Trees growing in their natural habitat represent a valuable resource for elucidating mechanisms of adaptation to environmental constraints. Along the Erqis river, there are various Populus forests, which provide 'natural laboratories' for studying tree ecophysiological responses to their habitat. Reproduction strategies and natural variation of the 'mosaic' distributed Populus canescens patches were studied using a proteomic approach and nuclear microsatellite markers. Clonal reproduction was the primary reproduction strategy of these P. canescens patches. Forty-eight percent of the locations represented in one or two P. canescens patches were identified. In total, 83 different proteins were identified in 118 of 119 protein spots, most of them involved in metabolism. Distinct proteomes and post-translational modifications were found in different P. canescens patches. The differences in the proteomes originate both from the expression of different protein isoforms with the same function and from the differential expression of proteins with different functions, suggesting that different patches might have a functional basis for their adaptation to their environments. Our studies provide a good example of applying proteomics to measure natural variation between patches and will provide a basis for understanding how trees survive through their responses to natural conditions.