryopreservation for

stable and cost effective long term storage of vegetatively propagated plant genetic resources. 2. To increase the use of the cryo-plate method, it is necessary to expand it to additional crop

species, to publish a manual with inclusive application examples and to publicize its merits to potential users.

Fig. 1. An aluminium cryo-plate for cryopreservation

References 1. Yamamoto S, Rafique T, Priyantha W.S, Fukui K, Matsumoto T, Niino T (2011) Development

of a cryopreservation procedure using aluminium cryo-plates CryoLetters 32(3):256-265 2. Sekizawa K, Yamamoto S, Rafique T, Fukui K, Niino T (2011) Cryopreservation of in vitro-grown

shoot tips of carnation (Dianthus caryophyllus L.) by vitrification method using aluminium cryo-plates Plant Biotechnology 28(4):401-405

3. Yamamoto S, Fukui K, Rafique T, Khan N.I, Castillo Martinez C.R, Sekizawa K, Matsumoto T, Niino T (2012) Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminium cryo-plates Plant Genetic Resources: Characterization and Utilization 10(01):14-19

Fig. 2. Procedures for cryopreservation of tropical mulberry using the V-crio-plate method

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Masaru Takeya1, Fukuhiro Yamasaki1, Shihomi Uzuhashi2, Hiroyuki Sawada2, Toshirou Nagai2, Keisuke Tomioka2, Toyozo Sato2, Takayuki Aoki2, Makoto Kawase3 1Genetic Resources Conservation Research Unit, NIAS, 2Classification and Evaluation Research Unit, NIAS, 3Genetic Resources Center, NIAS

A database of plant diseases in Japan was constructed and is available on the NIAS website. The database contains more than 10,000 common names of plant diseases and can be searched according to disease name, host plant, or pathogen name. The disease database is linked to the NIAS genetic resource databases for plants and microorganisms by connecting the host and pathogen names, respectively.

Key words: plant disease names, genetic resources, database, genebank Background

Results and Discussion The plant disease database has been developed based on hosts, disease names, pathogens, and

references recorded in the book, “Common Names of Plant Diseases in Japan, 1st ed.” Data on pathogen categories including fungi, bacteria, and viruses were added recently. The database includes 1,957 names of host plants and 11,406 common names of plant diseases as of Jan. 16, 2012. The Web-based data retrieval system provides candidate disease names based on the search criteria of disease name, host, and pathogen. Flexible retrieval is accomplished through user support functions such as partial match options, multiple selections, and auto-complete functions of the key-word input fields (Fig. 1). Matching of pathogen and microorganism genetic resources was checked by the collation of both scientific names and coincidence of the disease host and the isolation source of a strain. Host and plant matches were established by the collation of plant names and a set of evaluation data for resis-tance to the disease. Linkage of the disease database to the genetic resources database is provided as “Related strains” and “Related hosts of plant diseases” in the window of detailed information. When the characteristics in the window of detailed information indicated pathogenicity of the strain or when pathogenicity was reported in the literature cited, the link function to the corresponding disease name in the database was prepared in the microorganism search system. These link functions may provide an easy search and distribution system of genetic resources in relation to preparing an experi-mental sample set for plant pathological research works (Fig. 1).

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　Microorganism genetic resources preserved in the NIAS Genebank are frequently used for phyto-pathology and taxonomy research because many reference strains of Japanese plant pathogens have been preserved and prepared for distribution. Many accessions of plant and microorganism genetic resources also have records of attributes related to plant diseases, such as disease resistance and pathogenicity. The recorded data are open to users through the Web-based data retrieval system. Plant disease information has been managed as individual data sets of correlated plants and microorganisms. However, these important mutual relationships between hosts and pathogens were not always reflected within the genetic resources database. Therefore, we constructed a database of plant diseases in Japan based on the book, “Common Names of Plant Diseases in Japan, 1st ed.” edited by the Phytopathological Society of Japan, to realize an information system where the plant and microorganism databases were effectively connected through the plant disease names.

A database of plant diseases in Japan to provide unified linkage of information for plant and microorganism genetic resources

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Future prospects

References 1. Sato T, Yamasaki F, Takeya M (2009) Common name database of plant diseases in Japan. Plant

protection 63(9):587-591 . (in Japanese) 2. Takeya M, Yamasaki F, Uzuhashi S, Aoki T, Sawada H, Nagai T, Tomioka K, Tomooka N, Sato T,

Kawase M (2011) NIASGBdb: NIAS Genebank databases for genetic resources and plant disease information Nucleic Acids Research 39(suppl 1):D1108-D1113

3. Database of plant diseases in Japan : http://www.gene.affrc.go.jp/databases-micro_pl_diseases_en.php

Fig. 1. Database search interface

The database is provided with a search interface that allows a user to specity the search conditions for a particular plant disease and execute a search (upper panel). The search results include links to other pathogens related to the disease (lower left) and other plant hosts of the pathogen (lower right).

1.

2.

Since the plant disease database may facilitate access to pertinent genome information such as DNA sequence data of the barcode gene regions of microorganisms linked to the genetic resource database, the database can be expected to be utilized for various research purposes. The number of external linked sites will be increased in order to enrich the information on diseases and their prevention/protection. Preparation of original images and descriptions may be considered to provide additional information on useful characteristics about the unique genetic resources preserved in the NIAS Genebank.

4. The link function to the floricultural diseases was built in relation to the website of the NARO Insti-tute of Floricultural Science to enable users to refer to photographic pictures of the diseases and their prevention/protection methods.

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Takeshi Izawa1, Motohiro Mihara1, Meenu Gupta1, Hironori Itoh1, Ritsuko Motoyama2, Atsushi J. Nagano1, Masahiro Yano3, Yoshiaki Nagamura2, Yuji Suzuki4, Amane Makino4, Yuji Sawada5, Masami Yokota Hirai5

1Functional Plant Research Unit, NIAS 2Genome Resource Unit, NIAS, 3Argogenomics Research Center, NIAS, 4Tohoku University, 5RIKEN

We first identified a new circadian clock mutant of rice using flowering-time phenotypes. We next grew the mutant in a paddy and extensively analyzed temporal dynamics of transcriptomes, temporal changes of primary and secondary metabolites, photosynthetic performance, and yield-related traits, to elucidate the agricultural significance of the circadian clock of rice in field conditions.

Keywords: field environments, rice, transcriptome, metabolome, circadian clock, flowering-time

Background


2.

3.4.

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By screening for revertants of an early flowering mutant of rice (termed se5), which is a photoreceptor-deficient mutation, we identified a new mutant (osgi) and cloned the original gene OsGIGANTEA (OsGI). We further characterized osgi as the first circadian clock mutant iden-tified in rice. It has been reported that the GIGANTEA (GI) gene, the sole OsGI ortholog in Arabidopsis thaliana, also has a circadian clock function, but the role of OsGI to maintain the circadian clock of rice is more significant than that of GI. We grew the osgi mutant plants in a paddy field in Tsukuba City. Although osgi plants flow-ered later under short-day conditions than under long-day conditions, flowering of osgi was com-parable to the wild-type (WT) cultivar, suggesting the growing conditions in the paddy field mimic the long-day conditions in the laboratory. The photosynthetic rate of osgi is the same as that of WT rice plants. osgi plants had increased sugar content in leaves, increased grain numbers per panicle and panicle numbers per plant, but did not have increased yield due to the smaller grain size of osgi. We performed field transcriptome analysis at 2-hr intervals over 24 hr for four devel-opmental stages and monitored temporal expression patterns for 27,201 genes (Fig.1). More than half of the genes were affected primarily by the circadian clock and less so by environmental changes. In addition, no critical changes in primary metabolites were observed. A significant reduction in fertility under abnormal transplanting conditions indicated that osgi was more sensitive than WT to the changes of growing conditions (Fig.2).

　Extensive evaluations of genetically cloned genes are necessary before they are used for agro-nomic improvement. Because most evaluations so far have been done in controlled and relatively simple experimental conditions in laboratories, additional tests usually must be conducted in the appropriate target environments. In this study, therefore, we examined a newly isolated mutant affecting the circadian clock of rice growing in paddy fields. We analyzed the temporal transcrip-tomes, temporal metabolomes (both primary and secondary metabolites), photosynthetic perfor-mance, and yield-related traits. As a result, we revealed how the circadian clock contributes to the agronomic establishment of rice.

Agricultural significance of the circadian clock of rice in paddy fields

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Future prospects

References 1. Izawa T, Mihara M, Suzuki Y, Gupta M, Itoh H, Nagano A.J, Motoyama R, Sawada Y, Yano M,

Yokota Hirai M, Makino A, Nagamura Y (2011) Os-GIGANTEA confers robust diurnal rhythms on the global transcriptome of rice in the field The Plant Cell 23(5):1741-1755

Time of the sampling day Time of the sampling day

Pear

son’

s C

orre

latio

n

Fig. 1. Field transcriptome analysis

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2. Itoh H, Izawa T (2011) A study of phytohormone biosynthetic gene expression using a circadian clock-related mutant in rice Plant Signaling & Behavior 6(12):1932-1936

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It may be advantageous to monitor the circadian clock in rice as a means to develop new cultivars with broader cultivation areas and tolerance to fluctuating environments. New methods to monitor the circadian clock will be required in the near future. If we can utilize some mutation to reduce the cost to maintain the circadian rhythms some increase of grain yields will be expected when we grow the rice plant under stably controlled diurnal conditions. Experimental results obtained only in laboratory studies should be augmented with as much field data as possible.

We calculated the Pearson’s correleation coefficient using all combinations of 2-hr interval microarray data (27,201 genes) and plotted the changes for one-time point data as a line. 13 lines are shown. In the wild-type, clear sine-curve lines are observed primarily as two patterns; day-type and night-type. The changes between transcriptomes were less in osgi than those in the wild-type.

Fig. 2. Fertility of wild-type and osgi mutant plants When we transplanted at the normal time in May, a high fertility was observed in osgi mutant. When we transplanted in June, the fertility was more reduced in osgi mutant than in the wild-type. **: 1% significant

Although the circadian clock of rice is involved in regulation of photosynthesis and crop yields, the osgi circadian clock mutant plants were little affected by the typical paddy field conditions tested. However, osgi was very sensitive to atypical transplanting conditions due to reduced adaptability.

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Haruko Imaizumi-Anraku1, Yoshikazu Shimoda1, Teruyuki Hayashi2, Mari Banba1, Hiroshi Kouchi1, Lu Han1, Toshimasa Yamazaki3, Rintaro Suzuki3, Naoya Takeda4, Takaki Maekawa5, Makoto Hayashi1

1Plant Symbiosis Research Unit, NIAS, 2Nara Institute of Science and Technology, 3Biomolecular Research Unit, NIAS, 4National Institute for Basic Biology, 5Ludwig-Maximilians-Universität München

The function of CCaMK was investigated in order to reveal molecular mechanisms in symbiosis between plants and soil microbes. Interaction between CCaMK and calmodulin was found to be essential for symbiosis between plants and rhizobia but not between plants and arbuscular mycorrhizal (AM) fungi. CCaMK also induces formation of a cytosolic structure, which is important for infection by AM fungi.

Keywords: Plant-microbe symbiosis, rhizobia, arbuscular mycorrhizal fungi, CCaMK,

Lotus japonicus Background


2.

3.

　Nitrogen-fixing bacteria, collectively called rhizobia, infect legume plant roots to induce formation of nodules in which bacteria fix atmospheric nitrogen to ammonia that is supplied to host plants. This allows legume plants to survive in nitrogen-poor soil envi-ronments. AM fungi, on the other hand, can establish symbiosis with roughly 80% of land plant species and thereby supply host plants with phosphorus and other nutrients through hyphae developed between host roots and the rhizosphere. These symbioses are regulated by a series of host genes called common symbiosis genes. In order to better utilize the symbio-sis between host plants and soil microbes for agriculture, we are investigating the mo-lecular mechanisms involved in symbiosis. In this regard, we identified a gene, CCaMK, that plays central roles in symbiosis, among common symbiosis genes, and investigated its function.

A periodic change of cytosolic calcium concentration, calcium spiking, has been known to be one of the physiological responses of plants to symbionts. Common sym-biosis genes can be classified by their roles as “upstream genes” that are essential for the calcium spiking, and as “downstream genes” that are not required for calcium spiking. One of the “downstream genes”, CCaMK, causes host plants to form nodule structures even in the absence of rhizobia when the gene has been mutated to a gain-of-function state. We combined gain-of-function CCaMK with loss-of-function forms of other common sym-biosis genes to clarify their roles in symbiosis. We found that “upstream genes” were only required for activation of CCaMK. CCaMK consists of a kinase domain, a calmodulin-binding domain, and a calcium-binding domain. When we conferred a CCaMK mutation such that CCaMK failed to bind calmodulin, rhizobia could not infect plants, but AM fungi could. We concluded that binding between CCaMK and calmodulin is essential for symbiosis of rhizobia (Fig. 1).

A common symbiosis gene CCaMK plays a central role in symbiosis between plants and soil microbes

－ 17－

Future prospects 1.

2.

3.

References 1. Hayashi T, Banba M, Shimoda Y, Kouchi H, Hayashi M, Imaizumi-Anraku H (2010) A dominant

function of CCaMK in intracellular accommodation of bacterial and fungal endosymbionts The Plant Journal 63(1):141-154

2. Shimoda Y, Han L, Yamazaki T, Suzuki R, Hayashi M, Imaizumi-Anraku H (2012) Rhizobial and fungal symbioses show different requirements for calmodulin binding to calcium calmodulin-dependent protein kinase in Lotus japonicus The Plant Cell 24(1):304-321

Fig. 1. Activation of CCaMK

4.

Step 1: Calcium ion binds to the calcium-binding domain. Step 2: The kinase domain is phosphorylated. At this step, mycorrhiza can infect host plants. Step 3: Calmodulin binds to the calmodulin-binding domain (CaMBD). At this step, rhizobia can infect host plants.

Fig. 2. Induction of mycorrhiza-symbiosis host gene SbtM1 expression by the common symbiosis gene CCaMKLocalization of gain-of-function CCaMK in the cell nucleus resulted in more than 100 times expression of SbtM1 (black bar), whereas exclusion of gain-of-function CCaMK from the cell nucleus resulted in almost no induction (white bar).

Given the fact that roles of CCaMK are different between the two symbioses, we investigated function of CCaMK in AM symbiosis. We found that induction of host genes essential for infection of AM fungi required nuclear localization of CCaMK (Fig. 2). This fact strongly suggests that CCaMK is a decoder of calcium spiking. Furthermore, CCaMK was essential and sufficient to develop the pre-penetration apparatus, which is important for infection by AM fungi.

The hypothesis that CCaMK is important in symbiosis has been verified by our present studies. We can now regulate host-gene expression regardless of the type of symbiont through the action of gain-of-function CCaMK. This will facilitate identification of genes that are functioning downstream of CCaMK. It is expected that AM fungi and rhizobia can be more effectively utilized in agriculture by presenting the whole picture of symbiosis genes.

3. Takeda N, Maekawa T, Hayashi M (2012) Nuclear-localized and deregulated calcium- and calmodulin-dependent protein kinase activates rhizobial and mycorrhizal responses in Lotus japonicus The Plant Cell 24(2):810-822

－ 18－


Masaki Nishikiori1, Hideyasu Okamura2, Hongyu Xiang2, Masashi Mori3, Koji Dohi3, Etsuko Katoh2, Satoshi Naito4, Tetsuo Meshi5, Masayuki Ishikawa1

1Plant-Microbe Interactions Research Unit, NIAS, 2Biomolecular Research Unit, NIAS, 3Ishikawa Prefectural University; and 4Hokkaido University, 5Division of Plant Sciences, NIAS

We identified a host protein that co-purified with the replication proteins of Tomato mosaic virus (ToMV) as ARL8. Disruption of the ARL8 genes in plants completely inhibited ToMV multiplication without affecting plant growth. ARL8 played a role in enzymatic activation of ToMV replication proteins. In addition, we revealed the three-dimensional solution structures of ARL8 by NMR spectrometry. Keywords: plant virus, replication, host factor, NMR, solution structure Background


2.

3.

4.

Future prospects 1.

　Tomato mosaic virus (ToMV) and other related viruses (collectively called ‘Tobamoviruses’) infect tomato and many other crop plants and reduce the yield and qual-ity of agricultural products. Virions of Tobamoviruses are stable and remain active in soil for many years. While strong and durable resistance genes to Tombamoviruses are known for some crops, such genes are not available for others. In this study, we aimed to identify host factors required for tobamovirus multiplication to establish new strategies to control infection of the viruses.

We identified a protein that co-purified with ToMV replication proteins from infected plant cells as ARL8 by determining partial amino acid sequences. ARL8 is a small GTP-binding protein belonging to the ARF/ARL family, whose members are known to be involved in membrane trafficking. We established lines of Arabidopsis thaliana and Nicotiana tabacum in which three ARL8 genes were disrupted. These plants grew normally, but were completely resistant to ToMV and Tobacco mosaic virus (TMV)-Cg, another Tobamovirus (Fig. 1). Cucumber mosaic virus (CMV), which belongs to a genus other than Tobamovirus, multiplied normally in these plants. Like ARL8, the host protein TOM1 is necessary for ToMV multiplication. We found that ToMV replication proteins, TOM1 and ARL8 interact with each other. ToMV replica-tion proteins gained guanylyltransferase activity when TOM1 and ARL8 were co-expressed. These results suggest that TOM1 and ARL8 are components of the ToMV replication complex and play roles in enzymatic activation of the replication proteins (Fig. 2). ARF/ARL family proteins bind GTP or GDP and function as molecular switches. We determined the three-dimensional solution structures of the GTP-bound and GDP-bound forms of ARL8, and revealed the mechanism of structural interconversion (Fig. 3). This structural interconversion may be important for ToMV RNA replication.

It is expected that resistance to Tobamoviruses can be introduced to a variety of plants by suppressing ARL8 expression.

Identification of ARL8, a host protein that is required for tomato mosaic virus multiplication

－ 19－

2.

Fig. 1. Analysis of A. thaliana arl8 mutants

TOMARL

TOMARL

Replica�on complex

References 1. Nishikiori M, Mori M, Dohi K, Okamura H, Katoh E, Naito S, Meshi T and Ishikawa M (2011) A

host small GTP-binding protein ARL8 plays crucial roles in tobamovirus RNA replication PLoS Pathogens 7(12):e1002409

2. Okamura H, Nishikiori M, Xiang H, Ishikawa M and Katoh E (2011) Interconversion of two GDP-bound conformations and their selection in an Arf-family small G protein Structure 19(7):988-998

3. Patent application# JP-2007-14197 (Japan)

The solution structures and mechanism of structural interconversion have been uncovered for ARL8. We plan to utilize this information to develop anti-tobamovirus substances that inhibit the conformational interconversion in ARL8 or the interaction between ARL8 and ToMV replication proteins.

(A) Morphology of an arl8 triple mutant (right) compared with the wild type (left). (B) Multiplication of TMV-Cg and CMV in arl8 mutants. A. thaliana has three ARL8 genes. TMV-Cg or CMV RNAs were inocu-lated onto A. thaliana plants that lack either one (a, b, c), two (ac, bc, ab) or three (abc) ARL8 genes. Total proteins were prepared from the inoculated leaves two days after inoculation, and analyzed by SDS-PAGE and immunoblottiong to detect the coat proteins. The accumulation of TMV-Cg coat protein was specifi-cally inhibited in the mutants ab and abc.

