Development of a rice gene targeting with autonomous self-marker free system as a new breeding technique

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K07560
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Science in genetics and breeding
• Research Institution: Meijo University
• Principal Investigator: Terada Rie 名城大学, 農学部, 教授 (30137799)
• Research Collaborator: SHIMATANI zenpei ; KAWANO yoji ; ARAKAWA masao
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: ゲノム編集 / ターゲティング / イネ / トランスポゾン / OsRacGEF1 / 耐病性

Outline of Final Research Achievements

We have developed a gene targeting (GT) system with an autonomous self-marker free as a new breeding technique. In this system the positive selection marker of homologous recombination (HR) mediated GT is placed in maize DNA transposon, Ac/Ds and by induction of Ac expression just after GT the positive selection marker is eliminated from targeted gene locus. Since pseud-phosphorylation of OsRacGEF1 was found increase the resistance to blast fungus, we modified OsRacGEF1-S549D by GT and subsequently the positive selection marker was eliminated by Ac activation. TG rice lines of OsRacGEF1-S549D homozygote without any artificial marker sequences were obtained, which revealed increased resistance to both of blast fungus and sheath blight disease.

Free Research Field

遺伝育種学

Academic Significance and Societal Importance of the Research Achievements

イネ遺伝子ターゲティング法は標的遺伝子のみを自在に改変でき既に30以上の遺伝子改変に成功した。ターゲティング遺伝子改変とAc/Dsによる自律的選抜マーカー削除による迅速育種法では、ポジティブ選抜マーカーの削除によって複数遺伝子のターゲティング改変が可能で、分子遺伝子学の最新知見を活用した複数の遺伝子編集によるゲノムデザインが可能であり、圃場展開も視野に入れた信頼性の高い実用化イネを1年弱の短期間に作り出す迅速次世代育種法として利用促進が期待できる。