The size of the spores produced by Bacillus megaterium ATCC 19213 depended upon the nature of the carbon source present in the defined medium in which they were produced. Homogeneous preparations of small (0.38 mum(3)), nearly spherical spores were produced after batch culture in the presence of 2.8 mm citrate, and large (1.17 mum(3)), oblong spores were produced by replacement culture in the presence of 7.35 mm acetate. Large and small spores had approximately the same deoxyribonucleic acid content, density, and heat resistance. Large spores contained about 2.5 times the dipicolinic acid, glucosamine, ribonucleic acid, Mn(2+), and lipid and about 1.5 times the Mg(2+), Fe(2+), Ca(2+), and dry weight of small spores. Large spores were especially enriched in Zn(2+) (4.5-fold). More protein (1.5-fold) was extracted from small spores with 1 n NaOH than from large spores, possibly indicating a difference in the spore coats, but large spores contained about twice the Kjeldahl nitrogen of small spores. A difference in the coats may account for the fact that, unlike small spores, large spores showed improved germination with increased times and temperature of heat shocking. The possibility of determining the location of some of these substances within the spore by comparing the compositional ratios with estimated volumes of specific spore layers is discussed.