{"_buckets": {"deposit": "e6fd369e-728c-415b-bc8e-d425c78d841e"}, "_deposit": {"created_by": 21, "id": "1691", "owners": [21], "pid": {"revision_id": 0, "type": "depid", "value": "1691"}, "status": "published"}, "_oai": {"id": "oai:ir.soken.ac.jp:00001691", "sets": ["20"]}, "author_link": ["0", "0", "0"], "item_1_biblio_info_21": {"attribute_name": "書誌情報（ソート用）", "attribute_value_mlt": [{"bibliographicIssueDates": {"bibliographicIssueDate": "2010-03-24", "bibliographicIssueDateType": "Issued"}, "bibliographic_titles": [{}]}]}, "item_1_creator_2": {"attribute_name": "著者名", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "水多, 陽子"}], "nameIdentifiers": [{"nameIdentifier": "0", "nameIdentifierScheme": "WEKO"}]}]}, "item_1_creator_3": {"attribute_name": "フリガナ", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "ミズタ, ヨウコ"}], "nameIdentifiers": [{"nameIdentifier": "0", "nameIdentifierScheme": "WEKO"}]}]}, "item_1_date_15": {"attribute_name": "公開希望日", "attribute_value_mlt": [{"subitem_date_issued_datetime": "2010-09-01", "subitem_date_issued_type": "Created"}]}, "item_1_date_granted_11": {"attribute_name": "学位授与年月日", "attribute_value_mlt": [{"subitem_dategranted": "2010-03-24"}]}, "item_1_degree_grantor_5": {"attribute_name": "学位授与機関", "attribute_value_mlt": [{"subitem_degreegrantor": [{"subitem_degreegrantor_name": "総合研究大学院大学"}]}]}, "item_1_degree_name_6": {"attribute_name": "学位名", "attribute_value_mlt": [{"subitem_degreename": "博士（理学）"}]}, "item_1_description_1": {"attribute_name": "ID", "attribute_value_mlt": [{"subitem_description": "2010041", "subitem_description_type": "Other"}]}, "item_1_description_12": {"attribute_name": "要旨", "attribute_value_mlt": [{"subitem_description": "　　　Identification of genes responsible for barriers to gene flow between two species provides insight into molecular mechanisms of reproductive isolation and relationships between evolution of barrier genes and species diversification. To investigate the molecular mechanism of reproductive isolation and the status of evolutionary differentiation, Asian cultivated rice, \u003ci\u003eOryza sativa\u003c/i\u003e L., is an ideal species due to its population structure and genetic diversity. \u003ci\u003eO. sativa\u003c/i\u003e has diverged subspecies, \u003ci\u003eindica\u003c/i\u003e and \u003ci\u003ejaponica\u003c/i\u003e Inter-subspecific cross between them exhibits various hybrid incompatibilities, but the mechanisms are still largely unknown. Cross between \u003ci\u003eindica\u003c/i\u003e cultivar(cv.) Kasalath and \u003ci\u003ejaponica\u003c/i\u003e cv. Nipponbare showed almost no abnormal F\u003csmall\u003e1\u003c/small\u003e hybrid, although 33 reproductive barriers were mapped along whole chromosomes in their F\u003csmall\u003e2\u003c/small\u003e population. In these barriers, a prominent interactive barrier locus was detected on rice chromosomes 1 and 6. Based on the analysis of reciprocal backcrosses progenies, this interaction occurs only in the male gametophyte, pollen.\u003cbr /\u003e　　　To identify the causal genes at each locus, map-based cloning of a pair of reproductive barrier genes has been done. Using more than 10,000 individual plants, responsible genes were mapped within regions of 59 kb on Nipponbare chromosome 1 and 11 kb on Nipponbare chromosome 6. A pair of genes, one from each region shared a high degree of homology with each other, and both genes have different sequences between Nipponbare and Kasalath. These homologous genes were regarded as primary candidates, and these were designed as \u003ci\u003eDOPPELGANGER1 (DPL1)\u003c/i\u003e and \u003ci\u003eDOPPELGANGER2 (DPL2), \u003c/i\u003erespectively. Hybrid pollen carrying both alleles on Kasalath chromosome 1 (\u003ci\u003eDPL1-K\u003c/i\u003e) and Nipponbare chromosome 6 (\u003ci\u003eDPL2-N\u003c/i\u003e)together became non-functional, and did not germinate.