Metabolic engineering of fatty acidcomposition in soybean by using TILLING

• Project/Area Number: 22580006
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Breeding science
• Research Institution: Saga University
• Principal Investigator: ANAI Toyoaki 佐賀大学, 農学部, 准教授 (70261774)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2010: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 突然変異 / 逆遺伝学 / ダイズ / TILLING / 脂肪酸 / 突然変異体 / TILLING法

Research Abstract

TILLING is one of the most attractive methods for screening novel mutant lines with reverse genetics approach in recent years.Inthis study aimed at improving fatty acid composition of soybean oil by metabolic engineering, I have tried to isolate novel mutant lines and to combine their mutant alleles. As a result, I have isolated several mutant lines corresponding to acyl-ACP thioesterase, stearoyl-ACP desaturase and microsomal omega-3 fatty acid desaturase genes, and successfully developed novel fatty acid mutant lines with more than 80% oleic acid or less than 1.5% linolenic acid in seed oil.

Research Products

• [Journal Article] 2013 年 Genetic Variation in Soybean at the Maturity Locus E4 Is Involved in Adaptation to Long Days at High Latitudes Agronomy (2013)
  • Author(s): Y. Tsubokura, H. Matsumura, M. Xu, B. Liu, H. Nakashima, T. Anai, F. Kong, X. Yuan, H. Kanamori, Y. Katayose, R. Takahashi, K. Harada and J. Abe
  • Journal Title: Agronomy Volume: 3 Issue: 1 Pages: 117-134
  • DOI: 10.3390/agronomy3010117
  • Peer Reviewed
• [Journal Article] The National BioResource Project (NBRP) Lotus and Glycine in Japan (2012)
  • Author(s): Hashiguchi M, Abe J, Aoki T, Anai T, Suzuki A, Akashi R
  • Journal Title: Breeding Science Volume: 61 Issue: 5 Pages: 453-461
  • DOI: 10.1270/jsbbs.61.453
  • NAID: 130004057126
  • ISSN: 1344-7610, 1347-3735
  • Peer Reviewed
• [Journal Article] 2012 年 Positional cloning and characterization reveal the molecular basis for soybean maturity locus E1, which regulates photoperiodic flowering (2012)
  • Author(s): Z. Xia, S. Watanabe, T. Yamada, Y. Tsubokura, H. Nakashima, H. Zhai, T. Anai, S. Sato, T. Yamazaki, S. Lu, H. Wu, S. Tabata and K. Harada
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 109 Issue: 32 Pages: 2155-2164
  • DOI: 10.1073/pnas.1117982109
  • Peer Reviewed
• [Journal Article] Potential of a mutant-based reverse genetic approach for functional genomics and molecular breeding in soybean (2012)
  • Author(s): T. Anai
  • Journal Title: Breeding Science Volume: 61 Issue: 5 Pages: 462-467
  • DOI: 10.1270/jsbbs.61.462
  • NAID: 130004057127
  • ISSN: 1344-7610, 1347-3735
  • Peer Reviewed
• [Journal Article] Molecular characterization of two high-palmitic-acid mutant loci induced by X-ray irradiation in soybean (2012)
  • Author(s): T. Anai*, T. Hoshino, N. Imai and Y. Takagi
  • Journal Title: Breeding Science Volume: 61 Issue: 5 Pages: 631-638
  • DOI: 10.1270/jsbbs.61.631
  • NAID: 130004057143
  • ISSN: 1344-7610, 1347-3735
  • Peer Reviewed
• [Journal Article] Positional cloning and characterization reveal the molecular basis for soybean maturity locus E1, which regulates photoperiodic flowering (2012)
  • Author(s): Z.Xia, S.Watanabe, T.Yamada, Y.Tsubokura, H.Nakashima, H.Zhai, T.Anai. S.Sato, T.Yamazaki, S.Lu, H.Wu, S.Tabata, K.Harada
  • Journal Title: Natl.Acad.Sci.USA Volume: (in press)
  • Peer Reviewed
