Updated on 2025/09/30

写真a

 
USHIJIMA Koichiro
 
Organization
Faculty of Environmental, Life, Natural Science and Technology Professor
Position
Professor
External link

Degree

  • 博士（理学） ( 2002.3   横浜市立大学 )

  • 修士（理学） ( 1999.3   横浜市立大学 )

  • 学士（農学） ( 1997.3   東京農工大学 )

  • 博士（理学）

Research Areas

  • Environmental Science/Agriculture Science / Horticultural science

 

Papers

  • Effect of storage temperature and 1-MCP treatment on fruit ripening in Japanese apricot fruit (Prunus mume)

    C. Yano, T. Kashiwamoto, M. Muqadas, K. Ohashi, O.W. Mitalo, T. Akagi, Y. Kubo, T. Kawai, K. Ushijima, F. Fukuda

    Acta Horticulturae   ( 1404 )   717 - 724   2024.9

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    Publishing type：Research paper (scientific journal)   Publisher：International Society for Horticultural Science (ISHS)  

    DOI： 10.17660/actahortic.2024.1404.98

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  • Effect of sugar-ester edible coating on postharvest life and development of chilling injury in tomatoes during ambient and cold storage

    M. Muqadas, O. W. Mitalo, C. Yano, K. Ohashi, N. Hira, T. Akagi, K. Ushijima, Y. Kubo

    Acta Horticulturae   3 ( 1404 )   437 - 444   2024.9

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    Publishing type：Research paper (international conference proceedings)  

    DOI： 10.17660/ActaHortic.2024.1404.59

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  • Degenerate oligonucleotide primer MIG-seq: an effective PCR-based method for high-throughput genotyping International journal

    Kazusa Nishimura, Hiroyuki Kokaji, Ko Motoki, Akira Yamazaki, Kyoka Nagasaka, Rihito Takisawa, Yasuo Yasui, Takashi Kawai, Koichiro Ushijima, Masanori Yamasaki, Hiroki Saito, Ryohei Nakano, Tetsuya Nakazaki

    The Plant journal : for cell and molecular biology   118 ( 6 )   2296 - 2317   2024.5

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Cold Spring Harbor Laboratory  

    Summary

    Multiplexed inter-simple sequence repeats genotyping by sequencing (MIG-seq) is an next-generation sequencing library construction method developed for the analysis of DNA in ecology. Although MIG-seq can generate libraries from low-quality DNA, few polymorphisms can be obtained in species with small genomes. In this study, we developed degenerate oligonucleotide primer MIG-seq (dpMIG-seq) as an effective polymorphism discovery method that allows for variation in the number of polymorphisms while retaining the advantages of MIG-seq, including independence from DNA quality. In dpMIG-seq, a proportion of the simple sequence repeats in the primer sequence of the first PCR in MIG-seq was changed to degenerate oligonucleotides to enable annealing to a wider range of sequences. In tests of several crop species other than wheat, the number of loci that could be sequenced using dpMIG-seq with a data volume of 0.3 gigabases (Gb) was increased compared with that sequenced using MIG-seq. In wheat, the number of polymorphisms obtained via dpMIG-seq was higher than that obtained via MIG-seq when a data volume of about ≥2 Gb was obtained. In dpMIG-seq, different loci could be sequenced by changing the positions of the degenerate oligonucleotides. By applying dpMIG-seq, we constructed a linkage map consisting of 5,142 markers for the rice inter-subspecies F2 population, and we detected quantitative trait loci for heading date in the regions where known heading-related genes were located. Overall, our results show that dpMIG-seq is a useful tool for the genetic analysis of crop species.

    DOI： 10.1101/2022.08.25.504752

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  • Rapid and dynamic evolution of a giant Y chromosome inSilene latifolia

    Takashi Akagi, Naoko Fujita, Kanae Masuda, Kenta Shirasawa, Kiyotaka Nagaki, Ayano Horiuchi, Eriko Kuwada, Riko Kunou, Koki Nakamura, Yoko Ikeda, Koichiro Ushijima, Deborah Charlesworth

    2023.9

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  • Application of deep learning diagnosis for multiple traits sorting in peach fruit

    Kanae Masuda, Rika Uchida, Naoko Fujita, Yoshiaki Miyamoto, Takahiro Yasue, Yasutaka Kubo, Koichiro Ushijima, Seiichi Uchida, Takashi Akagi

    Postharvest Biology and Technology   201   112348 - 112348   2023.7

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.postharvbio.2023.112348

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  • Genetic basis of lineage-specific evolution of fruit traits in hexaploid persimmon. International journal

    Ayano Horiuchi, Kanae Masuda, Kenta Shirasawa, Noriyuki Onoue, Ryusuke Matsuzaki, Ryutaro Tao, Yasutaka Kubo, Koichiro Ushijima, Takashi Akagi

    DNA research : an international journal for rapid publication of reports on genes and genomes   2023.6

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    Language：English   Publishing type：Research paper (scientific journal)  

    Frequent polyploidization events in plants have led to the establishment of many lineage-specific traits representing each species. Little is known about the genetic bases for these specific traits in polyploids, presumably due to plant genomic complexity and their difficulties of applying genetic approaches. Hexaploid Oriental persimmon (Diospyros kaki) has evolved specific fruit characters, including wide variations in fruit shapes and astringency. In this study, using whole-genome diploidized/quantitative genotypes from ddRAD-Seq data of 173 persimmon cultivars, we examined their population structures and potential correlations between their structural transitions and variations in nine fruit traits. The population structures of persimmon cultivars were highly randomized and not substantially correlated with the representative fruit traits focused on in this study, except for fruit astringency. With genome-wide association analytic tools considering polyploid alleles, we identified the loci associated with the nine fruit traits; we mainly focused on fruit-shape variations, which have been numerically characterized by principal component analysis of elliptic Fourier descriptors. The genomic regions that putatively underwent selective sweep exhibited no overlap with the loci associated with these persimmon-specific fruit traits. These insights will contribute to understanding of the genetic mechanisms by which fruit traits are independently established, possibly due to polyploidization events.

    DOI： 10.1093/dnares/dsad015

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  • Recurrent neo-sex chromosome evolution in kiwifruit

    Takashi Akagi, Erika Varkonyi-Gasic, Kenta Shirasawa, Andrew Catanach, Isabelle M. Henry, Daniel Mertten, Paul Datson, Kanae Masuda, Naoko Fujita, Eriko Kuwada, Koichiro Ushijima, Kenji Beppu, Andrew C. Allan, Deborah Charlesworth, Ikuo Kataoka

    Nature Plants   2023.3

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    Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

    DOI： 10.1038/s41477-023-01361-9

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    Other Link： https://www.nature.com/articles/s41477-023-01361-9

  • 1-Methylcyclopropene Pretreatment in ‘La France’ Pears to Extend Postharvest Life and Maximize Fruit Quality Reviewed

    Abdul H. Kazimi, Oscar W. Mitalo, Azimullah Azimi, Kanae Masuda, Chikara Yano, Takashi Akagi, Koichiro Ushijima, Yasutaka Kubo

    2023.1

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  • ウメの青果流通期間拡大に向けた台湾ウメ，日本ウメの収穫後果実の低温応答と1-MCP処理効果の調査

    Tomoaki Kashiwamoto, Takashi Kawai, Takaaki Oe, Yasuhisa Tsuchida, Chikara Yano, Maqsood Muqadas, Takashi Akagi, Fumio Fukuda, Yasutaka Kubo, Koichiro Ushijima

    Horticultural Research (Japan)   22 ( 4 )   303 - 311   2023

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    Publishing type：Research paper (scientific journal)   Publisher：Japanese Society for Horticultural Science  

    DOI： 10.2503/hrj.22.303

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  • Ongoing rapid evolution of a post-Y region revealed by chromosome-scale genome assembly of a hexaploid monoecious persimmon (Diospyros kaki)

    Ayano Horiuchi, Kanae Masuda, Kenta Shirasawa, Noriyuki Onoue, Naoko Fujita, Koichiro Ushijima, Takashi Akagi

    2022.12

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  • Genome-wide cis-decoding for expression design in tomato using cistrome data and explainable deep learning. International journal

    Takashi Akagi, Kanae Masuda, Eriko Kuwada, Kouki Takeshita, Taiji Kawakatsu, Tohru Ariizumi, Yasutaka Kubo, Koichiro Ushijima, Seiichi Uchida

    The Plant cell   34 ( 6 )   2174 - 2187   2022.5

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    Language：English   Publishing type：Research paper (scientific journal)  

    In the evolutionary history of plants, variation in cis-regulatory elements (CREs) resulting in diversification of gene expression has played a central role in driving the evolution of lineage-specific traits. However, it is difficult to predict expression behaviors from CRE patterns to properly harness them, mainly because the biological processes are complex. In this study, we used cistrome datasets and explainable convolutional neural network (CNN) frameworks to predict genome-wide expression patterns in tomato (Solanum lycopersicum) fruit from the DNA sequences in gene regulatory regions. By fixing the effects of trans-acting factors using single cell-type spatiotemporal transcriptome data for the response variables, we developed a prediction model for crucial expression patterns in the initiation of tomato fruit ripening. Feature visualization of the CNNs identified nucleotide residues critical to the objective expression pattern in each gene, and their effects were validated experimentally in ripening tomato fruit. This cis-decoding framework will not only contribute to the understanding of the regulatory networks derived from CREs and transcription factor interactions, but also provides a flexible means of designing alleles for optimized expression.

    DOI： 10.1093/plcell/koac079

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  • Effect of storage temperature on development of chilling injury and its transcriptome analysis in ‘Fuyu’ persimmon fruit with or without 1-MCP treatment Reviewed

    T. Furuta, O.W. Mitalo, T. Otsuki, R. Okada, K. Ohashi, K. Masuda, T. Akagi, K. Ushijima, R. Nakano, Y. Kubo

    Acta Horticulturae   ( 1338 )   343 - 350   2022.4

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：International Society for Horticultural Science (ISHS)  

    DOI： 10.17660/actahortic.2022.1338.49

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  • Reinvention of hermaphroditism via activation of a RADIALIS-like gene in hexaploid persimmon. International journal

    Kanae Masuda, Yoko Ikeda, Takakazu Matsuura, Taiji Kawakatsu, Ryutaro Tao, Yasutaka Kubo, Koichiro Ushijima, Isabelle M Henry, Takashi Akagi

    Nature plants   8 ( 3 )   217 - 224   2022.3

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    Language：English   Publishing type：Research paper (scientific journal)  

    In flowering plants, different lineages have independently transitioned from the ancestral hermaphroditic state into and out of various sexual systems1. Polyploidizations are often associated with this plasticity in sexual systems2,3. Persimmons (the genus Diospyros) have evolved dioecy via lineage-specific palaeoploidizations. More recently, hexaploid D. kaki has established monoecy and also exhibits reversions from male to hermaphrodite flowers in response to natural environmental signals (natural hermaphroditism, NH), or to artificial cytokinin treatment (artificial hermaphroditism, AH). We sought to identify the molecular pathways underlying these polyploid-specific reversions to hermaphroditism. Co-expression network analyses identified regulatory pathways specific to NH or AH transitions. Surprisingly, the two pathways appeared to be antagonistic, with abscisic acid and cytokinin signalling for NH and AH, respectively. Among the genes common to both pathways leading to hermaphroditic flowers, we identified a small-Myb RADIALIS-like gene, named DkRAD, which is specifically activated in hexaploid D. kaki. Consistently, ectopic overexpression of DkRAD in two model plants resulted in hypergrowth of the gynoecium. These results suggest that production of hermaphrodite flowers via polyploidization depends on DkRAD activation, which is not associated with a loss-of-function within the existing sex determination pathway, but rather represents a new path to (or reinvention of) hermaphroditism.

    DOI： 10.1038/s41477-022-01107-z

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  • The effect of layer-by-layer edible coating on the shelf life and transcriptome of ‘Kosui’ Japanese pear fruit Reviewed

    Natsuki Hira, Oscar W. Mitalo, Rui Okada, Momoka Sangawa, Kanae Masuda, Naoko Fujita, Koichiro Ushijima, Takashi Akagi, Yasutaka Kubo

    Postharvest Biology and Technology   185   111787 - 111787   2022.3

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier {BV}  

    DOI： 10.1016/j.postharvbio.2021.111787

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  • MIG-seq is an effective method for high-throughput genotyping in wheat (Triticum spp.)

