Evaluation of an Alternative Small Stem Assay for Blight Resistance in American, Chinese, and Hybrid Chestnuts ( spp.).

We tested an alternative small stem assay (SSA) for blight resistance in chestnuts ( spp.). Whereas standard SSAs are done by inoculating small incisions in stems, we cut off stems (4 to 5 mm diameter), inoculated the cut ends with discs of inoculum, and covered them with plastic sleeves. This method was designed to be simple to implement, to consistently induce cankering, and to better enable seedlings to recover by developing shoots from the lower stem (standard SSAs delay removal of blighted stems until late in the growing season, if at all). We conducted six experiments with seedlings and orchard trees of (susceptible), (resistant), and hybrids expected to vary in resistance. Experiments with seedlings and two of the three orchard experiments showed clear differentiation between susceptible and resistant types, especially >90 days postinoculation and when the orange-colored zone of the canker was measured. One orchard experiment failed to give clear results but was ended earlier (60 days) than the other experiments. We observed only two failed inoculations out of >200 performed. Comparisons with other studies suggest that this SSA method performs at least as well as the standard SSA method in distinguishing resistant and susceptible types, at least in seedlings. Survivorship after 1 year for seedlings inoculated in 2018 ranged from 70% for to 100% for , and in 2019 they ranged from 40% in hybrids to 100% for Deaths of seedlings after SSAs were mostly unrelated to the inoculations (e.g., root rot).