2018 Fiscal Year Final Research Report

Development of efficient production technique for pine wood nematode-resistant seedlings derived from embryogenic cells

Research Project

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Project/Area Number	16K14949
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Forest science
Research Institution	Forest Research and Management Organization
Principal Investigator	Maruyama Tsuyoshi 国立研究開発法人森林研究・整備機構, 森林総合研究所, 主任研究員等 (20353865)
Co-Investigator(Kenkyū-buntansha)	細井佳久国立研究開発法人森林研究・整備機構, 森林総合研究所, 主任研究員等 (50353842)
Project Period (FY)	2016-04-01 – 2019-03-31
Keywords	胚性万能細胞 / マツノザイセンチュウ抵抗性苗 / 不定胚形成 / クロマツ / 胚培養 / 体細胞分化 / 外気順化 / プロトプラスト培養
Outline of Final Research Achievements	Fourteen lines of embryogenic cells were induced from 3 pine wood nematode-resistant black pine families, and lines with high somatic embryo differentiation ability were selected. The induction rate of mature somatic embryos was significantly different depending on the cell line, but in the top five lines with high induction rates, more than 180 somatic embryos were formed per plate. When the resulting mature somatic embryos were desiccated for about 10 to 20 days and then cultured in a germination medium, they exhibited a germination rate of 68 to 93%. Thereafter, about 80 to 90% of the germinated somatic embryos formed plants. By carrying out stepwise acclimatization of the in vitro seedlings to the external air environment, 90% or more survival rates were obtained. In addition, protoplast culture of embryogenic cells revealed that somatic embryo induction is possible.
Free Research Field	農学
Academic Significance and Societal Importance of the Research Achievements	マツノザイセンチュウによる松枯れ被害対策の一環として、抵抗性個体の大量増殖が望まれている。これまで、実生や挿し木による苗生産が主に行われているが、これらの方法では増殖に時間と手間がかかり、大規模植林への実用的な対応が難しいという問題がある。そこで、組織培養により、不定胚形成能力を持つ胚性万能細胞から苗の大量生産技術を開発することで、マツノザイセンチュウ抵抗性苗を安定的かつ効率的に作出するための条件を明らかにし、大きな問題となっている松枯れ被害の対策として、実用化に向けた基盤技術の開発に貢献できる。