    Kazusa Nishimura, Ko Motoki, Akira Yamazaki, Rihito Takisawa, Yasuo Yasui, Takashi Kawai, Koichiro Ushijima, Ryohei Nakano, Tetsuya Nakazaki

    DNA Research   29 ( 2 )   2022.2

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    Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

    Abstract

    MIG-seq (Multiplexed inter-simple sequence repeats genotyping by sequencing) has been developed as a low cost genotyping technology, although the number of polymorphisms obtained is assumed to be minimal, resulting in the low application of this technique to analyses of agricultural plants. We applied MIG-seq to 12 plant species that include various crops and investigated the relationship between genome size and the number of bases that can be stably sequenced. The genome size and the number of loci, which can be sequenced by MIG-seq, are positively correlated. This is due to the linkage between genome size and the number of simple sequence repeats (SSRs) through the genome. The applicability of MIG-seq to population structure analysis, linkage mapping, and quantitative trait loci (QTL) analysis in wheat, which has a relatively large genome, was further evaluated. The results of population structure analysis for tetraploid wheat showed the differences among collection sites and subspecies, which agreed with previous findings. Additionally, in wheat biparental mapping populations, over 3,000 SNPs/indels with low deficiency were detected using MIG-seq, and the QTL analysis was able to detect recognized flowering-related genes. These results revealed the effectiveness of MIG-seq for genomic analysis of agricultural plants with large genomes, including wheat.

    DOI： 10.1093/dnares/dsac011

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    Other Link： https://academic.oup.com/dnaresearch/article-pdf/29/2/dsac011/43439343/dsac011.pdf

  • Deep Learning Predicts Rapid Over-softening and Shelf Life in Persimmon Fruits

    Maria Suzuki, Kanae Masuda, Hideaki Asakuma, Kouki Takeshita, Kohei Baba, Yasutaka Kubo, Koichiro Ushijima, Seiichi Uchida, Takashi Akagi

    Horticulture Journal   91 ( 3 )   408 - 415   2022

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.2503/hortj.UTD-323

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  • Examining the Role of Low Temperature in Satsuma Mandarin Fruit Peel Degreening via Comparative Physiological and Transcriptomic Analysis. International journal

    Oscar W Mitalo, William O Asiche, Seung W Kang, Hiroshi Ezura, Takashi Akagi, Yasutaka Kubo, Koichiro Ushijima

    Frontiers in plant science   13   918226 - 918226   2022

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    Language：English   Publishing type：Research paper (scientific journal)  

    Peel degreening is the most conspicuous aspect of fruit ripening in many citrus fruits because of its importance for marketability. In this study, peel degreening in response to propylene (an ethylene analog) and at varying storage temperatures was characterized in Satsuma mandarin (Citrus unshiu Marc.) fruit. Propylene treatment triggered rapid peel degreening (within 4-6 days), indicated by an increase in the citrus color index (CCI) and chlorophyll loss. Peel degreening was also observed in fruit at 10°C and 15°C after 28-42 days, with gradual CCI increase and chlorophyll reduction. However, fruit at 5°C, 20°C, and 25°C remained green, and no substantial changes in peel CCI and chlorophyll content were recorded during the 42-day storage duration. The transcriptomes of peels of fruit treated with propylene for 4 days and those stored at varying temperatures for 28 days were then analyzed by RNA-Seq. We identified three categories of differentially expressed genes that were regulated by (i) propylene (and by analogy, ethylene) alone, (ii) low temperature (5°C, 10°C, or 15°C vs. 25°C) alone, and (iii) either propylene or low temperature. Gene-encoding proteins associated with chlorophyll degradation (such as CuSGR1, CuNOL, CuACD2, CuCAB2, and CuLHCB2) and a transcription factor (CuERF114) were differentially expressed by propylene or low temperature. To further examine temperature-induced pathways, we also monitored gene expression during on-tree fruit maturation vs. postharvest. The onset of on-tree peel degreening coincided with autumnal drops in field temperatures, and it was accompanied by differential expression of low temperature-regulated genes. On the contrary, genes that were exclusively regulated by propylene (such as CuCOPT1 and CuPOX-A2) displayed insignificant expression changes during on-tree peel degreening. These findings indicate that low temperatures could be involved in the fruit ripening-related peel degreening independently of ethylene.

    DOI： 10.3389/fpls.2022.918226

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  • Combined signal sequence trap and macroarray analysis identifies genes associated with differential fruit softening characteristics during ripening in European and Chinese pears Reviewed

    Mercy W. Mwaniki, Oscar W. Mitalo, Eric G. Mworia, Willis O. Owino, Kyoko Hiwasa-Tanase, Jocelyn K. C. Rose, Koh Aoki, Tomoya Esumi, Takashi Kawai, Ryohei Nakano, Koichiro Ushijima, Yasutaka Kubo

    POSTHARVEST BIOLOGY AND TECHNOLOGY   174   2021.4

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    DOI： 10.1016/j.postharvbio.2020.111436

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  • Association of T2/S-RNase With Self-Incompatibility of Japanese Citrus Accessions Examined by Transcriptomic, Phylogenetic, and Genetic Approaches. Reviewed International journal

    Chitose Honsho, Koichiro Ushijima, Misa Anraku, Shuji Ishimura, Qibin Yu, Frederick G Gmitter Jr, Takuya Tetsumura

    Frontiers in plant science   12   638321 - 638321   2021

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    Language：English   Publishing type：Research paper (scientific journal)  

    Several citrus varieties show gametophytic self-incompatibility (GSI), which can contribute to seedless fruit production in several cultivars. This study investigated the genes regulating this trait through RNA-seq performed using styles collected from the flowers of Japanese citrus cultivars 'Hyuganatsu,' 'Tosabuntan,' 'Hassaku,' 'Banpeiyu,' and 'Sweet Spring'. We screened the transcripts of putative T2 RNases, i.e., the protein family including all S-RNases from S-RNase-based GSI plants, and constructed a phylogenetic tree using the screened T2 RNases and S-RNases retrieved from citrus genome databases and a public database. Three major clusters (class I-III) were formed, among which, the class III cluster contained family specific subclusters formed by S-RNase and a citrus-specific cluster monophyletic to the S-RNase clusters. From the citrus class III cluster, six transcripts were consistent with the S haplotypes previously determined in Japanese citrus accessions, sharing characteristics such as isoelectric point, extracellular localization, molecular weight, intron number and position, and tissue-specific expression with S-RNases. One T2 RNase gene in self-incompatible Hyuganatsu was significantly down-regulated in the styles of a self-compatible mutant of Hyuganatsu in RNA-seq and qPCR analyses. In addition, the inheritance pattern of some T2 RNase genes was consistent with the pattern of the S haplotype in the progeny population of Hyuganatsu and Tosabuntan. As all results supported citrus self-incompatibility being based on S-RNase, we believe that six T2 RNase genes were S-RNases. The homology comparison between the six T2 RNases and S-RNases recently reported in Chinese citrus revealed that three out of six T2 RNases were identical to S-RNases from Chinese citrus. Thus, the other three T2 RNases were finally concluded to be novel citrus S-RNases involved in self-incompatibility.

    DOI： 10.3389/fpls.2021.638321

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  • Control of Fruit Softening during Marine Transport Bound for North America in Three Astringent-type Persimmons

    Takahiro Furuta, Takumi Otsuki, Rui Okada, Takashi Kawai, Fumio Fukuda, Koichiro Ushijima, Yasutaka Kubo, Ryohei Nakano

    Horticultural Research (Japan)   20 ( 4 )   455 - 461   2021

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    Publishing type：Research paper (scientific journal)   Publisher：Japanese Society for Horticultural Science  

    DOI： 10.2503/hrj.20.455

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  • Molecular Mechanism Underlying Derepressed Male Production in Hexaploid Persimmon Reviewed

    Kanae Masuda, Naoko Fujita, Ho-Wen Yang, Koichiro Ushijima, Yasutaka Kubo, Ryutaro Tao, Takashi Akagi

    FRONTIERS IN PLANT SCIENCE   11   2020.12

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.3389/fpls.2020.567249

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  • Low temperature modulates natural peel degreening in lemon fruit independently of endogenous ethylene Reviewed

    Oscar W Mitalo, Takumi Otsuki, Rui Okada, Saeka Obitsu, Kanae Masuda, Yuko Hojo, Takakazu Matsuura, Izumi C Mori, Daigo Abe, William O Asiche, Takashi Akagi, Yasutaka Kubo, Koichiro Ushijima

    Journal of Experimental Botany   71 ( 16 )   4778 - 4796   2020.8

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    Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1093/jxb/eraa206

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    Other Link： http://academic.oup.com/jxb/article-pdf/71/16/4778/33573305/eraa206.pdf

  • Genome-wide study on the polysomic genetic factors conferring plasticity of flower sexuality in hexaploid persimmon Reviewed

    Kanae Masuda, Eiji Yamamoto, Kenta Shirasawa, Noriyuki Onoue, Atsushi Kono, Koichiro Ushijima, Yasutaka Kubo, Ryutaro Tao, Isabelle M. Henry, Takashi Akagi

    DNA RESEARCH   27 ( 3 )   2020.6

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1093/dnares/dsaa012

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  • Characterization of Ethylene Production and Fruit Softening in ‘Tosui’ Peach Reviewed

    Takashi Kawai, Kagari Akita, Sakine Watanabe, Yosuke Fukamatsu, Daisuke Takata, Mamoru Sato, Koichiro Ushijima, Fumio Fukuda, Ryohei Nakano

    Horticulture Research   19 ( 1 )   61 - 67   2020.1

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    Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE  

    Flesh texture of peach (Prunus persica (L.) Batsch) is one of the most important traits regarding post-harvest preservation and consumer preference. Melting flesh peaches show a typical climacteric type of ripening characterized by a burst of ethylene production and rapid loss of firmness in the late maturing stage. In contrast, stony hard peaches produce little ethylene and maintain flesh firmness on the tree and after harvest. Here, we demonstrated that ‘Tosui’ peach, which is selected from open-pollinated seedlings of ‘Kawanakajimahakuto’, bears stony hard characteristics. ‘Tosui’ peach shows a long shelf life and crisp flesh; however, its ethylene production and softening characteristics remain unclear. We investigated the change of ethylene production and flesh firmness of ‘Tosui’ peach treated with or without exogenous propylene, an ethylene analogue. In non-treated control fruit, little ethylene production was detected and marked flesh firmness was maintained irrespective of the harvest season and production area. Although the flesh firmness was significantly reduced by propylene treatment, little ethylene production was detected during fruit softening. Analysis of the PpYUC11 gene, a strong candidate for the stony hard phenotype in peaches, revealed that the genotype of the transposon insertion of the 5′-flanking region and simple sequence repeat (SSR) of the first intron of ‘Tosui’ is the same as that of other stony hard peaches. These results collectively suggest that ‘Tosui’ is a stony hard peach whose ethylene production and resulting loss of firmness are suppressed.

    DOI： 10.2503/hrj.19.61

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    Other Link： https://www.jstage.jst.go.jp/article/hrj/19/1/19_61/_pdf

  • Postharvest Properties of Ultra-Late Maturing Peach Cultivars and Their Attributions to Melting Flesh (M) Locus: Re-evaluation of M Locus in Association With Flesh Texture. International journal

    Ryohei Nakano, Takashi Kawai, Yosuke Fukamatsu, Kagari Akita, Sakine Watanabe, Takahiro Asano, Daisuke Takata, Mamoru Sato, Fumio Fukuda, Koichiro Ushijima

    Frontiers in plant science   11   554158 - 554158   2020

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    Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    The postharvest properties of two ultra-late maturing peach cultivars, "Tobihaku" (TH) and "Daijumitsuto" (DJ), were investigated. Fruit were harvested at commercial maturity and held at 25°C. TH exhibited the characteristics of normal melting flesh (MF) peach, including rapid fruit softening associated with appropriate level of endogenous ethylene production In contrast, DJ did not soften at all during 3 weeks experimental period even though considerable ethylene production was observed. Fruit of TH and DJ were treated with 5,000 ppm of propylene, an ethylene analog, continuously for 7 days. TH softened rapidly whereas DJ maintained high flesh firmness in spite of an increase in endogenous ethylene production, suggesting that DJ but not TH lacked the ability to be softened in response to endogenous and exogenous ethylene/propylene. DNA-seq analysis showed that tandem endo-polygalacturonase (endoPG) genes located at melting flesh (M) locus, Pp-endoPGM (PGM), and Pp-endoPGF (PGF), were deleted in DJ. The endoPG genes at M locus are known to control flesh texture of peach fruit, and it was suggested that the non-softening property of DJ is due to the lack of endoPG genes. On the other hand, TH possessed an unidentified M haplotype that is involved in determination of MF phenotype. Structural identification of the unknown M haplotype, designated as M 0, through comparison with previously reported M haplotypes revealed distinct differences between PGM on M 0 haplotype (PGM-M0 ) and PGM on other haplotypes (PGM-M1 ). Peach M haplotypes were classified into four main haplotypes: M 0 with PGM-M0 ; M 1 with both PGM-M1 and PGF; M 2 with PGM-M1 ; and M 3 lacking both PGM and PGF. Re-evaluation of M locus in association with MF/non-melting flesh (NMF) phenotypes in more than 400 accessions by using whole genome shotgun sequencing data on database and/or by PCR genotyping demonstrated that M 0 haplotype was the common haplotype in MF accessions, and M 0 and M 1 haplotypes were dominant over M 2 and M 3 haplotypes and co-dominantly determined the MF trait. It was also assumed on the basis of structural comparison of M haplotypes among Prunus species that the ancestral haplotype of M 0 diverged from those of the other haplotypes before the speciation of Prunus persica.

