Development of infection structures of compatible (A–G) strain 98AG31 and incompatible strain 93ID6 (H) of M. larici-populina during time-course infection of leaves of P. trichocarpa × P. deltoides ‘Beaupré’. A, Aniline blue-stained urediniospores producing several ramified germ tubes at vicinity of stomata and hyphal penetration through stomata without appressorium formation at 2 hpi. B, Urediniospores producing germ tubes and appressoria at vicinity of stomata and primary infection hyphae penetrating through stomata at 6 hpi. C, Substomatal vesicle formed under the abaxial epidermis and infection hyphae developing in the spongy mesophyll at 12 hpi. D, Infection hyphae colonizing the spongy mesophyll at 48 hpi. E, Infection hyphae developing from the spongy mesophyll to the palisade mesophyll at 48 hpi. F, Spongy and palisade mesophyll colonized by infection hyphae at 96 hpi. G, Uredinium formed on the abaxial epidermis releasing newly formed urediniospores at 196 hpi. H, Limited development of hyphae of the incompatible M. larici-populina strain 93ID6 in the mesophyll at 96 hpi compared to compatible strain in F. The hyphae of M. larici-populina were painted in red in F and H to help with visualization of fungal hyphae in plant mesophyll. Fungal infection structures are pinpointed by arrowheads. ap, Appressorium; gt, germ tube; ih, infection hyphae; inf, infection site; nf sp, newly formed spore; pih, primary infection hyphae; sp, spore; ssv, substomatal vesicle; ur, uredinium.