Development and Validation of a SPME-GC-MS Method for In situ Passive Sampling of Root Volatiles from Glasshouse-Grown Broccoli Plants Undergoing Below-Ground Herbivory by Larvae of Cabbage Root Fly, Delia radicum L.
BACKGROUND
Research on plant root chemical ecology has benefited greatly from recent developments in analytical chemistry. Numerous reports document techniques for sampling root volatiles, although only a limited number describe in situ collection.


OBJECTIVE
To demonstrate a new method for non-invasive in situ passive sampling using solid phase micro extraction (SPME), from the immediate vicinity of growing roots.


METHODS
SPME fibres inserted into polyfluorotetrafluoroethylene (PTFE) sampling tubes located in situ which were either perforated, covered with stainless steel mesh or with microporous PTFE tubing, were used for non-invasive sub-surface sampling of root volatiles from glasshouse-grown broccoli. Sampling methods were compared with above surface headspace collection using Tenax TA. The roots were either mechanically damaged or infested with Delia radicum larvae. Principal component analysis (PCA) was used to investigate the effect of damage on the composition of volatiles released by broccoli roots.


RESULTS
Analyses by gas chromatography-mass spectrometry (GC-MS) with SPME and automated thermal desorption (ATD) confirmed that sulphur compounds, showing characteristic temporal emission patterns, were the principal volatiles released by roots following insect larval damage. Use of SPME with in situ perforated PTFE sampling tubes was the most robust method for out-of-lab sampling.


CONCLUSIONS
This study describes a new method for non-invasive passive sampling of volatiles in situ from 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. Copyright © 2016 John Wiley & Sons, Ltd.
The James Hutton Institute, Invergowrie, Dundee, DD2 5DA, UK.|Scotland's Rural College, Nicholas Kemmer Road, Edinburgh, EH9 3FH, UK.|School of Biological Sciences, The University of Edinburgh, Darwin Building, The King's Buildings, Max Born Crescent, Edinburgh, EH9 3BF, UK.
William
W
Deasy
The James Hutton Institute, Invergowrie, Dundee, DD2 5DA, UK. tom.shepherd@hutton.ac.uk.
Tom
T
Shepherd
Biomathematics and Statistics Scotland, Invergowrie, Dundee, DD2 5DA, UK.
Colin J
CJ
Alexander
The James Hutton Institute, Invergowrie, Dundee, DD2 5DA, UK.
A Nicholas E
AN
Birch
Scotland's Rural College, Nicholas Kemmer Road, Edinburgh, EH9 3FH, UK.
K Andrew
KA
Evans
D000818:Animals; D001937:Brassica / Q000737:chemistry*; D004175:Diptera / Q000254:growth & development*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D060434:Herbivory; D007814:Larva / Q000502:physiology*; D018517:Plant Roots / Q000737:chemistry*; D052617:Solid Phase Microextraction / Q000379:methods*; D014835:Volatilization
Brassica; Chemical ecology; Delia radicum; SPME-GC-MS; in situ root volatiles analysis