    DOI： 10.3389/fpls.2020.554158

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  • ‘Passe Crassane’ pear fruit (Pyrus communis L.) ripening: Revisiting the role of low temperature via integrated physiological and transcriptome analysis Reviewed

    Oscar W. Mitalo, Yasuaki Tosa, Sumire Tokiwa, Yuki Kondo, Azimullah Azimi, Yuko Hojo, Takakazu Matsuura, Izumi C. Mori, Ryohei Nakano, Takashi Akagi, Koichiro Ushijima, Yasutaka Kubo

    Postharvest Biology and Technology   158   110949 - 110949   2019.12

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    Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.postharvbio.2019.110949

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  • A Sequence Resource of Autosomes and Additional Chromosomes in the Peach Pathotype of Alternaria alternata Reviewed

    Koichiro Ushijima, Mikihiro Yamamoto

    MOLECULAR PLANT-MICROBE INTERACTIONS   32 ( 10 )   1273 - 1276   2019.10

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    Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1094/MPMI-04-19-0093-A

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  • Low temperature storage stimulates fruit softening and sugar accumulation without ethylene and aroma volatile production in kiwifruit Reviewed

    Oscar W. Mitalo, Sumire Tokiwa, Yuki Kondo, Takumi Otsuki, Ivan Galis, Katsuhiko Suezawa, Ikuo Kataoka, Anh T. Doan, Ryohei Nakano, Koichiro Ushijima, Yasutaka Kubo

    Frontiers in Plant Science   10   1 - 15   2019.7

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.3389/fpls.2019.00888

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  • Comparative analysis of fruit ripening and associated genes in two kiwifruit cultivars (‘Sanuki Gold’ and ‘Hayward’) at various storage temperatures Reviewed

    Oscar Witere Mitalo, William Olubero Asiche, Yuka Kasahara, Yasuaki Tosa, Sumire Tokiwa, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    Postharvest Biology and Technology   147   20 - 28   2019.1

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    DOI： 10.1016/j.postharvbio.2018.08.017

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  • Aberrant Stamen Development is Associated with Parthenocarpic Fruit Set through Up-regulation of Gibberellin Biosynthesis in Tomato Reviewed

    Okabe, Yoshihiro, Yamaoka, Tatsuya, Ariizumi, Tohru, Ushijima, Koichiro, Kojima, Mikiko, Takebayashi, Yumiko, Sakakibara, Hitoshi, Kusano, Miyako, Shinozaki, Yoshihito, Pulungan, Sri I, Kubo, Yasutaka, Nakano, Ryohei, Ezura, Hiroshi

    Plant & Cell Physiology   60 ( 1 )   38 - 51   2019.1

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    DOI： 10.1093/pcp/pcy184

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  • Determination of optimum temperature for long-term storage and analysis of ripening-related genes in 'Rainbow Red' kiwifruit Reviewed

    Mitalo, O.W, Tokiwa, S, Kasahara, Y, Tosa, Y, Kondo, Y, Asiche, W.O, Kataoka, I, Suezawa, K, Ushijima, K, Nakano, R, Kubo, Y

    Acta Hortic.   1218   517 - 524   2018.10

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    DOI： 10.17660/ActaHortic.2018.1218.71

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  • Comparative Mapping of the ASTRINGENCY Locus Controlling Fruit Astringency in Hexaploid Persimmon (Diospyros kaki Thunb.) with the Diploid D. lotus Reference Genome Reviewed

    Nishiyama Soichiro, Onoue Noriyuki, Kono Atsushi, Sato Akihiko, Ushijima Koichiro, Yamane Hisayo, Tao Ryutaro, Yonemori Keizo

    HORTICULTURE JOURNAL   87 ( 3 )   315 - 323   2018.7

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    DOI： 10.2503/hortj.OKD-140

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  • Comparative transcriptome analysis reveals distinct ethylene-independent regulation of ripening in response to low temperature in kiwifruit Reviewed

    William O. Asiche, Oscar W. Mitalo, Yuka Kasahara, Yasuaki Tosa, Eric G. Mworia, Willis O. Owino, Koichiro Ushijima, Ryohei Nakano, Kentaro Yano, Yasutaka Kubo

    BMC Plant Biology   18 ( 1 )   2018.3

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    DOI： 10.1186/s12870-018-1264-y

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  • Nondestructive detection of split pit in peaches using an acoustic vibration method Reviewed

    Ryohei Nakano, Hidemi Akimoto, Fumio Fukuda, Takashi Kawai, Koichiro Ushijima, Yosuke Fukamatsu, Yasutaka Kubo, Yuichiro Fujii, Ken Hirano, Kunihisa Morinaga, Naoki Sakurai

    Horticulture Journal   87 ( 2 )   281 - 287   2018

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    DOI： 10.2503/hortj.OKD-094

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  • Characterization of ripening-related genes involved in ethylene-independent low temperature-modulated ripening in ‘rainbow red’ kiwifruit during storage and on-vine Reviewed

    Oscar Witere Mitalo, William Olubero Asiche, Yuka Kasahara, Yasuaki Tosa, Willis Omondi Owino, Eric Gituma Mworia, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    Horticulture Journal   87 ( 3 )   421 - 429   2018

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    DOI： 10.2503/hortj.OKD-035

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  • Effect of Storage Temperature on Fruit Ripening in Three Kiwifruit Cultivars Reviewed

    William Olubero Asiche, Oscar Witere Mitalo, Yuka Kasahara, Yasuaki Tosa, Eric Gituma Mworia, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    HORTICULTURE JOURNAL   86 ( 3 )   403 - 410   2017.7

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    DOI： 10.2503/hortj.OKD-028

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  • Two Members of the Aluminum-Activated Malate Transporter Family, SlALMT4 and SlALMT5, are Expressed during Fruit Development, and the Overexpression of SlALMT5 Alters Organic Acid Contents in Seeds in Tomato (Solanum lycopersicum) Reviewed

    Takayuki Sasaki, Yoshiyuki Tsuchiya, Michiyo Ariyoshi, Ryohei Nakano, Koichiro Ushijima, Yasutaka Kubo, Izumi C. Mori, Emi Higashiizumi, Ivan Galis, Yoko Yamamoto

    PLANT AND CELL PHYSIOLOGY   57 ( 11 )   2367 - 2379   2016.11

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    DOI： 10.1093/pcp/pcw157

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  • Extension of Shelf-life by Limited Duration of Propylene and 1-MCP Treatments in Three Kiwifruit Cultivars Reviewed

    William Olubero Asiche, Eric Gituma Mworia, Chisato Oda, Oscar Witere Mitalo, Willis Omondi Owino, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    HORTICULTURE JOURNAL   85 ( 1 )   76 - 85   2016.1

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.2503/hortj.MI-066

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  • Effect of postharvest short-term radiation of near infrared light on transpiration of lettuce leaf Reviewed

    Ayako Kozuki, Yutaka Ishida, Kazumasa Kakibuchi, Toshihiro Mishima, Naoki Sakurai, Yoshiyuki Murata, Ryohei Nakano, Koichiro Ushijima, Yasutaka Kubo

    POSTHARVEST BIOLOGY AND TECHNOLOGY   108   78 - 85   2015.10

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    DOI： 10.1016/j.postharvbio.2015.05.010

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  • Genetic Control of Floral Morph and Petal Pigmentation in Linum grandiflorum Desf., a Heterostylous Flax Reviewed

    Koichiro Ushijima, Kazuo Ikeda, Ryohei Nakano, Miyo Matsubara, Yuri Tsuda, Yasutaka Kubo

    HORTICULTURE JOURNAL   84 ( 3 )   261 - 268   2015.7

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    Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.2503/hortj.MI-045

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  • Evaluation of Optimal Harvesting Time Based on Quality Attributes during Growth in Green Soybeans ‘Hukura’ and ‘Yuagari-musume’ Reviewed

    Hiroshi Mizuno, Saki Hashimoto, Nina Tanaka, Tatsuya Yamamoto, Ryohei Nakano, Kouichiro Ushijima, Yasutaka Kubo

    Horticultural Research (Japan)   14 ( 1 )   61 - 67   2015.1

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    Publishing type：Research paper (scientific journal)   Publisher：Japanese Society for Horticultural Science  

    DOI： 10.2503/hrj.14.61

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  • Effect of Storage Temperature and Configuration on Postharvest Quality of 'Hukura' and 'Yuagari-musume' Green Soybeans

    Mizuno Hiroshi, Tanaka Nina, Hashimoto Saki, Yamamoto Tatsuya, Nakano Ryohei, Ushijima Kouichiro, Kubo Yasutaka

    Hort. Res. (Japan)   14 ( 3 )   297 - 304   2015

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    As green soybeans, immature seeds of Glycine max (L.) Merr., are usually harvested during summer, their quality rapidly deteriorates after harvest. We determined several quality attributes of postharvest green soybeans stored at various temperatures with short intervals and the effects of configuration (with or without leaves and stems) in 'Hukura' and 'Yuagari-musume'. In both cultivars, green soybeans stored at 15°C or higher showed a 20% or greater decrease in total sugars and free amino acids only 3 h and 6 h after harvest respectively, compared with levels at harvest. Storage temperatures at 10°C or lower were effective in preserving the appearance of pods and resulted in an 80% or greater retention of the level at harvest of total sugars and free amino acids even after 10 h and 24 h storage, respectively. Storing green soybeans with leaves and stems at room temperature had a slightly beneficial effect on the retention of quality attributes; however, the effects of the storage temperature were much more marked than those of the configuration. These results indicate that cooling at 10°C or lower within 3 h after harvest is important for the quality control of green soybeans.

    DOI： 10.2503/hrj.14.297

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    Other Link： http://agriknowledge.affrc.go.jp/RN/2010892824

  • Effect of Gelatinization Degree before Freezing on the Physical Properties of Green Soybean 'Miryoku' and 'Yuagari-musume'

    MIZUNO Hiroshi, TANAKA Nina, HASHIMOTO Saki, YAMAMOTO Tatsuya, NAKANO Ryohei, USHIJIMA Kouichiro, KUBO Yasutaka

    Jpn. J. Food Eng.   16 ( 1 )   63 - 70   2015

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    Effects of ratio of first and second cooking periods before and after freeze on physical characteristics in green soybeans were determined to study effects of gelatinization level before freezing, in the condition that total cooking period was set at 210 seconds. The green soybeans gelatinized to 55-83% by first cooking exhibited higher firmness and higher crispness index (CI) after both they were thawed and fully gelatinized by second cooking, compared with other gelatinization degree at first cooking. The firmness and CI of green beans frozen with gelatinization level less than 41% were relatively high after thawing but decreased severely after second cooking. The green beans gelatinized over 92% by first cooking had considerably low CI after thawing. SEM imaging confirmed that cell structure of green soybeans gelatinized to 55-83% by first cooking was more intact even after they were thawed and fully then gelatinized than that of soybeans cooked with other condition.

