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The sex-determining gene in tiger pufferfish (fugu)

Research Project

Project/Area Number 24380102
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypePartial Multi-year Fund
Section一般
Research Field General fisheries
Research InstitutionThe University of Tokyo

Principal Investigator

KIKUCHI Kiyoshi  東京大学, 農学生命科学研究科, 准教授 (20292790)

Co-Investigator(Renkei-kenkyūsha) TASUMI Satoshi  東京大学, 大学院農学生命科学研究科, 特任助教 (90359646)
SUETAKE Hiroaki  福井県立大学, 海洋生物資源学部, 准教授 (00334326)
Research Collaborator MIZUNO Naoki  東京大学, 大学院農学生命科学研究科, 技術専門職員
JYO Yuka  東京大学, 大学院農学生命科学研究科, 技術専門職員
FUJITA Masashi  東京大学, 大学院農学生命科学研究科, 技術職員
Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥15,340,000 (Direct Cost: ¥11,800,000、Indirect Cost: ¥3,540,000)
Fiscal Year 2014: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2012: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
Keywords性決定 / 生殖 / 遺伝 / 遺伝的多様性
Outline of Final Research Achievements

Sex determination in fish is often genetic, where segregation of sex-determining (SD) locus assigns the phenotypic sex. In these species, SD genes can be used for sex identification, potentially providing important information regarding the ecology and conservation of many wild species, as well as management of cultured species. However, no SD gene had been identified in aquaculture species at the time when this project started. We investigated fugu (Tiger pufferfish) that is one of the most important aquaculture fish in Japan. We provide a line of evidences showing that a SNP in the Amhr2 gene determines phenotypic sex in fugu. In addition, for high-throughput sex identification of fugu, we developed an efficient assay for this SNP site. Moreover, we successfully produced all-male population of fugu through marker-assisted selection.

Report

(4 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Annual Research Report
  • 2012 Annual Research Report

Research Products

(60 results)

All 2015 2014 2013 2012 Other

All Journal Article (21 results) (of which Peer Reviewed: 15 results,  Acknowledgement Compliant: 3 results,  Open Access: 1 results) Presentation (34 results) (of which Invited: 6 results) Book (2 results) Remarks (3 results)

  • [Journal Article] Screening of candidate genes encoding proteins expressed in pectoral fins of fugu Takifugu rubripes, in relation to habitat site of parasitic copepod Caligus fugu, using suppression subtractive hybridization.2015

    • Author(s)
      Tasumi S, Norshida I, Boxshall GA, Kikuchi K, Suzuki Y, Ohtsuka S.
    • Journal Title

      Fish Shellfish Immunol.

      Volume: 44 Pages: 356-364

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A microsatellite-based genetic linkage map and putative sex-determining genomic regions in Lake Victoria cichlids.2015

    • Author(s)
      Kudo Y., Nikaido M., Kondo A., Suzuki H., Yoshida K., Kikuchi K., Okada N.
    • Journal Title

      GENE

      Volume: 560 Pages: 156-164

    • DOI

      10.1016/j.gene.2015.01.057

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] トラフグ凍結精子の家庭用冷蔵庫での二次保存2015

    • Author(s)
      細谷将・水野直樹・城夕香・藤田真志・鈴木譲・菊池潔
    • Journal Title

      水産技術

      Volume: 7 Pages: 85-88

    • NAID

      40020458374

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] An efficient molecular technique for sexing tiger pufferfish (fugu) and the occurrence of sex reversal in a hatchery population.2014

    • Author(s)
      Matsunaga, T., Ieda, R., Hosoya, S., Kuroyanagi, M., Suzuki, S., Suetake, H., Tasumi, S., Suzuki, Y., Miyadai, T., Kikuchi, K.
    • Journal Title

      Fish. Sci.

      Volume: 80 Pages: 933-942

    • DOI

      10.1007/s12562-014-0768-0

    • NAID

      130005091912

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Rearing Takifugu rubripes larvae in communal tanks: paternal genetic contribution to survivability.2014

    • Author(s)
      Hosoya, S., Mizono, N. Kikuchi, K., Kurokura, H.
    • Journal Title

      Fish. Sci.

      Volume: 80 Pages: 1037-1043

    • DOI

      10.1007/s12562-014-0795-x

    • NAID

      40020207484

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Development of a multiplex PCR assay for population genetic analysis of the tiger puffer Takifugu rubripes using 16 microsatellite DNA loci.2014

    • Author(s)
      Katamachi D., Ikeda M., Sato T., Suzuki S., Kikuchi, K., Ojima D.
    • Journal Title

      Aquaculture Sci.

