Species with widespread distributions that grow in varied habitats may consist of ecotypes adapted to a particular habitat, or may exhibit cross-tolerance that enables them to exploit a variety of habitats. Populations of Arabidopsis lyrata ssp. lyrata (L.) O’Kane & Al-Shehbaz grow in a wide variety of edaphic settings including serpentine soil, limestone sand, and alluvial flood plains. While all three of these environments share some stressors, a crucial difference among these environments is soil calcium to magnesium ratio, which ranges from 25∶1 in the limestone sand to 0.2∶1 in serpentine soil. The three populations found on these substrates were subjected to three different Ca to Mg ratios under controlled environmental conditions during germination and rosette growth. Response to Ca to Mg ratio was evaluated through germination success and radicle growth rate, rosette growth rate, and the content of Ca and Mg in the rosette. All three populations were particularly efficient in fueling growth under nutrient deficiency, with the highest nutrient efficiency ratio for Ca under Ca deficiency and for Mg under Mg deficiency. Although the serpentine population had significantly higher leaf Ca to Mg ratio than the limestone or flood plain populations under all three Ca to Mg ratios, this increase did not result in any advantage in growth or appearance of the serpentine plants, during early life stages before the onset of flowering, even in the high Mg substrate. The three populations showed no population by substrate interaction for any of the parameters measured indicating that these populations may have cross-tolerance to substrate Ca to Mg ratio.