Fig. 2. The role of ARL8 and TOM1 in ToMV RNA replication ToMV replication protein is produced in an inactive form, and acquires enzymatic activity necessary for viral RNA replication through interaction with TOM1 and ARL8.

Fig. 3. Solution structures of the ARL8 protein. The structure was determined by using NMR spectrom-etry. When Mg2+ concentration is low, GDP-bound ARL8 mainly takes so-called GDP form (right). In this condition, only a low population GDP-bound ARL8 takes the GTP-like form (left). On the other hand, in the presence of Mg2+, the GTP-like conformation is stabilized and a higher population of GDP-bound ARL8 adopts this confor-mation.

－ 20－


Rintaro Suzuki1, Zui Fujimoto1, Takahiro Shiotsuki2, Wataru Tsuchiya1, Mitsuru Momma1, Akira Tase1, Mitsuhiro Miyazawa3, Toshimasa Yamazaki1

1Biomolecular Research Unit, NIAS, 2Insect Growth Regulation Research Unit, NIAS, 3Insect Mimetics Research Unit, NIAS

We investigated structural mechanism of juvenile hormone (JH) delivery by JH-binding protein (JHBP) based upon the crystal and solution structures of unliganded and JH-bound JHBP. The structural information derived from our study opens the way to structure-based design of insect growth regulators (IGRs) which inhibit complex formation between JH and JHBP, and thus disrupt JH signaling in Lepidoptera.

Keywords: juvenile hormone (JH), JH-binding protein (JHBP),

insect growth regulator (IGR), structure-based drug design (SBDD) Background

Results and Discussion: 1.

2.

3.

4.

5.

　Juvenile hormone (JH) plays crucial roles in many aspects of the insect life. All the JH actions are initiated by transport of JH in the hemolymph as a complex with an insect-specific JH-binding protein (JHBP) to target tissues. Hence, JHBP is expected to be an excellent target for safe insect growth regulators (IGRs) and insecticides. However, the lack of structural knowledge of the JHBP-JH complex as well as the unliganded JHBP prevents our understanding of the JH-binding mode and the molecular mechanism of JH delivery by JHBP. To gain such structural information required for effective development of IGRs, we determined the crystal and solution structures of the JH-bound and unliganded forms of recombinant JHBP of silkworn, Bombyx mori.

The crystal and solution structures of the JHBP-JH complexes were determined by X-ray and NMR, respectively. As shown in Fig. 1A, the JH-bound JHBP adopts a gate-closed conformation where the bound JH is completely buried inside the carrier protein and thus is protected from unfavorable nonspecific absorption and enzymatic degradation during its transport in the hemolymph. JHBP recognizes the characteristic shape and functional groups of JH with high speci-ficity by means of hydrogen-bond networks and hydrophobic contacts (Fig. 2). The func-tional relevance of the hormone-protein contacts was confirmed by JH-binding assays of 23 JHBP single-mutants. We also determined the crystal structure of the unliganded JHBP (Fig. 1B). In this struc-ture, the gate-helix resides in an open conformation which permits access of JH to the hormone binding pocket (Fig. 1B). The JH-bound JHBP opens the gate-helix to release the bound hormone likely by sens-ing reduction of the dielectric constant at the membrane surface of target cells. Our structural biology study enables us to outline the molecular mechanism of JH deliv-ery by JHBP in hemolymph from the corpora allata (CA) cells where JH is synthesized to the target cells where JH functions as shown in Fig. 3.

Structural mechanism of juvenile hormone delivery in hemolymph by juvenile hormone-binding protein of silkworm

－ 21－

Future prospects 1.

Fig. 1. The crystal structure of JHBP-JH complex (A) and unliganded JHBP (B)

Corpora allata cell membrane　　　　　　Target cell membrane

Hemolymph

CytosolCytosol

JH

C-tail N

-arm

Gate

C-tail

N-arm

Gate

C-tail

N-arm

Gate

C-tail

N-armGate

C-tail

N-arm

GateJH

JH

JHBPJHBPa

dcb

JH Transporter

Reference 1. Suzuki R, Fujimoto Z, Shiotsuki T, Tsuchiya W, Momma M, Tase A, Miyazawa M, Yamazaki T

(2011) Structural mechanism of JH delivery in hemolymph by JHBP of silkworm, Bombyx mori Scientific Reports 1:133

Chemicals that disrupt JH signaling by inhibiting JHBP-JH complex formation could be used as effective IGRs and insecticides against household and agricultural pests. The detailed molecular mechanism of JH recognition by JHBP provides the first opportu-nity to rationalize computer-assisted design of such chemicals. This is the first report that provides structural insight into JH signaling.

(A) The crystal structure of JHBP-JH complex (left) where the bound JH molecule is shown as a space-filling model (pink) and the gate-helix on the JH-binding pocket is highlighted in blue. Close-up view of the JH-binding pocket of the complex (right). (B) The crystal structure of the unliganded JHBP. The gate-helix shown in blue resides in an open conformation which permits access of JH to the binding pocket.

Fig. 3. Proposed mechanism for JHBP-mediated JH transport in hemolymph from the CA cells to target cells

(a) The free JHBP assumes various forms of the gate-helix. (b) JH binds to the open form at the CA cell surface. (c) In the fully closed form of the complex, the bound JH is completely buried inside the protein and is protected. (d) When the complex reaches the target cells, JHBP opens the gate to release the bound JH.

Fig. 2. Interactions of JHBP with (A) the epoxide moiety and (B) the ester moiety of JH shown in ball-and-stick model

Hydrogen bonds and stacking interactions are indicated by light blue and orange dotted lines, respectively.

JHBP JHBP JHBP

－ 22－

Bioindustry

Jun Ishibashi1, Megumi Kasashima1, Makoto Nakamura2, Seiji Tokino2, Hideaki Suzuki3, Tomio Nagano3

1Insect Mimetics Research Unit, NIAS, 2Industrial Technology Center of Wakayama Prefecture, 3JITSUBO Co. Ltd.

We developed a novel fiber-processing technique to produce antimicrobial materials using antimicrobial peptide-polymer conjugates.

Keywords: antimicrobial peptide, polymer, antimicrobial fiber Background


2.

3.

4.

　Prevention of infectious diseases is a major concern. Because people strongly prefer a clean environment, antimicrobial materials are widely used in Japan. The market for antimicro-bial materials is estimated to be 800 billion yen per year. We isolated and identified a beetle defensin and developed defensin-derived antimicrobial peptides (AMP) which have potent antimicrobial activity. We then developed AMP-immobilized antimicrobial cotton fibers that have high durability by conjugation of the AMPs via covalent bonding. Immobilization is a complicated procedure, however, and the fibers could suffer chemical damage from reagents during the processing. Therefore, by using AMP and less severe conditions, we developed a novel technique to obtain fibers with an antimicrobial finish.

We synthesized polymethacrylate-PEG-NH2 by radical polymerization of methacrylate conjugated with NH2-PEG as a spacer molecule (Fig. 1). Then maleimide groups were introduced to the spacer (polymer 1). A defensin-derived AMP “peptideA” with addi-tion of an N-terminal cysteine residue (Cys-peptideA) was conjugated to the maleimide groups to obtain polymer 2. The C-terminal cysteine addition variant poly-mer 3 was also synthesized. These peptide-polymer conjugates were coated onto cotton fibers to obtain an antimicrobial finish. The antimicrobial activities of the fibers were determined by absorption methods. The fibers were washed and sterilized by autoclaving and the antimicrobial assays were repeated. Both polymer 2 and 3 coated fibers showed potent antimicrobial activities (Table 1). The antimicrobial finish was durable for at least 10 cycles of washing and autoclaving.

This method for producing antimicrobial cotton fibers is similar to standard proce-dures in textile polymer-processing and would not require investment in novel facilities. It is expected that these antimicrobial materials would be widely used after reducing the cost of peptide synthesis.

Development of a processing technique for antimicrobial fibers using beetle defensin-derived antimicrobial peptide-polymer conjugates

2.

Future prospects 1.

－ 23－

References 1. Nakamura M, Iwasaki T, Tokino S, Asaoka A, Yamakawa M, Ishibashi J (2011) Development

of a bioactive fiber with immobilized synthetic peptides designed from the active site of a beetle defensing Biomacromolecules 12(5):1540-1545

2. Patent application# JP-2011-202173 (Japan)

Fig. 1. Synthesis of antimicrobial peptide-polymer conjugates

washing 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

polymer1 0.1 1.1 - - - - - - - - - - - -

polymer2 >5.7 >7.1 >6.9 >6.6 >6.9 >7.0 6.2 >8.2 >8.4 >8.3 >9.0 >8.6 2.6 >8.9 0.1 0.0

polymer3 >5.7 >7.1 >6.9 >6.6 >6.9 3.1 >8.2 3.9 >8.3 3.6 4.6 0.2 3.8 0.2 0.1 -

Japan Textile Evaluation Technology Council’s certification standard for antimicrobial finished textile:Bacteriostatic activity value >2.2 (bold)

Table 1. Bacteriostatic activity values of antimicrobial peptide-polymer conjugates processed cotton fiber

－ 24－


Keywords: juvenile hormone, P450, metamorphosis, insect growth regulator, silkworm

Background

Results and Discussion

1.

2.

3.

4.

5.

1Insect Growth Regulation Research Unit, NIAS, 2University of Tsukuba, 3Transgenic Silkworm Research Unit, NIAS, 4Kyushu University Graduate School, 5The University of Tokyo, 6Insect Genome Research Unit, NIAS, 7Institute of Floricultural Sciences, National Agriculture and Food Research Orga-nization, 8Meiji University

　Silkworms typically have five larval instars, but the mod (dimolting) mutation causespreco-cious larval-pupal metamorphosis in the third or fourth instar. Most mod larvae form larval-pupal inter-mediates, but some individuals can become miniature moths with normal fertility. To elucidate the molecu-lar mechanism that underlies plasticity in the number of larval instars, we identified and characterized the gene responsible for the mod mutation.

　 This study identified the gene responsible for the dimolting (mod) mutant that under-goes precocious metamorphosis with fewer larval–larval molts. We showed that a cyto-chrome P450 gene CYP15C1 is responsible for mod and is essential for juvenile hormone (JH) biosynthesis. Precocious metamorphosis in mod larvae is caused by complete loss of JHs in the larval hemolymph.

Takaaki Daimon1, Toshinori Kozaki1, Ryusuke Niwa2, Isao Kobayashi3, Kenjiro Furuta1, Toshiki Namiki1, Keiro Uchino3, Yutaka Banno4, Susumu Katsuma5, Toshiki Tamura1, Kazuei Mita6, Hideki Sezutsu3, Masayoshi Nakayama7, Kyo Itoyama8, Toru Shimada5, Tetsuro Shinoda1

We performed positional cloning of mod and found a null mutation in the cytochrome P450 gene CYP15C1 in the mod allele. A biochemical experiment showed that CYP15C1 epoxidizes farnesoic acid to JH acid in a highly stereospecific manner. We showed that the mod mutation results in complete loss of JHs in the larval hemolymph and that the mutant phenotype can be rescued when the wild-type allele of CYP15C1 was expressed in transgenic mod larvae using the GAL4/UAS system. We demonstrated that CYP15C1 plays an indispensable role in JH biosynthesis, and its molecular defect results in the loss of JHs in the hemolymph, thereby causing precocious metamorphosis in the mod strain. Precocious larval-pupal transitions in mod larvae always occur after the larval third instar, but not in the first or second instar. Our data provide evidence that authentic (epoxidized) JHs are essential for the classic “status quo” molting in late larval stages (third and fourth instar), but not in early larval stages (first and second instar) of B. mori.

This JH–deficient mutation is the first described to date in any insect species and, therefore, the mod strain will serve as a useful model for elucidating the molecular mechanism of JH action. The JH biosynthetic reaction catalyzed by CYP15C1 is insect-specific and is therefore a potential target for biorational insecticides.

Future prospects 1.

2.

Identification of the gene responsible for the dimolting mutant of the silkworm Bombyx mori

－ 25－

Reference 1.

Fig. 2. Characterization of CYP15C1

CYP15C1 is specifically expressed in corpora allata (left). In the JH biosyn-thetic pathway (right), CYP15C1 epoxidizes ethyl-branched farnesoic acids (homo-FAs) produce JH acids (JHA).

Fig. 3. Transgenic rescue experiments of mod

Larvae carrying both ET14 (enhancer trap line 14) and UAS-CYP151 constructs entered the fifth larval instar and eventually formed larger adults.

Daimon T, Kozaki T, Niwa R, Kobayashi I, Furuta K, Namiki T, Uchino K, Banno Y, Katsuma S, Tamura T, Mita K, Sezutsu H, Nakayama M, Itoyama K, Shimada T, Shinoda T (2012) Precocious metamorphosis in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori PLoS Genetics 8(3):e1002486

Fig. 1. Precocious metamorphosis observed in mod larvae mod individuals form small cocoons and are able to develop into small but normal adults with normal fertility. Upper, wild type; lower, mod.

Lists of Publications (In order of the subject number of the Third Five-year plan)

1-11 Conservation of genetic resources for food and agriculture and intensification

of their use

Original Papers

1. Aoki T, Scandiani M.M, OÕDonnell K (2012) Phenotypic, molecular phylogenetic, and pathogenetic

characterization of Fusarium crassistipitatum sp. nov., a novel soybean sudden death syndrome pathogen

from Argentina and Brazil Mycoscience 53(3):167-186

2. Araki M, Numaoka A, Kawase M, Fukunaga K (2012) Origin of waxy common millet, Panicum miliaceum

L. in Japan Genetic Resources and Crop Evolution 59(7):1303-1308

3. Djedidi S, Yokoyama T, Tomooka N, Ohkama-Ohtsu N, Risal C.P, Abdelly C, Sekimoto H (2011)

Phenotypic and genetic characterization of rhizobia associated with alfalfa in the Hokkaido and Ishigaki

regions of Japan Systematic and Applied Microbiology 34(6):453-461

4. Fujinaga M, Yamagishi N, Ogiso H, Takeuchi J, Moriwaki J, Sato T (2011) First report of celery stunt

anthracnose caused by Colletotrichum simmondsii in Japan Journal of General Plant Pathology 77(4):243-

247

5. Hirano R, Naito K, Fukunaga K, Watanabe K.N, Ohsawa R, Kawase M (2011) Genetic structure of

landraces in foxtail millet (Setaria italica (L.) P. Beauv.) revealed with transposon display and interpretation

to crop evolution of foxtail millet Genome 54(6):498-506

6. Hirokawa M, Tatematsu K (2011) The order of four homeotic-mutant genes, Nc, ECw, EKp and EMu, on the 6th-

linkage group of Bombyx mori Journal of Insect Biotechnology and Sericology 80(2):41-48

7. Hsiang E-H, Tsai W-T, Hsieh T-F, Matsumoto T, Niino T (2011) Effects of Drying Treatment and

Cryopreservation on Seed Viability of Phalaenopsis amabilis and Phalaenopsis Aphrodite Journal of Taiwan

Agricultural Research 60(4):309-317 (in Chinese with English Summary)

8. Javadi F, Tun Tun Y, Kawase M, Guan K, Yamaguchi H (2011) Molecular phylogeny of the subgenus

Ceratotropis (genus Vigna, Leguminosae) reveals three eco-geographical groups and Late

PlioceneÐPleistocene diversification: evidence from four plastid DNA region sequences Annals of Botany

108(2):367-380

9. Kaga A, Shimizu T, Watanabe S, Tsubokura Y, Katayose Y, Harada K, Vaughan D.A, Tomooka N (2012)

Evaluation of soybean germplasm conserved in NIAS genebank and development of mini core collections

Breeding Science 61(5):566-592

10. Kondo N, Tomooka N (2012) New sources of resistance to Cadophora gregata f. sp. adzukicola and

Fusarium oxysporum f. sp. adzukicola in Vigna spp. Plant Disease 96(4):562-568

11. Kongjaimun A, Kaga A, Tomooka N, Somta P, Shimizu T, Shu Y, Isemura T, Vaughan D.A, Srinives P

(2012) An SSR-based linkage map of yardlong bean (Vigna unguiculata (L.) Walp. subsp. unguiculata

Sesquipedalis Group) and QTL analysis of pod length Genome 55(2):81-92

12. Kongjaimun A, Kaga A, Tomooka N, Somta P, Vaughan D.A, Srinives P (2012) The genetics of

domestication of yardlong bean, Vigna unguiculata (L.) Walp. ssp. unguiculata cv.-gr. sesquipedalis Annals

of Botany 109(6):1185-1200

－ 26 －

13. Kusano M, Fukushima A, Fujita N, Okazaki Y, Kobayashi M, Fujita Oitome N, Ebana K, Saito K (2012)

Deciphering starch quality of rice kernels using metabolite profiling and pedigree network analysis

Molecular Plant 5(2):442-451

14. Kushiro M, Saitoh H, Sugiura Y, Aoki T, Kawamoto S, Sato T (2012) Experimental infection of Fusarium

proliferatum in Oryza sativa plants; fumonisin B1 production and survival rate in grains International

Journal of Food Microbiology 156(3):204-208

15. Matsuda F, Okazaki Y, Oikawa A, Kusano M, Nakabayashi R, Kikuchi J, Yonemaru J, Ebana K, Yano M,

Saito K (2012) Dissection of genotype-phenotype associations in rice grains using metabolome quantitative

trait loci analysis The Plant Journal 70(4):624-636

16. Matsuura T, Mizuno A, Tsukamoto T, Shimizu Y, Saito N, Sato S, Kikuchi S, Uzuki T, Azegami K, Sawada

H (2012) Erwinia uzenensis sp. nov., a novel pathogen that affects European pear trees (Pyrus communis L.)

