Introduction - Research on the chemical ecology of plant roots has benefited greatly from recent developments in analytical chemistry. Numerous reports have documented techniques for sampling root volatiles, although only a limited number of studies describe in situ collection. Objectives - To demonstrate the feasibility of non-invasive in situ passive sampling using SPME from the immediate vicinity of growing roots. Methods - SPME fibres inserted into perforated PTFE sampling tubes, stainless steel mesh covered PTFE sampling tubes and microporous tubing covered PTFE sampling tubes located in situ were used for non-invasive sub-surface sampling of root volatiles from glasshouse-grown broccoli plants. An above surface headspace collection technique using Tenax TA was used to compare sampling methods. The roots were either mechanically damaged or infested with Delia radicum larvae. PCA was applied to investigate the effect of damage on the composition of volatiles released by broccoli roots. Results - SPME-GC-MS and ATD-GC-MS analyses revealed that sulfur compounds, showing characteristic temporal emission patterns, were the principal volatiles released by roots in response to insect larval damage. Use of SPME with perforated PTFE sampling tubes positioned in situ was the most robust of the methods for out-of-lab sampling. Conclusion - This study is the first to describe non-invasive passive sampling of volatiles in situ from the immediate vicinity of intact and insect damaged roots using SPME. The method is highly suitable for remote sampling and has potential for wide application in chemical ecology/root/soil research.