Establishing microspore culture for efficient genome editing in apple.

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K07636
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Horticultural science
• Research Institution: Iwate University
• Principal Investigator: KOMORI Sadao 岩手大学, 農学部, 教授 (00333758)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 果樹園芸学 / リンゴ / 小胞子培養 / 葯培養 / カルス / 成長点 / 珠心細胞 / シュート再分化

Outline of Final Research Achievements

Microspore culture, anther culture, and experiments of shoot regeneration from calli derived from apical meristem and nucellar cells were conducted to achieve efficient genome editing in apple. For microspore culture, four cultivars included ‘Fuji’ formed callus and/or a heart-shaped embryo. From anther culture, ‘95P6’, which is doubled haploid cultivar, showed a higher embryo formation rate and a higher shoot regeneration rate than those of other cultivars. Results suggest ‘95P6’ as suitable for experiments such as genome editing.
In experiments using calli in ‘Fuji’ and other cultivars, the calli derived from apical meristem formed shoots directly. The calli derived from nucellar cells excised from seeds at 30-40 days after anthesis formed adventitious embryos. From results of these experiments, we established two cultural systems: direct shoot regeneration from calli and shoot regeneration from calli via nucellar cells.

Free Research Field

農学

Academic Significance and Societal Importance of the Research Achievements

ゲノム編集技術の発展には様々な培養系の確立が必須であるが、これまでリンゴで形質転換実験・ゲノム編集実験に適用できる培養系は葉切片からの直接再分化系のみであった。今回の研究成果によってアグロバクテリウム法などの形質転換によらないゲノム編集の実現に一歩近いた。ゲノム編集技術の発展は、リンゴ育種の効率化を通してリンゴ生産者および消費者の利益につながり、学術的には研究の国際競争力の向上に資するものである。