International Journal of Systematic and Evolutionary Microbiology 62(Pt 8):1799-1803

17. Miyoshi T, Shimizu S, Shinozaki T, Sawada H (2011) Detection and quantification of Ceratocystis fimbriata

in fig plants by real-time PCR Japanese Journal of Phytopathology 77(2):96-104 (in Japanese with English

Summary)

18. Morishita T, Shimizu A (2011) The effects of chronic gamma ray irradiation on the growth of soybean

Japanese Journal of Crop Science 80(3):341-345 (in Japanese with English Summary)

19. Msalya G, Shimogiri T, Ohno S, Okamoto S, Kawabe K, Minezawa M, Maeda Y (2011) Evaluation of

PRNP expression based on genotypes and alleles of two indel loci in the medulla oblongata of Japanese

Black and Japanese Brown cattle PLoS ONE 6(5):e18787

20. Muramatsu N, Yamamoto M, Nakano H, Yamanouchi H (2011) Analysis of factors governing water flow

traits in fruiting plants twigs Scientia Horticulturae 130(1):175-180

21. Muramatsu N, Yamanouchi H, Ito Y, Masuda T, Yoshioka T, Takyu T (2011) Characters of the leaf color

mutant of Japanese pear (Pyrus pyrifolia (Burm. f.) Nakai) found in gamma ray irradiation field (gamma

field) Horticultural Research (Japan) 10(2):143-149 (in Japanese with English Summary)

22. Noguchi T, Hosobuchi S, Iizuka H, Kato A, Takamiya T, Yamashita H, Murakami Y, Okuizumi H (2011)

Development of the cultivar-discrimination markers in Amorphophallus konjac DNA Polymorphism 19:75-

81 (in Japanese with English Summary)

23. Nomura K, Ishii K, Dadi H, Takahashi Y, Minezawa M, Cho C.Y, Sutopo, Faruque M.O, Nyamsamba D,

Amano T (2012) Microsatellite DNA markers indicate three genetic lineages in East Asian indigenous goat

populations Animal Genetics (Published Online):Early View (DOI: 10.1111/j.1365-2052.2012.02334.x)

24. Ooshiro A, Hiyane S, Takushi T, Takaesu Y, Sato T (2011) Charcoal rot of bushkiller (Cayratia spp.) caused

by Macrophomina phaseolina (Tassi) Goidanich Japanese Journal of Phytopathology 77(3):143-147 (in

Japanese with English Summary)

25. Okawa M, Niino T, Shirata K, Nagamine T (2011) International situation of farmers' rights and proposals for

improving current circumstances in Japan Breeding Research 14(1):1-8 (in Japanese with English

Summary)

26. OÕDonnell K, Humber R.A, Geiser D.M, Kang S, Park B, Robert V, Crous P.W, Johnston P, Aoki T, Rooney

A.P, Rehner S.A (2012) Phylogenetic diversity of insecticolous fusaria inferred from multilocus DNA

－ 27 －

sequence data and their molecular identification via FUSARIUM-ID and Fusarium MLST Mycologia

104(2):427-445

27. Park Y-J, Nemoto K, Nishikawa T, Matsushima K, Minami M, Kawase M (2011) Genetic diversity and

expression analysis of granule bound starch synthase I gene in the new world grain amaranth (Amaranthus

cruentus L.) Journal of Cereal Science 53(3):298-305

28. Park Y-J, Nishikawa T, Tomooka N, Nemoto K (2012) The molecular basis of mutations at the Waxy locus

from Amaranthus caudatus L.: evolution of the waxy phenotype in three species of grain amaranth

Molecular Breeding 30(1):511-520

29. Redestig H, Kusano M, Ebana K, Kobayashi M, Oikawa A, Okazaki Y, Matsuda F, Arita M, Fujita N, Saito

K (2011) Exploring molecular backgrounds of quality traits in rice by predictive models based on high-

coverage metabolomics BMC Systems Biology 5:176

30. Sarver B.A.J, Ward T.J, Gale L.R, Broz K, Corby Kistler H, Aoki T, Nicholson P, Carter J, OÕDonnell K

(2011) Novel fusarium head blight pathogens from Nepal and Louisiana revealed by multilocus genealogical

concordance Fungal Genetics and Biology 48(12):1096-1107

31. Sawada H, Watauchi K, Kunugi Y (2011) Polyphasic comparison of bacterial spot pathogen of grapevine

(Xanthomonas arboricola) with that of stone fruits (X. arboricola pv. pruni) Japanese Journal of

Phytopathology 77(4):265-277 (in Japanese with English Summary)

32. Scandiani M.M, Ruberti D.S, Giorda L.M, Pioli R.N, Luque A.G, Bottai H, Ivancovich J.J, Aoki T,

OÕDonnell K (2011) Comparison of inoculation methods for characterizing relative aggressiveness of two

soybean sudden-death syndrome pathogens, Fusarium virguliforme and F. tucumaniae Tropical Plant

Pathology 36(3):133-140

33. Sekizawa K, Yamamoto S, Rafique T, Fukui K, Niino T (2011) Cryopreservation of in vitro-grown shoot

tips of carnation (Dianthus caryophyllus L.) by vitrification method using aluminium cryo-plates Plant

Biotechnology 28(4):401-405

34. Shimomoto Y, Morita Y, Takeuchi S, Hikichi Y, Kiba A, Sat T (2011) Reidentification of the causal fungus

of black blight of eggplant Japanese Journal of Phytopathology 77(2):105-108 (in Japanese with English

Summary)

35. Tabuchi H, Zhang Y, Hattori S, Omae M, Shimizu-Sato S, Oikawa T, Qian Q, Nishimura M, Kitano H, Xie

H, Fang X, Yoshida H, Kyozuka J, Chen F, Sato Y (2011) LAX PANICLE2 of rice encodes a novel nuclear

protein and regulates the formation of axillary meristems The Plant Cell 23(9):3276-3287

36. Takeya M, Yamasaki F, Uzuhashi S, Kumagai M, Sawada H, Nagai T, Tomioka K, Sato T, Aoki T, Kawase

M (2012) Development of a database of plant diseases in Japan and a system for making microorganism

genetic resources and their DNA sequence data available to the research community Japan Agricultural

Research Quarterly 46(2):193-198

37. Tanaka K, Endo M, Hirayama K, Okane I, Hosoya T, Sato T (2011) Phylogeny of Discosia and

Seimatosporium, and introduction of Adisciso and Immersidiscosia genera nova Persoonia 26:85-98

38. Tomioka K, Hirooka Y, Sawada H, Aoki T, Sato T (2011) Plectosporium blight of monkshood caused by

Plectosporium tabacinum Journal of General Plant Pathology 77(4):266-268

39. Tomioka K, Sato T (2011) Fruit rot of sweet pepper caused by Stemphylium lycopersici in Japan Journal of

General Plant Pathology 77(6):342-344

－ 28 －

40. Tomioka K, Sato T, Moriwaki J, Terasawa Y, Koganezawa H (2012) Anthracnose of bacopa caused by

Colletotrichum destructivum Journal of General Plant Pathology 78(2):133-135

41. Watanabe K, Niino T, Murayama T, Nanzyo M (2011) Effects of the pre-transplanting phosphorus

application on dry matter production, photosynthesis, root activities and nutrient absorption of cabbage and

maize at initial growth stage Japanese Journal of Crop Science 80(4):391-402 (in Japanese with English

Summary)

42. Yamamoto J, Tanaka K, Ohtaka N, Sato T (2012) Black leaf spot of Japanese persimmon (Diospyros kaki), a

new disease caused by Adisciso kaki sp. nov. Journal of General Plant Pathology 78(2):99-105

43. Yamamoto S, Fukui K, Rafique T, Khan N.I, Castillo Martinez C.R, Sekizawa K, Matsumoto T, Niino T

(2012) Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using

aluminium cryo-plates Plant Genetic Resources: Characterization and Utilization 10(01):14-19

44. Yamamoto S, Moe Kyaw Aung, Watanabe K, Wunna, Kawase M (2012) Third field survey collecting

traditionally grown crops in northern areas of Myanmar, 2011 Annual Report on Exploration and

Introduction of Plant Genetic Resources 27:95-109

45. Yamamoto S, Rafique T, Fukui K, Sekizawa K, Niino T (2012) V-Cryo-plate procedure as an effective

protocol for cryobanks: Case study of mint cryopreservation CryoLetters 33(1):12-23

46. Yamamoto S, Rafique T, Priyantha W.S, Fukui K, Matsumoto T, Niino T (2011) Development of a

cryopreservation procedure using aluminium cryo-plates CryoLetters 32(3):256-265

47. Yonemaru J, Yamamoto T, Ebana K, Yamamoto E, Nagasaki H, Shibaya T, Yano M (2012) Genome-wide

haplotype changes produced by artificial selection during modern rice breeding in Japan PLoS ONE

7(3):e32982

Reviews and Monographs (in English)

1. Fukui K, Shirata K, Niino T, Kashif I.M (2011) Cryopreservation of mulberry winter buds in Japan Acta

Horticulturae 908:483-488

2. Nagai T (2012) Bacteriophages of Bacillus subtilis (natto) and their contamination in natto factories

Bacteriophages (6):95-110

3. Okuizumi H, Takamiya T, Okazaki Y, Hayashizaki Y (2011) Restriction landmark genome scanning

Methods in Molecular Biology 791(8):101-112

4. Takamiya T, Hosobuchi S, Seetharam K, Murakami Y, Okuizumi H (2012) Inheritance of DNA methylation

in plant genomeby DNA Methylation - From Genomics to Technology (4):69-92

5. Takeya M, Yamasaki F, Hattori S, Kawase M (2011) Experiences on PGR training from the perspective of

genebank management Reviewing Plant Genetic Resources Education in East and Southeast Asia :65-68

6. Tanaka D, Nishiuchi A, Niino T, Matsumoto T (2011) A comparison of vitrification and droplet vitrification

procedures for the cryopreservation of in vitro grown black chokeberry shoot tips Acta Horticulturae

908:325-330

7. Yamamoto S, Fukui K, Niino T (2012) Cryostorage facilitation by the NIAS Genebank NIAS-FAO

International Symposium Plant genetic resources for food and agriculture in Asia and the Pacific:

impacts and future directions :75-90

－ 29 －

8. Yano T, Shimizu S, Miyoshi T, Miyata N, Immon K, Shinozaki T, Sawada H, Kageyama K (2011) Tolerant

Actinidia species to Pythium helicoides and P. vexans causing root rot Acta Horticulturae 913:517-523

1-21 Enhancing the potential of the genome sequence and resources from

agriculturally important organisms

Original Papers

1. Asano K, Yamasaki M, Takuno S, Miura K, Katagiri S, Ito T, Doi K, Wu J, Ebana K, Matsumoto T, Innan

H, Kitano H, Ashikari M, Matsuoka M (2011) Artificial selection for a green revolution gene during

japonica rice domestication Proceedings of the National Academy of Sciences of the United States of

America 108(27):11034-11039

2. Asano T, Hayashi N, Kobayashi M, Aoki N, Miyao A, Mitsuhara I, Ichikawa H, Komatsu S, Hirochika H,

Kikuchi S, Ohsugi R (2012) A rice calcium-dependent protein kinase OsCPK12 oppositely modulates salt-

stress tolerance and blast disease resistance The Plant Journal 69(1):26-36

3. Ashikawa I, Hayashi N, Abe F, Wu J, Matsumoto T (2012) Characterization of the rice blast resistance gene

Pik cloned from Kanto51 Molecular Breeding 30(1):485-494

4. Aslam A.F.M, Kiya T, Mita K, Iwami M (2011) Identification of novel bombyxin genes from the genome of

the silkmoth Bombyx mori and analysis of their expression Zoological Science 28(8):609-616

5. Aya K, Suzuki G,Suwabe K, Hobo T, Takahashi H, Shiono K, Yano K, Tsutsumi N, Nakazono M,

Nagamura Y, Matsuoka M, Watanabe M (2011) Comprehensive network analysis of anther-expressed genes

in rice by the combination of 33 laser microdissection and 143 spatiotemporal microarrays PLoS ONE

6(10):e26162

6. Bahrini I, Ogawa T, Kobayashi F, Kawahigashi H, Handa H (2011) Overexpression of the pathogen-

inducible wheat TaWRKY45 gene confers disease resistance to multiple fungi in transgenic wheat plants


7. Bahrini I, Sugisawa M, Kikuchi R, Ogawa T, Kawahigashi H, Ban T, Handa H (2011) Characterization of a

wheat transcription factor, TaWRKY45, and its effect on Fusarium head blight resistance in transgenic wheat

plants Breeding Science 61(2):121-129

8. Bashir K, Ishimaru Y, Shimo H, Kakei Y, Senoura T, Takahashi R, Sato Y, Sato Y, Uozumi N, Nakanishi

H, Nishizawa N.K (2011) Rice phenolics efflux transporter 2 (PEZ2) plays an important role in solubilizing

apoplasmic iron Soil Science & Plant Nutrition 57(6):803-812

9. Buchmann J.P, Matsumoto T, Stein N, Keller B, Wicker T (2012) Inter-species sequence comparison of

Brachypodium reveals how transposon activity corrodes genome colinearity The Plant Journal 71(4):550-

563

10. Chen G, Komatsuda T, Ma J.F, Li C, Yamaji N, Nevo E (2011) A functional cutin matrix is required for

plant protection against water loss Plant Signaling & Behavior 6(9):1297-1299

11. Chen G, Komatsuda T, Ma J.F, Nawrath C, Pourkheirandish M, Tagiri A, Hu Y-G, Sameri M, Li X, Zhao X,

Liu Y, Li C, Ma X, Wang A, Nair S, Wang N, Miyao A, Sakuma S, Yamaji N, Zheng X, Nevo E (2011) An

－ 30 －

ATP-binding cassette subfamily G full transporter is essential for the retention of leaf water in both wild

barley and rice Proceedings of the National Academy of Sciences of the United States of America

108(30):12354-12359

12. Chen H, Wei T, Saotome A, Sakaguchi K, Shikamoto Y, Mizuno H, Akita F, Shimizu T, Uehara-Ichiki T,

Omura T (2012) Aggregation ability of virus-specific antibodies is correlated with their capacity to

neutralize Rice dwarf virus Japan Agricultural Research Quarterly 46(1):65-71

13. De Vleesschauwer D, Van Buyten E, Satoh K, Balidion J, Mauleon R, Choi I-R, Vera-Cruz C, Kikuchi S,

Hıfte M (2012) Brassinosteroids antagonize gibberellin- and salicylate-mediated root immunity in rice

Plant Physiology 158(4):1833-1846

14. Endo M, Nakayama S, Umeda-Hara C, Ohtsuki N, Saika H, Umeda M, Toki S (2012) CDKB2 is involved in

mitosis and DNA damage response in rice The Plant Journal 69(6):967-977

15. Fujii T, Abe H, Yamamoto K, Katsuma S, Shimada T (2011) Interspecies linkage analysis of mo, a Bombyx

mori locus associated with mosaicism and gynandromorphism Genetica 139(10):1323-1329

16. Hamada H, Kurusu T, Okuma E, Nokajima H, Kiyoduka M, Koyano T, Sugiyama Y, Okada K, Koga J, Saji

H, Miyao A, Hirochika H, Yamane H, Murata Y, Kuchitsu K (2012) Regulation of a proteinaceous elicitor-

induced Ca2+ influx and production of phytoalexins by a putative voltage-gated cation channel, OsTPC1, in

cultured rice cells Journal of Biological Chemistry 287(13):9931-9939

17. Ibrahim N, Handa H, Cosset A, Koulintchenko M, Konstantinov Y, Lightowlers R.N, Dietrich A, Weber-

Lotfi F (2011) DNA delivery to mitochondria: Sequence specificity and energy enhancement

Pharmaceutical Research 28(11):2871-2882

18. Ishimaru Y, Kakei Y, Shimo H, Bashir K, Sato Y, Sato Y, Uozumi N, Nakanishi H, Nishizawa N.K (2011) A

rice phenolic efflux transporter is essential for solubilizing precipitated apoplasmic iron in the plant stele

Journal of Biological Chemistry 286(28):24649-24655

19. Izawa T, Mihara M, Suzuki Y, Gupta M, Itoh H, Nagano A.J, Motoyama R, Sawada Y, Yano M, Yokota

Hirai M, Makino A, Nagamura Y (2011) Os-GIGANTEA confers robust diurnal rhythms on the global

transcriptome of rice in the field The Plant Cell 23(5):1741-1755

20. Katayose Y, Kanamori H, Shimomura M, Ohyanagi H, Ikawa H, Minami H, Shibata M, Ito T, Kurita K, Ito

K, Tsubokura Y, Kaga A, Wu J, Matsumoto T, Harada K, Sasaki T (2012) DaizuBase, an integrated soybean

genome database including BAC-based physical maps Breeding Science 61(5):661-664

21. Kawahigashi H, Kasuga S, Okuizumi H, Kanamori H, Ando T, Matsumoto T (2011) Classification of

genotypes of the target leaf spot-resistant gene (ds1) in a sorghum collection Crop Science 51(5):2095-2103

22. Kawaoka S, Kadota K, Arai Y, Suzuki Y, Fujii T, Abe H, Yasukochi Y, Mita K, Sugano S, Shimizu K,

Tomari Y, Shimada T, Katsuma S (2011) The silkworm W chromosome is a source of female-enriched

piRNAs RNA 17(12):2144-2151

23. Kawaura K, Wu J, Matsumoto T, Kanamori H, Katagiri S, Ogihara Y (2012) Genome change in wheat

observed through the structure and expression of α/β-gliadin genes Functional & Integrative Genomics

12(2):341-355

24. Khodayari H, Saeidi H, Rahiminejad M.R, Komatsuda T (2011) Transferability and polymorphism of barley

microsatelite markers across H-genome containing species in the genus Hordeum (H. vulgare and H.

bulbosum) The Iranian Journal of Botany 17(2):200-211

－ 31 －

25. Kikuchi R, Kawahigashi H, Oshima M, Ando T, Handa H (2012) The differential expression of HvCO9, a

member of the CONSTANS-like gene family, contributes to the control of flowering under short-day

conditions in barley Journal of Experimental Botany 63(2):773-784

26. Kim C, Kim J, Kikuchi S, Choi J, Kim Y, Park H, Seol Y, Park D, Hahn J, Kim Y (2011) Computational

identification of Chinese cabbage anthocyaninspecific genes BioChip Journal 5(2):184-192

27. Mase K, Hirayama C, Okada E, lizuka T, Kadono-Okuda K, Narukawa J, Yamamoto K (2011)

Reinstatement of the 27th linkage group in the classical linkage map of Bombyx mori by molecular linkage

analysis of faint-green cocoon gene Journal of Insect Biotechnology and Sericology 80(2):71-77

28. Mayer K.F.X, Martis M, Hedley P.E, ÿSimkov⁄ H, Liu H, Morris J.A, Steuernagel B, Taudien S, Roessner S,

Gundlach H, Kubal⁄kov⁄ M, Such⁄nkov⁄ P, Murat F, Felder M, Nussbaumer T, Graner A, Salse J, Endo T,

Sakai H, Tanaka T, Itoh T, Sato K, Platzer M, Matsumoto T, Scholz U, Doleÿzel J, Waugh R, Stein N (2011)

Unlocking the barley genome by chromosomal and comparative genomics The Plant Cell 23(4):1249-1263

29. Miyao A, Nakagome M, Ohnuma T, Yamagata H, Kanamori H, Katayose Y, Takahashi A, Matsumoto T,

Hirochika H (2012) Molecular spectrum of somaclonal variation in regenerated rice revealed by whole-

genome sequencing Plant and Cell Physiology 53(1):256-264

30. Mizuno H, Kawahara Y, Wu J, Katayose Y, Kanamori H, Ikawa H, Itoh T, Sasaki T, Matsumoto T (2011)

Asymmetric distribution of gene expression in the centromeric region of rice chromosome 5 Frontiers in

Plant Science 2:16

31. Morita Y, Saito R, Ban Y, Tanikawa N, Kuchitsu K, Ando T, Yoshikawa M, Habu Y, Ozeki Y, Nakayama

M (2012) Tandemly arranged chalcone synthase A genes contribute to the spatially regulated expression of

siRNA and the natural bicolor floral phenotype in Petunia hybrida The Plant Journal 70(5):739-749