    DOI： 10.11301/jsfe.16.63

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    Other Link： http://agriknowledge.affrc.go.jp/RN/2010891565

  • Double flower formation induced by silencing of C-class MADS-box genes and its variation among petunia Cultivars Reviewed

    Siti Hajar Noor, Koichiro Ushijima, Ayaka Murata, Kaori Yoshida, Miki Tanabe, Tomoki Tanigawa, Yasutaka Kubo, Ryohei Nakano

    SCIENTIA HORTICULTURAE   178   1 - 7   2014.10

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    DOI： 10.1016/j.scienta.2014.07.029

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  • Isolation of the floral morph-related genes in heterostylous flax (Linum grandiflorum): the genetic polymorphism and the transcriptional and post-transcriptional regulations of the S locus Reviewed

    Koichiro Ushijima, Ryohei Nakano, Mayu Bando, Yukari Shigezane, Kazuo Ikeda, Yuko Namba, Saori Kume, Toshiyuki Kitabata, Hitoshi Mori, Yasutaka Kubo

    PLANT JOURNAL   69 ( 2 )   317 - 331   2012.1

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    DOI： 10.1111/j.1365-313X.2011.04792.x

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  • Low-temperature-modulated fruit ripening is independent of ethylene in 'Sanuki Gold' kiwifruit Reviewed

    Eric G. Mworia, Takashi Yoshikawa, Nadiah Salikon, Chisato Oda, William O. Asiche, Naoki Yokotani, Daigo Abe, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    JOURNAL OF EXPERIMENTAL BOTANY   63 ( 2 )   963 - 971   2012.1

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    DOI： 10.1093/jxb/err324

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  • The Arabidopsis Na+/H+ Antiporters NHX1 and NHX2 Control Vacuolar pH and K+ Homeostasis to Regulate Growth, Flower Development, and Reproduction (vol 23, pg 3482, 2011) Reviewed

    Elias Bassil, Hiromi Tajima, Yin-Chih Liang, Masa-aki Ohto, Koichiro Ushijima, Ryohei Nakano, Tomoya Esumi, Ardian Coku, Mark Belmonte, Eduardo Blumwald

    PLANT CELL   23 ( 12 )   4526 - 4526   2011.12

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  • Sequence divergence and loss-of-function phenotypes of S locus F-box brothers genes are consistent with non-self recognition by multiple pollen determinants in self-incompatibility of Japanese pear (Pyrus pyrifolia) Reviewed

    Hiroyuki Kakui, Masaki Kato, Koichiro Ushijima, Miyoko Kitaguchi, Shu Kato, Hidenori Sassa

    PLANT JOURNAL   68 ( 6 )   1028 - 1038   2011.12

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    DOI： 10.1111/j.1365-313X.2011.04752.x

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  • Effect of MA Storage and 1-MCP on Storability and Quality of 'Sanuki Gold' Kiwifruit Harvested at Two Different Maturity Stages Reviewed

    Eric Gituma Mworia, Takashi Yoshikawa, Nadiah Salikon, Chisato Oda, Tetsuo Fukuda, Katsuhiko Suezawa, William Olubero Asiche, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    JOURNAL OF THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE   80 ( 3 )   372 - 377   2011.7

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    DOI： 10.2503/jjshs1.80.372

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  • Characterization of ethylene biosynthesis and its regulation during fruit ripening in kiwifruit, Actinidia chinensis 'Sanuki Gold' Reviewed

    Eric G. Mworia, Takashi Yoshikawa, Naoki Yokotani, Tetsuo Fukuda, Katsuhiko Suezawa, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo

    POSTHARVEST BIOLOGY AND TECHNOLOGY   55 ( 2 )   108 - 113   2010.2

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    DOI： 10.1016/j.postharvbio.2009.08.007

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  • Identification and functional analysis of pistil self-incompatibility factor HT-B of Petunia Reviewed

    Alejandro Raul Puerta, Koichiro Ushijima, Takato Koba, Hidenori Sassa

    JOURNAL OF EXPERIMENTAL BOTANY   60 ( 4 )   1309 - 1318   2009.3

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    DOI： 10.1093/jxb/erp005

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  • Characterization of Cell Wall Proteins in Pears by Two-dimensional Gel Electrophoresis

    BANDO Mayu, USHIJIMA Koichiro, NAKANO Ryohei, INABA Akitsugu, KUBO Yasutaka

    Food Preservation Science   34 ( 3 )   127 - 132   2008.5

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    'La France' European pear fruit undergoes dramatic softening during ripening in an ethylene-dependent manner, whereas 'Yali' Chinese pear does not exhibit softening in spite of its massive ethylene production during ripening. To identify the key factors involved in fruit softening, the two-dimensional gel electrophoresis profiles of cell wall proteins extracted from ripening flesh tissues of 'La France' and 'Yali' pears were compared. The accumulation levels of 19 protein spots changed significantly during the ripening in 'La France' pear. When softening pears of this variety were treated with 1-MCP, a strong ethylene inhibitor, fruit softening was retarded. In parallel with this retardation, the levels of 15 of the 19 protein spots were restored or decreased to those at preclimacteric stages. For 'Yali' pear, the accumulation levels of 8 of the 15 protein spots remained unchanged during ripening, suggesting the involvement of these 8 proteins in the nonsoftening feature of 'Yali' pear. We therefore selected 2 of the 8 protein spots as potential key factors of fruit softening, since their accumulation was up-regulated during the ripening in 'La France' pear.

    DOI： 10.5891/jafps.34.127

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  • Differential expression of ethylene biosynthetic genes in climacteric and non-climacteric Chinese pear fruit Reviewed

    Miki Yamane, Daigo Abe, Sayaka Yasui, Naoki Yokotani, Wataru Kimata, Koichiro Ushijima, Ryohei Nakano, Yasutaka Kubo, Akitsugu Inaba

    POSTHARVEST BIOLOGY AND TECHNOLOGY   44 ( 3 )   220 - 227   2007.6

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    DOI： 10.1016/j.postharvbio.2006.12.010

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  • S locus F-Box brothers: Multiple and pollen-specific F-box genes with S haplotype-specific polymorphisms in apple and Japanese pear Reviewed

    Hidenori Sassa, Hiroyuki Kakui, Mayu Miyamoto, Yusuke Suzuki, Toshio Hanada, Koichiro Ushijima, Makoto Kusaba, Hisashi Hirano, Takato Koba

    GENETICS   175 ( 4 )   1869 - 1881   2007.4

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    DOI： 10.1534/genetics.106.068858

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  • beta-galactosidase and alpha-L-arabinofuranosidase activities and gene expression in European and Chinese pear fruit during ripening Reviewed

    Mercy W. Mwaniki, Francis M. Mathooko, Kyoko Hiwasa, Akira Tateishi, Naoki Yokotani, Koichiro Ushijima, Ryohei Nakano, Akitsugu Inaba, Yasutaka Kubo

    JOURNAL OF THE JAPANESE SOCIETY FOR HORTICULTURAL SCIENCE   76 ( 1 )   85 - 90   2007.1

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    DOI： 10.2503/jjshs.76.85

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  • Ethylene regulation of fruit softening and cell wall disassembly in Charentais melon Reviewed

    Kiyomi Nishiyama, Monique Guis, Jocelyn K. C. Rose, Yasutaka Kubo, Kristen A. Bennett, Wangjin Lu, Kenji Kato, Koichiro Ushijima, Ryohei Nakano, Akitsugu Inaba, Mondher Bouzayen, Alain Latche, Jean-Claude Pech, Alan B. Bennett

    JOURNAL OF EXPERIMENTAL BOTANY   58 ( 6 )   1281 - 1290   2007

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    DOI： 10.1093/jxb/erl283

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  • Expression characteristics of seven members of the beta-galactosidase gene family in 'La France' pear (Pyrus communis L.) fruit during growth and their regulation by 1-methylcyclopropene during postharvest ripening Reviewed

    MW Mwaniki, FM Mathooko, M Matsuzaki, K Hiwasa, A Tateishi, K Ushijima, R Nakano, A Inaba, Y Kubo

    POSTHARVEST BIOLOGY AND TECHNOLOGY   36 ( 3 )   253 - 263   2005.6

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    DOI： 10.1016/j.postharvbio.2005.02.002

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  • Linkage and physical distances between the S-haplotype S-RNase and SFB genes in sweet cherry Reviewed

    K Ikeda, K Ushijima, H Yamane, R Tao, NR Hauck, AM Sebolt, AF Iezzoni

    SEXUAL PLANT REPRODUCTION   17 ( 6 )   289 - 296   2005.4

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    DOI： 10.1007/s00497-004-0240-x

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  • Primary structural features of the S haplotype-specific F-box protein, SFB, in Prunus Reviewed

    K Ikeda, B Igic, K Ushijima, H Yamane, NR Hauck, R Nakano, H Sassa, AF Iezzoni, Kohn, JR, R Tao

    SEXUAL PLANT REPRODUCTION   16 ( 5 )   235 - 243   2004.1

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    DOI： 10.1007/s00497-003-0200-x

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  • The use of the S haplotype-specific F-box protein gene, SFB, as a molecular marker for S-haplotypes and self-compatibility in Japanese apricot (Prunus mume). International journal

    Hisayo Yamane, Koichiro Ushijima, Hidenori Sassa, Ryutaro Tao

    TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik   107 ( 8 )   1357 - 61   2003.11

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    Japanese apricot ( Prunus mume) exhibits the S-RNase-based gametophytic self-incompatibility system as do other self-incompatible Prunus species. This report identifies the S haplotype-specific F-box protein gene ( SFB), a candidate gene for pollen- S, of Japanese apricot, which leads to the development of a molecular typing system for S-haplotype in this fruit species. Both 5'- and 3'-RACE (rapid amplification of cDNA ends) were performed with SFB gene-specific oligonucleotide primers to clone Pm-SFB(1) and Pm-SFB(7) of 'Nanko ( S(1) S(7))'. As in the case of SFB of other Prunus species, Pm-SFB(1) and Pm-SFB(7) showed a high level of S-haplotype-specific sequence polymorphism and their expression was specific to pollen. Genomic DNA-blot analyses of 11 Japanese apricot cultivars with the Pm-SFB probes under low stringency conditions yielded RFLP bands specific to the S(1)- to S(8)-haplotypes as well as a self-compatible S(f)-haplotype. A practical usage of SFB as a molecular marker for S-haplotypes and self-compatibility in Japanese apricot is discussed.

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  • A pollen-expressed gene for a novel protein with an F-box motif that is very tightly linked to a gene for S-RNase in two species of cherry, Prunus cerasus and P. avium.

    Hisayo Yamane, Kazuo Ikeda, Koichiro Ushijima, Hidenori Sassa, Ryutaro Tao

    Plant & cell physiology   44 ( 7 )   764 - 9   2003.7

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    This study describes a novel F-box protein gene in the S-locus of sour cherry (Prunus cerasus) and sweet cherry (P. avium). The gene showed an S-haplotype-specific sequence polymorphism and the expression was specific to pollen. Genomic DNA blot analysis of eight sweet cherry cultivars with the probe for the F-box protein gene under low stringency conditions yielded RFLP bands specific to the S-haplotypes of each cultivar. We discuss the possibility of the gene for the F-box protein being a candidate for the male determinant of gametophytic self-incompatibility in PRUNUS:

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  • Structural and transcriptional analysis of self-incompatibility (S) locus of almond (Prunus dulcis) : identification of a pollen-expressed F-box gene with haplotype-specific polymorphism

    USHIJIMA K.

    Plant Cell   15   771 - 781   2003

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  • A pistil-specific thaumatin/PR5-like protein gene of Japanese pear (Pyrus serotina): sequence and promoter activity of the 5' region in transgenic tobacco. International journal

    Hidenori Sassa, Koichiro Ushijima, Hisashi Hirano

    Plant molecular biology   50 ( 3 )   371 - 7   2002.10

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    The genomic clone encoding the pistil-specific thaumatin/PR5-like protein (PsTL1) was isolated from Japanese pear (Pyrus serotina). Sequence analysis showed that the genomic clone contained the 5'-flanking sequence of 2.4 kb, the 3'-flanking sequence of 648 bp and the coding region interrupted by a intron of 351 bp. A sequence motif conserved in some pistil self-incompatibility gene promoters of solanaceous and brassicaceous species was located in the 5'-flanking region of the PsTL1 gene. The 2.4 kb 5'-flanking region was fused to the GUS coding sequence and transferred to tobacco. Transgenic tobacco showed GUS activity in pistil and, at low level, in anther, but not in other floral organs and leaf. Histochemical analysis localized GUS activity to stigma, transmitting tissue, anther and pollen of transgenic tobacco.