• [Journal Article] 2011 年 Molecular characterization and marker development of mid-oleic-acid mutant M23 for the development of high-oleic cultivars of soybean (2011)
  • Author(s): T. Hoshino, N. Kawashita, Y. Takagi and T. Anai
  • Journal Title: Plant Breeding Volume: 130 Issue: 5 Pages: 544-550
  • DOI: 10.1111/j.1439-0523.2011.01871.x
  • Peer Reviewed
• [Journal Article] 2011 年 A map-based cloning strategy employing a residual heterozygous line reveals that the GIGANTEA gene is involved in soybean maturity and flowering (2011)
  • Author(s): S. Watanabe, Z. Xia, R. Hideshima, Y. Tsubokura, S. Sato, N. Yamanaka, R. Takahashi, T. Anai, S. Tabata, K. Kitamura, K. Harada
  • Journal Title: Genetics Volume: 188 Issue: 4 Pages: 395-407
  • DOI: 10.1534/genetics.108.098772
  • Peer Reviewed
• [Journal Article] A mapbased cloning strategy employing a residual heterozygous line reveals that the GIGANTEA gene is involved in soybean maturity and flowering (2011)
  • Author(s): S.Watanabe, Z.Xia, R.Hideshima, Y.Tsubokura, S.Sato, N.Yamanaka, R.Takahashi, T.Anai, S.Tabata, K.Kitamura, K.Harada
  • Journal Title: Genetics Volume: 188 Issue: 2 Pages: 395-407
  • DOI: 10.1534/genetics.110.125062
  • Peer Reviewed
• [Journal Article] A map-based cloning strategy employing a residual heterozygous line reveals that the GIGANTEA gene is involved in soybean maturity and flowering. (2011)
  • Author(s): S.Watanabe, Z.Xia, R.Hideshima, Y.Tsubokura, S.Sato, N.Yamanaka, R.Takahashi, T.Anai, S.Tabata, K.Kitamura, K.Harada
  • Journal Title: Genetics Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] 2010 年 Phylogenetic relationships of Citrus and its relatives based on rbcL gene sequences (2010)
  • Author(s): T. Penjor, T. Anai, Y. Nagano, R. Matsumoto, M. Yamamoto
  • Journal Title: Tree Genetics and Genomes Volume: 6 Issue: 6 Pages: 931-939
  • DOI: 10.1007/s11295-010-0302-1
  • Peer Reviewed
• [Journal Article] Agrobacterium-Mediated Transformation of Euphorbia tirucalli Callus (2010)
  • Author(s): H. Uchida, H. Yamashita, T. Anai, T. Muranaka, K. Ohyama
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 74 Issue: 4 Pages: 851-853
  • DOI: 10.1271/bbb.90783
  • NAID: 10027554214
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Novel GmFAD2-1b mutant alleles created by reverse genetics induce marked elevation of oleic acid content in soybean seeds in combination with GmFAD2-1a mutant alleles (2010)
  • Author(s): T. Hoshino, Y. Takagi, T. Anai
  • Journal Title: Breeding Science Volume: 60 Issue: 4 Pages: 419-425
  • DOI: 10.1270/jsbbs.60.419
  • NAID: 130004146252
  • ISSN: 1344-7610, 1347-3735
  • Peer Reviewed
• [Journal Article] Novel GmFAD2-1b mutant alleles created by reverse genetics induce drastic elevation of oleic acid content in soybean seeds in combination with GmFAD2-1a mutant alleles. (2010)
  • Author(s): T.Hoshino, Y.Takagi, T.Anai
  • Journal Title: Breeding Science Volume: 60 Pages: 419-425
  • Peer Reviewed
• [Journal Article] Agrobacterium-mediated transformation of Euphorbia tirucalli callus. (2010)
  • Author(s): H.Uchida, H.Yamashita, T.Anai, T.Muranaka, K.Ohyama
  • Journal Title: Bioscience.Biotechnol.Biochem Volume: 74 Pages: 851-853
  • NAID: 10027554214
  • Peer Reviewed
• [Journal Article] Phylogenetic relationships of Citrus and its relatives based on rbcL gene sequences. (2010)
  • Author(s): T.Penjor, T.Anai, Y.Nagano, R.Matsumoto, M.Yamamoto