32. Moumeni A, Satoh K, Kondoh H, Asano T, Hosaka A, Venuprasad R, Serraj R, Kumar A, Leung H, Kikuchi

S (2011) Comparative analysis of root transcriptome profiles of two pairs of drought-tolerant and susceptible

rice near-isogenic lines under different drought stress BMC Plant Biology 11:174

33. Nakamura S, Abe F, Kawahigashi H, Nakazono K, Tagiri A, Matsumoto T, Utsugi S, Ogawa T, Handa H,

Ishida H, Mori M, Kawaura K, Ogihara Y, Miura H (2011) A wheat homolog of MOTHER OF FT AND

TFL1 acts in the regulation of germination The Plant Cell 23(9):3215-3229

34. Nakayama S (2012) Inter-MITE polymorphisms of a newly identified MITE show relationships among

sugarcane (Saccharum) species Genetic Resources and Crop Evolution 59(7):1389-1396

35. Ogawa D, Abe K, Miyao A, Kojima M, Sakakibara H, Mizutani M, Morita H, Toda Y, Hobo T, Sato Y,

Hattori T, Hirochika H, Takeda S (2011) RSS1 regulates the cell cycle and maintains meristematic activity

under stress conditions in rice Nature Communications 2:278

36. Oono Y, Kawahara Y, Kanamori H, Mizuno H, Yamagata H, Yamamoto M, Hosokawa S, Ikawa H,

Akahane I, Zhu Z, Wu J, Itoh T, Matsumoto T (2011) mRNA-seq reveals a comprehensive transcriptome

profile of rice under phosphate stress Rice 4(2):50-65

37. Ozawa K, Wakasa Y, Ogo Y, Matsuo K, Kawahigashi H, Takaiwa F (2012) Development of an efficient

Agrobacterium-mediated gene targeting system for rice and analysis of rice knockouts lacking granule-

bound starch synthase (Waxy) and β1,2-xylosyltransferase Plant and Cell Physiology 53(4):755-761

－ 32 －

38. Saika H, Oikawa A, Matsuda F, Onodera H, Saito K, Toki S (2011) Application of gene targeting to

designed mutation breeding of high-tryptophan rice Plant Physiology 156(3):1269-1277

39. Sakai H, Mizuno H, Kawahara Y, Wakimoto H, Ikawa H, Kawahigashi H, Kanamori H, Matsumoto T, Itoh

T, Gaut B.S (2011) Retrogenes in rice (Oryza sativa L. ssp. japonica) exhibit correlated expression with

their source genes Genome Biology and Evolution 3:1357-1368

40. Sameri M, Pourkheirandish M, Chen G, Tonooka T, Komatsuda T (2011) Detection of photoperiod

responsive and non-responsive flowering time QTL in barley Breeding Science 61(2):183-188

41. Sharoni A.M, Nuruzzaman M, Satoh K, Moumeni A, Attia K, Venuprasad R, Serraj R, Kumar A, Leung H,

Rafiul Islam A.K.M, Kikuchi S (2012) Comparative transcriptome analysis of AP2/EREBP gene family

under normal and hormone treatments, and under two drought stresses in NILs setup by Aday Selection and

IR64 Molecular Genetics and Genomics 287(1):1-19

42. Takahashi H, Saika H, Matsumura H, Nagamura Y, Tsutsumi N, Nishizawa N.K, Nakazono M (2011) Cell

division and cell elongation in the coleoptile of rice alcohol dehydrogenase 1-deficient mutant are reduced

under complete submergence Annals of Botany 108(2):253-261

43. Takamizo T, Nakatsu S, Nagamura Y, Fujimori M, Tarumoto I (2012) Mapping of DNA markers linked to a

thermosensitivity gene in sorghum Bulletin of NARO Institute of Livestock and Grassland Science 12:1-8

44. Takehisa H, Sato Y, Igarashi M, Abiko T, Antonio B.A, Kamatsuki K, Minami H, Namiki N, Inukai Y,

Nakazono M, Nagamura Y (2012) Genome-wide transcriptome dissection of the rice root system:

implications for developmental and physiological functions The Plant Journal 69(1):126-140

45. Uraguchi S, Kamiya T, Sakamoto T, Kasai K, Sato Y, Nagamura Y, Yoshida A, Kyozuka J, Ishikawa S,

Fujiwara T (2011) Low-affinity cation transporter (OsLCT1) regulates cadmium transport into rice grains

Proceedings of the National Academy of Sciences of the United States of America 108(52):20959-20964

46. Yamamoto-Kihara M, Kotani E, Mori H (2011) Characterization of a novel C-type lectin cDNA, CLEM 20

cDNA, specifically expressed in mouthparts of the flesh fly Sarcophaga peregrina Journal of Insect

Biotechnology and Sericology 80(1):31-39

47. Yang C, Kawahara Y, Mizuno H, Wu J, Matsumoto T, Itoh T (2012) Independent domestication of Asian

rice followed by gene flow from japonica to indica Molecular Biology and Evolution 29(5):1471-1479

48. Yasukochi Y, Miura N, Nakano R, Sahara K, Ishikawa Y (2011) Sex-linked pheromone receptor genes of the

European corn borer, Ostrinia nubilalis, are in tandem arrays PLoS ONE 6(4):e18843

49. Yuo T, Shiotani K, Shitsukawa N, Miyao A, Hirochika H, Ichii M, Taketa S (2011) Root hairless 2 (rth2)

mutant represents a loss-of-function allele of the cellulose synthase-like gene OsCSLD1 in rice (Oryza sativa

L.) Breeding Science 61(3):225-233


1. Arita K, Kanno T, Yoshikawa M, Habu Y (2011) Mechanisms linking cytosine methylation to histone

modification in Arabidopsis thaliana Non Coding RNAs in Plants :237-249

2. Endo M, Shimizu T, Toki S (2012) Selection of transgenic rice plants using a herbicide tolerant form of the

acetolactate synthase gene Transgenic Plants: Methods and Protocols 3(6):59-66

－ 33 －

3. Kanno T, Habu Y (2011) siRNA-mediated chromatin maintenance and its function in Arabidopsis thaliana

Biochimica et Biophysica Acta - Gene Regulatory Mechanisms 1809(8):444-451

4. Saika H, Onodera H, Toki S (2012) Visual selection in rice: A strategy for the efficient identification of

transgenic calli accumulating transgene products Transgenic Plants: Methods and Protocols 3(7):67-74

1-22 Bioinformatics approach for advancement of agrobiological research

Original Papers

1. Aslam A.F.M, Kiya T, Mita K, Iwami M (2011) Identification of novel bombyxin genes from the genome of

the silkmoth Bombyx mori and analysis of their expression Zoological Science 28(8):609-616

2. Kawaoka S, Kadota K, Arai Y, Suzuki Y, Fujii T, Abe H, Yasukochi Y, Mita K, Sugano S, Shimizu K,

Tomari Y, Shimada T, Katsuma S (2011) The silkworm W chromosome is a source of female-enriched

piRNAs RNA 17(12):2144-2151

3. Leroy P, Guilhot N, Sakai H, Bernard A, Choulet F, Theil S, Reboux S, Amano N, Flutre T, Pelegrin C,

Ohyanagi H, Seidel M, Giacomoni F, Reichstadt M, Alaux M, Gicquello E, Legeai F, Cerutti L, Numa H,

Tanaka T, Mayer K, Itoh T, Quesneville H, Feuillet C (2012) TriAnnot:a versatile and high performance

pipeline for the automated annotation of plant genomes Frontiers in Plant Science 3:5

4. Mayer K.F.X, Martis M, Hedley P.E, ÿSimkov⁄ H, Liu H, Morris J.A, Steuernagel B, Taudien S, Roessner S,

Gundlach H, Kubal⁄kov⁄ M, Such⁄nkov⁄ P, Murat F, Felder M, Nussbaumer T, Graner A, Salse J, Endo T,

Sakai H, Tanaka T, Itoh T, Sato K, Platzer M, Matsumoto T, Scholz U, Doleÿzel J, Waugh R, Stein N (2011)

Unlocking the barley genome by chromosomal and comparative genomics The Plant Cell 23(4):1249-1263

5. Oono Y, Kawahara Y, Kanamori H, Mizuno H, Yamagata H, Yamamoto M, Hosokawa S, Ikawa H,

Akahane I, Zhu Z, Wu J, Itoh T, Matsumoto T (2011) mRNA-seq reveals a comprehensive transcriptome

profile of rice under phosphate stress Rice 4(2):50-65

6. Rice WRKY Working Group (2012) Nomenclature report on rice WRKY's -Conflict regarding gene names

and its solution Rice 5:3

7. Sakai H, Mizuno H, Kawahara Y, Wakimoto H, Ikawa H, Kawahigashi H, Kanamori H, Matsumoto T, Itoh

T, Gaut B.S (2011) Retrogenes in rice (Oryza sativa L. ssp. japonica) exhibit correlated expression with

their source genes Genome Biology and Evolution 3:1357-1368

8. Yang C, Kawahara Y, Mizuno H, Wu J, Matsumoto T, Itoh T (2012) Independent domestication of Asian

rice followed by gene flow from japonica to indica Molecular Biology and Evolution 29(5):1471-1479

1-23 Genomics approach for advancement of research in crop improvement

Original Papers

1. Adachi S, Nito N, Kondo M, Yamamoto T, Arai-Sanoh Y, Ando T, Ookawa T, Yano M, Hirasawa T (2011)

Identification of chromosomal regions controlling the leaf photosynthetic rate in rice by using a progeny

from Japonica and high-yielding Indica varieties Plant Production Science 14(2):118-127

－ 34 －

2. Ebitani T, Hayashi N, Omoteno M, Ozaki H, Yano M, Morikawa M, Fukuta Y (2011) Characterization of

Pi13, a blast resistance gene that maps to chromosome 6 in indica rice (Oryza sativa L. variety, Kasalath)


3. Fukuoka S, Mizobuchi R, Saka N, Ivan S, Matsumoto T, Okuno K, Yano M (2012) A multiple gene

complex on rice chromosome 4 is involved in durable resistance to rice blast Theoretical and Applied

Genetics 125(3):551-559

4. Hirota T, Sayama T, Yamasaki M, Sasama H, Sugimoto T, Ishimoto M, Yoshida S (2012) Diversity and

population structure of black soybean landraces originating from Tanba and neighboring regions Breeding

Science 61(5):593-601

5. Ishimaru Y, Takahashi R, Bashir K, Shimo H, Senoura T, Sugimoto K, Ono K, Yano M, Ishikawa S, Arao

T, Nakanishi H, Nishizawa N.K (2012) Characterizing the role of rice NRAMP5 in manganese, iron and

cadmium transport Scientific Reports 2:286

6. Kaga A, Shimizu T, Watanabe S, Tsubokura Y, Katayose Y, Harada K, Vaughan D.A, Tomooka N (2012)

Evaluation of soybean germplasm conserved in NIAS genebank and development of mini core collections


7. Kasajima I, Ebana K, Yamamoto T, Takahara K, Yano M, Kawai-Yamada M, Uchimiya H (2011)

Molecular distinction in genetic regulation of nonphotochemical quenching in rice Proceedings of the

National Academy of Sciences of the United States of America 108(33):13835-13840

8. Katayose Y, Kanamori H, Shimomura M, Ohyanagi H, Ikawa H, Minami H, Shibata M, Ito T, Kurita K, Ito

K, Tsubokura Y, Kaga A, Wu J, Matsumoto T, Harada K, Sasaki T (2012) DaizuBase, an integrated soybean

genome database including BAC-based physical maps Breeding Science 61(5):661-664

9. Kato T, Segami S, Toriyama M, Kono I, Ando T, Yano M, Kitano H, Miura K, Iwasaki Y (2011) Detection

of QTLs for grain length from large grain rice (Oryza sativa L.) Breeding Science 61(3):269-274

10. Kitamoto N, Kaga A, Kuroda Y, Ohsawa R (2012) A model to predict the frequency of integration of fitness-

related QTLs from cultivated to wild soybean Transgenic Research 21(1):131-138

11. Komatsu K, Hwang T-Y, Takahashi M, Sayama T, Funatsuki H, Oki N, Ishimoto M (2012) Identification of

QTL controlling post-flowering period in soybean Breeding Science 61(5):646-652

12. Marzougui S, Sugimoto K, Yamanouchi U, Shimono M, Hoshino T, Hori K, Kobayashi M, Ishiyama K,

Yano M (2012) Mapping and characterization of seed dormancy QTLs using chromosome segment

substitution lines in rice Theoretical and Applied Genetics 124(5):893-902

13. Matsubara K, Ebana K, Mizubayashi T, Itoh S, Ando T, Nonoue Y, Ono N, Shibaya T, Ogiso E, Hori K,

Fukuoka S, Yano M (2011) Relationship between transmission ratio distortion and genetic divergence in

intraspecific rice crosses Molecular Genetics and Genomics 286(5-6):307-319

14. Matsubara K, Ogiso-Tanaka E, Hori K, Ebana K, Ando T, Yano M (2012) Natural variation in Hd17, a

homolog of Arabidopsis ELF3 that is involved in rice photoperiodic flowering Plant and Cell Physiology

53(4):709-716

15. Matsuda F, Okazaki Y, Oikawa A, Kusano M, Nakabayashi R, Kikuchi J, Yonemaru J, Ebana K, Yano M,

Saito K (2012) Dissection of genotype-phenotype associations in rice grains using metabolome quantitative

trait loci analysis The Plant Journal 70(4):624-636

－ 35 －

16. Ohnishi S, Miyake N, Takeuchi T, Kousaka F, Hiura S, Kanehira O, Saito M, Sayama T, Higashi A,

Ishimoto M, Tanaka Y, Fujita S (2012) Fine mapping of foxglove aphid (Aulacorthum solani) resistance

gene Raso1 in soybean and its effect on tolerance to Soybean dwarf virus transmitted by foxglove aphid


17. Oki N, Komatsu K, Sayama T, Ishimoto M, Takahashi M, Takahashi M (2012) Genetic analysis of

antixenosis resistance to the common cutworm (Spodoptera litura Fabricius) and its relationship with

pubescence characteristics in soybean (Glycine max (L.) Merr.) Breeding Science 61(5):608-617

18. Saisho D, Ishii M, Hori K, Sato K (2011) Natural variation of barley vernalization requirements: Implication

of quantitative variation of winter growth habit as an adaptive trait in East Asia Plant and Cell Physiology

52(5):738-749

19. Saito H, Ogiso-Tanaka E, Okumoto Y, Yoshitake Y, Izumi H, Yokoo T, Matsubara K, Hori K, Yano M,

Inoue H, Tanisaka T (2012) Ef7 encodes an ELF3-like protein and promotes rice flowering by negatively

regulating the floral repressor gene Ghd7 under both short- and long-day conditions Plant and Cell

Physiology 53(4):717-728

20. Segami S, Kono I, Ando T, Yano M, Kitano H, Miura K, Iwasaki Y (2012) Small and round seed 5 gene

encodes alpha-tubulin regulating seed cell elongation in rice Rice 5:4

21. Shibaya T, Nonoue Y, Ono N, Yamanouchi U, Hori K, Yano M (2011) Genetic interactions involved in the

inhibition of heading by heading date QTL, Hd2 in rice under long-day conditions Theoretical and Applied

Genetics 123(7):1133-1143

22. Sugimoto T, Yoshida S, Kaga A, Hajika M, Watanabe K, Aino M, Tatsuda K, Yamamoto R, Matoh T,

Walker D.R, Biggs A.R, Ishimoto M (2011) Genetic analysis and identification of DNA markers linked to a

novel Phytophthora sojae resistance gene in the Japanese soybean cultivar Waseshiroge Euphytica

182(1):133-145

23. Takada Y, Tayama I, Sayama T, Sasama H, Saruta M, Kikuchi A, Ishimoto M, Tsukamoto C (2012) Genetic

analysis of variations in the sugar chain composition at the C-3 position of soybean seed saponins Breeding

Science 61(5):639-645

24. Takagi K, Nishizawa K, Hirose A, Kita A, Ishimoto M (2011) Manipulation of saponin biosynthesis by RNA

interference-mediated silencing of β-amyrin synthase gene expression in soybean Plant Cell Reports

30(10):1835-1846

25. Tamura K, Kiyoshi T, Yonemaru J (2012) The development of highly transferable intron-spanning markers

for temperate forage grasses Molecular Breeding 30(1):1-8

26. Tsubokura Y, Hajika M, Kanamori H, Xia Z, Watanabe S, Kaga A, Katayose Y, Ishimoto M, Harada K

(2012) The β-conglycinin deficiency in wild soybean is associated with the tail-to-tail inverted repeat of the

α-subunit genes Plant Molecular Biology 78(3):301-309

27. Uga Y, Hanzawa E, Nagai S, Sasaki K, Yano M, Sato T (2012) Identification of qSOR1, a major rice QTL

involved in soil-surface rooting in paddy fields Theoretical and Applied Genetics 124(1):75-86

28. Wadahama H, Iwasaki K, Matsusaki M, Nishizawa K, Ishimoto M, Arisaka F, Takagi K, Urade R (2012)

Accumulation of β-conglycinin in soybean cotyledon through the formation of disulfide bonds between α’-

and α-subunits Plant Physiology 158(3):1395-1405

－ 36 －

29. Yamada T, Hajika M, Yamada N, Hirata K, Okabe A, Oki N, Takahashi K, Seki K, Okano K, Fujita Y, Kaga

A, Shimizu T, Sayama T, Ishimoto M (2012) Effects on flowering and seed yield of dominant alleles at

maturity loci E2 and E3 in a Japanese cultivar, Enrei Breeding Science 61(5):653-660

30. Yonemaru J, Yamamoto T, Ebana K, Yamamoto E, Nagasaki H, Shibaya T, Yano M (2012) Genome-wide

haplotype changes produced by artificial selection during modern rice breeding in Japan PLoS ONE

7(3):e32982

31. Yoshida A, Ohmori Y, Kitano H, Taguchi-Shiobara F, Hirano H (2012) ABERRANT SPIKELET AND

PANICLE1, encoding a TOPLESS-related transcriptional co-repressor, is involved in the regulation of

meristem fate in rice The Plant Journal 70(2):327-339


1. Funatsuki H, Hajika M, Yamada T, Suzuki M, Hagihara S, Tanaka Y, Fujita S, Ishimoto M, Fujino K (2012)

Mapping and use of QTLs controlling pod dehiscence in soybean Breeding Science 61(5):554-558

2. Harada K, Watanabe S, Xia Z, Tsubokura Y, Yamanaka N, Anai T (2011) Positional cloning of the

responsible genes for maturity loci E1, E2 and E3 in soybean Soybean - Genetics and Novel Techniques for

Yield Enhancement (3):51-76

3. Ishimoto M, Harada K (2012) Breeding Science Special Issue: Soybean breeding in genomic era Breeding

Science 61(5):435

4. Sugimoto T, Kato M, Yoshida S, Matsumoto I, Kobayashi T, Kaga A, Hajika M, Yamamoto R, Watanabe

K, Aino M, Matoh T, Walker D.R, Biggs A.R, Ishimoto M (2012) Pathogenic diversity of Phytophthora

sojae and breeding strategies to develop Phytophthora-resistant soybeans Breeding Science 61(5):511-522

