A palynological record spanning the last glacial–interglacial period was derived from high-resolution, deep-sea core MD03-2607, located near Kangaroo Island in South Australia. The core site lies opposite the mouth of the River Murray that, together with the Darling River, drains the extensive (∼1.6 × 10⁶ km²) Murray-Darling Basin (MDB). The record comprises 120 samples and is compared with detailed records of sea-surface temperature (SST), the C₃/C₄ plant ratio obtained from the δ¹³C of n-alkanes from leaf waxes, the fluvial clay fraction and its neodymium isotopic composition, airborne dust and the biomass-burning component levuglosan. The chronology of the core is robust; it is built on 24 radiocarbon dates derived from planktic foraminifera, 16 optically stimulated luminescence dates, plus 12 tie points linked to the astronomically tuned marine isotopic record. Algal remains are found in nearly all samples supporting our postulation that the palynoflora is predominantly waterborne. Major findings are that the gymnosperm Callitris, together with high percentages of herb pollen (mostly C₃ plants), is predominant during cold, arid phases, whereas Eucalyptus, is predominant during warmer and wetter periods. High charcoal concentration coincides with high percentages of Eucalyptus, mostly during wet and warm periods. Using the geochemistry of the core’s fluvial sediments, it has been possible to identify when water-transported palynoflora and charcoal originated from the Murray sub-basin (consisting of the River Murray and its main tributaries but not from central or western South Australia). During those periods, rainfall principally originated from the southeastern Indian Ocean. When the Darling sub-basin was the main source of the palynoflora, rainfall must have instead originated from northern Australia. The eolian dust record from the core shows that the dust signal generally coincides with the increased values in herb pollen, in particular duringthe Last Glacial Maximum (LGM) when, in addition to high herb percentages, Callitris representation also increased. This dry landscape taxon likely colonised the then-exposed Lacepede Shelf during this period of extreme low sea-level. There is a good correspondence between SST and mean annual precipitation reconstructed from the pollen counts. During warm phases in the ocean, Eucalyptus was the dominant tree taxon, especially for the entirety of Marine Isotope Stages (MIS) 5, plus MIS 3 and MIS 1. Charcoal levels were particularly low during the dry phases MIS 4 and 2, and even more so during the LGM.