      Volume: 62 Pages: 55-63

    • NAID

      130005062545

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] トラフグにおける1塩基の性決定-性決定遺伝子の誕生と死(特集 愛と性の科学)2014

    • Author(s)
      菊池潔
    • Journal Title

      科学

      Volume: 84 Pages: 770-772

    • Related Report
      2014 Annual Research Report
  • [Journal Article] 移り変わる性決定遺伝子―魚類の性決定遺伝子同定からみえてきた風景―2014

    • Author(s)
      菊池潔
    • Journal Title

      生物科学

      Volume: 65 Pages: 136-145

    • Related Report
      2014 Annual Research Report 2013 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] ホルモンによる性転換と遺伝マーカーによる超オスの作出2014

    • Author(s)
      菊池潔, 細谷将
    • Journal Title

      養殖ビジネス

      Volume: 51 Pages: 36-39

    • Related Report
      2014 Annual Research Report 2013 Annual Research Report
  • [Journal Article] Development of a multiplex PCR assay for population genetic analysis of the tiger puffer Takifugu rubripes using 16 microsatellite DNA loci2014

    • Author(s)
      Daisuke KATAMACHI, Minoru IKEDA, Takafumi SATO, Shigenori SUZUKI, Kiyoshi KIKUCHI and Daisuke OJIMA
    • Journal Title

      Aquaculture Science

      Volume: 62

    • NAID

      130005062545

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] New approach for fish breeding by chemical mutagenesis: establishment of TILLING method in fugu (Takifugu rubripes) with ENU mutagenesis.2013

    • Author(s)
      Kuroyanagi M, Katayama T, Imai T, Yamamoto Y, Chisada S, Yoshiura Y, Ushijima T, Matsushita T, Fujita M, Nozawa A, Suzuki Y, Kikuchi K, Okamoto H.
    • Journal Title

      BMC Genomics

      Volume: 14 Pages: 786-786

    • DOI

      10.1186/1471-2164-14-786

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Genomic regions of pufferfishes responsible for host specificity of a monogenean parasite Heterobothrium okamotoi.2013

    • Author(s)
      Hosoya S, Kido S, Hirabayashi Y, Kai W, Kinami R, Yoshinaga T, Ogawa K, Suetake H, Kikuchi K, Suzuki Y.
    • Journal Title

      Int. J. Parasitol.

      Volume: 43 Pages: 909-915

    • DOI

      10.1016/j.ijpara.2013.06.006

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Novel sex-determining genes in fish and sex chromosome evolution2013

    • Author(s)
      Kikuchi K, Hamaguchi S.
    • Journal Title

      Developmental Dynamics

      Volume: 242 Pages: 339-353

    • DOI

      10.1002/dvdy.23927

    • Related Report
      2013 Annual Research Report 2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 魚類の性統御2013

    • Author(s)
      菊池潔,細谷将,田角聡志
    • Journal Title

      アクアネット

      Volume: 16 Pages: 42-47

    • Related Report
      2013 Annual Research Report
  • [Journal Article] Sex determination in fish2013

    • Author(s)
      菊池潔,細谷将,田角聡志
    • Journal Title

      The Journal of Animal Genetics

      Volume: 41 Issue: 1 Pages: 37-48

    • DOI

      10.5924/abgri.41.37

    • NAID

      10031190360

    • ISSN
      1345-9961, 1884-3883
    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The genetic architecture of growth rate in juvenile Takifugu species2013

    • Author(s)
      Hosoya, S., et al.
    • Journal Title

      Evolution

      Volume: 67 Pages: 590-598

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 魚類性決定遺伝子の多様性と進化2013

    • Author(s)
      菊池潔, 濱口哲
    • Journal Title

      細胞工学

      Volume: 32 Pages: 164-169

    • Related Report
      2012 Annual Research Report
  • [Journal Article] A Trans-species Missense SNP in Amhr2 is Associated with Sex Determination in the Tiger Pufferfish, Takifugu rubripes (fugu)2012

    • Author(s)
      Kamiya, T., Kai, W., Tasumi, S., Oka, A., Matsunaga, T., Mizuno, N., Fujita, M., Suetake, H., Suzuki, S., Hosoya, S., Tohari, S., Brenner, S., Miyadai, T., Venkatesh, B., Suzuki, Y., and Kikuchi, K.
    • Journal Title

      PLOS Genetics

      Volume: 8

    • DOI

      10.1371/journal.pgen.1002798

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Early T-cell and thymus development in Japanese eel Anguilla japonica2012

    • Author(s)
      Kawabe, M., et al.
    • Journal Title

      Fisheries Science

      Volume: 78 Pages: 539-547

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] フグのゲノム地図作製と性統御および育種への応用2012

