Glyphosate is an important component of herbicide programs in orchard crops in California. It can be applied alone or in tank-mix combinations under the crop rows or to the entire field and often is used multiple times each year. There has been speculation about the potential impacts of repeated use of glyphosate in perennial crop systems, because of uptake from shallow root systems or indirectly because of effects on nutrient availability in soil. To address these concerns, research was conducted from 2013 to 2020 on key orchard crops to evaluate tree response to glyphosate regimens. Almond, cherry, and prune were evaluated in separate experiments. In each crop, the experimental design was a factorial arrangement of two soil types, four glyphosate rates (0, 1.1, 2.2, and 4.4 kg ae ha−1, applied three times annually), and two post-glyphosate application irrigation treatments. In the first 2 yr of the study, there was no clear impact of the glyphosate regimens on shikimate accumulation or leaf chlorophyll content, which suggested no direct effect on the crop. In the seventh year of the study, after six consecutive years of glyphosate application to the orchard floors, there were no negative impacts of glyphosate application on leaf nutrient concentration or on cumulative trunk growth in any of the three orchard crops. Lack of a negative growth impact even at the highest treatment rate, which included 18 applications of glyphosate totaling nearly 80 kg ae ha−1 glyphosate over the course of the experiment suggest there is not likely a significant risk to tree health of judicious use of the herbicide in these production systems. Given the economic importance of orchard crops in California, and grower and industry concerns about pesticides generally and specifically about glyphosate, these findings are timely contributions to weed management concerns in perennial specialty crops.