Development of genome-wide screening method for induced mutations

• Project/Area Number: 25660006
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Science in genetics and breeding
• Research Institution: Kyoto University
• Principal Investigator: YASUI Yasuo 京都大学, (連合)農学研究科(研究院), 助教 (70293917)
• Co-Investigator(Kenkyū-buntansha): 相井 城太郎 新潟薬科大学, 応用生命科学部, 助教 (10391591)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2013: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: 突然変異育種 / 変異検出 / 次世代シークエンシング / ソバ / 育種 / 変異スクリーニング / NGS

Outline of Final Research Achievements

We aimed to develop the genome-wide-mutation-detection method using a next generation sequencer. Total DNAs were extracted from 960 of M2 plants which were treated by mutation-induced processing. All total DNAs were arranged in 96 well plates, and bulked in 30 PCR template pools (3D pools). Using these 30 pools, six genomic regions (the total length = 12Kb) were amplified by PCR and mutations on the PCR products were screened. We succeeded in the signal detection regarded as SNPs in four PCR regions. Furthermore, it turned out that much more genomic regions will be handled in this method. The method developed in this study will be great help for mutation breedings.

Research Products

• [Journal Article] Diversification of 13S globulins, allergenic seed storage proteins, of common buckwheat (2014)
  • Author(s): Sano, M., Nakagawa, M., Oishi, A., Yasui, Y., Katsube-Tanaka, T
  • Journal Title: Food Chemistry Volume: 155 Pages: 192-198
  • DOI: 10.1016/j.foodchem.2014.01.047
  • Peer Reviewed
• [Presentation] Intact S-ELF 3 is exclusive to heteromorphic SI species in Fagopyrum (2013)
  • Author(s): Yasuo YASUI, Masashi MORI, Jotaro AII, Shingo SATO, Ohmi OHNISHI, Tomoko ABE, Yoriko HAYASHI, Daiki MATSUMOTO, Tatsuya OTA
  • Organizer: 12th International Symposium on Buckwheat
  • Place of Presentation: Lasko, Slovenia
• [Presentation] ソバの異形花型自家不和合性機構 (2013)
  • Author(s): 相井城太郎
  • Organizer: 日本育種学会
  • Place of Presentation: 鹿児島大学