    • Author(s)
      菊池潔, 甲斐渉
    • Journal Title

      日本水産学会誌

      Volume: 78 Pages: 269-269

    • NAID

      10030124122

    • Related Report
      2012 Annual Research Report
  • [Journal Article] ゲノム育種によりトラフグの優良品種作出をめざす2012

    • Author(s)
      鈴木譲, 菊池潔
    • Journal Title

      日本水産学会誌

      Volume: 78 Pages: 88-88

    • NAID

      10030024794

    • Related Report
      2012 Annual Research Report
  • [Presentation] Amhr2Y型遺伝子はトラフグを雄化する-トランスジェニック海産魚による機能解析-2015

    • Author(s)
      家田梨櫻・神谷隆史・渥美九郁・藤田真志・細谷将・鈴木重則・鈴木譲・田角聡志・菊池潔
    • Organizer
      平成27年度日本水産学会春季大会
    • Place of Presentation
      東京
    • Year and Date
      2015-03-27 – 2015-03-31
    • Related Report
      2014 Annual Research Report
  • [Presentation] ブリ性決定遺伝子座の連鎖不平衡解析2015

    • Author(s)
      小山喬・吉田一範・尾崎照遵・青木純哉・荒木和男・細谷将・菊池潔・坂本 崇
    • Organizer
      平成27年度日本水産学会春季大会
    • Place of Presentation
      東京
    • Year and Date
      2015-03-27 – 2015-03-31
    • Related Report
      2014 Annual Research Report
  • [Presentation] ゲノム情報から推定する種分化と交雑の歴史-トラフグ近縁12魚類の全ゲノム比較-2015

    • Author(s)
      細谷将・田角聡志・小林久人・菊池潔
    • Organizer
      平成27年度日本水産学会春季大会
    • Place of Presentation
      東京
    • Year and Date
      2015-03-27 – 2015-03-31
    • Related Report
      2014 Annual Research Report
  • [Presentation] 集団ゲノミクスで探るゲノム上に残された家魚化の痕跡-宮城県産ギンザケ品種の解析-2015

    • Author(s)
      細谷将・菊池潔・永島宏・小野寺淳一・杉本晃一・佐藤 好・松崎圭佑・熊谷明・上田賢一・八杉公基・永野 惇・黒川忠英
    • Organizer
      平成27年度日本水産学会春季大会
    • Place of Presentation
      東京
    • Year and Date
      2015-03-27 – 2015-03-31
    • Related Report
      2014 Annual Research Report
  • [Presentation] 有用遺伝子の探索・同定・選抜・編集2015

    • Author(s)
      菊池潔
    • Organizer
      平成 27 年度日本水産学会水産増殖懇話会第 1 回講演会
    • Place of Presentation
      東京
    • Year and Date
      2015-03-27
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 魚類の性決定遺伝子たちーフグ近縁種群を中心としてー2014

    • Author(s)
      菊池潔
    • Organizer
      新学術領域研究「複合適応形質進化」公開シンポジウム
    • Place of Presentation
      千葉
    • Year and Date
      2014-12-12
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 魚類の多様な性決定遺伝子について2014

    • Author(s)
      家田梨櫻・青木勇磨・細谷将・田角聡志・菊池潔
    • Organizer
      浜名湖をめぐる研究者の会
    • Place of Presentation
      静岡
    • Year and Date
      2014-12-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] 連鎖および連鎖不平衡マッピングによる表現型原因遺伝子の同定2014

    • Author(s)
      菊池潔・細谷将
    • Organizer
      2014 年度日本魚類学会年会シンポジウム
    • Place of Presentation
      神奈川
    • Year and Date
      2014-11-17
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] フグの性決定遺伝子 最近の研究報告2014

    • Author(s)
      菊池潔
    • Organizer
      第9会Dmyの会
    • Place of Presentation
      栃木
    • Year and Date
      2014-11-15 – 2014-11-16
    • Related Report
      2014 Annual Research Report
  • [Presentation] ゲノム情報の整備と活用による新しい性決定遺伝子の同定2014

    • Author(s)
      菊池潔
    • Organizer
      名古屋大学リーディングプログラム学生企画セミナー「バイオインフォマティクスへの招待」
    • Place of Presentation
      愛知
    • Year and Date
      2014-10-31
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 海産魚のゲノム編集-不透明で油球をもつ粘着卵を産むトラフグの場合-2014

