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The American Chestnut Project
Enhanced Blight Resistance in Transgenic American Chestnut

Small Stem Assay Method Instructions

American chestnut test for enhanced blight resistance

small stem assay crankers

very small stem blight resistance assay shoiwng the significant blight resistance enhancement using the oxo gene

From left to right, the trees are a blight-susceptible wild-type American chestnut (C. dentata) called Ellis 1, a blight-resistant Chinese chestnut (C. mollissima) tree called 'Qing,' and two transgenic American chestnut lines called Darling 215 and 311. The Chinese and American trees are "controls" that show the difference between susceptible and resistant trees typically seen in the wild. The transgenic American chestnut lines are derived from the Ellis 1 line, so they are identical to the American control, except for the oxalate oxidase gene from wheat that we added to enhance blight resistance. Watch as the wild-type American chestnut dies from chestnut blight, while the Darling 215 and 311 trees survive just as well as the blight-resistant Chinese chestnut control. Make sure your sound is turned on for the full effect.

Previous assay on even smaller stems

The possibility that the transgenic American chestnut trees may have higher levels of blight resistance than the Chinese chestnut control is supported by another experiment using trees with smaller stems (figure below) and using a novel leaf inoculation assay developed in our lab. In the "very-small-stem-assay", both the Ellis American control and the Chinese chestnut died, while both the Darling American chestnut trees survived. The Chinese chestnut died in that assay because the stem was so small the fungus could overwhelm it more easily. But the oxalate oxidase gene in the Darling trees was apparently sufficient to block the fungus, even on these very small stems. Therefore, these American chestnut trees might have higher levels of resistance than the Qing Chinese control.


1. Powell, W. A., P. Morley, M. King and C. A. Maynard. 2007. Small stem chestnut blight resistance assay. Journal of The American chestnut Foundation 21(2): 34-38

2. Zhang B, AD Oakes, AE Newhouse, KM Baier, CA Maynard and WA Powell. 2013. A threshold level of oxalate oxidase transgene expression reduces Cryphonectria parasitica - induced necrosis in a transgenic American chestnut (Castanea dentata) leaf bioassay. Transgenic Research 22, Issue 5 (2013), Page 973-982

American chestnut test for enhanced blight resistance

Time-lapse blight-resistance demonstration video

chestnut saplings