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Chestnut, American (Castanea dentata (Marsh.) Borkh.)
Charles A Maynard1, Linda D McGuigan, Allison D Oakes
1Department of Forest and Natural Resources Management, SUNY College of Environmental Science and Forestry, 1 Forestry Dr., Syracuse, NY, 13210, USA, cmaynard@syr.edu.
Methods in Molecular Biology (Clifton, N.J.)
|November 23, 2014
Summary
Developing a robust protocol for transgenic American chestnut (Castanea dentata) involves optimizing explant tissue, DNA delivery, and regeneration. This enhanced method significantly increases transformation efficiency and yields blight-resistant trees.
Area of Science:
- Plant Biotechnology
- Forest Genetics
- Molecular Biology
Background:
- The American chestnut (Castanea dentata) is a vital forest species facing significant decline due to chestnut blight.
- Developing blight-resistant American chestnut trees is crucial for ecological restoration and preserving biodiversity.
Purpose of the Study:
- To establish a highly efficient and robust transformation protocol for American chestnut.
- To identify and develop transgenic American chestnut with enhanced resistance to chestnut blight.
Main Methods:
- Utilized somatic embryos for transformation, employing gentle handling during Agrobacterium inoculation and cocultivation.
- Implemented desiccation plates for cocultivation and temporary-immersion bioreactors for selection.
- Regenerated transgenic events into whole plants and conducted field trials to assess blight resistance.
Main Results:
- Achieved an average transformation efficiency of approximately 170 stable embryogenic transformation events per gram of somatic embryo tissue.
- Successfully regenerated nearly 100 transgenic events into whole plants, with 1,275 planted in field trials by fall 2013.
- Identified three transgenes conferring intermediate to higher resistance to chestnut blight compared to non-transgenic American chestnut and Chinese chestnut.
Conclusions:
- The developed protocol represents a significant advancement in American chestnut transformation, improving efficiency and success rates.
- Transgenic American chestnut with enhanced blight resistance has been successfully generated and field-tested.
- This research provides a pathway for restoring the American chestnut population through genetic resistance to blight.
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