    • Author(s)
      家田梨櫻・藤田真志・松田勝・木南竜平・鈴木重則・末武弘章・細谷将・田角聡志・菊池潔
    • Organizer
      平成26年度日本水産学会秋期大会
    • Place of Presentation
      福岡
    • Year and Date
      2014-09-20 – 2014-09-21
    • Related Report
      2014 Annual Research Report
  • [Presentation] 数百万年でゲノムはどれほど分化するのだろうか? -近縁6魚種間の全ゲノム比較-2014

    • Author(s)
      細谷将,田角聡志,菊池潔
    • Organizer
      2013年度水産学会春季大会
    • Place of Presentation
      北海道
    • Related Report
      2013 Annual Research Report
  • [Presentation] 性転換を利用したクサフグ性決定遺伝子座の解析2014

    • Author(s)
      家田梨櫻,田島祥太,藤田真志,細谷将,田角聡志,菊池潔
    • Organizer
      2013年度水産学会春季大会
    • Place of Presentation
      北海道
    • Related Report
      2013 Annual Research Report
  • [Presentation] 複数個体の全ゲノム解読による表現型原因遺伝子の同定の試み2014

    • Author(s)
      細谷将,八谷剛史,田島祥太,家田梨櫻,藤田真志,田角聡志,菊池潔
    • Organizer
      2013年度水産学会春季大会
    • Place of Presentation
      北海道
    • Related Report
      2013 Annual Research Report
  • [Presentation] トラフグ凍結精子における家庭用冷蔵庫での二次保存2014

    • Author(s)
      水野直樹,細谷将,城夕香,藤田真志,菊池潔
    • Organizer
      2013年度水産学会春季大会
    • Place of Presentation
      北海道
    • Related Report
      2013 Annual Research Report
  • [Presentation] 移り変わる性決定遺伝子ーフグ近縁種群を材料としてー2014

    • Author(s)
      菊池潔
    • Organizer
      日本動物学中部支部会会公開シンポジウム
    • Place of Presentation
      愛知
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Evolution of : sex determining mechanisms in fugu fish2013

    • Author(s)
      Kiyoshi Kikuch
    • Organizer
      International workshop: Sex determination in dragonas, frogs and fugu fish (invited)
    • Place of Presentation
      広島
    • Year and Date
      2013-03-06
    • Related Report
      2012 Annual Research Report
  • [Presentation] フグ類の表現型多様化の遺伝基盤 : 遡河回遊性の獲得など2013

    • Author(s)
      菊池潔
    • Organizer
      日本生態学会第60回全国大会自由集会
    • Place of Presentation
      静岡(招待)
    • Year and Date
      2013-03-05
    • Related Report
      2012 Annual Research Report
  • [Presentation] フグ近縁種群における性決定遺伝子の移り変わり2013

    • Author(s)
      菊池潔
    • Organizer
      第7回Dmyの会/第2回新潟大学系統生物研究センター研究集会
    • Place of Presentation
      栃木(招待)
    • Year and Date
      2013-03-05
    • Related Report
      2012 Annual Research Report
  • [Presentation] 200万年でどれほどゲノムは分化するのだろうか?-近縁魚種間の全ゲノム比較-2013

    • Author(s)
      家田梨櫻,細谷将,田角聡志,菊池潔
    • Organizer
      日本動物遺伝育種学会第15回大会
    • Place of Presentation
      東京
    • Related Report
      2013 Annual Research Report
  • [Presentation] フグでやっていることとやりたいこと2013

    • Author(s)
      細谷将,家田梨櫻,甲斐渉,山野上祐介,菊池潔
    • Organizer
      次世代シーケンサー現場の会2013
    • Place of Presentation
      兵庫
    • Related Report
      2013 Annual Research Report
  • [Presentation] 表現型多様化のゲノム基盤解明を目指した近縁複数種のゲノム比較:フグを材料として2013

    • Author(s)
      細谷将,山野上祐介,甲斐渉,菊池潔
    • Organizer
      日本進化学会第15回東京大会
    • Place of Presentation
      東京
    • Related Report
      2013 Annual Research Report
  • [Presentation] 性決定遺伝子はなぜ移り変わるのか?-その遷移を促す遺伝基盤の探求-2013

    • Author(s)
      家田梨櫻,細谷将,田角聡志,菊池潔
    • Organizer
      特定領域ゲノム支援拡大班会議2013
    • Place of Presentation
      兵庫
    • Related Report
      2013 Annual Research Report
  • [Presentation] Rapid turnover of sex chromosomes in fugu and related species2013

    • Author(s)
      Kiyoshi Kikuchi
    • Organizer
      International mini symposium: Mechanisms and functions of sex chromosome turnover.
    • Place of Presentation
      静岡
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] トラフグの雄型性決定遺伝子はクサフグを雄化する2013

