Several photochemical and spectral properties of maize (Zea mays) bundle sheath and mesophyll chloroplasts are reported that provide a better understanding of the photosynthetic apparatus of C4 plants. The difference absorption spectrum at 298 K and the fluorescence excitation and emission spectra of chlorophyll at 298 K and 77 K provide new information on the different forms of chlorophyll a in bundle sheath and mesophyll chloroplasts: the former contain, relative to short wavelength chlorophyll a forms, more long wavelength chlorophyll a form (e.g. chlorophyll a 693 and chlorophyll a 705) and less chlorophyll b than the latter. The degree of polarization of chlorophyll a fluorescence is 6% in bundle sheath and 4% in mesophyll chloroplasts. This result is consistent with the presence of relatively high amounts of oriented long wavelength forms of chlorophyll a in bundle sheath compared to mesophyll chloroplasts. The relative yield of variable, with respect to constant, chorophyll a fluorescence in mesophyll chloroplasts is more than twice that in bundle sheath chloroplast. Furthermore, the relative yield of total chlorophyll a fluorescence is 40% lower in bundle sheath compared to that in mesophyll chloroplasts. This is in agreement with the presence of the higher ratio of the weakly fluorescent pigment system I to pigment system II in bundle sheath than in mesophyll chloroplast. The efficiency of energy transfer from chlorophyll b and carotenoids to chlorophyll a are calculated to be 100 and 50%, respectively, in both types of chloroplasts. Fluorescence quenching of atebrin, reflecting high energy state of chloroplasts, is 10 times higher in mesophyll chloroplasts than in bundle sheath chloroplasts during noncyclic electron flow but is equal during cyclic flow. The entire electron transport chain is shown to be present in both types of chloroplasts, as inferred from the antagonistic effect of red (650 nm pending on the species, age, and growth conditions. Several papers have been published on the photochemical activities of the two types of chloroplasts (2,5,20,24). In spite of some of the controversial reports, it is now clear that BS Chip2 are enriched in PS I relative to PS II: there is about 3-fold higher PS I/PS II ratio in these chloroplasts than in M Chlp.In this study, we provide further evidence, in addition to the new information on the different forms of Chl a, for enrichment of BS Chlp in PS I relative to PS II by new types of measurements: the difference absorption spectrum at 298 K, the excitation spectra of Chl a fluorescence at 77 K and 298 K, and the degree of polarization of Chl a fluorescence. Furthermore, new information is presented on the relative efficiencies of energy transfer from the accessory pigments to Chl a. We also provide new and direct evidence, aside from enzymatic assay for system II activity, for the relative deficiency in BS Chlp of system II from the measurements of the time course of Chl a fluorescence, and of the fluorescence quenching of the membrane probe ateb...