The lychee tree is native to China and produce small fleshy fruit up to 5 cm in diameter. Lychee production in Australia is worth > $20 million annually. Pruning of trees encourages new growth, has a positive effect on fruiting of lychee, makes fruit-picking easier, and may increase yield, as it increases light interception and tree crown surface area. The objective of this research was to assess changes in tree structure, i.e. tree crown circumference, width, height and Plant Projective Cover (PPC) using multi-spectral Remotely Piloted Aircraft System (RPAS) imagery collected before and after pruning of a lychee plantation. A secondary objective was to assess any variations in the results as a function of various flying heights (30, 50 and 70 m). Pre- and post-pruning results showed significant differences in all measured tree structural parameters, including an average decrease in: tree crown circumference of 1.94 m; tree crown width of 0.57 m; tree crown height of 0.62 m; and PPC of 14.8 %. The different flying heights produced similar measurements of tree crown width and PPC, whereas tree crown circumference and height measurements decreased with increasing flying height. These results show that multi-spectral RPAS imagery can provide a suitable means of assessing pruning efforts undertaken by contractors based on changes in tree structure of lychee plantations and that it is important to collect imagery in a consistent manner, as varying flying heights may cause changes to tree structural measurements.