{"created":"2023-04-24T06:51:38.791762+00:00","id":2000946,"links":{},"metadata":{"_buckets":{"deposit":"bd54d1e8-ada5-4503-86ad-fb6730b5399f"},"_deposit":{"id":"2000946","owners":[1],"pid":{"revision_id":0,"type":"depid","value":"2000946"},"status":"published"},"_oai":{"id":"oai:niigata-u.repo.nii.ac.jp:02000946","sets":["453:455","468:563:564"]},"author_link":[],"item_6_date_granted_51":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"2022-09-20"}]},"item_6_degree_grantor_49":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_language":"ja","subitem_degreegrantor_name":"新潟大学"},{"subitem_degreegrantor_language":"en","subitem_degreegrantor_name":"Niigata University"}],"subitem_degreegrantor_identifier":[{"subitem_degreegrantor_identifier_name":"13101","subitem_degreegrantor_identifier_scheme":"kakenhi"}]}]},"item_6_degree_name_48":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（学術）","subitem_degreename_language":"ja"}]},"item_6_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Understanding how genes work together and interact is the focus of functional genomics, a branch of genomics that attempts to find the correlation between genome and phenotype. Changes in gene expression have a significant impact on phenotypic responses. In the present study, two distinct gene expression patterns were demonstrated in two different model plants in order to investigate the effect of gene expression on phenotypic responses. A floral homeotic gene from dicotyledonous Arabidopsis thaliana (AtLFY) was transferred to a monocot liliaceous ornamental plant Tricyrtis sp. by Agrobacterium-mediated transformation. Nine independent transgenic plants with overexpressing AtLFY have been obtained, and all of them exhibited dwarf phenotypes compared with the vector control. These transgenic plants could be classified into three types according to the degree of dwarfism: one showed an extremely dwarf phenotype with smaller leaves (Type I); two showed moderately dwarf phenotypes (Type II); and six showed slightly dwarf phenotypes (Type III). All of Type I, Type II and Type III transgenic plants produced flower buds 1-3 weeks earlier than the vector control. Vector control and Type III transgenic plants produced 1-4 apical flower buds, whereas Type I and Type II transgenic plants produced only a single apical flower bud. Type I and Type II transgenic plants rarely produced fully-opened flowers. Quantitative real-time reverse transcription-polymerase chain reaction analysis showed that the AtLFY expression level generally correlated with the degree of dwarfism. These results indicate that morphological alterations observed in the transgenic plants were induced by overexpression of AtLFY. Lower levels of ectopic expression of LFY may be valuable for producing dwarf and early flowering ornamental plants. Homologous expression of metabolic genes was demonstrated in a dicotyledonous medicinal plant, Artemisia annua L, to examine the role of key enzyme genes on biochemical production. A multifunctional sesquiterpene oxidase gene, CYP71AV1, which is responsible for the conversion of artemisinin precursors, was successfully cloned into an RNA silencing plasmid using the gateway pANDA vector. Three independent RNAi suppression of CYP71AV1 gene (siCYP) transgenic plants were obtained and used for biochemical analysis, in comparison with transgenic A. annua with sense overexpression of CYP71AV1 gene (oxCYP) and wild type. Results revealed variations in terpenoid compounds among transgenic lines. Reduced numbers of terpenoid compounds were detected in siCYP transgenic lines. Some useful sesquiterpenes such as copaene, trans-β-farnesene and aromadendrene were also absent. On the other hand, the levels of some terpenoids significantly increased in oxCYP transgenic lines. The contents of artemisinin derivatives were observed at relatively high levels in oxCYP transgenic lines, while only slightly decreased in siCYP transgenic lines. Overall results suggest the diverse effect of different CYP71AV1 constructs on alteration of metabolites in A. annua. The present study demonstrated the phenotypic response of model plants to different patterns of gene expression. Overexpression of AtLFY successfully induced dwarfism and early flowering in transgenic Tricyrtis sp., which are attractive traits, especially for ornamental plants. Transgenic A. annua plants with over- and suppressed expressions of CYP71AV1 exhibited variations in metabolic compositions, which may be valuable materials for functional genomic studies and pharmaceutical applications.","subitem_description_language":"en","subitem_description_type":"Abstract"},{"subitem_description":"遺伝子がどのように連携して相互作用するかを理解することは，ゲノムと表現型の関係を解明する機能ゲノミクスにおいて重要である．