5. Watanabe S, Harada K, Abe J (2012) Genetic and molecular bases of photoperiod responses of flowering in

soybean Breeding Science 61(5):531-543

6. Yamada T, Takagi K, Ishimoto M (2012) Recent advances in soybean transformation and their application

to molecular breeding and genomic analysis Breeding Science 61(5):480-494

1-24 Genomics approach for advancement of research in livestock production

Original Papers

1. Ando A, Shigenari A, Ota M, Sada M, Kawata H, Azuma F, Kojima-Shibata C, Nakajoh M, Suzuki K,

Uenishi H, Kulski J.K, Inoko H (2011) SLA-DRB1 and -DQB1 genotyping by the PCRÐSSOPÐLuminex

method Tissue Antigens 78(1):49-55

2. Gotoh H, Hirawatari K, Hanzawa N, Miura I, Wakana S (2012) QTL on mouse chromosomes 1 and 4

causing sperm-head morphological abnormality and male subfertility Mammalian Genome 23(7-8):399-403

3. Katayama M, Fukuda T, Okamuara T, Suda Y, Suzuki E, Uenishi H, Suzuki K (2011) Immunophenotype

characterization for swine selected line, which is resistant for the Mycoplasma Pneumonia Asian-

Australasian Journal of Animal Sciences 24(7):889-897

4. Kojima M, Ashino T, Yoshida T, Iwakura Y, Degawa M (2011) Interleukin-1 controls the constitutive

－ 37 －

expression of the Cyp7a1 gene by regulating the expression of Cyp7a1 transcriptional regulators in the

mouse liver Biological & Pharmaceutical Bulletin 34(10):1644-1647

5. Kojima M, Ashino T, Yoshida T, Iwakura Y, Degawa M (2012) Involvement of interleukin-1 in lead nitrate-

induced hypercholesterolemia in mice Biological & Pharmaceutical Bulletin 35(2):246-250

6. Matsumoto T, Nakajima I, Eguchi-Ogawa T, Nagamura Y, Hamasima N, Uenishi H (2012) Changes in gene

expression in a porcine preadipocyte cell line during differentiation Animal Genetics 43(5):535-544

7. Matsumoto T, Okumura N, Uenishi H, Hayashi T, Hamasima N, Awata T (2012) Population structure of

pigs determined by single nucleotide polymorphisms observed in assembled expressed sequence tags

Animal Science Journal 83(1):14-22

8. Matsumoto T, Okumura N, Uenishi H, Hayashi T, Hamasima N, Awata T (2012) Porcine single nucleotide

polymorphism (SNP) development and population structure of pigs assessed by validated SNPs

Biochemical Genetics 50(5-6):428-439

9. Morozumi T, Toki D, Eguchi-Ogawa T, Uenishi H (2011) A rapid and cost-effective method for sequencing

pooled cDNA clones by using a combination of transposon insertion and Gateway technology

Biotechniques 51(3):195-197

10. Muneta Y, Minagawa Y, Kusumoto M, Shinkai H, Uenishi H, Splichal I (2012) Development of allele-

specific primer PCR for a swine TLR2 SNP and comparison of the frequency among several pig breeds of

Japan and the Czech Republic Journal of Veterinary Medical Science 74(5):553-559

11. Muneta Y, Minagawa Y, Kusumoto M, Shinkai H, Uenishi H, Splichal I (2012) Allele-specific primer

polymerase chain reaction for a single nucleotide polymorphism (C1205T) of swine Toll-like receptor 5 and

comparison of the allelic frequency among several pig breeds in Japan and the Czech Republic

Microbiology and Immunology 56(6):385-391

12. Nemoto K, Ito S, Yoshida C, Miyata M, Kojima M, Degawa M (2011) Hepatic expression of spermatogenic

genes and their transiently remarkable downregulations in Wistar-Kyoto rats in response to lead-nitrate

administration: strain-difference in the gene expression patterns The Journal of Toxicological Sciences

36(3):357-364

13. Nemoto K, Yasuda T, Hikida T, Ito S, Kojima M, Degawa M (2011) Effects of lead nitrate on liver weight

and serum total cholesterol amounts in stroke-prone spontaneously hypertensive and Wistar-Kyoto rats

Journal of Health Science 57(2):192-196

14. Sato M, Ohtsuka M, Miura H, Miyoshi K, Watanabe S (2012) Determination of the optimal concentration of

several selective drugs useful for generating multi-transgenic porcine embryonic fibroblasts Reproduction in

Domestic Animals 47(5):759-765

15. Sato M, Akasaka E, Saitoh I, Ohtsuka M, Watanabe S (2012) Development of a technique for efficient gene

transfer to antral follicular cells in the mouse ovary Systems Biology in Reproductive Medicine 58(3):136-

141

16. Sato S, Hayashi T, Kobayashi E (2011) Characterization of porcine autism susceptibility candidate 2 as a

candidate gene for the number of corpora lutea in pigs Animal Reproduction Science 126(3-4):211-220

17. Suto J (2011) Identification of the modifier locus that suppresses neonatal lethality in (♀DDD×♂DH-

Dh/+) F1-Dh/+ male mice Journal of Veterinary Medical Science 73(10):1327-1332

－ 38 －

18. Suto J (2011) Genetic dissection of testis weight in mice: quantitative trait locus analysis using F2

intercrosses between strains with extreme testis weight, and association study using Y-consomic strains

Mammalian Genome 22(11-12):648-660

19. Suto J (2012) Quantitative trait loci that control plasma lipid levels in an F2

intercross between C57BL/6J and

DDD.Cg-Ay inbred mouse strains Journal of Veterinary Medical Science 74(4):449-456

20. Suzuki K, Kobayashi E, Yamashita H, Uenishi H, Churkina I, Plastow G, Hamasima N, Mitsuhashi T (2012)

Structural analysis of MHC alleles in an RSV tumour regression chicken using a BAC library Animal

Genetics 43(3):348-351

Monograph (in English)

1. Lunney J.K, Eguchi-Ogawa T, Uenishi H, Wertz N, Butler J.E (2011) Immunogenetics The Genetics of the

Pig, 2nd Edition (6):101-133

1-25 Structural and functional analysis of biomolecules related to agriculture

Original Papers

1. Fujimoto Z, Kimura K (2012) Crystal structure of bacteriophage ΦNIT1 zinc peptidase PghP that

hydrolyzes γ-glutamyl linkage of bacterial poly-γ-glutamate Proteins: Structure, Function, and

Bioinformatics 80(3):722-732

2. Ichinose H, Diertavitian S, Fujimoto Z, Kuno A, Leggio L.L, Kaneko S (2012) Structure-based engineering

of glucose specificity in a family 10 xylanase from Streptomyces olivaceoviridis E-86 Process Biochemistry

47(3):358-365

3. Kim Y-M, Kiso Y, Muraki T, Kang M-S, Nakai H, Saburi W, Lang W, Kang H-K, Okuyama M, Mori H,

Suzuki R, Funane K, Suzuki N, Momma M, Fujimoto Z, Oguma T, Kobayashi M, Kim D, Kimura A (2012)

Novel dextranase catalyzing cycloisomaltooligosaccharide-formation and identification of catalytic amino

acids and their functions using chemical rescue approach Journal of Biological Chemistry 287(24):19927-

19935

4. Kim Y-M, Shimizu R, Nakai H, Mori H, Okuyama M, Kang M-S, Fujimoto Z, Funane K, Kim D, Kimura A

(2011) Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic

activity Applied Microbiology and Biotechnology 91(2):329-339

5. Kitamoto K.H, Shinozaki Y, Cao X-h, Morita T, Konishi M, Tago K, Kajiwara H, Koitabashi M, Yoshida S,

Watanabe T, Sameshima-Yamashita Y, Nakajima-Kambe T, Tsushima S (2011) Phyllosphere yeasts rapidly

break down biodegradable plastics AMB Express 1:44

6. Kiyota S, Xie X, Takano M (2012) Phytochromes A and C cooperatively regulate early and transient gene

expression after red-light irradiation in rice seedlings Plant Physiology and Biochemistry 51:10-17

7. Matsuura Y, Takehira M, Sawano M, Ogasahara K, Tanaka T, Yamamoto H, Kunishima N, Katoh E, Yutani

K (2012) Role of charged residues in stabilization of Pyrococcus horikoshii CutA1, which has a

denaturation temperature of nearly 150 ¡C FEBS Journal 279(1):78-90

－ 39 －

8. Michikawa M, Ichinose H, Momma M, Biely P, Jongkees S, Yoshida M, Kotake T, Tsumuraya Y, Withers

S, Fujimoto Z, Kaneko S (2012) Structural and biochemical characterization of glycoside hydrolase family

79 β-glucuronidase from Acidobacterium capsulatum Journal of Biological Chemistry 287(17):14069-

14077

9. Nishikiori M, Mori M, Dohi K, Okamura H, Katoh E, Naito S, Meshi T, Ishikawa M (2011) A host small

GTP-binding protein ARL8 plays crucial roles in tobamovirus RNA replication PLoS Pathogens

7(12):e1002409

10. Okamura H, Nishikiori M, Xiang H, Ishikawa M, Katoh E (2011) Interconversion of two GDP-bound

conformations and their selection in an Arf-family small G protein Structure 19(7):988-998

11. Shimoda Y, Han L, Yamazaki T, Suzuki R, Hayashi M, Imaizumi-Anraku H (2012) Rhizobial and fungal

symbioses show different requirements for calmodulin binding to calcium calmodulinÐdependent protein

kinase in Lotus japonicus The Plant Cell 24(1):304-321

12. Shinkai H, Okumura N, Suzuki R, Muneta Y, Uenishi H (2012) Toll-like receptor 4 polymorphism impairing

lipopolysaccharide signaling in Sus scrofa, and its restricted distribution among Japanese wild boar

populations DNA and Cell Biology 31(4):575-581

13. Shinya S, Ohnuma T, Kawamura S, Torikata T, Nishimura S, Katoh E, Fukamizo T (2011) Interaction of a

goose-type lysozyme with chitin oligosaccharides as determined by NMR spectroscopy The Journal of

Biochemistry 150(5):569-577

14. Suzuki N, Kim Y-M, Fujimoto Z, Momma M, Kang H-K, Funane K, Okuyama M, Mori H, Kimura A (2011)

Crystallization and preliminary crystallographic analysis of dextranase from Streptococcus mutans Acta

Crystallographica Section F 67(12):1542-1544

15. Suzuki N, Kim Y-M, Fujimoto Z, Momma M, Okuyama M, Mori H, Funane K, Kimura A (2012) Structural

elucidation of dextran degradation mechanism by Streptococcus mutans dextranase belonging to glycoside

hydrolase family 66 Journal of Biological Chemistry 287(24):19916-19926

16. Suzuki R, Fujimoto Z, Shiotsuki T, Tsuchiya W, Momma M, Tase A, Miyazawa M, Yamazaki T (2011)

Structural mechanism of JH delivery in hemolymph by JHBP of silkworm, Bombyx mori Scientific Reports

1:133

17. Suzuki R, Tsuchiya W, Shindo H, Yamazaki T (2011) NMR assignments of ubiquitin fold domain (UFD) in

SUMO-activating enzyme subunit 2 from rice Biomolecular NMR Assignments 5(2):245-248

18. Teramoto H, Kojima K, Kajiwara H, Ishibashi J (2012) Expansion of the amino acid repertoire in protein

biosynthesis in silkworm cells ChemBioChem 13(1):61-65

19. Xiang H, Ishibashi K, Nishikiori M, Jaudal M.C, Ishikawa M, Katoh E (2012) Expression, purification, and

functional characterization of a stable helicase domain from a tomato mosaic virus replication protein

Protein Expression and Purification 81(1):89-95

20. Xiang H, Niiyama M, Sugiyama S, Adachi H, Takano K, Murakami S, Inoue T, Mori Y, Ishikawa M,

Matsumura H, Katoh E (2011) Crystallization and preliminary X-ray crystallographic analysis of a helicase-

like domain from a tomato mosaic virus replication protein Acta Crystallographica Section F 67(12):1649-

1652

－ 40 －

2-11 Elucidation of the mechanisms involved in biomass production, growth,

differentiation, and environmental response of agricultural crops

Original Papers

1. Akasaka M, Konishi S, Izawa T, Ushiki J (2011) Histological and genetic characteristics associated with the

seed-shattering habit of weedy rice (Oryza sativa L.) from Okayama, Japan Breeding Science 61(2):168-173




3. Endo-Higashi N, Izawa T (2011) Flowering-time genes Heading date 1 and Early heading date 1 together

control panicle development in rice Plant and Cell Physiology 52(6):1083-1094

4. Fukami R, Yamamoto K, Okushima L, Kuriyama A, Ishikawa M (2011) Deep supercooling and freezing

behaviors in the leaf blade and leaf sheath of a cold hardy bamboo, Sasa kurilensis, analyzed using

cryomicroscopy Cryobiology and Cryotechnology 57(1):71-76 (in Japanese with English Summary)

5. Fukuda M, Satoh-Cruz M, Wen L, Crofts A.J, Sugino A, Washida H, Okita T.W, Ogawa M, Kawagoe Y,

Maeshima M, Kumamaru T (2011) The small GTPase Rab5a is essential for intracellular transport of

proglutelin from the Golgi apparatus to the protein storage vacuole and endosomal membrane organization

in developing rice endosperm Plant Physiology 157(2):632-644

6. Ikeda-Kawakatsu K, Maekawa M, Izawa T, Itoh J, Nagato Y (2012) ABERRANT PANICLE

ORGANIZATION 2/RFL, the rice ortholog of Arabidopsis LEAFY, suppresses the transition from

inflorescence meristem to floral meristem through interaction with APO1 The Plant Journal 69(1):168-180

7. Itoh H, Izawa T (2011) A study of phytohormone biosynthetic gene expression using a circadian clock-

related mutant in rice Plant Signaling & Behavior 6(12):1932-1936

8. Iwamoto M, Kiyota S, Hanada A, Yamaguchi S, Takano M (2011) The multiple contributions of

phytochromes to the control of internode elongation in rice Plant Physiology 157(3):1187-1195

9. Izawa T, Mihara M, Suzuki Y, Gupta M, Itoh H, Nagano A.J, Motoyama R, Sawada Y, Yano M, Yokota

Hirai M, Makino A, Nagamura Y (2011) Os-GIGANTEA confers robust diurnal rhythms on the global

transcriptome of rice in the field The Plant Cell 23(5):1741-1755

10. Kiyota S, Xie X, Takano M (2012) Phytochromes A and C cooperatively regulate early and transient gene

expression after red-light irradiation in rice seedlings Plant Physiology and Biochemistry 51:10-17

11. Nagamine A, Matsusaka H, Ushijima T, Kawagoe Y, Ogawa M, Okita T.W, Kumamaru T (2011) A role for

the Cysteine-rich 10 kDa prolamin in protein body I formation in rice Plant and Cell Physiology

52(6):1003-1016

12. Osugi A, Itoh H, Ikeda-Kawakatsu K, Takano M, Izawa T (2011) Molecular dissection of the roles of

phytochrome in photoperiodic flowering in rice Plant Physiology 157(3):1128-1137

13. Tanaka N, Itoh H, Sentoku N, Kojima M, Sakakibara H, Izawa T, Itoh J, Nagato Y (2011) The COP1

ortholog PPS regulates the juvenile-adult and vegetative-reproductive phase changes in rice The Plant Cell

23(6):2143-2154

14. Tanaka W, Toriba T, Ohmori Y, Yoshida A, Kawai A, Mayama-Tsuchida T, Ichikawa H, Mitsuda N, Ohme-

－ 41 －

Takagi M, Hirano H (2012) The YABBY gene TONGARI-BOUSHI1 is involved in lateral organ development

and maintenance of meristem organization in the rice spikelet The Plant Cell 24(1):80-95

15. Ujiie K, Okuizumi H, Takano M, Ishimaru K (2011) Natural variations in photosynthesis-related traits at an

early vegetative stage of grain sorghum (Sorghum bicolor (L.) Moench) Japanese Journal of Crop Science

80(4):441-447 (in Japanese with English Summary)

16. Yamazaki H, Yoshida S, Ishikawa M (2011) Freezing behavior in blueberry stems analyzed using

differential infra-red thermography and differential thermal analysis Cryobiology and Cryotechnology

57(1):77-81 (in Japanese with English Summary)

17. Yun M-S, Umemoto T, Kawagoe Y (2011) Rice debranching enzyme isoamylase3 facilitates starch

metabolism and affects plastid morphogenesis Plant and Cell Physiology 52(6):1068-1082

Reviews (in English)

1. Muramatsu M, Hihara Y (2012) Acclimation to high-light conditions in cyanobacteria: from gene expression

to physiological responses Journal of Plant Research 125(1):11-39

2. Onda Y, Kawagoe Y (2011) Oxidative protein folding: selective pressure for prolamin evolution in rice

Plant Signaling & Behavior 6(12):1966-1972

2-12 Elucidation of the regulatory mechanisms involved in insect growth,

development and differentiation

Original Papers

1. Daimon T, Fujii T, Yago M, Hsu Y-F, Nakajima Y, Fujii T, Katsuma S, Ishikawa Y, Shimada T (2012)

Female sex pheromone and male behavioral responses of the bombycid moth Trilocha varians: comparison

with those of the domesticated silkmoth Bombyx mori Naturwissenschaften 99(3):207-215

2. Daimon T, Kozaki T, Niwa R, Kobayashi I, Furuta K, Namiki T, Uchino K, Banno Y, Katsuma S, Tamura

T, Mita K, Sezutsu H, Nakayama M, Itoyama K, Shimada T, Shinoda T (2012) Precocious metamorphosis

in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori PLoS Genetics 8(3):e1002486

3. Ichikawa A, Ono H, Echigo T, Mikata Y (2011) Crystal structures and chiral recognition of the diastereomeric

salts prepared from 2-methoxy-2-(1-naphthyl)propanoic acid CrystEngComm 13(14):4536-4548

4. Ishikawa A, Ogawa K, Gotoh H, Walsh T.K, Tagu D, Brisson J.A, Rispe C, S Jaubert-Possamai S, Kanbe T,

Tsubota T, Shiotsuki T, Miura T (2012) Juvenile hormone titre and related gene expression during the

change of reproductive modes in the pea aphid Insect Molecular Biology 21(1):49-60

5. Kaihara K, Kotaki T, Numata H, Ohfune Y, Shinada T (2012) Structure-activity relationship of novel

juvenile hormone, JHSB3, isolated from the stink bug, Plautia stali Tetrahedron 68(1):106-113

6. Kamimura M, Saito H, Niwa R, Niimi T, Toyoda K, Ueno C, Kanamori Y, Shimura S, Kiuchi M (2012)

Fungal ecdysteroid-22-oxidase: a new tool for manipulating ecdysteroid signaling and insect development



－ 42 －


1:133


1. Kotaki T, Shinada T, Numata H (2012) Structure determination of a natural juvenile hormone isolated from

a heteropteran insect Psyche 2012:981475

2. Tanaka Y (2011) Recent topics on the regulatory mechanism of ecdysteroidogenesis by the prothoracic

glands in insects Frontiers in Endocrinology 2:107

2-13 Elucidation of the molecular mechanisms involved in the development and

differentiation of germ cell and stem cell of livestock

Original Papers

1. Akagi S, Yamaguchi D, Matsukawa K, Mizutani E, Hosoe M, Adachi N, Kubo M, Takahashi S (2011)

Developmental ability of somatic cell nuclear transferred embryos aggregated at the 8-cell stage or 16- to

