Plant germplasm collections are important reservoirs of diversity for plant breeders and adding to the knowledge of the diversity among and within germplasm accessions helps plant breeders to use them more efficiently. The annual legume, bladder clover (Trifolium spumosum L.), was the model plant in this study. The hypothesis that ecogeographical variables are important sources of diversity in the germplasm collection of bladder clover and some variables are more important than others has been studied. Three themes have been addressed here: firstly, analysis of the existing passport data to determine a minimum set of ecogeographical descriptors required for capturing ecological aspects of each collection site; secondly, illustration of the ecogeographical regions and subregions using GIS techniques to identify areas with the greatest likelihood of filling gaps in the collection; thirdly, development of an approach for the evaluation of genetic diversity by grouping accessions with a focus on outliers within the collection using principal component and cluster analyses. In this paper the importance of longitude as a geographical factor has been documented and the overall ecogeographical diversity in the germplasm collection of bladder clover has been screened.