  • Journal Title: Tree Genet.Genomes Volume: 6 Pages: 931-939
  • Peer Reviewed
• [Presentation] Soybean Mutant Resource for Reverse Genetic Study (2012)
  • Author(s): T. Anai
  • Organizer: 第53回日本植物生理学会シンポジウム
  • Place of Presentation: 京都産業大学
  • Year and Date: 2012-03-16
• [Presentation] Soybean mutant resource for reverse genetic study (2012)
  • Author(s): Toyoaki Anai
  • Organizer: 第53回日本植物生理学会
  • Place of Presentation: 京都産業大学
  • Year and Date: 2012-03-16
• [Presentation] ダイズ突然変異リソースの開発と利用 (2012)
  • Author(s): 穴井豊昭
  • Organizer: 日本遺伝学会第84回大会ワークショップ
  • Place of Presentation: 九州大学
• [Presentation] ダイズ変異体ライブラリーの作出と利用 (2012)
  • Author(s): 穴井豊昭
  • Organizer: 日本育種学会第122回講演会シンポジウム
  • Place of Presentation: 京都産業大学
• [Presentation] ILLING法に利用できるダイズ突然変異体リソース (2011)
  • Author(s): 穴井豊昭
  • Organizer: 日本植物分子細胞生物学会第29回大会シンポジウム
  • Place of Presentation: 九州大学
  • Year and Date: 2011-09-07
• [Presentation] TILLING法に利用できるダイス突然変異体リソース (2011)
  • Author(s): 穴井豊昭
  • Organizer: 第29回日本植物細胞分子生物学会
  • Place of Presentation: 九州大学
  • Year and Date: 2011-09-07
• [Presentation] X線により誘発されたダイズ脂肪酸突然変異体におけるDNA変異形態の特徴 (2010)
  • Author(s): 星野友紀、川下宣久、高木胖、穴井豊昭
  • Organizer: 育種学研究12(別2)
  • Place of Presentation: 秋田県立大学
  • Year and Date: 2010-09-25
• [Presentation] ダイズの矮性突然変異遺伝子についての解析 (2010)
  • Author(s): 穴井豊昭、北原阿子
  • Organizer: 育種学研究12(別2)
  • Place of Presentation: 秋田県立大学
  • Year and Date: 2010-09-25
• [Presentation] 2010年 TILLING法を用いたダイズ品種「フクユタカ」からのFT3変異体の単離 (2010)
  • Author(s): 西美友紀、木下剛仁、穴井豊昭、中島寿亀
  • Organizer: 育種学研究12(別2)
  • Place of Presentation: 秋田県立大学
  • Year and Date: 2010-09-25
• [Presentation] X線により誘発されたダイズ脂肪酸突然変異体におけるDNA変異形態の特徴 (2010)
  • Author(s): 星野友紀、川下宣久、高木胖、穴井豊昭
  • Organizer: 日本育種学会第118回講演会
  • Place of Presentation: 秋田県立大学
  • Year and Date: 2010-09-25
• [Presentation] ダイズの矮性突然変異遺伝子についての解析 (2010)
  • Author(s): 穴井豊昭、北原阿子
  • Organizer: 日本育種学会第118回講演会
  • Place of Presentation: 秋田県立大学
  • Year and Date: 2010-09-25
• [Presentation] TILLING法を用いたダイズ品種「フクユタカ」からのFT3突然変異体の単離 (2010)
  • Author(s): 西美友紀、木下剛仁、穴井豊昭、中島寿亀
  • Organizer: 日本育種学会第118回講演会
  • Place of Presentation: 秋田県立大学
  • Year and Date: 2010-09-24
• [Book] 2012年 Development of novel fatty acid composition in soybean oil through induced mutation (2012)
  • Author(s): Y. Takagi, T. Anai, H. Nakagawa
  • Publisher: CABI Publishers
• [Book] Development of novel fatty acid composition in soybean oil through induced mutation (2012)
  • Author(s): Y.Takagi, T.Anai, H.Nakagawa
  • Publisher: IAEA Publication Series(in press)
• [Book] 2011 年 Mutation breeding of soybean in Japan. Breeding of Pulse Crops (2011)
  • Author(s): H. Nakagawa, T. Anai, A. Okabe, K. Takahashi, M. Hajika, Y. Takagi
  • Publisher: Kalyani Publishers
• [Book] 植物生理化学研究と遺伝子解析法 (2011)
  • Author(s): 星野友紀, 穴井豊昭
  • Publisher: 大学教育出版
• [Book] 2011 年 Positional cloning of the responsible genes for maturity loci E1, E2 and E3 in soybean (2011)
  • Author(s): K. Harada, S. Watanabe, Z. Xia, Y. Tsubokura, N. Yamanaka, T. Anai
  • Publisher: InTech Publishers
• [Book] 最新植物生理化学 (2011)
  • Author(s): 星野友紀, 穴井豊昭
  • Publisher: 大学教育出版
• [Book] Soybean (2011)
  • Author(s): K.Harada, S.Watanabe, Z.Xia, Y.Tsubokura, N.Yamanaka, T.Anai
  • Publisher: InTech Publishers
• [Book] Breeding of Pulse Crops (2010)
  • Author(s): Mutation breeding of soybean in Japan.
  • Total Pages: 249
  • Publisher: Kalyani Publishers