遺伝子発現の変化は表現型に大きな影響を及ぼす．本研究では，表現型応答に対する遺伝子発現の影響を明らかにするために，2つの異なるモデル植物における2つの異なる遺伝子発現パターンを調査した．まず，双子葉植物シロイヌナズナの花のホメオティック遺伝子(AtLFY)を，アグロバクテリウム法により単子葉鑑賞植物のホトトギス(Tricyrtis sp.)に導入した．AtLFYを過剰発現する9系統の形質転換体が得られ，それらの全てがベクターコントロールと比較して矮性の表現型を示した．形質転換系統は，矮化の程度により3つのタイプに分類することができた：1系統は著しく矮化しており小さな葉をつけていた(タイプI)；2系統は中程度の矮化を示した(タイプII)，残りの6系統はわずかな矮化を示した(タイプIII)．タイプI〜IIIいずれの形質転換体も，ベクターコントロールより1〜3週間早期に花蕾を形成した．ベクターコントロールおよびタイプIII形質転換体は1〜4の頂生花蕾を形成したが，タイプIおよびII形質転換体は単一の頂生花蕾のみを形成した．タイプIおよびII形質転換体の花は，ほとんど展開しなかった．リアルタイム定量RT-PCRを行ったところ，AtLFYの発現レベルと矮化の程度の間には相関がみられた．これらの結果は，形質転換体における形態変化がAtLFYの過剰発現によって誘導されたことを示している．また，外来LFYの低発現レベルが矮性および早咲きの観賞植物の作出に有効であることも示している．次に，薬効成分の生合成における重要な酵素遺伝子の役割を調査するために，双子葉薬用植物のクソニンジン(A. annua)を用いた実験を行なった．アルテミシニン前駆体の変換に関与する多機能セスキテルペンオキシダーゼ遺伝子(CYP71AV1)を過剰発現する形質転換体(oxCYP系統)，RNAiによりCYP71AV1の発現が抑制された形質転換体(siCYP系統)および野生型の個体について，テルペノイド化合物の分析を行った．siCYP系統では，テルペノイド化合物の数の減少がみられ，copaeneやtrans-β-farnesene，aromadendreneのような有用なセスキテルペンも存在していなかった．一方，oxCYP系統においては，一部のテルペノイドのレベルが有意に増加した．アルテミシニン誘導体の含有量は，oxCYP系統では比較的高レベルで検出されたが，siCYP系統ではわずかに減少していた．これらの結果は，CYP71AV1の発現レベルの変化により代謝産物の変化が誘導されたことを示している．本研究は，2種類のモデル植物において異なる遺伝子発現レベルによる表現型の変化を示すものである．AtLFYの過剰発現は，ホトトギスにおいて矮化および早期開花を誘導した．これにより，鑑賞植物の育種におけるLFYを用いた形質転換の有用性が示された．一方，CYP71AV1の過剰発現および発現抑制は，クソニンジンにおける代謝産物の変動を誘導した．CYP71AV1の発現が変化した形質転換体は，機能ゲノムの研究および製薬に関して有用な材料となる可能性がある．","subitem_description_language":"ja","subitem_description_type":"Abstract"}]},"item_6_description_53":{"attribute_name":"学位記番号","attribute_value_mlt":[{"subitem_description":"新大院博(学)第226号","subitem_description_language":"ja","subitem_description_type":"Other"}]},"item_6_dissertation_number_52":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"甲第5090号"}]},"item_access_right":{"attribute_name":"アクセス権","attribute_value_mlt":[{"subitem_access_right":"open access","subitem_access_right_uri":"http://purl.org/coar/access_right/c_abf2"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Sankhuan, Darunmas","creatorNameLang":"en"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_access","date":[{"dateType":"Available","dateValue":"2023-04-24"}],"displaytype":"detail","filename":"r4fik226.pdf","filesize":[{"value":"2.81MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"本文","objectType":"fulltext","url":"https://niigata-u.repo.nii.ac.jp/record/2000946/files/r4fik226.pdf"},"version_id":"f52a1807-5abd-4c14-9816-83908424f768"},{"accessrole":"open_access","date":[{"dateType":"Available","dateValue":"2023-04-24"}],"displaytype":"detail","filename":"r4fik226_a.pdf","filesize":[{"value":"341KB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"要旨","objectType":"abstract","url":"https://niigata-u.repo.nii.ac.jp/record/2000946/files/r4fik226_a.pdf"},"version_id":"c569177f-aefc-49ed-bba3-195025d37f69"}]},"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":"doctoral thesis","resourceuri":"http://purl.org/coar/resource_type/c_db06"}]},"item_title":"Effects of overexpression or suppression of heterologous or homologous genes on phenotypic alteration in transgenic plants","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Effects of overexpression or suppression of heterologous or homologous genes on phenotypic alteration in transgenic plants","subitem_title_language":"en"},{"subitem_title":"外来および内生遺伝子の過剰発現および発現抑制が形質転換体における表現型の変化に及ぼす影響","subitem_title_language":"ja"}]},"item_type_id":"6","owner":"1","path":["455","564"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2023-04-24"},"publish_date":"2023-04-24","publish_status":"0","recid":"2000946","relation_version_is_last":true,"title":["Effects of overexpression or suppression of heterologous or homologous genes on phenotypic alteration in transgenic plants"],"weko_creator_id":"1","weko_shared_id":-1},"updated":"2023-04-24T06:51:41.623155+00:00"}