\u003cbr /\u003e　　　\u003ci\u003eDPL\u003c/i\u003e genes encode plant specific protein with unknown functions, which are highly conserved among angiosperms. Sequence analysis of the Nipponbare and Kasalath genomes and their transcripts suggested that alleles on Nipponbare chromosome 1(\u003ci\u003eDPLI-N\u003c/i\u003e) and Kasalath chromosome 6 (\u003ci\u003eDPL2-K\u003c/i\u003e) had the same coding sequence structure. In contrast, alleles on Kasalath chromosome 1 (\u003ci\u003eDPLI-K\u003c/i\u003e) and Nipponbare chromosome 6 (\u003ci\u003eDPL2-N\u003c/i\u003e) had structural differences from the above two alleles. \u003ci\u003eDPL1-K\u003c/i\u003e had an insertion of a predicted transposable element (TE) in the coding sequence and the transcript could not be detected in any tissues. The transcript of \u003ci\u003eDPL2-N\u003c/i\u003e was a read through product of the second intron generating a premature stop codon. Higher expression of \u003ci\u003eDPLs\u003c/i\u003e in pollen was also observed in \u003ci\u003ein situ\u003c/i\u003e hybridization experiments. Anti-DPL antibodies recognized proteins of DPL1-N and DPL2-K in extracts from Nipponbare and Kasalath mature anthers, respectively. However, DPL2-N protein was not detected in extracts from Nipponbare. The lack of \u003ci\u003eDPL1-K\u003c/i\u003e transcript and the absence of DPL2-N protein suggested that both \u003ci\u003eDPL1-K\u003c/i\u003e and \u003ci\u003eDPL2-N\u003c/i\u003e were loss of function alleles. Phenorype observation also indicated that \u003ci\u003eDPL1-K\u003c/i\u003e and \u003ci\u003eDPL2-N\u003c/i\u003e were loss of function alleles, due to the hybrid pollen carrying both of them became non-functional, and did not germinate.\u003cbr /\u003e　　　The relatively high expression of  both \u003ci\u003eDPL1-N\u003c/i\u003e and \u003ci\u003eDPL2-K\u003c/i\u003e were observed in pollen at the late stage of pollen development. Both \u003ci\u003eDPL1-N\u003c/i\u003e and \u003ci\u003eDPL2-K\u003c/i\u003e were thought to have normal functions, because they were normally transmitted to progenies. Complementation tests of \u003ci\u003eDPLs\u003c/i\u003e using near isogenic lines also indicated that \u003ci\u003eDPLs\u003c/i\u003e are responsible genes for this reproductive isolation, and either \u003ci\u003eDPL1-N\u003c/i\u003e or \u003ci\u003eDPL2-K\u003c/i\u003e are necessary for pollen transmission. These results clearly showed that a functional \u003ci\u003eDPL1-N\u003c/i\u003e or \u003ci\u003eDPL2-K\u003c/i\u003e allele is essential for pollen transmission, whereas \u003ci\u003eDPL1-K\u003c/i\u003e and \u003ci\u003eDPL2-N\u003c/i\u003e are loss-of-function alleles that act as a pair of reproductive barrier genes by their combination in hybrid pollen.\u003cbr /\u003e　　　In this study, the molecular mechanism of male gametophytic reproductive isolation by the combination of disrupted \u003ci\u003eDPLs\u003c/i\u003e in rice was identified. After gene duplication of \u003ci\u003eDPL\u003c/i\u003e, an ancestral population seems to have diverged forming the Kasalath ancestral population, which subsequently lost the function of \u003ci\u003eDPL1\u003c/i\u003e by TE insertion, and the Nipponbare ancestral population, which lost the function of \u003ci\u003eDPL2\u003c/i\u003e by means of a splicing defect. When they met again by crossing, hybrid pollen having the loss of function alleles together became non-functional and failed to transmit themselves to the next generation. This is a typical case of the Dobzhansky-Muller model for barier formation by genetic incompatibility between species.\u003cbr /\u003e　　　To discuss when this reproductive isolation mechanism was established, the duplications and disruptions of \u003ci\u003eDPLs\u003c/i\u003e were also investigated along with flowering species differentiation.