Insects that depend on microbial mutualists evolved a variety of organs to transport the microsymbionts while dispersing.The ontogeny and variability of such organs is rarely studied,and the microsymbiont’s effects on the animal tissue development remain unknown in most cases.Ambrosia beetles (Coleoptera:Curculionidae:Scolytinae or Platypodinae) and their mutualistic fungi are an ideal system to study the animalfungus interactions.While the interspecific diversity of their fungus transport organ—mycangia—is well-known,their developmental plasticity has been poorly described.To determine the ontogeny of the mycangium and the influence of the symbiotic fungus on the tissue development,we dissected by hand or scanned with micro-CT the mycangia in various developmental stages in five Xylosandrus ambrosia beetle species that possess a large,mesonotal mycangium:Xylosandrus amputatus,Xylosandrus compactus,Xylosandrus crassiusculus,Xylosandrus discolor,and Xylosandrus germanus.We processed 181 beetle samples from the United States and China.All five species displayed three stages of the mycangium development:(1) young teneral adults had an empty,deflated and cryptic mycangium without fungal mass;(2) in fully mature adults during dispersal,the promesonotal membrane was inflated,and most individuals developed a mycangium mostly filled with the symbiont,though size and symmetry varied;and (3) after successful establisent of their new galleries,most females discharged the bulk of the fungal inoculum and deflated the mycangium.Experimental aposymbiotic individuals demonstrated that the pronotal membrane invaginated independently of the presence of the fungus,but the fungus was required for inflation.Mycangia are more dynamic than previously thought,and their morphological changes correspond to the phases of the symbiosis.Importantly,studies of the fungal symbionts or plant pathogen transmission in ambrosia beetles need to consider which developmental stage to sample.We provide illustrations of the different stages,including microphotography of dissections and micro-CT scans.