AlGaN/GaN high-electron mobility transistor (HEMT) structures were grown by metal–organic chemical vapor deposition on SiC, hydride vapor phase epitaxy (HVPE) GaN, and ammonothermal GaN substrates to achieve HEMTs with over five orders of magnitude variation in extended defect density. This enables a direct comparison of the effect of extended defects on device performance to achieve the best possible reliability. As-grown material was characterized by atomic force microscopy, electron channeling contrast imaging, and Raman spectroscopy. Devices were characterized by Hall, dc $I$ – $V$ , and pulsed $I$ – $V$ behavior. Reduced threading dislocation density provides an increased 2-D electron gas mobility, but inhibits ohmic contact formation resulting in high contact resistance. Transistor characteristics were nominally identical, with higher OFF-state leakage in the HEMTs on ammonothermal GaN. The pulsed $I$ – $V$ response indicated significantly reduced current collapse in the HEMT on HVPE GaN due to reduced buffer trapping.