    • Author(s)
      家田梨櫻, ら
    • Organizer
      2012年度水産学会春季大会
    • Place of Presentation
      東京
    • Related Report
      2012 Annual Research Report
  • [Presentation] トラフグ属魚類における性決定遺伝子座の変遷 : 新しい性決定遺伝子たちの抬頭2013

    • Author(s)
      田島祥太, ら
    • Organizer
      2012年度水産学会春季大会
    • Place of Presentation
      東京
    • Related Report
      2012 Annual Research Report
  • [Presentation] ゲノム情報の水産育種への活用 : フグ性決定遺伝子の話を中心に2012

    • Author(s)
      菊池潔
    • Organizer
      本動物遺伝育種学会第18回動物遺伝育種シンポジウム
    • Place of Presentation
      宮城(招待)
    • Year and Date
      2012-10-07
    • Related Report
      2012 Annual Research Report
  • [Presentation] 性決定遺伝子が移りかわる現象の解明 : フグ近縁種群を用いて表現型多様化の遺伝的基盤を探る2012

    • Author(s)
      菊池潔, ら
    • Organizer
      新学術領域研究「複合適応形質進化の遺伝子基盤解明」平成24年度公開シンポジウム
    • Place of Presentation
      東京
    • Year and Date
      2012-09-26
    • Related Report
      2012 Annual Research Report
  • [Presentation] トラフグ稚魚の雌雄判別と雄の作出について2012

    • Author(s)
      菊池潔
    • Organizer
      第9回アクアカルチャーネットワーク懇話会
    • Place of Presentation
      佐賀(招待)
    • Year and Date
      2012-08-23
    • Related Report
      2012 Annual Research Report
  • [Presentation] 組み換え抑制がない性決定遺伝子座における選択の痕跡 : フグの場合2012

    • Author(s)
      渥美九郁, ら
    • Organizer
      日本進化学会第14回東京大会
    • Place of Presentation
      東京
    • Year and Date
      2012-08-21
    • Related Report
      2012 Annual Research Report
  • [Presentation] 性決定遺伝子が移りかわる現象の理解に向けて : トラフグの性決定遺伝子の同定2012

    • Author(s)
      菊池潔, ら
    • Organizer
      日本進化学会第14回東京大会
    • Place of Presentation
      東京
    • Year and Date
      2012-08-21
    • Related Report
      2012 Annual Research Report
  • [Presentation] 超雄トラフグ(YY型魚)の作出とその生存率2012

    • Author(s)
      菊池潔, ら
    • Organizer
      2012年度水産学会春季大会
    • Place of Presentation
      東京
    • Related Report
      2012 Annual Research Report
  • [Presentation] Fine mapping of the sex-determining locus in fugu2012

    • Author(s)
      Kiyoshi Kikuchi, et al.
    • Organizer
      The International Symposium on Aquaculture Genetics XI
    • Place of Presentation
      Auburn, USA
    • Related Report
      2012 Annual Research Report
  • [Presentation] THE SEX-DETERMINING LOCI IN FUGU AND ITS RELATED SPECIES2012

    • Author(s)
      Kiyoshi Kikuchi, et al.
    • Organizer
      SIXTH INTERNATIONAL SYMPOSIUM ON VERTEBRATE SEX DETERMINATION
    • Place of Presentation
      Kona, USA
    • Related Report
      2012 Annual Research Report
  • [Book] 魚類遺伝育種学(10章担当、水産育種におけるゲノム情報の利用)2014

    • Author(s)
      菊池潔
    • Publisher
      京都大学出版
    • Related Report
      2013 Annual Research Report
  • [Book] フグ研究とトラフグ生産技術の最前線(第6章の執筆)2012

    • Author(s)
      鈴木讓, 菊池潔
    • Total Pages
      15
    • Publisher
      恒星社厚生閣
    • Related Report
      2012 Annual Research Report
  • [Remarks] 男と女はどれくらい違う? たった1つのDNA塩基で性別が決まるトラフグの事例

    • URL

      http://www.u-tokyo.ac.jp/ja/utokyo-research/editors-choice/fugu-sex-determined-by-single-nucleotide/

    • Related Report
      2014 Annual Research Report
  • [Remarks] 東京大学大学院農学生命科学研究科附属水産実験所

    • URL

      http://www.se.a.u-tokyo.ac.jp/research.html

    • Related Report
      2013 Annual Research Report
  • [Remarks]

    • URL

      http://www.se.a.u-tokyo.ac.jp/research.html

    • Related Report
      2012 Annual Research Report

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Published: 2012-04-23   Modified: 2019-07-29  

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