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  • Identification of self-incompatibility genotypes of almond by allele-specific PCR analysis

    M. Tamura, K. Ushijima, H. Sassa, H. Hirano, R. Tao, T. M. Gradziel, A. M. Dandekar

    Theoretical and Applied Genetics   101 ( 3 )   344 - 349   2000

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    DOI： 10.1007/s001220051489

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  • Characterization of the flanking regions of the S-RNase genes of Japanese pear (Pyrus serotina) and apple (Malus x domestica)

    Koichiro Ushijima, Hidenori Sassa, Hisashi Hirano

    Gene   211 ( 1 )   159 - 167   1998.4

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    DOI： 10.1016/S0378-1119(98)00105-X

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  • Cloning and characterization of cDNAs encoding S-RNases from almond (Prunus dulcis): Primary structural features and sequence diversity of the S-RNases in Rosaceae

    K. Ushijima, H. Sassa, R. Tao, H. Yamane, A. M. Dandekar, T. M. Gradziel, H. Hirano

    Molecular and General Genetics   260 ( 2-3 )   261 - 268   1998

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    DOI： 10.1007/s004380050894

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  • Evolutionary dynamics of giant sex chromosomes in the genus Silene

    赤木剛士, 藤田尚子, 増田佳苗, 白澤健太, 長岐清孝, 堀内綾乃, 桑田恵理子, 久能理子, 中村光希, 池田陽子, 牛島幸一郎, CHARLESWORTH Deborah

    育種学研究   26   2024

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  • Population structure analysis and GWAS of fruit shape traits in hexaploid persimmon cultivars

    堀内綾乃, 堀内綾乃, 尾上典之, 松崎隆介, 南川舞, 西村和紗, 白澤健太, 久保康隆, 牛島幸一郎, 赤木剛士

    育種学研究   26   2024

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  • GWASによる六倍体カキ品種群の果実形状多様性機構の解明に向けて

    堀内綾乃, 堀内綾乃, 尾上典之, 南川舞, 西村和紗, 白澤健太, 久保康隆, 牛島幸一郎, 赤木剛士

    日本分子生物学会年会プログラム・要旨集(Web)   47th   2024

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  • Chromosome-scale genome assembly of a persimmon cv. Taishu revealed rapid evolution of a post-Y chromosome

    堀内綾乃, 増田佳苗, 白澤健太, 尾上典之, 藤田尚子, 牛島幸一郎, 赤木剛士

    園芸学研究 別冊   22 ( 1 )   2023

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  • Recurrent neo-sex chromosome evolution in the genus Actinidia (II)

    赤木剛士, 赤木剛士, VARKONYI-GASIC Erika, 白澤健太, CATANACH Andrew, HENRY Isabelle M., MERTTEN Daniel, DATSON Paul, 増田佳苗, 藤田尚子, 桑田恵理子, 牛島幸一郎, 別府賢治, ALLAN Andrew C., CHARLESWORTH Deborah, 片岡郁雄

    園芸学研究 別冊   22 ( 1 )   2023

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  • Silene属植物の全ゲノム比較解析から性の成立過程を紐解く

    藤田尚子, 白澤健太, 牛島幸一郎, 赤木剛士

    Plant Morphology   35 ( 1 )   2023

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  • Silene属の全ゲノム比較進化解析から紐解く性染色体と性決定因子群の成立

    藤田尚子, 白澤健太, 牛島幸一郎, 赤木剛士

    日本植物学会大会研究発表記録(CD-ROM)   86th   2022

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  • Recurrent neo-sex chromosome evolution in the genus Actinidia (I)

    赤木剛士, 赤木剛士, VARKONYI-GASIC Erika, 白澤健太, CATANACH Andrew, HENRY Isabelle M., DATSON Paul, 増田佳苗, 藤田尚子, 桑田恵理子, 牛島幸一郎, 別府賢治, ALLAN Andrew C., COMAI Luca, CHARLESWORTH Deborah, 片岡郁雄

    園芸学研究 別冊   21 ( 2 )   2022

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  • Population genetic analysis for fruit shape diversity specific to hexaploid persimmon cultivars

    堀内綾乃, 増田佳苗, 増田佳苗, 尾上典之, 松崎隆介, 白澤健太, 久保康隆, 牛島幸一郎, 赤木剛士

    園芸学研究 別冊   21 ( 2 )   2022

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  • Comparative genome analysis of two Silene species with and without sex chromosomes

    藤田尚子, 白澤健太, 牛島幸一郎, 赤木剛士

    園芸学研究 別冊   21 ( 2 )   2022

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  • Prediction and premonitory symptoms characterization for rapid over-softening in persimmon fruit, with deep learning

    鈴木茉莉亜, 増田佳苗, 杉浦真由, 鈴木哲也, 新川猛, 久保康隆, 牛島幸一郎, 内田誠一, 赤木剛士, 赤木剛士

    園芸学研究 別冊   21 ( 2 )   2022

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  • Variety differentiation in hexaploid persimmon, with a wide fruit shape diversity

    堀内綾乃, 増田佳苗, 増田佳苗, 尾上典之, 松崎隆介, 白澤健太, 久保康隆, 牛島幸一郎, 赤木剛士

    育種学研究   24   2022

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  • Comparative analysis on somaclonal mutants for fruit shape determination in persimmon

    堀内綾乃, 増田佳苗, 松崎隆介, 尾上典之, 久保康隆, 牛島幸一郎, 赤木剛士

    育種学研究   24   2022

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  • Characterization of Fruit Enlargement and Ripening in Extremely Late Maturing Peach

    20 ( 1 )   65 - 71   2021.1

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  • The genetic control and transcriptome analyses of the slow softening strain of melon

    合田駿平, 池田和生, 小林和真, 赤木剛士, 加藤鎌司, 久保康隆, 中野龍平, 牛島幸一郎

    園芸学研究 別冊   20 ( 1 )   2021

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  • A candidate gene responsible for susceptibility to the host-specific toxin of Alternaria alternata pathotype peach

    浅野貴洋, 河井崇, 鵜木悠治郎, 日原誠介, 白澤健太, 赤木剛士, 久保康隆, 高田大輔, 中野龍平, 福田文夫, 山本幹博, 牛島幸一郎

    園芸学研究 別冊   20 ( 2 )   2021

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  • Prediction of enriched small RNA sequences with deep learning for application to horticultural crops

    榎那津美, 増田佳苗, 久保康隆, 牛島幸一郎, 内田誠一, 赤木剛士, 赤木剛士

    園芸学研究 別冊   20 ( 2 )   2021

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  • Genome-wide survey of cis-motifs responsible for fruit ripening in kiwifruit, with explainable deep learning

    桑田恵理子, 竹下孔喜, 藤田尚子, 牛島幸一郎, 久保康隆, 別府賢治, 片岡郁雄, 内田誠一, 赤木剛士, 赤木剛士

    園芸学研究 別冊   20 ( 2 )   2021

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  • Genome-wide cis-decoding for expression designing in tomato using explainable deep learning

    赤木剛士, 赤木剛士, 増田佳苗, 桑田恵理子, 竹下孔喜, 川勝泰二, 有泉亨, 久保康隆, 牛島幸一郎, 内田誠一

    園芸学研究 別冊   20 ( 2 )   2021

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  • Prediction and premonitory symptoms characterization for rapid over-softening in persimmon fruit, with deep learning

    鈴木茉莉亜, 増田佳苗, 竹下孔喜, 朝隈英昭, 杉浦真由, 鈴木哲也, 新川猛, 久保康隆, 牛島幸一郎, 内田誠一, 赤木剛士, 赤木剛士

    園芸学研究 別冊   20 ( 2 )   2021

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  • Comparative analysis for persimmon fruit shape determination in ‘Hiratanenashi’ and the bud sport ‘Koushimaru’

    堀内綾乃, 増田佳苗, 松崎隆介, 尾上典之, 久保康隆, 牛島幸一郎, 赤木剛士

    園芸学研究 別冊   20 ( 2 )   2021

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  • Reinvention of hermaphroditism triggered by activation of a RADIALIS-like gene in hexaploid persimmon

    増田佳苗, 池田陽子, 松浦恭和, 川勝泰二, 田尾龍太郎, 久保康隆, 牛島幸一郎, HENRY Isabelle M., 赤木剛士, 赤木剛士

    園芸学研究 別冊   20 ( 2 )   2021

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  • Inheritance of ultra-late maturing trait of ‘Fuyumomogatari’ peach in F1 progeny

    河井崇, 牛島幸一郎, 三宅春菜, 草加芽依, 深松陽介, 原美由紀, 鵜木悠治郎, 日原誠介, 中野龍平, 小田賢司, 福田文夫

    園芸学研究 別冊   19 ( 1 )   2020

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  • 性の可塑化を規定する遺伝子ネットワーク:六倍体カキにおける両性花派生機構

    増田佳苗, 牛島幸一郎, 久保康隆, 赤木剛士

    日本植物学会大会研究発表記録(CD-ROM)   84th   2020

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  • Polyploid-specific reinvention of hermaphroditism in persimmon is triggered by stress-induced NGATHA

    増田佳苗, 牛島幸一郎, 久保康隆, 赤木剛士

    育種学研究   22   2020

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  • カキ品種の果実生育・成熟に関する倍数体用ゲノムワイドアソシエーション解析

    赤木剛士, 増田佳苗, 牛島幸一郎, 久保康隆

    園芸学研究 別冊   18 ( 2 )   2019

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  • カキの両性花派生に関する共発現ネットワーク解析

    増田佳苗, 牛島幸一郎, 久保康隆, 田尾龍太郎, 赤木剛士

    園芸学研究 別冊   18 ( 2 )   2019

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  • ‘香粋’キウイフルーツにおける芳香成分と一次代謝産物のGC/MSによる解析

    常盤淑玲, 笠原有加, 土佐康彰, MITALO Witere Oscar, 近藤憂紀, 大槻巧, 大地菜摘, AZIMI Azimullah, DOAN Anh Thu, GALIS Ivan, 中野龍平, 中野龍平, 牛島幸一郎, 牛島幸一郎, 久保康隆, 久保康隆

    園芸学研究 別冊   17 ( 2 )   2018

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  • 棚持ち性の良いメロン系統の交配後代における表現型の調査

    岡本健, 上山美穂, 中西諒祐, 牛島幸一郎, 村上賢治, 加藤鎌司, 久保康隆, 中野龍平

    園芸学研究 別冊   15 ( 1 )   2016

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  • モモ晩生品種の香港中秋節時期に向けた輸出試験に関して

    中野龍平, 岡村憲一, 荒木克也, 志水基修, 長谷川圭則, 藤井雄一郎, 荒木有朋, 樋野友之, 牛島幸一郎, 久保康隆, 森永邦久, 福田文夫

    園芸学研究 別冊   15 ( 1 )   2016

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  • マメガキを用いたカキの甘渋性決定遺伝子座の比較ゲノム解析

    西山総一郎, 佐藤明彦, 河野淳, 尾上典之, 牛島幸一郎, 山根久代, 田尾龍太郎, 米森敬三

    園芸学研究 別冊   14 ( 2 )   2015

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  • ハミウリおよびその交配後代の棚持ち性に関する研究

    岡本健, 中村智哉, 上山美穂, 山岡達也, 牛島幸一郎, 久保康隆, 加藤鎌司, 中野龍平

    園芸学会中四国支部研究発表要旨   ( 54 )   2015

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  • 棚持ち性の良いメロン系統のエチレン応答性に関する研究

    岡本健, 中村智哉, 上山美穂, 浜田佳代子, 山岡達也, 牛島幸一郎, 村上賢治, 加藤鎌司, 久保康隆, 中野龍平

    園芸学研究 別冊   14 ( 2 )   2015

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  • メロン果実成熟過程におけるエチレンシグナル伝達系遺伝子の発現パターン

    山岡達也, 中野龍平, 加藤鎌司, 久保康隆, 牛島幸一郎

    園芸学研究 別冊   11 ( 2 )   2012

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  • Subtraction analysis for cloning of hetelostyle related genes in Linum grandiflorum

    BANDO M., KUBO Y., NAKANO R., USHIJIMA K.

    7 ( 1 )   406 - 406   2008.3

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  • Coupling functional screen and macroarray for a comprehensive analysis of the secretomes associated with ripening-related changes in pear fruits

    MWANIKI M. W., NAKANO R., USHIJIMA K., AOKI K., ROSE J. K. C., KUBO Y.

