the marine pelagic archaeal community is dominated by three major groups, the marine group i (MGi) thaumarchaeota, and the marine groups ii and iii (MGii and MGiii) euryarchaeota. Studies of both MGi cultures and the environment have shown that the MGi core membrane lipids are predominantly composed of glycerol dibiphytanyl glycerol tetraether (GDGt) lipids and the diether lipid archaeol. However, there are no cultured representatives of MGii and iii archaea and, therefore, both their membrane lipid composition and potential contribution to the marine archaeal lipid pool remain unknown. Here, we show that GDGts present in suspended particulate matter of the (sub)surface waters of the north Atlantic ocean and the coastal north Sea are derived from MGi archaea, and that MGII archaea do not significantly contribute to the pool of GDGTs and archaeol. This implies, in contrast to previous suggestions, that their lipids do not affect the widely used sea surface temperature proxy teX 86. These findings also indicate that MGII archaea are not able to produce any known archaeal lipids, implying that our understanding of the evolution of membrane lipid biosynthesis in Archaea is far from complete. The dominance of archaeal communities in the marine pelagic ocean by marine group I (MGI) Thaumarchaeota and the marine group II and III (MGII and MGIII) Euryarchaeota has been well established by numerous studies 1-3. It is known from culture and environmental studies that the Thaumarchaeota are capable of oxidizing ammonia 4,5 and that some members are able to use urea as an alternative substrate 6. The metabolism of MGII and MGIII archaea is thought to be (photo)heterotrophic and these Archaea are potentially able to degrade proteins, carbohydrates, fatty acids and other lipids 1,7-9. However, these suggestions are based solely on metagenomic data since pure cultures of MGII and MGIII archaea have as yet not been obtained. In the marine environment, archaeal membrane lipids are used as biomarkers for the presence of Archaea in microbial ecology studies 10,11 but also in the paleotemperature proxy TEX 86 12 , commonly used in paleoclimatological studies. The membrane lipid composition of the MGI archaea has been widely studied 13-20 and found to include the diether lipid archaeol, glycerol dibiphytanyl glycerol tetraether (GDGT) lipids with zero to 4 cyclopentane moieties, and crenarchaeol, a GDGT with four cyclopentane moieties and a cyclohexane moiety, so far exclusively found in Thaumarchaeota 21. In living and intact cells, GDGTs are mostly bound to polar head groups, termed intact polar lipid GDGTs (IPL-GDGTs). These occur mostly with sugar head groups such as monohexose (MH), dihexose (DH) and hexose phosphohexose (HPH) 13,16,18,19,22. Despite their importance in the marine water column as evidenced by metagenomic surveys 1,7-9 , it is not possible to directly determine the lipid membrane composition of MGII and MGIII archaea as pure cultures are lacking. A previous study based on a combination of metagenomics, sequencing...