Biocontrol of Ailanthus altissima: inoculation protocol and risk assessment for Verticillium nonalfalfae (Plectosphaerellaceae: Phyllachorales)

Verticillium nonalfalfae has been proposed as a biocontrol for invasive Ailanthus altissima (tree-of-heaven) in Ohio, Pennsylvania and Virginia. However, previous studies evaluating this potential biocontrol utilised a conidial suspension with a short shelf life as inoculum. Anticipating future expanded use of V. nonalfalfae, we evaluated other inoculum formulations, inoculation protocols and sensitivity of non-target (non-Ailanthus) plant species within Pennsylvania. The most effective inoculum formulation, with an extended shelf life, was prepared by mixing water with stored, refrigerated soil containing V. nonalfalfae. Less successful, but positive infections were obtained by simply using infected Ailanthus wood and leaves as inoculum. Monthly inoculation of Ailanthus trees demonstrated that the optimal time for successful inoculations was April to May, but limited infections were achieved during all months, including the winter. The health of Ailanthus and non-target species was evaluated within a decade-old natural Verticillium wilt epicentre, where all mature Ailanthus trees had been killed by V. nonalfalfae. Verticillium wilt was observed on a few small Ailanthus trees, likely newly established seedlings, whereas non-target species were asymptomatic. Our findings reveal that soil formulated and natural inocula are effective biocontrols against Ailanthus, and V. nonalfalfae appears to pose little threat to non-target plants.