    7 ( 1 )   243 - 243   2008.3

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  • Analysis of ripening-associated genes using tomato macroarray

    KAMITAKA D., HARANO T., YOKOTANI N., NAKANO R., USIJIMA K., INABA A., KUBO Y.

    6 ( 2 )   405 - 405   2007.9

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  • Studies on long-term storage of 'Sanuki-Gold' kiwi fruit

    YOSHIKAWA T., MWORIA E., NAKANO R., USHIJIMA K., FUKUDA T., SUEZAWA K., KUBO Y.

    6 ( 2 )   401 - 401   2007.9

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  • Characterization of ethylene biosynthesis and its regulation fruit ripening in Actinidia chinensis var. 'Sanuki Gold'

    MWORIA E., YOSHIKAWA T., NAKANO R., USHIJIMA K., FUKUDA T., SUEZAWA K., KUBO Y.

    6 ( 2 )   403 - 403   2007.9

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  • Expression pattern of ethylene responsible genes in 'Honey Dew' melon

    NISHIYAMA K., NAKANO R., KATO K., INABA A., KUBO Y., USHIJIMA K.

    6 ( 2 )   404 - 404   2007.9

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  • Phenotype analysis of transgenic tomato fruit with suppressed EIN3/EIL

    HARANO T., CHAMITA D., YUCATAN N., NAKANO R., USHIJIMA K., INDABA A., KUBO Y.

    6 ( 2 )   649 - 649   2007.9

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  • Characterization of cell wall proteins related with fruit softening in European pear

    BANDO M., USHIJIMA K., NAKANO R., INABA A., KUBO Y.

    6 ( 1 )   289 - 289   2007.3

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  • β-Galactosidase and α-L-Arabinofuranosidase Activities and Gene Expression in European and Chinese Pear Fruit During Ripening :

    Mwaniki Mercy W., Mathooko Francis M., Hiwasa Kyoko, Tateishi Akira, Yokotani Naoki, Ushijima Koichiro, Nakano Ryohei, Inaha Akitsugu, Kubo Yasutaka

    Journal of the Japanese Society for Horticultural Science   76 ( 1 )   85 - 90   2007

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    Language：English   Publisher：Japanese Society for Horticultural Science  

    Substantial decreases in cell wall bound galactosyl and arabinosyl residues are two of the most evident cell wall compositional changes that occur during fruit ripening. The roles of β-galactosidase (β-Gal) and α-L-arabinofuranosidase (α-Af), the enzymes responsible for these respective losses, were investigated and compared in European (Pyrus communis L. 'La France') and Chinese (Pyrus bretschneideri Rehd. 'Yali') pear fruits which exhibit different softening characteristics during ripening. The increase in the activities of β-Gal and α-Af during ripening in both types of pear fruit correlated well with an increase in climacteric ethylene production, and a concomitant decrease in flesh firmness in 'La France' fruit. However, there was no noticeable decrease in 'Yali' flesh firmness even after 28 days of storage at room temperature. In both fruit types, enzyme activity and the accumulation of transcripts hybridizing with PpGAL1, PpGAL4, PpARF2, and PcARF1 increased with fruit ripening. Increases in gene expression and enzyme activities in 'Yali' fruit with no detectable softening during ripening indicate that β-Gal and α-Af may not mediate difference in fruit softening between two pears, but that they could play some role(s) in cell wall changes, perhaps in cooperation with other cell wall-modifying enzymes such as polygalacturonase.

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    Other Link： http://dl.ndl.go.jp/info:ndljp/pid/10471864

  • Comparative characterization of fruit development in 'Nakataniwase', 'Tonewase' and 'Hiratanenashi' Japanese persimmons and inhibition of postharvest fruit softening

    SAGARA H., HARIMA S., NAKANO R., USHIJIMA K., KUBO Y., INABA A.

    75 ( 2 )   409 - 409   2006.9

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  • The responsibility of ripening-related characteristics to ethylene in 'Honey Dew' melon

    NISHIYAMA K., USHIJIMA K., KATO K., NAKANO R., INABA A., KUBO Y.

    75 ( 2 )   610 - 610   2006.9

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  • メロン品種における果実成熟特性とエチレン関連遺伝子の発現

    西山精視, 牛島幸一郎, 加藤鎌司, 稲葉昭次, 久保康隆

    園芸学会雑誌 別冊   75 ( 1 )   2006

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  • Comparison of fruit softening and expression pattern of polygalacturonase gene among four melon lines

    NISHIYAMA K., MIYAZAKI T., USHIJIMA K., KATO K., INABA A., KUBO Y.

    74 ( 2 )   553 - 553   2005.10

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  • Expression of softening-related genes in melon fruit during ripening

    NISHIYAMA K., MIYAZAKI T., USHIJIMA K., KATO K., INABA A., KUBO Y.

    74 ( 1 )   401 - 401   2005.3

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  • Cloning and Expression of Ripening-related MADS Family Genes in Grape Berry

    YAMANE M., OZAKI M., NAKANO R., USHIJIMA K., KUBO Y., INABA A.

    74 ( 1 )   383 - 383   2005.3

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  • Suppression of genetic recombination at S locus of sweet cherry

    IKEDA K, USHIJIMA K, YAMANE H, IEZZONI Amy, TAO R

    園芸学会雑誌. 別冊, 園芸学会大会研究発表   73 ( 2 )   557 - 557   2004.9

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  • Analysis of self-compatible S^f haplotype of Japanese apricot

    KAKEHI E, USHIJIMA K, YAMANE H, TAO R

    園芸学会雑誌. 別冊, 園芸学会大会研究発表   73 ( 2 )   558 - 558   2004.9

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  • β-galactosidase and arabinofuranosidase activities and gene expression in three types of pear fruit

    MWANIKI M. W., MATHOOKO F. M., HIWASA K., TATEISHI A., USHIJIMA K., INABA A., KUBO Y.

    73 ( 2 )   517 - 517   2004.9

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  • Molecular markers for the self-compatible S-4 '-haplotype, a pollen-part mutant in sweet cherry (Prunus avium L.)

    K Ikeda, A Watari, K Ushijima, H Yamane, NR Hauck, AF Iezzoni, R Tao

    JOURNAL OF THE AMERICAN SOCIETY FOR HORTICULTURAL SCIENCE   129 ( 5 )   724 - 728   2004.9

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  • Primary structural features of the S haplotype-specific protein, SFB, in Prunus

    IKEDA K, USHIJIMA K, YAMANE H, SASSA H, IEZZONI A. F, KOHN J. R, TAO R

    園芸学会雑誌. 別冊, 園芸学会大会研究発表   73 ( 1 )   180 - 180   2004.4

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  • dCAPS markers for the self-compatible S^<4'>-haplotype of sweet cherry

    WATARI A., USHIJIMA K., TAO R.

    73 ( 1 )   369 - 369   2004.4

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  • A mode of mutation in SFB of the self-compatible S^<4'> haplotype in sweet cherry

    USHIJIMA K., IKEDA K., TAO R.

    73 ( 1 )   368 - 368   2004.4

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  • The S haplotype-specific F-box protein gene, SFB, is defective in self-compatible haplotypes of Prunus avium and P. mume

    USHIJIMA K

    Plant J   39   573 - 586   2004

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  • Characterization of self-compatible S^4' haplotype of sweet cherry : 1. DNA blot analysis for the S haplotype-specific F-box protein gene (SFB)

    IKEDA K, WATARI A, USHIJIMA K, YAMANE H, IEZZONI A. F, TAO R

    園芸学会雑誌. 別冊, 園芸学会大会研究発表   72 ( 2 )   291 - 291   2003.9

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  • Linked to a Gene for S-RNase in Two Species of Cherry, Prununs cerasus and P. avium

    YAMANE Hisayo, IKEDA Kazuo, USHIJIMA Koichiro, SASSA Hidenori, TAO Ryutaro

    Plant and Cell Physiology   44 ( 7 )   764 - 769   2003.7

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  • Structural features of the Prunus Slocus

    USHIJIMA K, SASSA H, YAMANE H, TAO R

    園芸学会雑誌. 別冊, 園芸学会大会研究発表   72 ( 1 )   155 - 155   2003.4

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  • Identification of the candidate gene for pollen S in cherry

    YAMANE H., IKEDA K., USHIJIMA K., SASSA H., TAO R.

    72 ( 1 )   342 - 342   2003.4

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  • Identification of SFB, a candidate gene for the pollen-S, in self-incompatible Japanese apricot cultivars

    YAMANE H., NAMBA A., USHIJIMA K., SASSA H., TAO R.

    72 ( 1 )   340 - 340   2003.4

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  • Isolation and expression pattern analysis of the candidate for almond pollen S gene

    USHIJIMA K., SASSA H., GRADZIEL Thomas M., DANDEKAR Abhaya M., TAO R.

    71 ( 2 )   489 - 489   2002.10

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  • Characterization of the S-locus region of almond (Prunus dulcis): analysis of a somaclonal mutant and a cosmid contig for an S haplotype

    USHIJIMA K.

    Genetics   158   379 - 386   2001

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Research Projects

  • Transcriptome/Cistrome analysis of ethylene synthesis mutants to elucidate the regulation of fruit ripening

    Grant number：24K01747  2024.04 - 2029.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    牛島 幸一郎, 柏本 知晟, 赤木 剛士, 中野 龍平, 池田 和生, 河井 崇

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    Grant amount：\18590000 （ Direct expense: \14300000 、 Indirect expense：\4290000 ）

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  • Construction of a fine reference genome of Hyuganatsu applied for identification of causal genes inducing mutate traits

    Grant number：23K26898  2023.04 - 2027.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    本勝 千歳, 稲葉 丈人, 牛島 幸一郎

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    Grant amount：\13910000 （ Direct expense: \10700000 、 Indirect expense：\3210000 ）

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  • 黒毛和種の受胎率低下に関わるロバートソン転座の構造解明と遺伝子診断法の確立

    Grant number：23K05556  2023.04 - 2026.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    辻 岳人, 牛島 幸一郎

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    Grant amount：\4680000 （ Direct expense: \3600000 、 Indirect expense：\1080000 ）

    本年度はA県とB県の黒毛和種繁殖雌牛から採取した血液をもちいて染色体標本を作成し、ロバートソン転座の有無を調査した。合計62頭を調査した結果、ロバートソン転座は認められなかった。黒毛和種におけるロバートソン転座に関する報告としては、30年ほど前のもので近年の調査報告はない。また30年前の報告では、調査された複数の県すべてにおいてロバートソン転座が約5%から15%の個体で認めらたことが示されている。したがって、本年度の我々の結果は、現在の黒毛和種においてロバートソン転座の頻度はかなり低下している可能性を示している。しかし、地域集団ごとにロバートソン転座に頻度に大きな違いがあることも考えられるため、今後さらに調査対象を拡大する必要がある。

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  • Construction of the peach pan-genome and its application to the identification of genes that control the major traits of peach

    Grant number：22H00368  2022.04 - 2027.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (A)

    中野 龍平, 牛島 幸一郎, 山本 幹博, 福田 文夫, 西村 和紗, 高田 大輔, 鵜木 悠治郎, 森本 拓也, 小田 賢司

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    Grant amount：\41860000 （ Direct expense: \32200000 、 Indirect expense：\9660000 ）

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  • New Frontier of Fruit Postharvest Physiology-Low Temperature modulated Ripening and its application

    Grant number：20H02977  2020.04 - 2024.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    久保 康隆, 矢野 健太郎, 阿部 大吾, 加藤 雅也, 牛島 幸一郎, 赤木 剛士, 河井 崇

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    Grant amount：\17810000 （ Direct expense: \13700000 、 Indirect expense：\4110000 ）