32-cell stage in cattle Journal of Reproduction and Development 57(4):500-506

2. Hasegawa H, Noguchi J, Yamashita M, Okada R, Sugimoto R, Furuya M, Unoki T, Funakoshi Y, Baba T,

Kanaho Y (2012) Phosphatidylinositol 4-phosphate 5-kinase is indispensable for mouse spermatogenesis

Biology of Reproduction 86(5):136

3. Kaedei Y, Naito M, Naoi H, Sato Y, Taniguchi M, Tanihara F, Kikuchi K, Nagai T, Otoi T (2012) Effects of

(－)-epigallocatechin gallate on the motility and penetrability of frozenÐthawed boar spermatozoa incubated

in the fertilization medium Reproduction in Domestic Animals (Published Online):Early View (DOI:

10.1111/j.1439-0531.2012.01984.x)

4. Miyashita N, Kubo Y, Yonai M, Kaneyama K, Saito N, Sawai K, Minamihashi A, Suzuki T, Kojima T,

Nagai T (2011) Cloned cows with short telomeres deliver healthy offspring with normal-length telomeres

Journal of Reproduction and Development 57(5):636-642

5. Naito M (2011) Spermatogonial stem cells Avian Biology Research 4(2):66-69

6. Naito M, Harumi T, Kuwana T (2012) Expression of GFP gene in cultured PGCs isolated from embryonic

blood and incorporation into gonads of recipient embryos The Journal of Poultry Science 49(2):116-123

7. Nakagawa S, Maedomari N, Kikuchi K, Nagai T, Miyano T, Fulka J.Jr, Manabe N (2011) Vitrification of

fully grown and growing porcine oocytes using germinal vesicle transfer Journal of Reproduction and

Development 57(3):335-341

8. Nakai M, Ito J, Sato K, Noguchi J, Kaneko H, Kashiwazaki N, Kikuchi K (2011) Pre-treatment of sperm

reduces success of ICSI in the pig Reproduction 142(2):285-293

9. Noguchi M, Yoshioka K, Suzuki C, Itoh S, Kaneko H (2011) An efficient protocol for inducing

pseudopregnancy using estradiol dipropionate and follicular development associated with changes in

reproductive hormones after prostaglandin F2alpha treatment in pseudopregnant sows Reproductive Biology

and Endocrinology 9:157

－ 43 －

10. Okada K, Kimura M, Moriyama Y, Nakai M, Kikuchi K, Kaneko H, Kunieda T, Baba T, Noguchi J (2011)

Expression analysis of MIF4GD in the rat testis Journal of Reproduction and Development 57(2):256-261

11. Somfai T, Kaneda M, Akagi S, Watanabe S, Haraguchi S, Mizutani E, Dang-Nguyen T.Q, Geshi M, Kikuchi

K, Nagai T (2011) Enhancement of lipid metabolism with L-carnitine during in vitro maturation improves

nuclear maturation and cleavage ability of follicular porcine oocytes Reproduction, Fertility and

Development 23(7):912-920

12. Somfai T, Kikuchi K, Kaneda M, Akagi S, Watanabe S, Mizutani E, Haraguchi S, Dang-Nguyen T.Q, Inaba

Y, Geshi M, Nagai T (2011) Cytoskeletal abnormalities in relation with meiotic competence and ageing in

porcine and bovine oocytes during in vitro maturation Anatomia, Histologia, Embryologia 40(5):335-344

13. Viet Linh N, Kikuchi K, Nakai M, Noguchi J, Kaneko H, Dang-Nguyen T.Q, Maedomari N, Nguyen B.X,

Nagai T, Manabe N (2011) Improvement of porcine oocytes with low developmental ability after fusion of

cytoplasmic fragments prepared by serial centrifugation Journal of Reproduction and Development

57(5):620-626


1. Dang-Nguyen T.Q, Somfai T, Haraguchi S, Kikuchi K, Tajima A, Kanai Y, Nagai T (2011) In vitro

production of porcine embryos: current status, future perspectives and alternative applications Animal

Science Journal 82(3):374-382

2. Kikuchi K, Nakai M, Kashiwazaki N, Kaneko H (2011) Xenografting of gonadal tissues into mice as a

possible method for conservation and utilization of porcine genetic resources Animal Science Journal

82(4):495-503

3. Nakai M, Kashiwazaki N, Ito J, Maedomari N, Ozawa M, Shino M, Noguchi J, Kaneko H, Kikuchi K

(2011) Factors affecting fertilization and embryonic development during intracytoplasmic sperm injection in

pigs Journal of Reproduction and Development 57(2):183-187

4. Somfai T, Kikuchi K, Nagai T (2012) Factors affecting cryopreservation of porcine oocytes Journal of

Reproduction and Development 58(1):17-24

2-14 Elucidation of the mechanisms involved in the control of the behavior and

reproduction of livestock

Original Papers

1. Hagiwara S, Ohta S, Okamoto C, Shibata T, Yayou K, Ito S (2011) Effects of sawdust or grass-sod floor

environments on behaviour of meat-type chickens, broilers and Kumamoto breed Journal of Warm

Regional Society of Animal Science, Japan 54(1):5-14 (in Japanese with English Summary)

2. Hayashi K, Ushizawa K, Hosoe M, Takahashi T (2011) Differential gene expression of serine protease

inhibitors in bovine ovarian follicle: possible involvement in follicular growth and atresia Reproductive

Biology and Endocrinology 9:72

－ 44 －

3. Kasuya E, Kushibiki S, Yayou K, Hodate K, Ogino M, Sutoh M (2012) The effect of lighting conditions on

the rhythmicity of growth hormone secretion in Holstein steers Animal Science Journal 83(3):238-244

4. Komatsu M, Kojima M, Okamura H, Nishio M, Kaneda M, Kojima T, Takeda H, Malau-Aduli A.E.O,

Takahashi H (2012) Age-related changes in gene expression of the growth hormone secretagogue and

growth hormone-releasing hormone receptors in Holstein-Friesian cattle Domestic Animal Endocrinology

42(2):83-93

5. Koshi K, Ushizawa K, Kizaki K, Takahashi T, Hashizume K (2011) Expression of endogenous retrovirus-

like transcripts in bovine trophoblastic cells Placenta 32(7):493-499

6. Matsuyama S, Ohkura S, Mogi K, Wakabayashi Y, Mori Y, Tsukamura H, Maeda K, Ichikawa M, Okamura

H (2011) Morphological evidence for direct interaction between kisspeptin and gonadotropin-releasing

hormone neurons at the median eminence of the male goat: An immunoelectron microscopic study

Neuroendocrinology 94(4):323-332

7. Mishra B, Kizaki K, Koshi K, Ushizawa K, Takahashi T, Hosoe M, Sato T, Ito A, Hashizume K (2012)

Expression of extracellular matrix metalloproteinase inducer (EMMPRIN) and its expected roles in the

bovine endometrium during gestation Domestic Animal Endocrinology 42(2):63-73

8. Murata K, Wakabayashi Y, Sakamoto K, Tanaka T, Takeuchi Y, Mori Y, Okamura H (2011) Effects of brief

exposure of male pheromone on multiple-unit activity at close proximity to kisspeptin neurons in the goat

arcuate nucleus Journal of Reproduction and Development 57(2):197-202

9. Ogino M, Matsuura A, Yamazaki A, Irimajiri M, Kushibiki S, Shingu H, Kasuya E, Hodate K (2011)

Maximum entropy method for the spectral analysis of secretory rhythms of plasma growth hormone and

prolactin in cattle Nihon Chikusan Gakkaiho 83(1):35-46 (in Japanese with English Summary)

10. Sakumoto R, Vermehren M, Kenngott R.A, Okuda K, Sinowatz F (2011) Localization of gene and protein

expressions of tumor necrosis factor-α and tumor necrosis factor receptor types I and II in the bovine corpus

luteum during the estrous cycle Journal of Animal Science 89(10):3040-3047

11. Suzuki Y, Koshi K, Imai K, Takahashi T, Kizaki K, Hashizume K (2011) Bone morphogenetic protein-4

accelerates the establishment of bovine trophoblastic cell lines Reproduction 142(5):733-743

12. Tanaka T, Ohkura S, Wakabayashi Y, Okamura H (2012) Effect of peripherally administered kisspeptin-10

on GnRH neurosecretion into the hypophyseal portal circulation in ovariectomized goat does Small

Ruminant Research 105(1-3):273-276

13. Uenoyama Y, Inoue N, Pheng V, Homma T, Takase K, Yamada S, Ajiki K, Ichikawa M, Okamura H, Maeda

K.I, Tsukamura H (2011) Ultrastructural evidence of kisspeptin-gonadotrophin-releasing hormone (GnRH)

interaction in the median eminence of female rats: Implication of axo-axonal regulation of GnRH release

Journal of Neuroendocrinology 23(10):863-870

14. Yayou K, Kitagawa S, Ito S, Kasuya E, Sutoh M (2011) Effect of oxytocin, prolactin-releasing peptide, or

corticotropin-releasing hormone on feeding behavior in steers General and Comparative Endocrinology

174(3):287-291

－ 45 －

Review (in English)

1. Yayou K (2012) Central regulation of stress response in cattle Japan Agricultural Research Quarterly

46(1):27-34

2-21 Elucidation of the mechanisms involved in plant pathogenic microbe infections

and development of innovative technologies for their applications

Original Papers

1. Abe H, Tomitaka Y, Shimoda T, Seo S, Sakurai T, Kugimiya S, Tsuda S, Kobayashi M (2012) Antagonistic

plant defense system regulated by phytohormones assists interactions among vector insect, thrips and a

tospovirus Plant and Cell Physiology 53(1):204-212

2. Akimoto-Tomiyama C, Furutani A, Tsuge S, Washington E.J, Nishizawa Y, Minami E, Ochiai H (2012)

XopR, a type III effector secreted by Xanthomonas oryzae pv. oryzae, suppresses microbe-associated

molecular pattern-triggered immunity in Arabidopsis thaliana Molecular Plant-Microbe Interactions

25(4):505-514




4. Bogdanove A.J, Koebnik R, Lu H, Furutani A, Angiuoli S.V, Patil P.B, Van Sluys M-A, Ryan R.P, Meyer

D.F, Han S-W, Aparna G, Rajaram M, Delcher A.L, Phillippy A.M, Puiu D, Schatz M.C, Shumway M,

Sommer D.D, Trapnell C, Benahmed F, Dimitrov G, Madupu R, Radune D, Sullivan S, Jha G, Ishihara H,

Lee S-W, Pandey A, Sharma V, Sriariyanun M, Szurek B, Vera-Cruz C.M, Dorman K.S, Ronald P.C,

Verdier V, Dow J.M, Sonti R.V, Tsuge S, Brendel V, Rabinowicz P.D, Leach J.E, White F.F, Salzberg S.L

(2011) Two new complete genome sequences offer insight into host and tissue specificity of plant

pathogenic Xanthomonas spp. Journal of Bacteriology 193(19):5450-5464

5. Iki T, Yoshikawa M, Meshi T, Ishikawa M (2012) Cyclophilin 40 facilitates HSP90-mediated RISC

assembly in plants The EMBO Journal 31(2):267-278

6. Imai T, Ohashi Y, Mitsuhara I, Seo S, Toshima H, Hasegawa M (2012) Identification of a degradation

intermediate of the momilactone a rice phytoalexin by the rice blast fungus Bioscience, Biotechnology and

Biochemistry 76(2):414-416

7. Kawazu K, Mochizuki A, Sato Y, Sugeno W, Murata M, Seo S, Mitsuhara I (2012) Different expression

profiles of jasmonic acid and salicylic acid inducible genes in the tomato plant against herbivores with

various feeding modes Arthropod-Plant Interactions 6(2):221-230

8. Marui J, Yamane N, Ohashi-Kunihiro S, Ando T, Terabayashi Y, Sano M, Ohashi S, Ohshima E, Tachibana

K, Higa Y, Nishimura M, Koike H, Machida M (2011) Kojic acid biosynthesis in Aspergillus oryzae is

regulated by a Zn(II)2Cys

6transcriptional activator and induced by kojic acid at the transcriptional level

Journal of Bioscience and Bioengineering 112(1):40-43

9. Murota K, Hagiwara-Komoda Y, Komoda K, Onouchi H, Ishikawa M, Naito S (2011) Arabidopsis cell-free

－ 46 －

extract, ACE, a new in vitro translation system derived from Arabidopsis callus cultures Plant and Cell

Physiology 52(8):1443-1453

10. Nishikiori M, Mori M, Dohi K, Okamura H, Katoh E, Naito S, Meshi T, Ishikawa M (2011) A host small

GTP-binding protein ARL8 plays crucial roles in tobamovirus RNA replication PLoS Pathogens

7(12):e1002409

11. Okamura H, Nishikiori M, Xiang H, Ishikawa M, Katoh E (2011) Interconversion of two GDP-bound

conformations and their selection in an Arf-family small G protein Structure 19(7):988-998

12. Xiang H, Ishibashi K, Nishikiori M, Jaudal M.C, Ishikawa M, Katoh E (2012) Expression, purification, and

functional characterization of a stable helicase domain from a tomato mosaic virus replication protein

Protein Expression and Purification 81(1):89-95

13. Xiang H, Niiyama M, Sugiyama S, Adachi H, Takano K, Murakami S, Inoue T, Mori Y, Ishikawa M,

Matsumura H, Katoh E (2011) Crystallization and preliminary X-ray crystallographic analysis of a helicase-

like domain from a tomato mosaic virus replication protein Acta Crystallographica Section F 67(12):1649-

1652

2-22 Elucidation of the mechanisms involved in crop response to microbial infection

and development of crop strains with multiple pathogen resistance

Original Papers




2. Ashikawa I, Hayashi N, Abe F, Wu J, Matsumoto T (2012) Characterization of the rice blast resistance gene

Pik cloned from Kanto51 Molecular Breeding 30(1):485-494

3. Fukumoto T, Kano A, Ohtani K, Yamasaki-Kokudo Y, Kim B-G, Hosotani K, Saito M, Shirakawa C,

Tajima S, Izumori K, Ohara T, Shigematsu Y, Tanaka K, Ishida Y, Nishizawa Y, Tada Y, Ichimura K, Gomi

K, Akimitsu K (2011) Rare sugar D-allose suppresses gibberellin signaling through hexokinase-dependent

pathway in Oryza sativa L. Planta 234(6):1083-1095

4. Fukushima S, Yoshida M, Takatsuji H (2012) Role of linkers between zinc fingers in spacing recognition by

plant TFIIIA-type zinc-finger proteins Journal of Amino Acids 2012:848037

5. Higuchi-Takeuchi M, Ichikawa T, Kondou Y, Matsui K, Hasegawa Y, Kawashima M, Sonoike K, Mori M,

Hirochika H, Matsui M (2011) Functional analysis of two isoforms of leaf-type ferredoxin-NADP+-

oxidoreductase in rice using the heterologous expression system of Arabidopsis Plant Physiology 157(1):96-

108

6. Kano A, Hosotani K, Gomi K, Yamasaki-Kokudo Y, Shirakawa C, Fukumoto T, Ohtani K, Tajima S,

Izumori K, Tanaka K, Ishida Y, Nishizawa Y, Ichimura K, Tada Y, Akimitsu K (2011) D-Psicose induces

upregulation of defense-related genes and resistance in rice against bacterial blight Journal of Plant

Physiology 168(15):1852-1857

－ 47 －

7. Kim S-H, Oikawa T, Kyozuka J, Wong H.L, Umemura K, Kishi-Kaboshi M, Takahashi A, Kawano Y,

Kawasaki T, Shimamoto K (2012) The bHLH rac immunity1 (RAI1) is activated by OsRac1 via OsMAPK3

and OsMAPK6 in rice immunity Plant and Cell Physiology 53(4):740-754

8. Konishi T, Aohara T, Igasaki T, Hayashi N, Miyazaki Y, Takahashi A, Hirochika H, Iwai H, Satoh S, Ishii T

(2011) Down-regulation of UDP-arabinopyranose mutase reduces the proportion of arabinofuranose present

in rice cell walls Phytochemistry 72(16):1962-1968

9. Kouzai Y, Mochizuki S, Saito A, Ando A, Minami E, Nishizawa Y (2012) Expression of a bacterial

chitosanase in rice plants improves disease resistance to the rice blast fungus Magnaporthe oryzae Plant

Cell Reports 31(4):629-636

10. Mentlak T.A, Kombrink A, Shinya T, Ryder L.S, Otomo I, Saitoh H, Terauchi R, Nishizawa Y, Shibuya N,

Thomma B.P.H.J, Talbota N.J (2012) Effector-mediated suppression of chitin-triggered immunity by

Magnaporthe oryzae is necessary for rice blast disease The Plant Cell 24(1):322-335

11. Miyamoto K, Shimizu T, Mochizuki S, Nishizawa Y, Minami E, Nojiri H, Yamane H, Okada K (2012)

Stress-induced expression of the transcription factor RERJ1 is tightly regulated in response to jasmonic acid

accumulation in rice Protoplasma (Online First) (DOI: 10.1007/s00709-012-0400-z)

12. Nakagawa H, Tanaka A, Tanabata T, Ohtake M, Fujioka S, Nakamura H, Ichikawa H, Mori M (2012)

SHORT GRAIN 1 decreases organ elongation and brassinosteroid response in rice Plant Physiology

158(3):1208-1219

13. Seo E-K, Nakamura H, Mori M, Asami T (2012) Screening and characterization of a chemical regulatorfor

plant disease resistance Bioorganic & Medicinal Chemistry Letters 22(4):1761-1765

14. Shimono M, Koga H, Akagi A, Hayashi N, Goto S, Sawada M, Kurihara T, Matsushita A, Sugano S, Jiang

C-J, Kaku H, Inoue H, Takatsuji H (2012) Rice WRKY45 plays important roles in fungal and bacterial

disease resistance Molecular Plant Pathology 13(1):83-94

15. Yamaguchi K, Imai K, Akamatsu A, Mihashi M, Hayashi N, Shimamoto K, Kawasaki T (2012) SWAP70

functions as a Rac/Rop guanine nucleotide-exchange factor in rice The Plant Journal 70(3):389-397

16. Yazawa K, Jiang C-J, Kojima M, Sakakibara H, Takatsuji H (2012) Reduction of abscisic acid levels or

inhibition of abscisic acid signaling in rice during the early phase of Magnaporthe oryzae infection

decreases its susceptibility to the fungus Physiological and Molecular Plant Pathology 78:1-7

Monographs (in English)

1. Higuchi M, Kondou Y, Mori M, Ichikawa T, Matsui M (2012) Characterization of rice genes using a

heterologous full-length cDNA expression system Transgenic Plants: Methods and Protocols 3(8):75-90

2. Nishizawa Y, Mochizuki S, Saitoh K, Kishimoto K, Kouzai Y, Minami E (2011) Defense mechanisms

mediated by chitin in rice-blast interactions Genome-Enabled Analysis of Plant-Pathogen Interactions

