This article presents experimental data describing the morphology and the cell wall monosaccharide content of rosettes and flower stems of five Arabidopsis thaliana ecotypes grown at two contrasted temperatures. Besides, cell wall polysaccharides are reconstructed from data of monosaccharide quantification. The well-described and sequenced Columbia (Col) ecotype and four newly-described Pyrenees ecotypes (Duruflé et al., 2019) have been grown at two different temperatures (15 °C and 22 °C). For macrophenotyping, we provide dataset regarding (i) rosettes such as measurement of diameter and fresh mass as well as number of leaves just before bolting and (ii) floral stems at the first flower stage such as length, number of cauline leaves, mass and diameter at its base. Regarding cell wall composition, we provide data of quantification of seven monosaccharides and the reconstruction three polysaccharides. All these data are markers to differentiate both growth temperatures and the different ecotypes. They constitute a valuable resource for the community to study the adaptation of A. thaliana ecotypes to sub-optimal temperature growth conditions.