    本研究は、低温誘導成熟機構を、温帯性果実種横断的にゲノム・トランスクリプトーム解析、メタボローム解析、遺伝子組換え技術などを駆使して解析し、その統合的理解を目指す先駆的学術的取り組みである。低温誘導成熟果はエチレン誘導果よりも品質保持期間が長く、低温感受性の品種間差異は成熟の早生・晩生性に密接に関与している。本研究は、収穫後ロス低減に直結する新品質保持技術開発と分子マーカー開発による育種への貢献を目的とする。
    本年度は、研究の第２段階として、ウンシュウミカン、カキ、セイヨウナシ、リンゴ果実でのRNAseq解析、LC解析に基づいて成熟制御システムの全体像を把握するとともに、低温応答遺伝子および代謝物を抽出を試みた。今回解析したいずれの果実でもエチレン処理および低温遭遇によって、細胞壁分解系遺伝子、クロロフィル分解系遺伝子発現が刺激されることが明らかになった。さらに、ウンシュウミカンとレモン果実の低温応答に関するRNAseqデータの解析によって、カンキツでは、共通してERF3遺伝子が低温応答に重要な転写因子候補であることが示された。また、 ‘小原紅早生’ミカンではエチレンシグナルと低温シグナルによりCCD4遺伝子が活性化され、アポカロテノイド類が特異的に蓄積して成熟時に紅色に着色することが示された。また、‘富有’カキでは５℃貯蔵によってエチレン生成が刺激され果肉軟化と水浸状化を特徴とする低温障害症状がが進行するが、20℃以上または０℃では果肉軟化が抑制された。ただし、０℃貯蔵では常温に移行すると急激にエチレン生成の誘導と果肉軟化が進行した。１ヶ月程度なら20℃の常温貯蔵が可能であることが示された

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  • セイヨウナシの周縁キメラ分離カルスを利用した果皮の全面赤着色メカニズムの解明

    Grant number：20K06030  2020.04 - 2023.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    池田 和生, 牛島 幸一郎

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    Grant amount：\4420000 （ Direct expense: \3400000 、 Indirect expense：\1020000 ）

    本年度は、（１）周縁キメラであるセイヨウナシ果実の果皮からL-1層のみを分離したカルスからシュートおよび根を再分化させ個体として独立させること目的として、前年度作成したカルスを材料として、カルスからの発根培養条件の検討（２）周縁キメラ品種の果皮から脱分化させた赤着色カルスと非着色カルスで発現している遺伝子についてトランスクリプトーム解析、を行った。（１）では、セイヨウナシ‘リーガル・レッド・コミス’の果皮培養からカルスを作成し、カルスからの発根に適した培養条件について基本培地およびオーキシン濃度を検討した。基本培地については、MS基本培地もしくは窒素濃度を半減させた１/２MS培地を検討した結果、継代回数が少ない段階ではMS培地と１/２MS培地のカルス生存率に差は認められなかった．しかしながら，継代３回目以降では基本培地はMS培地と比較して1/2MS培地のカルス生存率が高い傾向が見られた。これより，発根培養時のカルスの生存には培地の窒素濃度が低い方がよいと考えられた。一方、（２）では、赤着色（Red）カルスと非着色（Green）カルスからtotal RNAを抽出し、それぞれのカルスについて3サンプルずつ次世代シークエンスに供した結果、平均2000万リードのデータを得た。 ‘バートレット’のゲノム情報（GDR,www.rosaceae.org）を基にマッピングを行い，約3万3000遺伝子について遺伝子発現解析を行った。．赤着色（Red）カルスにおける遺伝子発現量が非着色（Green）カルスの4倍以上の遺伝子が約1700遺伝子，8倍以上の遺伝子が約600遺伝子、16倍以上の遺伝子が270遺伝子であることが明らかとなり、これらの遺伝子が赤着色形質に関与している可能性が高いと考えられた。

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  • Genetic control of long term storage in melons

    Grant number：19H02948  2019.04 - 2024.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    牛島 幸一郎, 門田 有希, 中野 龍平, 池田 和生

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    Grant amount：\17160000 （ Direct expense: \13200000 、 Indirect expense：\3960000 ）

    メロンは一般に急激な呼吸量の増加とともにエチレンの生成と果実軟化を示すクライマクテリック型果実であるが，実際にはエチレンを感受しないノンクライマクテリック型果実や，エチレン合成量が少ない，軟化速度が遅いなど，様々な成熟様相を示す品種が存在する.メロン品種のハネデュとB2は共に棚持ち性の良い品種であるが，前者はエチレン生合成，後者は果肉軟化に変異が生じている.R3年度はB2の棚持ち性に関する研究を中心に解析を行った．
    B2についてはR2年度に軟化遅延形質がF５世代で分離した．これにより１遺伝子座支配である可能性が示唆されているが，本来の予定であればF6世代で分離する予定であり，想定外の結果であった．この結果がただしいか否かを再度検証するとともにGWAS解析を行うことにした．R2年度と同じF5集団に加えてあらたにF6集団を栽培し，果肉の形質を調査した．F5に関しては前年と同様に軟化遅延形質が1:1で，F6では3:1で分離した．これらの結果から，やはり軟化遅延形質は１遺伝子座支配であると考えられた．そこで，GRAS-di解析を実施しSNPをコールして，GWAS解析を行った．その結果，高い相関を示すSNPは検出されなかった．僅かに相関が観察される遺伝子座もあったが，遺伝子型の分離比が歪んでおり信頼性が低いと考えられ，遺伝子座の特定には至らなかった．
    ハネデュのエチレン合成不全については，R2年度までに典型的なクライマクテリック型果実のシャランテとのF2集団でGWAS解析やRNA-seq解析などから制御因子の特定を試みて，8番染色体に相関は見られたが候補遺伝子の特定には至っていない．他の形質と比べると相関が高くないため再度集団を作り解析を試みることにした．R3年度はF1の栽培及びF2世代の種子の回収を行い，次年度の解析のための材料を得た．

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  • Is self-incompatibility in Citrus spp. governed by S-RNase?

    Grant number：19H02949  2019.04 - 2023.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    本勝 千歳, 稲葉 丈人, 牛島 幸一郎

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    Grant amount：\17290000 （ Direct expense: \13300000 、 Indirect expense：\3990000 ）

    近年カンキツ属のいくつかの品種で自家不和合性Sハプロタイプが決定され，またこれまでに行った次世代シークエンス解析の結果から，SハプロタイプとT2RNase型との対応関係が示唆された．このことから，T2RNaseがS遺伝子である可能性が高いと考えられる．そこで本研究では，組換えタンパク質によるT2RNaseの機能解析，T2RNase型とSハプロタイプとの対応関係の調査と新規T2RNase遺伝子の獲得，そしてSハプロタイプとT2RNase遺伝子の遺伝解析を行い，T2RNase遺伝子とカンキツ属植物の自家不和合性の関連性について知見を得ようとするものである．
    本年度は，自家不和合性カンキツ属植物のヒュウガナツにおいて，枝変わりによって生じた自家和合性突然変異体の解析を行った．自家和合性ヒュウガナツである’井原日向’の花柱からRNAを抽出し，RNAseqデータを取得して，発現解析を行った．その結果，ヒュウガナツの自家不和合性遺伝子であるS15-RNaseが’井原日向’において，有意に発現が低下する発現変動遺伝子（DEG）に含まれていることが明らかとなった．この遺伝子については，PCRによって’井原日向’内に遺伝子が存在することを確認した．一方でRT-qPCRにより’井原日向’での発現低下が認められた．
    昨年度に取得したクレメンティンの花粉より獲得したRNAseqデータの解析を行った．その結果，クレメンティンのS-RNase遺伝子と思われるCiclev10027322m.gの周辺に存在する複数のF-Box遺伝子が花粉特異的に発現しており，これまでに報告されているS-RNase型自家不和合性植物と類似した発現パターンを示すことを明らかにした．

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  • Studies on regulatory mechanisms of ultra-late maturing trait in 'Gento' peach

    Grant number：19K06033  2019.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    KAWAI Takashi

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    Grant amount：\4420000 （ Direct expense: \3400000 、 Indirect expense：\1020000 ）

    In order to elucidate the mechanisms regulating ultra-late maturing trait in ‘Gento’ peach, its inheritance in F1 progeny as well as physiological and histological changes during fruit development were investigated. Although F1 individuals that matured as late as ‘Gento’ were not observed, maturity date in F1 progeny exhibited a binominal distribution and 9 bp indel in NAC gene on LG4 co-segregated with the maturity date of all the individuals. Cell size and ABA content in the flesh of ‘Gento’ increased much later than common peach cultivar, corresponding to their fruit growth patterns. These results, combined with transcriptome analysis by RNA-seq and future genetic analysis using F2 progeny, will help us to fully understand the molecular basis for the ultra-late maturing trait in ‘Gento’.

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  • Characterization of long shelf-life and pulp disorder-alleviating mechanisms using late-harvest rare peach cultivars bred private growers as genetic resource

    Grant number：18H02200  2018.04 - 2023.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    中野 龍平, 牛島 幸一郎, 福田 文夫, 高田 大輔, 河井 崇

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    Grant amount：\17030000 （ Direct expense: \13100000 、 Indirect expense：\3930000 ）

    １）桃水と大寿蜜桃を用いたエチレン生成およびエチレン応答に関する解析：桃水ではエチレン生成能を持たず、大寿蜜桃はエチレンに応答した軟化力を失っている。RNA-seq解析の結果、桃水では他の２品種と比べて、91のDEGが検出され、２遺伝子は転写因子をコードしており、これらの自己触媒的エチレン生成能の欠損性との関連が示唆された。大寿蜜桃では、441のDEGが検出され、16遺伝子は転写因子をコードしていた。本研究で欠損が証明されたPG遺伝子以外にも、軟化へ関与するExpansinなどの軟化関連遺伝子やAATなどの香り成分合成関連遺伝子の発現が抑制されていた。PG遺伝子に加えてエチレン応答性に関わる転写因子の抑制により、軟化や香り成分生成が抑えられていると考えられた。さらなる証明と関連遺伝子の探索のために、大寿蜜桃と川中島白桃の交配後代を育成したところ、大苗育苗などにより開花が促進され、約半数の後代において開花が認められた。
    ２）紅博桃、枝変わり系統およびあかつきの解析：紅博桃枝変わりに生じたLOH（Loss of heterozygosity）により、熟期以外にも果肉障害、異形果や落下果実の多発など様々な不良形質が現れることが分かった。このLOHによりホモ化すると不良形質を生む染色体領域は本白桃由来であること、本白桃、あかつき、川中島白桃など日本のモモ育種における中心的な品種がヘテロに保持していることが分かり、育種過程において不良形質を示す後代を選抜する有効な遺伝情報を得た。
    ３）ワッサーなど特徴的な日持ち性や肉質を示すとされる他の民間育成品種：ワッサー、CX、ネクタリン系のサンライズなど、最近、育成された新品種に桃水と同様の硬肉モモ系の品種が存在することが明らかとなった。一方、大寿蜜桃と同様の軟化形質やPGの遺伝子型を持つ品種は、缶モモも含め近年の国内栽培品種には見つからなかった。

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  • Studies on ripening and scenesense inducing-mechanism(s) controlled by factors at downstream of signaling pathway in non-climactericl fruit

    Grant number：16K14853  2016.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Exploratory Research

    Nakano Ryohei, Kubo Yasutaka

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    Grant amount：\3640000 （ Direct expense: \2800000 、 Indirect expense：\840000 ）

    Candidate factors that could work at down-streams on ripening related signaling pathways were studied using strawberry wild species F. vesca in an attempt to reveal the mechanism(s) that regulate fruit ripening and senescence in non-climacteric type of fruits. For EIN3/EILs transcription factors, RNAi transformant were generated using conserved region and strains with suppressed expression of Fv-EIL1 and 2 were obtained, though they ripened and senesced normally, suggesting the possible involvement of other isogenes such as Fv-EIL3. For NAC transcription factors, within isogene with target sequence of miRNA164, expression level of NAC-SF was dramatically increased during ripening suggesting its’ involvement. In virus induced gene silencing (vigs), infecting agrobacterium with trv-originated vigs constructs to seeds of F. vesca effectively induced gene silencing, showing increased possibility to perform functional analysis of target genes in strawberry.

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  • Iso-Seq and RNA-seq based transcriptome analyses associated with root-knot nematode resistance in sweetpotato

    Grant number：16K07557  2016.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Monden Yuki

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    Grant amount：\4810000 （ Direct expense: \3700000 、 Indirect expense：\1110000 ）

    Sweetpotato is one of the most important crop species in the world, and it has been required to breed superior cultivars with disease resistance, high yield, and nutrient richness etc. However, as a result of the crop’s complex genomic architecture, which results from its hexaploidy (2n = 6x = 90), high heterozygosity, huge genome (2~3Gb), and outcrossing nature, its genetic analysis and marker-assisted breeding has been challenging. In addition, the lack of whole genome sequence or gene annotations has hindered the isolation of agronomically important genes or validation of QTL regions. In this study, we performed an Iso-Seq to obtain full length transcripts without the assembly, and also conducted an RNA-seq analysis to understand the mechanism for nematode resistance in sweetpotato. As a result, more than 54,000 isoforms were produced by the Iso-seq, and we could identify candidate gene(s) for controlling nematode resistance.