:121-129

－ 48 －

2-23 Elucidation of the mechanisms involved in plant and soil microbe symbioses

Original Papers

1. Fukai E, Soyano T, Umehara Y, Nakayama S, Hirakawa H, Tabata S, Sato S, Hayashi M (2012)

Establishment of a Lotus japonicus gene tagging population using the exon-targeting endogenous

retrotransposon LORE1 The Plant Journal 69(4):720-730

2. Hakoyama T, Niimi K, Yamamoto T, Isobe S, Sato S, Nakamura Y, Tabata S, Kumagai H, Umehara Y,

Brossuleit K, Petersen T.R, Sandal N, Stougaard J, Udvardi M.K, Tamaoki M, Kawaguchi M, Kouchi H,

Suganuma N (2012) The integral membrane protein SEN1 is required for symbiotic nitrogen fixation in

Lotus japonicus nodules Plant and Cell Physiology 53(1):225-236

3. Ikeda S, Okubo T, Takeda N, Banba M, Sasaki K, Imaizumi-Anraku H, Fujihara S, Ohwaki Y, Ohshima K,

Fukuta Y, Eda S, Mitsui H, Hattori M, Sato T, Shinano T, Minamisawa K (2011) The genotype of the

calcium/calmodulin-dependent protein kinase gene (CCaMK) determines bacterial community diversity in

rice roots under paddy and upland field conditions Applied and Environmental Microbiology 77(13):4399-

4405

4. Murakami E, Nagata M, Shimoda Y, Kucho K, Higashi S, Abe M, Hashimoto M, Uchiumi T (2011) Nitric

oxide production induced in roots of Lotus japonicus by lipopolysaccharide from Mesorhizobium loti Plant

and Cell Physiology 52(4):610-617

5. Okamoto S, Nakagawa T, Kawaguchi M (2011) Expression and functional analysis of CLV3-like gene in the

model legume Lotus japonicus Plant and Cell Physiology 52(7):1211-1221

6. Shimoda Y, Han L, Yamazaki T, Suzuki R, Hayashi M, Imaizumi-Anraku H (2012) Rhizobial and fungal

symbioses show different requirements for calmodulin binding to calcium calmodulinÐdependent protein

kinase in Lotus japonicus The Plant Cell 24(1):304-321

7. Shiro S, Yamamoto A, Umehara Y, Hayashi M, Yoshida N, Nishiwaki A, Yamakawa T, Saeki Y (2012)

Effect of Rj-genotype and cultivation temperature on the community structure of soybean-nodulating

bradyrhizobia Applied and Environmental Microbiology 78(4):1243-1250

8. Takeda N, Maekawa T, Hayashi M (2012) Nuclear-localized and deregulated calcium- and calmodulin-

dependent protein kinase activates rhizobial and mycorrhizal responses in Lotus japonicus The Plant Cell

24(2):810-822

Review (in English)

1. Hayashi M, Saeki Y, Haga M, Harada K, Kouchi H, Umehara Y (2012) Rj (rj) genes involved in nitrogen-

fixing root nodule formation in soybean Breeding Science 61(5):544-553

－ 49 －

2-24 Elucidation of the mechanisms involved in insect pest infestation and plant

resistance to insects

Original Papers

1. Abe H, Tomitaka Y, Shimoda T, Seo S, Sakurai T, Kugimiya S, Tsuda S, Kobayashi M (2012) Antagonistic

plant defense system regulated by phytohormones assists interactions among vector insect, thrips and a

tospovirus Plant and Cell Physiology 53(1):204-212

2. Asaoka K (2011) Effects of food source and experience on meal characteristics in the feeding behavior of

the monophagous caterpillar, Bombyx mori International Journal of Wild Silkmoth and Silk 16:69-75

3. Fu Q, Matsumoto Y, Matsumura M, Hirai Y, Sato Y, Noda H (2012) Presence of a short repeat sequence in

internal transcribed spacer (ITS) 1 of the rRNA gene of Sogatella furcifera (Hemiptera: Delphacidae) from

geographically different populations in Asia Applied Entomology and Zoology 47(2):95-101

4. Hattori M, Nakamura M, Komatsu S, Tsuchihara K, Tamura Y, Hasegawa T (2012) Molecular cloning of a

novel calcium-binding protein in the secreted saliva of the green rice leafhopper Nephotettix cincticeps

Insect Biochemistry and Molecular Biology 42(1):1-9

5. Honda K, ïmura H, Chachin M, Kawano S, Inoue T.A (2011) Synergistic or antagonistic modulation of

oviposition response of two swallowtail butterflies, Papilio maackii and P. protenor, to Phellodendron

amurense by its constitutive prenylated flavonoid, phellamurin Journal of Chemical Ecology 37(6):575-581

6. Kawazu K, Mochizuki A, Sato Y, Sugeno W, Murata M, Seo S, Mitsuhara I (2012) Different expression

profiles of jasmonic acid and salicylic acid inducible genes in the tomato plant against herbivores with

various feeding modes Arthropod-Plant Interactions 6(2):221-230

7. Matsumoto Y, Matsumura M, Hoshizaki S, Sato Y, Noda H (2011) The strepsipteran parasite Elenchus

japonicus (Strepsiptera, Elenchidae) of planthoppers consists of three genotypes Applied Entomology and

Zoology 46(3):435-442

8. Mita T, Matsumoto Y (2012) First description of the male of Gonatopus javanus (R.C.L. Perkins)

determined by mitochondrial COI sequence (Hymenoptera: Dryinidae) Entomological Science 15(2):214-

218

2-25 Elucidation of insect-insect, insect-plant and insect-microbe interactions and

their applications

Original Papers

1. Arakawa T, Namba O, Tateishi K (2011) Effects of coadministration of chemical insecticides with

nucleopolyhedrovirus SpltNPV on the dietary intake of the common cutworm Spodoptera litura

(Lepidoptera: Noctuidae) Applied Entomology and Zoology 46(3):399-405

2. Arakawa T, Yukuhiro F, Noda H (2011) Subacute and delayed toxicity of iminoctadine liquid formulation,

which contains iminoctadine triacetate as an antifungal component on a nontarget domesticated insect, the

silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) Pesticide Biochemistry and Physiology 100(3):239-

243

－ 50 －

3. Asano S, Miyamoto K, Wada S, Mitsuhashi W (2011) Influence of sticker-spreader surfactants on the

stability of Bacillus thuringiensis solution under ultraviolet radiation Annual Report of the Ibaraki Plant

Protection Society (50):27-32 (in Japanese with English summary)

4. Ben Hamouda A, Tanaka S, Ben Hamouda M.H, Bouain A (2011) Density-dependent phenotypic plasticity

in body coloration and morphometry and its transgenerational changes in the migratory locust, Locusta

migratoria Journal of Entomology and Nematology 3(7):105-116

5. Boerjan B, De Loof A, Tanaka S, Schoofs L (2011) Dispersion of peptides in vegetable oil as a simple slow

release formula for both injection and oral uptake in insects: A case study with [His7]-corazonin in an albino

Locusta migratoria deficient in corazonin Peptides 32(7):1536-1539

6. Fujiwara-Tsujii N, Yasui H, Wakamura S, Hashimoto I, Minamishima M (2012) The white-spotted

longicorn beetle, Anoplophora malasiaca (Coleoptera: Cerambycidae), with a blueberry as host plant,

utilizes host chemicals for male orientation Applied Entomology and Zoology 47(2):103-110

7. Fujiwara-Tsujii N, Yasui H, Wakamura S, Tanaka S, Arakaki N (2011) Determination of the amount of sex

pheromone emitted by individual virgin females of the white grub beetle, Dasylepida ishigakiensis

(Coleoptera:Scarabaeidae) at different stages of reproductive life Applied Entomology and Zoology

46(4):527-532

8. Hinomoto N, Higaki T, Maeda T, Nagamori S, Todokoro Y (2011) Evaluation of natural enemies using

microsatellite DNA markers Plant Protection 65(5):274-279 (in Japanese with English summary)

9. Hokama Y, Arakaki N, Nagayama A, Kobayashi A, Ando T, Tanizaki S, Yasui H, Akino T, Wakamura S

(2011) Presence of allochronic populations of the white grub beetle Dasylepida ishigakiensis (Coleoptera:

Scarabaeidae) on Ishigaki Island, Japan Applied Entomology and Zoology 46(4):471-476

10. Kabir K.E, Koga D, Konno K, Takanashi T (2012) Fluorescein-5 isothiocyanate conjugated-chitin-binding

domain probe (FITC-CBD)-coupled detection of chitin in the peritrophic membrane of Monochamus

alternatus (Coleoptera: Cerambycidae) Journal of Asia-Pacific Entomology 15(3):397-400

11. Kishimoto H, Maeda T, Wright L.C, James D.G (2011) Identification of prey consumed by Stethorus

punctum picipes (Casey) (Cleoptera: Coccinellidae) in tree fruit and vines in Washington State, USA

International Journal of Acarology 37(Suppl. 1):216-220

12. Kobayashi H, Kawasaki K, Takeishi K, Noda H (2011) Symbiont of the stink bug Plautia stali synthesizes

rough-type lipopolysaccharide Microbiological Research 167(1):48-54

13. Kobayashi T, Takada M, Takagi S, Yoshioka A, Washitani I (2011) Spider predation on a mirid pest in

Japanese rice fields Basic and Applied Ecology 12(6):532-539

14. Maeda T (2011) Foraging experiences affect the importance of chemical cues in patch-leaving decision of

Neoseiulus womersleyi Schicha (Acari, Phytoseiidae) Journal of the Acarological Society of Japan 20(1):9-

18

15. Maeno K, Tanaka S (2012) Adult female desert locusts require contact chemicals and light for progeny

gregarization Physiological Entomology 37(2):109-118

16. Maeno K, Tanaka S (2011) Phase-specific responses to different qualities of food in the desert locust,

Schistocerca gregaria: Developmental, morphological and reproductive characteristics Journal of Insect


－ 51 －

17. Mitsuhashi W, Ikeda H, Muraji M (2011) Fifty-year trend towards suppression of Wolbachia-induced male-

killing by its butterfly host, Hypolimnas bolina Journal of Insect Science 11:92

18. Muraji M, Wakamura S, Yasui H, Arakaki N, Sadoyama Y, Ohno S, Matsuhira K (2011) Genetic variation

of the white-spotted longicorn beetle Anoplophora spp. (Coleoptera: Cerambycidae) in Japan detected by

mitochondrial DNA sequence Applied Entomology and Zoology 46(3):363-373

19. Narita S, Kageyama D, Hiroki M, Sanpei T, Hashimoto S, Kamitoh T, Kato Y (2011) Wolbachia-induced

feminisation newly found in Eurema hecabe, a sibling species of Eurema mandarina (Lepidoptera: Pieridae)

Ecological Entomology 36(3):309-317

20. Sugiyama K, Hinomoto N, Ohya T, Futamura T (2011) Species discrimination of three commercially

available whitefly parasitoids, Encarsia formosa Gahan, Eretmocerus eremicus Rose and Zolnerowich and

Er. mundus (Mercet) (Hymenoptera: Aphelinidae), by DNA markers Japanese Journal of Applied

Entomology and Zoology 55(4):258-260 (in Japanese with English summary)

21. Takahashi A, Fujiwara-Tsujii N, Yamaoka R, Itoh M, Ozaki M, Takano-Shimizu T (2012) Cuticular

hydrocarbon content that affects male mate preference of Drosophila melanogaster from west Africa

International Journal of Evolutionary Biology 2012:278903

22. Tanaka S, Harano K, Nishide Y (2012) Re-examination of the roles of environmental factors in the control of

body-color polyphenism in solitarious nymphs of the desert locust Schistocerca gregaria with special

reference to substrate color and humidity Journal of Insect Physiology 58(1):89-101

23. Wakamura S, Yasui H, Shimatani M, Tokashiki T, Okuhira T, Fujiwara-Tsujii N, Ishikawa M, Uesato T,

Zukeyama H, TŽbaru H (2011) Sex pheromone of the African armyworm Spodoptera exempta (Lepidoptera:

Noctuidae): identification of components of the Okinawan population and formulation for population

monitoring Applied Entomology and Zoology 46(3):415-420

24. Watanabe M, Tagami Y, Miura K, Kageyama D, Stouthamer R (2012) Distribution patterns of Wolbachia

endosymbionts in the closely related flower bugs of the genus Orius: Implications for coevolution and

horizontal transfer Microbial Ecology 64(2):537-545

25. Yamazaki T, Ishikawa T, Pandian G.N., Okazaki K, Haginoya K, Tachikawa Y, Mitsui T, Miyamoto K,

Angusthanasombat C, Hori H (2011) Midgut juice of Plutella xylostella highly resistant to Bacillus

thuringiensis Cry1Ac contains a three times larger amount of glucosinolate sulfatase which binds to Cry1Ac

compared to that of susceptible strain Pesticide Biochemistry and Physiology 101(2):125-131

26. Yanagawa A, Fujiwara-Tsujii N, Akino T, Yoshimura T, Yanagawa T, Shimizu S (2011) Musty odor of

entomopathogens enhances disease-prevention behaviors in the termite Coptotermes formosanus Journal of

Invertebrate Pathology 108(1):1-6

27. Yasui H, Inouchi J, Wakamura S, Fukaya M, Akino T, Arakaki N (2012) Male mate searching behavior in

the black chafer Holotrichia loochooana loochooana (Sawada) (Coleoptera: Scarabaeidae): response to

pheromone lures of various colors and intensities Journal of Ethology 30(2):233-238

28. Yasui H, Wakamura S, Fujiwara-Tsujii N, Arakaki N, Nagayama A, Hokama Y, Mochizuki F, Fukumoto T,

Oroku H, Harano K, Tanaka S (2012) Mating disruption by a synthetic sex pheromone in the white grub

beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) in the laboratory and sugarcane fields Bulletin of

Entomological Research 102(02):157-164

－ 52 －


1. Chen Y.P, Nakashima N, Christian P.D, Bakonyi T, Bonning B.C, Valles S.M, Lightner D (2011) Family

Iflaviridae Virus Taxonomy Ninth Report of the International Committee on Taxonomy of Viruses :846-

849

2. Chen Y.P, Nakashima N, Christian P.D, Bakonyi T, Bonning B.C, Valles S.M, Lightner D (2011) Family

Dicistroviridae Virus Taxonomy Ninth Report of the International Committee on Taxonomy of Viruses

:840-845

3. Kageyama D, Narita S, Watanabe M (2012) Insect sex determination manipulated by their endosymbionts:

Incidences, mechanisms and implications Insects 3(1):161-199

4. Nakashima N (2011) SINV-1-like viruses, Dicistroviridae The Springer Index of Viruses (23):445-449

2-26 Elucidation of molecular mechanisms in animal immune systems

Original Papers

1. Aoki S, Ikeda S, Takezawa T, Kishi T, Makino J, Uchihashi K, Matsunobu A, Noguchi M, Sugihara H,

Toda S (2011) Prolonged effect of fluid flow stress on the proliferative activity of mesothelial cells after

abrupt discontinuation of fluid streaming Biochemical and Biophysical Research Communications 416(3-

4):391-396

2. Aoki S, Makino J, Nagashima A, Takezawa T, Nomoto N, Uchihashi K, Matsunobu A, Sanai T, Sugihara H,

Toda S (2011) Fluid flow stress affects peritoneal cell kinetics: Possible pathogenesis of peritoneal fibrosis

Peritoneal Dialysis International 31(4):466-476

3. Kawano A, Tsukimoto M, Mori D, Noguchi T, Harada H, Takenouchi T, Kitani H, Kojima S (2012)

Regulation of P2X7-dependent inflammatory functions by P2X4 receptor in mouse macrophages

Biochemical and Biophysical Research Communications 420(1):102-107

4. Kawano A, Tsukimoto M, Noguchi T, Hotta N, Harada H, Takenouchi T, Kitani H, Kojima S (2012)

Involvement of P2X4 receptor in P2X7 receptor-dependent cell death of mouse macrophages Biochemical

and Biophysical Research Communications 419(2):374-380

5. Kita Y.F, Ando A, Tanaka K, Hosomichi K, Uenishi H, Inoko H, Shiina T (2011) Development of MHC

typing method and gene expression method using by next generation sequencer Roche 454 FLX DNA

Polymorphism 19:223-229 (in Japanese with English summary)

6. Kita Y.F, Ando A, Tanaka K, Suzuki S, Ozaki Y, Uenishi H, Inoko H, Kulski J.K, Shiina T (2012)

Application of high-resolution, massively parallel pyrosequencing for estimation of haplotypes and gene

expression levels of swine leukocyte antigen (SLA) class I genes Immunogenetics 64(3):187-199

7. Kitani H, Takenouchi T, Sato M, Yoshioka M, Yamanaka N (2011) A simple and efficient method to isolate

macrophages from mixed primary cultures of adult liver cells Journal of Visualized Experiments (51):2757

8. Kotani N, Maeda K, Watanabe T, Hiramatsu M, Gong L-k, Bi Y-a, Takezawa T, Kusuhara H, Sugiyama Y

(2011) Culture-period-dependent changes in the uptake of transporter substrates in sandwich-cultured rat and

human hepatocytes Drug Metabolism and Disposition 39(9):1503-1510

－ 53 －

9. Otani Y, Yamaguchi Y, Sato Y, Furuichi T, Ikenaka K, Kitani H, Baba H (2011) PLD4 is involved in

phagocytosis of microglia: Expression and localization changes of PLD4 are correlated with activation state

of microglia PLoS ONE 6(11):e27544

10. Sakuma C, Sato M, Takenouchi T, Chiba J, Kitani H (2012) Critical roles of the WASP N-terminal domain

and Btk in LPS-induced inflammatory response in macrophages PLoS ONE 7(1):e30351

11. Sato M, Sawahata R, Takenouchi T, Kitani H (2011) Identification of Fyn as the binding partner for the

WASP N-terminal domain in T cells International Immunology 23(8):493-502

12. Sekiyama K, Fujita M, Sekigawa A, Takamatsu Y, Waragai M, Takenouchi T, Sugama S, Hashimoto M

(2012) Ibuprofen ameliorates protein aggregation and astrocytic gliosis, but not cognitive dysfunction, in a

transgenic mouse expressing dementia with Lewy bodies-linked P123H β-synuclein Neuroscience Letters

515(1):97-101

13. Sekiyama K, Nakai M, Fujita M, Takenouchi T, Waragai M, Wei J, Sekigawa A, Takamatsu Y, Sugama S,

Kitani H, Hashimoto M (2012) Theaflavins stimulate autophagic degradation of α-synuclein in neuronal

cells Open Journal of Neuroscience 2:1

14. Shinkai H, Okumura N, Suzuki R, Muneta Y, Uenishi H (2012) Toll-like receptor 4 polymorphism impairing

lipopolysaccharide signaling in Sus scrofa, and its restricted distribution among Japanese wild boar

populations DNA and Cell Biology 31(4):575-581

15. Sugama S, Takenouchi T, Sekiyama K, Kitani H, Hashimoto M (2011) Immunological responses of astroglia

in the rat brain under acute stress: interleukin 1 beta co-localized in astroglia Neuroscience 192:429-437

