EPR signals arising from components in oriented multilayers of Photosystem II (PS II) membranes have been studied and the following results have been obtained. (1) The EPR signals arising from the primary semiquinone-iron complex (Q − AFe) were highly oriented, with features at g = 1.90, g = 1.82 and g = 1.66 showing maxima when the membranes were perpendicular to the magnetic field. (2) The EPR signal, arising from the reduced pheophytin acceptor interacting with Q − AFe, showed an orientation-dependent splitting, ranging from 39 G when the membranes were parallel to the magnetic field to 27 G when the membranes were perpendicular to the magnetic field. (3) The S 2 multiline signal associated with the O 2-evolving enzyme showed an orientation dependence. This was most marked as position shifts in the low-field wings of the spectrum. These effects indicate that the component is oriented within the membrane and has some magnetically anisotropic character. (4) The component at g ⋍ 4, though to be due to an oxidized charge carrier close to S 2, showed a slight orientation dependence in its amplitude, but a significant orientation-dependent field-position shift was present, indicating that this is a magnetically anisotropic centre with a fixed geometry in the membrane. (5) Cytochrome b-559 in its oxidized form showed large highly oriented signals. The g z 2.97 feature was maximum when the membranes were oriented parallel to the magnetic field, while the g y 2.22 was maximum when the membrane plane was perpendicular to the magnetic field. This indicates that the haem plane is perpendicular to the plane of the membrane, in agreement with previous reports using chloroplasts. Ageing of the sample brings about a change from low- to high-spin state accompanied by a change in orientation of the haem relative to the membrane (from perpendicular to approximately 45°). (6) Signal II slow, which is present in the dark and which arises from a component which acts as an electron donor in PS II, is highly oriented. The signal becomes resolved into two different symmetrical four-line spectra. When the membranes were parallel to the magnetic field a narrow signal centred at g ≈ 2.0032 was present, while when the membranes were perpendicular a wider signal centred at g ≈ 2.0061 was present. The g-shift may be taken as an indication that the semiquinone ring is perpendicular to the membrane. (7) The spin-polarized triplet state of P-680, the primary donor chlorophyll, can be photoinduced in oriented PS II multilayers. The Z transition was maximum when the membranes were oriented perpendicular to the magnetic field, while the X and Y transitions were maximum when the membranes were parallel to the magnetic field. This indicates that the plane of the chorophyll ring is parallel to the plane of the membrane.