A putative male-produced pheromone has recently been described for the global pest of pines, Sirex noctilio, but field-activity has not been demonstrated. This study aimed to investigate the pheromone biology of S. noctilio in more detail. Specifically, we i) analysed effluvia and extracts for additional compounds by gas chromatography coupled with electro-antennographic detection (GC-EAD), mass spectrometry (GC-MS) and two dimensional time of flight mass spectrometry (GC X GC TOF MS), ii) conducted dose-response experiments for putative pheromone components, iii) determined the site of synthesis/ storage of the putative pheromone and iv) determined the release rate of the putative pheromone from males and three types of lures. A blend of four compounds was identified, including the previously described (Z)-3-decenol and (Z)-4-decenol, and two new compounds (Z)-3-octenol and (Z)-3-dodecenol. All compounds elicited a response from both male and female antennae, but the strength of the response varied according to sex, compound and dose tested. (Z)-3-Decenol and (Z)-3-octenol at lower and higher doses, respectively, elicited larger responses in males and females than the other two compounds. (Z)-3-Octenol and (Z)-4-decenol generally elicited larger female than male antennal responses. The site of synthesis and/or storage in males was determined to be the hind legs, likely in the leg-tendon gland. The relative release rate of the major compound by male wasps was shown to be 90 ± 12.4 ng/min, which is between 4 and 15 times greater than that observed from typical lures used previously. These observations are consistent with the hypothesis that these compounds may mediate lek formation in S. noctilio males and lek location in females.