16. Tada Y, Takezawa T, Tani A, Nakamura T, Omori K (2012) Collagen vitrigel scaffold for regenerative

medicine of the trachea: Experimental study and quantitative evaluation Acta Oto-Laryngologica

132(4):447-452

17. Takano T, Yamaguchi S, Matsunuma E, Komiya S, Shinkai M, Takezawa T, Nagamune T (2012) Cell

transfer printing from patterned poly(ethylene glycol)-oleyl surfaces to biological hydrogels for rapid and

efficient cell micropatterning Biotechnology and Bioengineering 109(1):244-251

18. Takezawa T, Nishikawa K, Wang P-C (2011) Development of a human corneal epithelium model utilizing a

collagen vitrigel membrane and the changes of its barrier function induced by exposing eye irritant

chemicals Toxicology in Vitro 25(6):1237-1241

19. Tanemoto T, Enosawa S, Takezawa T, Kitagawa M, Chiba T, Tanaka T (2011) Non-suture skin closure

using collagen vitrigel membrane for future repair of endoscopic fetal myelomeningocele Organ Biology

18(3):292-296

20. Uchino T, Takezawa T, Ikarashi Y, Nishimura T (2011) Development of an alternative test for skin

sensitization using a three-dimensional human skin model consisting of dendritic cells, keratinocytes and

fibroblasts Alternatives to Animal Testing and Experimentation 16(1):1-8

21. Uenishi H, Shinkai H, Morozumi T, Muneta Y, Jozaki K, Kojima-Shibata C, Suzuki E (2011)

Polymorphisms in pattern recognition receptors and their relationship to infectious disease susceptibility in

pigs BMC Proceedings 5(Suppl 4):S27

22. Yanagitai M, Itoh S, Kitagawa T, Takenouchi T, Kitani H, Satoh T (2012) Carnosic acid, a pro-electrophilic

compound, inhibits LPS-induced activation of microglia Biochemical and Biophysical Research

Communications 418(1):22-26

－ 54 －


1. Haga S, Takezawa T, Ozawa T, Remington S.J, Morita N, Ozaki M (2012) ÔCollagen vitrigel sheetÕ as a

novel drug delivery bio-material IFMBE Proceedings 37(1):1354-1357

2. Sekiyama K, Sugama S, Fujita M, Sekigawa A, Takamatsu Y, Waragai M, Takenouchi T, Hashimoto M

(2012) Neuroinflammation in Parkinson's disease and related disorders: A lesson from genetically

manipulated mouse models of α-synucleinopathies Parkinson’s Disease 2012:271732

3. Takezawa T, Aoki S, Oshikata A, Okamoto C, Yamaguchi H, Narisawa Y, Toda S (2012) A novel material

of high density collagen fibrils: a collagen xerogel membrane and its application to transplantation in vivo

and a culture chamber in vitro 24th European Conference on Biomaterials :181-185

4. Uenishi H, Shinkai H, Morozumi T, Muneta Y (2012) Genomic survey of polymorphisms in pattern

recognition receptors and their possible relationship to infections in pigs Veterinary Immunology and

Immunopathology 148(1-2):69-73

3-01 Innovation of technologies for development of genetically modified crops and

intensification of their use

Original Papers

1. Hayashi S, Wakasa Y, Takahashi H, Kawakatsu T, Takaiwa F (2012) Signal transduction by IRE1-mediated

splicing of bZIP50 and other stress sensors in the endoplasmic reticulum stress response of rice The Plant

Journal 69(6):946-956

2. Md. Ali E, Tabei Y, Kobayashi K, Yamaoka N, Nishiguchi M (2012) Molecular analysis of transgenic

melon plants showing virus resistance conferred by direct repeat of movement gene of Cucumber green

mottle mosaic virus Plant Cell Reports 31(8):1371-1377

3. Ozawa K, Wakasa Y, Ogo Y, Matsuo K, Kawahigashi H, Takaiwa F (2012) Development of an efficient

Agrobacterium-mediated gene targeting system for rice and analysis of rice knockouts lacking granule-

bound starch synthase (Waxy) and β1,2-xylosyltransferase Plant and Cell Physiology 53(4):755-761

4. Seo M-S, Takahashi S, Kadowaki K, Kawamukai M, Takahara M, Takamizo T (2011) Expression of

CoQ10-producing ddsA transgene by efficient Agrobacterium-mediated transformation in Panicum

meyerianum Plant Cell, Tissue and Organ Culture 107(2):325-332

5. Suzuki K, Yang L, Takaiwa F (2012) Transgenic rice accumulating modified cedar pollen allergen Cry j 2

derivatives Journal of Bioscience and Bioengineering 113(2):249-251

6. Takahashi H, Kawakatsu T, Wakasa Y, Hayashi S, Takaiwa F (2012) A Rice transmembrane bZIP

transcription factor, OsbZIP39, regulates the endoplasmic reticulum stress response Plant and Cell


7. Tsuda M, Konagaya K, Okuzaki A, Kaneko Y, Tabei Y (2011) Occurrence of metaxenia and false hybrids in

Brassica juncea L. cv. Kikarashina × B. napus Breeding Science 61(4):358-365

8. Tucker M.R, Okada T, Johnson S.D, Takaiwa F, Koltunow A.M.G (2012) Sporophytic ovule tissues modulate

the initiation and progression of apomixis in Hieracium Journal of Experimental Botany 63(8):3229-3241

－ 55 －

9. Wakasa Y, Hirano K, Urisu A, Matsuda T, Takaiwa F (2011) Generation of transgenic rice lines with reduced

contents of multiple potential allergens using a null mutant in combination with an RNA silencing method

Plant and Cell Physiology 52(12):2190-2199

10. Yang L, Hirose S, Suzuki K, Hiroi T, Takaiwa F (2012) Expression of hypoallergenic Der f 2 derivatives

with altered intramolecular disulphide bonds induces the formation of novel ER-derived protein bodies in

transgenic rice seeds Journal of Experimental Botany 63(8):2947-2959


1. Hiroi T, Kaminuma O, Takaiwa F (2011) Vaccination with transgenic rice seed expressing mite allergen: a

new option for asthma sufferers? Expert Review of Vaccines 10(9):1249-1251

2. Maruyama N, Motoyama T, Yoshikawa M, Takaiwa F, Utsumi S (2011) Seed storage proteins; strategies for

developing crops promoting human health Soybean - Applications and Technology :243-254

3. Ozawa K (2012) A high-efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.)

Transgenic Plants: Methods and Protocols 3(5):51-57

4. Wakasa Y, Takaiwa F (2012) Use of a callus-specific selection system to develop transgenic rice seed

accumulating a high level of recombinant protein Transgenic Plants: Methods and Protocols 7(36):467-479

3-02 Development of novel technologies for efficient use of genetically modified

silkworm

Original Papers

1. Daimon T, Kozaki T, Niwa R, Kobayashi I, Furuta K, Namiki T, Uchino K, Banno Y, Katsuma S, Tamura

T, Mita K, Sezutsu H, Nakayama M, Itoyama K, Shimada T, Shinoda T (2012) Precocious metamorphosis

in the juvenile hormone-deficient mutant of the silkworm, Bombyx mori PLoS Genetics 8(3):e1002486

2. Hirokawa M, Tatematsu K (2011) The order of four homeotic-mutant genes, Nc, ECw, EKp and EMu, on the 6th-

linkage group of Bombyx mori Journal of Insect Biotechnology and Sericology 80(2):41-48

3. Kameda T, Kojima K, Zhang Q, Sezutsu H (2012) Identification of hornet silk gene with a characteristic

repetitive sequence in Vespa simillima xanthoptera Comparative Biochemistry and Physiology Part B:

Biochemistry and Molecular Biology 161(1):17-24

4. Kobayashi I, Tsukioka H, KŽmoto N, Uchino K, Sezutsu H, Tamura T, Kusakabe T, Tomita S (2012) SID-1

protein of Caenorhabditis elegans mediates uptake of dsRNA into Bombyx cells Insect Biochemistry and

Molecular Biology 42(2):148-154

5. KŽmoto N, Yukuhiro K, Tomita S (2012) Novel gene rearrangements in the mitochondrial genome of a

webspinner, Aposthonia japonica (Insecta: Embioptera) Genome 55(3):222-233




7. Mon H, Kobayashi I, Ohkubo S, Tomita S, Lee J, Sezutsu H, Tamura T, Kusakabe T (2012) Effective RNA

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interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1 RNA

Biology 9(1):40-46

8. Nakajima K, Kinoshita H, Miura M (2011) Effects of raw silk size and twist number on characteristics of

twisted silk yarn Journal of Insect Biotechnology and Sericology 80(1):9-13

9. Nakajima K, Kinoshita H, Miura M (2011) Analysis of a sensory test with thin and thick fabric and a

comparison of the KES hand value Sanshi-Konchu Biotec 80(1):53-58 (in Japanese with English summary)

10. Ngernyuang N, Kobayashi I, Promboon A, Ratanapo S, Tamura T, Ngernsiri L (2011) Cloning and

expression analysis of the Bombyx mori α-amylase gene (Amy) from the indigenous Thai silkworm strain,

Nanglai Journal of Insect Science 11:38

11. Sakurai T, Mitsuno H, Haupt S.S, Uchino K, Yokohari F, Nishioka T, Kobayashi I, Sezutsu H, Tamura T,

Kanzaki R (2011) A single sex pheromone receptor determines chemical response specificity of sexual

behavior in the silkmoth Bombyx mori PLoS Genetics 7(6):e1002115

12. Sezutsu H, Uchino K, Kobayashi I, Tamura T, Yukuhiro K (2010) Extensive sequence rearrangements and

length polymorphism in fibroin genes in the wild silkmoth, Antheraea yamamai (Lepidoptera, Saturniidae)

International Journal of Wild Silkmoth and Silk 15:35-50

13. Takabayashi C, Kumai T, Miyazaki E, Kenichi N, Tomizawa J (2011) Development of ÒSilkfillÓ bed pad and

its physical properties The Journal of Silk Science and Technology of Japan 19:45-49 (in Japanese with

English summary)

14. Yonemura N, Sehnal F, Konik P, Ajimura M, Tamura T, Mita K (2012) Conservation of a pair of serpin 2

genes and their expression in Amphiesmenoptera Insect Biochemistry and Molecular Biology 42(5):371-

380

15. Yukuhiro K, Sezutsu H, Tamura T, Kosegawa E, Kiuchi M (2011) Nucleotide sequence variation in

mitochondrial COI gene among 147 silkworm (Bombyx mori) strains from Japanese, Chinese, European and

moltinism classes Genes & Genetic Systems 86(5):315-323

Review (in English)

1. Sezutsu H, Yamada H, Rochmijati J, Kameda T, Mita K, Tamura T, Yukuhiro K (2011) Silk genes in wild

silk moths International Journal of Wild Silkmoth and Silk 16:1-8

3-03 Development of novel technologies for efficient use of genetically modified

animals

Original Papers

1. Fuchimoto D, Senbon S, Suzuki S, Onishi A (2011) Effects of preservation of porcine oocytes by dibutyryl

cyclic AMP on in vitro maturation, fertilization and development Japan Agricultural Research Quarterly

45(3):295-300

2. Iwasaki K, Miwa Y, Ogawa H, Yazaki S, Iwamoto M, Furusawa T, Onishi A, Kuzuya T, Haneda M,

Watarai Y, Uchida K, Kobayashi T (2012) Comparative study on signal transduction in endothelial cells

－ 57 －

after anti-A/B and human leukocyte antigen antibody reaction: Implication of accommodation

Transplantation 93(4):390-397

3. Sembon S, Fuchimoto D, Iwamoto M, Suzuki S, Yoshioka K, Onishi A (2011) A simple method for

producing tetraploid porcine parthenogenetic embryos Theriogenology 76(4):598-606

4. Sembon S, Iwamoto M, Hashimoto M, Oishi T, Fuchimoto D, Suzuki S, Yazaki S, Onishi A (2012) Porcine

androgenetic embryos develop to fetal stage in recipient mothers Theriogenology 78(1):225-231

5. Takeda K, Tasai M, Iwamoto M, Oe M, Chikuni K, Nakamura Y, Tagami T, Nirasawa K, Hanada H, Pinkert

C.A, Onishi A (2012) Comparative proteomic analysis of liver mitochondrial proteins derived from cloned

adult pigs reconstructed with Meishan pig fibroblast cells and European pig enucleated oocytes Journal of

Reproduction and Development 58(2):248-253

Review (in English)

1. Merkwitz C, Lochhead P, Tsikolia N, Koch D, Sygnecka K, Sakurai M, Spanel-Borowski K, Ricken A.M

(2011) Expression of KIT in the ovary, and the role of somatic precursor cells Progress in Histochemistry

and Cytochemistry 46(3):131-184

3-04 Development of novel technologies using biomaterials based on silk proteins

Original Papers

1. Aratani E, Kameda T, Tamada Y, Kojima K (2011) Gelation control of silk fibroin solution during the

dialysis process The Journal of Silk Science and Technology of Japan 20:85-87 (in Japanese with English

summary)

2. Hattori S, Terada D, Teramoto H, Kameda T, Tamada Y, Kobayashi H, Honda T, Yoshikawa C (2012)

Influence of sterilisations on silk protein-based materials Bioinspired, Biomimetic and Nanobiomaterials

1(3):195-199

3. Kameda T, Hashimoto T, Tamada Y (2011) Effects of supercooling and organic solvent on the formation of

a silk sponge with porous 3-D structure, and its dynamical and structural characterization using solid-state

NMR Journal of Materials Science 46(24):7923-7930

4. Kameda T, Kojima K, Zhang Q, Sezutsu H (2012) Identification of hornet silk gene with a characteristic

repetitive sequence in Vespa simillima xanthoptera Comparative Biochemistry and Physiology Part B:

Biochemistry and Molecular Biology 161(1):17-24

5. Kaneko A, Tamada Y, Hirai S, Kuzuya T, Hashimoto T (2012) Characterization of a silk-resinified compact

fabricated using a pulse-energizing sintering device Macromolecular Materials and Engineering

297(3):272-278

6. Kawakami M, Tomita N, Shimada Y, Yamamoto K, Tamada Y, Kachi N, Suguro T (2011) Chondrocyte

distribution and cartilage regeneration in silk fibroin sponge Bio-medical Materials and Engineering

21(1):53-61

7. Kuwana Y, Kojima K, Tamada Y (2011) Tensile properties of transgenic cocoon fibers containing foreign

－ 58 －

proteins :effect of EGFP and spider (Araneus ventricosus) dragline silk proteins The Journal of Silk

Science and Technology of Japan 20:81-83 (in Japanese with English summary)

8. Murakami M, Tamada Y, Kojima K (2011) Dissolving whole cocoon silk proteins by using a pH-adjusted

buffered LiBr solution The Journal of Silk Science and Technology of Japan 20:89-94 (in Japanese with

English summary)

9. Terada D, Hattori S, Teramoto H, Kameda T, Tamada Y, Kobayashi H (2012) Silk nanofibre arranging

improves its fibre mat transparency Bioinspired, Biomimetic and Nanobiomaterials 1(1):57-61



3-05 Elucidation of insect-specific biological functions and development of novel

technologies for their applications

Original Papers

1. Furuki T, Shimizu T, Kikawada T, Okuda T, Sakurai M (2011) Salt effects on the structural and

thermodynamic properties of a Group 3 LEA protein model peptide Biochemistry 50(33):7093-7103

2. Furuki T, Shimizu T, Charkbortee S, Hatanaka R, Takahashi T, Kikawada T, Okuda T, Mihara H,

Tunnacliffe A, Sakurai M (2011) Anti-aggregation effects on proteins by 22-mer model peptides of Group3

LEA proteins Cryobiology and Cryotechnology 57(2):139-142 (in Japanese with English summary)

3. Gotoh H, Cornette R, Koshikawa S, Okada Y, Lavine L.C, Emlen D.J, Miura T (2011) Juvenile hormone

regulates extreme mandible growth in male stag beetles PLoS ONE 6(6):e21139

4. Hatanaka R, Hagiwara-Komoda Y, Kanamori Y, Fujita M, Cornette R, Okuda T, Kikawada T (2011)

Characterization of the chironomid LEA protein, PvLEA4 Cryobiology and Cryotechnology 57(2):135-138

(in Japanese with English summary)

5. Kamimura M, Saito H, Niwa R, Niimi T, Toyoda K, Ueno C, Kanamori Y, Shimura S, Kiuchi M (2012)

Fungal ecdysteroid-22-oxidase: a new tool for manipulating ecdysteroid signaling and insect development


6. Lacey M.J, S”mon E, Krasulov⁄ J, Sillam-Duss‘s D, Robert A, Cornette R, Hoskovec M, Zÿ⁄cÿek P,

Valterov⁄ I, Bordereau C (2011) Chemical communication in termites: syn-4,6-Dimethylundecan-1-ol as

trail-following pheromone, syn-4,6-dimethylundecanal and (5E)-2,6,10-trimethylundeca-5,9-dienal as the

respective male and female sex pheromones in Hodotermopsis sjoestedti (Isoptera, Archotermopsidae)

Journal of Insect Physiology 57(12):1585-1591




8. Sabree Z.L, Huang C.Y, Arakawa G, Tokuda G, Lo N, Watanabe H, Moran N.A (2012) Genome shrinkage

and loss of nutrient-providing potential in the obligate symbiont of the primitive termite Mastotermes

darwiniensis Applied and Environmental Microbiology 78(1):204-210


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1:133

10. Tanaka H, Sagisaka A, Fujita K, Furukawa S, Ishibashi J, Yamakawa M (2012) BmEts upregulates promoter

activity of lebocin in Bombyx mori Insect Biochemistry and Molecular Biology 42(7):474-481



12. Tokuda G, Watanabe H, Hojo M, Fujita A, Makiya H, Miyagi M, Arakawa G, Arioka M (2012) Cellulolytic

environment in the midgut of the wood-feeding higher termite Nasutitermes takasagoensis Journal of Insect


13. Tomimoto K, Fujita K, Ishibashi J, Imanishi S, Yamakawa M, Tanaka H (2012) A novel method to convert a

DNA fragment inserted into a plasmid to an inverted repeat structure Molecular Biotechnology 50(1):18-27

14. Tomimoto K, Yamakawa M, Tanaka H (2012) Construction of a long hairpin RNA expression library using

Cre recombinase Journal of Biotechnology 160(3-4):129-139


1. Cornette R, Kikawada T (2011) The induction of anhydrobiosis in the sleeping chironomid: Current status of

our knowledge IUBMB Life 63(6):419-429

2. Kikawada T (2011) Mechanisms underpinning induction of anhydrobiosis in Polypedilum vanderplanki

Cryobiology and Cryotechnology 57(1):33-43

3. Okuda T (2012) A novel dry-preservation technology inspired by desiccation tolerant insect, Polypedilum

vanderplanki Design for Innovative Value Towards a Sustainable Society :679-680

4. Tanaka H, Yamakawa M (2011) Regulation of the innate immune responses in the silkworm, Bombyx mori

Invertebrate Survival Journal 8(1):59-69

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(Apr. 2011HZ - NARO

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