\u003ci\u003e DPL\u003c/i\u003e was highly conserved among 43 angiosperms. Database search indicated that not only rice, but also other four bangiosperms, i\u003eSorghum bicolor, Zea mays, Glycine max\u003c/i\u003e and \u003ci\u003eMedicago truncatula\u003c/i\u003e have two \u003ci\u003eDPL\u003c/i\u003e orthrologs. Using syntenic information of them, it was suggested that the duplication of \u003ci\u003eDPL\u003c/i\u003e occurred at least three times, twice in Poaceae and once in Leguminosae. The syntenic conservation around the region of the \u003ci\u003eDPL2\u003c/i\u003e on rice chromosome 6 among grass species suggested that \u003ci\u003eDPL2\u003c/i\u003e is the most ancient in Poaceae.\u003cbr /\u003e　　　In the genus of \u003ci\u003eOryza\u003c/i\u003e, all examined 42 accessions or varieties belonging to eight closely related species had both \u003ci\u003eDPL1\u003c/i\u003e and\u003ci\u003eDPL2.\u003c/i\u003e To investigate relationships between the disruption of \u003ci\u003eDPLs\u003c/i\u003e and \u003ci\u003eOryza\u003c/i\u003e differentiation, it was investigated when the disruptions of \u003ci\u003eDPLs\u003c/i\u003e occurred in these species including both \u003ci\u003eO. sativa\u003c/i\u003e and its ancestral species, \u003ci\u003eO. rufipogon.\u003c/i\u003e Based on the nucleotide variations in the coding region of \u003ci\u003eDPLs\u003c/i\u003e, \u003ci\u003eO. sativa\u003c/i\u003e and \u003ci\u003eO. rufipogon\u003c/i\u003e accessions or varieties could be classified into following four groups; group I, tropical and temperate \u003ci\u003ejaponica\u003c/i\u003e and \u003ci\u003eO. rufipogon;\u003c/i\u003egroup II, \u003ci\u003eindica\u003c/i\u003e and \u003ci\u003eO. rufipogon;\u003c/i\u003e group III, \u003ci\u003eIndica\u003c/i\u003e and \u003ci\u003eO. rufipogon\u003c/i\u003e and group IV, \u003ci\u003eO. rufipogon.\u003c/i\u003e The insertion of TE in \u003ci\u003eDPL1\u003c/i\u003e was only observed both \u003ci\u003eindica\u003c/i\u003e and \u003ci\u003eO. rufipogon\u003c/i\u003e belonging to group III, whereas the read through product of \u003ci\u003eDPL2\u003c/i\u003e was only observed in \u003ci\u003ejaponica\u003c/i\u003e cultivars belonging to group I. \u003ci\u003eDPL1-K\u003c/i\u003e was only observed in the partial indica varieties, suggesting the loss-of-function of \u003ci\u003eDPL1\u003c/i\u003e in \u003ci\u003eindica\u003c/i\u003e and that of \u003ci\u003eDPL2\u003c/i\u003e in \u003ci\u003ejaponica\u003c/i\u003e occurred after \u003ci\u003ejaponica-indica\u003c/i\u003e differentiation. Reproductive isolation by the combination of disrupted \u003ci\u003eDPL1\u003c/i\u003e and \u003ci\u003eDPL2\u003c/i\u003e was not initially act as barriers between \u003ci\u003eindica\u003c/i\u003e and \u003ci\u003ejaponica,\u003c/i\u003e due to the occurrence of these disruptions after the \u003ci\u003ejaponica-indica\u003c/i\u003e differentiation. Probably this isolation reinforced the population integrity of \u003ci\u003ejaponica\u003c/i\u003e and \u003ci\u003eindica\u003c/i\u003e in group III, when the populations came into contact with each other. Our results also suggested that \u003ci\u003eindica\u003c/i\u003e rice is polyphyletically domesticated from the different ancestors of \u003ci\u003eO.\u003c/i\u003e \u003ci\u003erufipogon\u003c/i\u003e, whereas \u003ci\u003ejaponica\u003c/i\u003e rice is monophyletic population. Other gene disruptions of \u003ci\u003eDPLs\u003c/i\u003e also occurred at least three times independently in rice.