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  • Comprehensive analysis of full length cDNA and alternative splicing for the genes related to fruit development and ripening

    Grant number：16K14854  2016.04 - 2018.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Exploratory Research

    Ushijima Koichiro

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    Grant amount：\3640000 （ Direct expense: \2800000 、 Indirect expense：\840000 ）

    3rd generation sequences promotes not only genome sequencing but also transcript analysis. Iso-seq analysis using PacBio are very useful method for reading full length cDNA and detecting alternative splicing. In this analysis, we carried out Iso-seq analysis for melon fruits. We used three different methods for construction of full length cDNA library. The libraries were sequenced by PacBio sequel and Nanopore MinION. Isoforms were mapped on the reference genome of melon and compared them. PacBio and Nanopore isoforms fell into ~ 6,500 and 12,000 loci respectively. 509 loci of them were located at intergenic region and predicted novel loci.

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  • Molecular biological studies on shelf-life in melon using various genetic resources

    Grant number：26292018  2014.04 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    Nakano Ryohei, Kubo Yasutaka

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    Grant amount：\15860000 （ Direct expense: \12200000 、 Indirect expense：\3660000 ）

    In melon genetic resources with long self-life, their physiological characteristics were studied as well as hereditary pattern of their traits and related genes. 1) B2 melon produced ripening ethylene, while lose ability to soften in response to ethylene. That trait was linked to suppressed expression of cell wall degradation related-genes, Cm-PG1 and Cm-EXP5. The trait showed dominant inheritance in cross between B2 and Haru3-melon. 2) Haneydew-melon lose ability to produce ripening ethylene. Linkage analysis in F2 population of corss between haneydew and charentais-melon suggest chromosome regions linked to this trait. 3) In progeny of cross between hami-melon and charentais, two types of strains, one lacking ability to produce ripening ethylene and the other lacking ability to soften in response to ethylene were found. Both traits were recessive and through repeated backcrosses against charentais-melon, strains possessing each trait was identified at BC4F2 generation.

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  • Molecular Analysis for heterostyly by using genomics and transcriptmics approches

    Grant number：25292023  2013.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    Ushijima Koichiro

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    Grant amount：\18980000 （ Direct expense: \14600000 、 Indirect expense：\4380000 ）

    Heterostyly is one of self-incompatibility that is a genetic system to reject the self-pollen and be linked to floral morph polymorphism．In this study, to elucidate the molecular mechanism of Linum heterostyly, we isolated the candidate gene controlling heterostyly and characterized the S locus region. Four candidate gene were newly isolated by transcriptome analysis. Genome analysis revealed the S haplotype specific region at least reached more that 1 Mbp. All candidate genes were included in this region but the relative locations were unclear. We further shotgun sequenced the genome of Linum grandiflorum and de novo assembly. The assembled contigs will give new insights on the study of heterostyly.

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  • Attempt for development of the novel method using JAZ, a hub factor for ethylene and JA signal crosstalk, to regulate fruit ripening and senescence

    Grant number：25660028  2013.04 - 2016.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Exploratory Research

    Ushijma Koichiro, NAKANO Ryohei

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    Grant amount：\3900000 （ Direct expense: \3000000 、 Indirect expense：\900000 ）

    Ripening in the climacteric fruits is regulated by ethylene. Many strategies have been proposed to modulate the ripening to elongate storage term and keep it moderate. Transgenic approaches and dose treatment can inhibit ethylene synthesis or perception completely, but the treated fruit is not suitable for food because they cannot ripe to be edible. We attempt to regulate ethylene signaling cascade by using hormone cross talking pathway to regulate fruit ripening moderately. From recent report, JAZ gene was expected to be a good candidate for the signaling hub between ethylene and JA. We isolate all JAZ genes from tomato and melon database and classify them. RNA-seq analysis demonstrated that the expression level of JAZ1 is drastically reduced at the ripening stage and affected by ethylene related mutant/transgenic plant. This fact suggested a possibility that JAZ1 is related to the ethylene signaling and fruit ripening.

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  • DIfferent mechanisms of self-incompatibility in Rosaceae: non-self recognition by multiple factors type ans self recognition by single factor type

    Grant number：24380004  2012.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    SASSA Hidenori

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    Grant amount：\19240000 （ Direct expense: \14800000 、 Indirect expense：\4440000 ）

    Pollen-expressed apple homologs of SBP1 and SSK1 were cloned and named MdSBP1 and MdSSK1, respectively, and subjected to further analyses. Both MdSBP1 and MdSSK1 interacted with SFBB. Further analyses showed that interaction of MdSSK1 with SFBB is stronger than that of MdSBP1, and that mRNA of MdSSK1 is much abundant than MdSBP1. Together, the results suggest that MdSSK1 containing SCFSFBB complex would play a major role in self-incompatibility in apple.

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  • Regulation of IPT gene under control of senescence activated promoter

    Grant number：23780025  2011 - 2013

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (B)

    NAKANO Ryohei, KUBO Yasutaka, USHIJIMA Koichiro

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    Grant amount：\3770000 （ Direct expense: \2900000 、 Indirect expense：\870000 ）

    Chimeric gene that induced cytokinin biosynthesis regulating gene IPT under the control of senescence activated promoter PSARK (PSARK-IPT) was introduced into floricultural plants such as petunia and chrysanthemum and their shelf-life were observed. As results, in 'Seikonomakoto' chrysanthemum which has a problem of leaf-yellowing during storage and transportation, cut flowers harvested from PSARK-IPT introduced trangenic plants showed markedly delayed leaf-yellowing. They also showed delayed leaf-senescence even under dark condition and ethylene exposure. These results provide valuable information for breeding floricultural plants with stress-tolerance and long-self life that would lead to low cost, energy saving and economical transportation.

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  • Analysis of molecular mechanism, origin and diversity of gametophytic self-incompatibility systems

    Grant number：20380003  2008 - 2011

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    SASSA Hidenori, USHIJIMA Koichiro, KOTODA Nobuhiro

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    Grant amount：\18980000 （ Direct expense: \14600000 、 Indirect expense：\4380000 ）

    Analysis of candidates for pollen-determinant in Japanese pear and apple of Rosaceae suggested that multiple and related F-box genes, SFBB, are collaboratively detoxify non-self pistil determinant, S-RNase. Isolation and functional analysis of HT-B gene, a non-S-specific self-incompatibility factor, of petunia showed that, unlike other solanaceous species, even a dramatic suppression of HT-B transcript resulted in partial breakdown of self-incompatibility in petunia. This may suggest slight functional divergence of HT-B gene in different species of Solanaceae.

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  • Establishment of transient RNAi methods in tree fruit and it's application on evaluation of genes regulating fruit ripening

    Grant number：20780021  2008 - 2010

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (B)

    NAKANO Ryouhei, KUBO Yasutaka, USHIJIMA Koichiro

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    Grant amount：\3640000 （ Direct expense: \2800000 、 Indirect expense：\840000 ）

    Transient RNA silencing technique was suggested to be possible in developing grape berry. When tomato is used as model plant for screening of fruit ripening-associated genes, AP3, TM6, AGL1, GRAS were assumed to be involved in ripening. Involvements of these genes in ripening of grape berry were investigated using the transient RNA silencing technique, although their involvements were not clear. It is suggested that the transient RNA silencing technique would be useful for analysis of ripening-associated genes in grape in future and that deeper investigation of function of these genes for fruit ripening will provide new insight into the study for fruit physiology.

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  • Studies on transcriptional regulation for ethylene biosynthesis genes in fruit ripening

    Grant number：19688003  2007 - 2009

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (A)

    USHIJIMA Koichiro

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    Grant amount：\25740000 （ Direct expense: \19800000 、 Indirect expense：\5940000 ）

    Tomatoes and melons are classified as climacteric fruits, whose ripening was regulated by ethylene. A mutant melon cultivar 'Honey dew' cannot synthesize ethylene at ripening stage and then be stored with a long storage life. Based on characterization of the mutation of 'Honey dew', I tried to identify the transcriptional regulator for ethylene synthesize genes and elucidate the mechanism for ethylene signaling and transcription. Then, 2 type genes were cloned as candidate genes. To date, the preliminary experiments, gene silencing and expression manipulation, suggested a possibility that that they and their homologues might repress fruits ripening.

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  • Identification of key factor fur cell wall disassembly in fruit by proteome analysis and transgenic technology

    Grant number：17380024  2005 - 2007

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    KUBO Yasutaka, NAKANO Ryohei, EZURA Hiroshi, USHIJIMA Khoichiro, INABA Akitsugu

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    Grant amount：\16950000 （ Direct expense: \15600000 、 Indirect expense：\1350000 ）

    Cell wall is main factor for physical structure of plant and the main limiting factors limiting their storage, transport and marketability. Previous studies indicated importance of plant hormone ethylene for fruit ripening and softening. However, key factor (s) in cell wall disassembly have not been identified. 'La France' European pear fruit and melon fruit undergo dramatic softening during ripening in an ethylene-dependent manner, whereas Chinese pear does not exhibit softening in spite of its massive ethylene production during ripening. To broad-scale and comprehensive screening for factors involved in fruit softening, in addition to gene expression analysis of polygalacturonase, α-arabinofuranosidase and β-galactosidase, the two-dimensional gel electrophoresis profiles and YSST analysis of cell wall proteins were employed. The accumulation levels of 200 protein spots were isolated in the ripening 'La France' pear. When softening pears of this variety were treated with 1-MCP, a strong ethylene inhibitor, fruit softening was retarded. On the comparison between treated and non-treated fruit, 3 protein spots were selected as candidates for key factors of fruit softening, since their accumulation showed specific pattern for softening.
    On YSST analysis, 170 clones (70 contigs and 100 singletons) encoding secreted proteins including clones which have not been previously reported were isolated. Based on the macroarray analysis, 28 clones showed elevated or down-regulated expression with pear fruit development and ripening. On comparison in 'La France' and the non-softening 'Yali' pear fruit, 11 potential new cell wall modifying proteins among 28 clones were selected for candidates for key factor of fruit softening.

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  • ノンクライマクテリック型果実の成熟制御因子の同定

    Grant number：17780025  2005 - 2006

    日本学術振興会  科学研究費助成事業  若手研究(B)

    牛島 幸一郎

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    Grant amount：\3600000 （ Direct expense: \3600000 ）

    果実はその成熟特性により、クライマクテリック型果実とノンクライマクテリック型果実とに分類される。クライマクテリック型果実はその成熟がエチレンによって制御されているタイプの果実であり、トマトやメロン、バナナなどが分類される。これまでに果実成熟の研究はクライマクテリック型果実を中心に行われており、ノンクライマクテリック型果実についての知見はわずかである。
    昨年度の研究において、様々なメロン品種の成熟特性の解析を行ったところ、クライマクテリック型果実とノンクライマクテリック型果実の中間のような特性を示す‘ハネデュ'という品種があることを示した。この品種はほとんどエチレンを生成しないため、ほとんど成熟が進行しない。そこで、エチレン生成以外の成熟形質に関して解析を行った。果肉軟化と香気成分生成について解析を行ったところ、空気条件下で保存した‘ハネデュ'果実は高い果肉硬度を維持し、ほとんど香気成分を生成しなかった。しかし、外生エチレンを処理すると果肉軟化、香気成分生成が促進された。これらの結果は、‘ハネデュ'ではエチレン信号の受容と伝達がなされていることを示している。従って、‘ハネデュ'果実ではエチレン信号伝達系のうち成熟エチレン生成を制御する系に何らかの変異があることが示唆された。
    ‘ハネデュ'の変異がどの程度の因子に影響を及ぼすのか、また変異遺伝子特定を目的として、メロンの転写因子の発現パターンの解析を行った。データベースには100ほどの転写因子が登録されているので、それぞれにプライマーを設計しリアルタイムPCRをおこなった。その結果27のエチレン応答性遺伝子のうち4割の遺伝子の発現パターンが他のメロン品種とは明らかに異なっていた。その中にはエチレンとの関連が報告されているものや他のホルモンとの関連が示唆されているものもあり、エチレン生成を制御する因子の候補となると考えられた。

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