Development of DNA-free genome editing technology for producing a new breed of citrus

2019 Fiscal Year Final Research Report

• Project/Area Number: 19K21150
• Project/Area Number (Other): 18H06010 (2018)
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund (2019); Single-year Grants (2018)
• Review Section: 0602:Agricultural and environmental biology and related fields
• Research Institution: Ehime University
• Principal Investigator: Kaya Hidetaka 愛媛大学, 農学研究科, 准教授 (80398825)
• Project Period (FY): 2018-08-24 – 2020-03-31
• Keywords: ゲノム編集 / カンキツ / カラタチ / DNAフリー / Cas9

Outline of Final Research Achievements

We have been researching for the purpose of developing a technology base for developing new citrus cultivars using DNA-free genome editing technology. First, we confirmed the sequence of the PDS gene of Poncirus trifoliata as the target gene, prepared sgRNA, and confirmed its function in vitro. Next, we examined the conditions for applying the particle bombardment method to the callus of P. trifoliata. After confirming the transduction efficiency using the GUS gene, RNP (Ribonucleoprotein) was injected and CAPS analysis was performed. However, so far no mutations due to genome editing have been confirmed. At present, we are also studying particles used in particle bombardment.

Free Research Field

植物細胞分子生物学

Academic Significance and Societal Importance of the Research Achievements

DNAフリー・ゲノム編集技術を柑橘類において確立することで，遺伝子組換え体の規制の対象外となるゲノム編集柑橘品種を作出できる．愛媛大学では，愛媛県オリジナル柑橘品種のゲノム解析が進められており，各品種における果実特性に関するゲノム情報が明らかになりつつある．DNAフリー・ゲノム編集技術を柑橘を適用することで，ゲノム情報と果実特性の関連性を検証し，圃場での特性評価実験も可能になる．本研究により柑橘類の機能ゲノム研究が進むことで，新品種や機能性柑橘の開発につながると期待される．