Apple proliferation (AP), a phytoplasma-induced disease of apple trees, was proven to be transmitted through infected grafting material and sap-sucking insects. To date there are little firm data on disease propagation in the field via natural root grafts. This question was thus addressed in the present case study by investigating trees of a 24-year old commercial apple orchard (‘Red Chief’ on MM 111), where the existence of root connections was discovered accidentally. After having displayed specific AP symptoms, nine trees were cut down and the stubs were infiltrated or brushed with glyphosate. Herbicide injury, however, remained not only restricted to the treated stubs, but also spread to approximately 50 neighbouring trees. Surprisingly, none of the pollinators (‘Granny Smith’ on M 9) growing interjacently and alternating between herbicide-damaged main crop trees was affected. Respective to the position of the nine AP-infected and glyphosate-treated cut stumps, four sections in the orchard were defined, from which a total of 122 trees was sampled and analysed using qualitative real-time PCR for detection of AP phytoplasma. The pathogen was found in 71.4% of ‘Red Chief’ trees with severe herbicide damage and 18.8% of trees with partial herbicide damage. None of the 31 investigated pollinators was AP-infected. Our data indicate that root connections seem to play a role for the spread of AP phytoplasma at least in older orchards and between trees on vigorous rootstocks.