Environmental characteristics (for example, temperature, photoperiod) as seasonal cues can affect the offspring sex ratio and reproductive tactics of many hymenopteran insects. Leptocybe invasa Fisher & La Salle is the most critical invasive insect pest of Eucalyptus spp. in the world and displays thelytokous reproduction. In the current study, we studied the effects of temperature and photoperiod on offspring sex ratio and reproductive tactics in L. invasa. Results show that sex ratio (female: male) of L. invasa was under 15, 25 and 35 °C with both L 12: D 12 and L 16: D 8, and cold and thermal acclimation were 74.5:1, 71.0:1, 59.0:0, 17.3:1, 53.0:0, 64.0:0, 47.0:1 and 56.0:0, respectively, which was highly significantly female biased and with no significant difference due to temperature or photoperiod. Offspring virgin females oviposited and induced the bump-shaped galls on plants under the same conditions as described above. Constant temperature, photoperiod and their interaction, and cold and thermal acclimation had no significant effect on the infestation rates of Eucalyptus branches induced by offspring virgin females. Thus, temperature, photoperiod and cold and thermal acclimation did not influence female-biased sex ratio and tactics with thelytokous reproduction of offspring females in L. invasa.