\u003cbr /\u003e　　　These findings showed the molecular mechanisms of reproductive isolation by the combination of disrupted \u003ci\u003eDPLs\u003c/i\u003e, and a comprehensive story of the evolution of \u003ci\u003eDPLs\u003c/i\u003e in plant. It remains unknown whether duplications and disruptions of \u003ci\u003eDPL\u003c/i\u003e genes resulted from adaptive selection or random drift in plant speciation. Further studies of \u003ci\u003eDPL\u003c/i\u003e function and analyses of reproductive isolation events in \u003ci\u003eOryza\u003c/i\u003e will provide fundamental understanding of molecular functions in plant reproduction and the mechanisms of species diversification.", "subitem_description_type": "Other"}]}, "item_1_description_18": {"attribute_name": "フォーマット", "attribute_value_mlt": [{"subitem_description": "application/pdf", "subitem_description_type": "Other"}]}, "item_1_description_7": {"attribute_name": "学位記番号", "attribute_value_mlt": [{"subitem_description": "総研大甲第1345号", "subitem_description_type": "Other"}]}, "item_1_select_14": {"attribute_name": "所蔵", "attribute_value_mlt": [{"subitem_select_item": "有"}]}, "item_1_select_16": {"attribute_name": "複写", "attribute_value_mlt": [{"subitem_select_item": "公開希望日以降"}]}, "item_1_select_17": {"attribute_name": "公開状況", "attribute_value_mlt": [{"subitem_select_item": "公開希望日以降"}]}, "item_1_select_8": {"attribute_name": "研究科", "attribute_value_mlt": [{"subitem_select_item": "生命科学研究科"}]}, "item_1_select_9": {"attribute_name": "専攻", "attribute_value_mlt": [{"subitem_select_item": "18 遺伝学専攻"}]}, "item_1_text_10": {"attribute_name": "学位授与年度", "attribute_value_mlt": [{"subitem_text_value": "2009"}]}, "item_creator": {"attribute_name": "著者", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "MIZUTA, Yoko", "creatorNameLang": "en"}], "nameIdentifiers": [{"nameIdentifier": "0", "nameIdentifierScheme": "WEKO"}]}]}, "item_files": {"attribute_name": "ファイル情報", "attribute_type": "file", "attribute_value_mlt": [{"accessrole": "open_date", "date": [{"dateType": "Available", "dateValue": "2016-02-17"}], "displaytype": "simple", "download_preview_message": "", "file_order": 0, "filename": "甲1345_要旨.pdf", "filesize": [{"value": "437.3 kB"}], "format": "application/pdf", "future_date_message": "", "is_thumbnail": false, "licensetype": "license_11", "mimetype": "application/pdf", "size": 437300.0, "url": {"label": "要旨・審査要旨", "url": "https://ir.soken.ac.jp/record/1691/files/甲1345_要旨.pdf"}, "version_id": "9622301f-38a7-4803-b6f3-c0c2c6065268"}, {"accessrole": "open_date", "date": [{"dateType": "Available", "dateValue": "2016-02-17"}], "displaytype": "simple", "download_preview_message": "", "file_order": 1, "filename": "甲1345_本文.pdf", "filesize": [{"value": "15.2 MB"}], "format": "application/pdf", "future_date_message": "", "is_thumbnail": false, "licensetype": "license_11", "mimetype": "application/pdf", "size": 15200000.0, "url": {"label": "本文", "url": "https://ir.soken.ac.jp/record/1691/files/甲1345_本文.pdf"}, "version_id": "6836116b-951e-4c7f-b5e3-23fdb3eb0f47"}]}, "item_language": {"attribute_name": "言語", "attribute_value_mlt": [{"subitem_language": "eng"}]}, "item_resource_type": {"attribute_name": "資源タイプ", "attribute_value_mlt": [{"resourcetype": "thesis", "resourceuri": "http://purl.org/coar/resource_type/c_46ec"}]}, "item_title": "Analysis of a pair of genes, DOPPELGANGER 1 (DPL1) and DOPPELGANGER 2 (DPL2) responsible for reproductive isolation between two rice subspecies", "item_titles": {"attribute_name": "タイトル", "attribute_value_mlt": [{"subitem_title": "Analysis of a pair of genes, DOPPELGANGER 1 (DPL1) and DOPPELGANGER 2 (DPL2) responsible for reproductive isolation between two rice subspecies"}, {"subitem_title": "Analysis of a pair of genes, DOPPELGANGER 1 (DPL1) and DOPPELGANGER 2 (DPL2) responsible for reproductive isolation between two rice subspecies", "subitem_title_language": "en"}]}, "item_type_id": "1", "owner": "21", "path": ["20"], "permalink_uri": "https://ir.soken.ac.jp/records/1691", "pubdate": {"attribute_name": "公開日", "attribute_value": "2011-01-19"}, "publish_date": "2011-01-19", "publish_status": "0", "recid": "1691", "relation": {}, "relation_version_is_last": true, "title": ["Analysis of a pair of genes, DOPPELGANGER 1 (DPL1) and DOPPELGANGER 2 (DPL2) responsible for reproductive isolation between two rice subspecies"], "weko_shared_id": -1}