Plant material
In previous studies, natural variation of Arabidopsis thalianapopulations from Catalonia [20] were tested in a multi-year small
scale common garden under carbonated and non-carbonated soils [5].
Seeds of 6 extreme phenotype plants from A1(c+), a soil
carbonate tolerant deme, and 6 extreme phenotype plants from
T6(c-), a soil carbonate sensitive deme, were collected
from the last reciprocal transplant experiment performed in 2015 and
stored under fresh (4ยบ C) and dry conditions until the beginning of
hydroponic and soil cultivation experiments. A1xT6 and T6xA1 crosses
were generated using the 6 A1(c+) and 6
T6(c-) lines (generating 12 crossing lines) and
self-crossed for three generations (F1, F2, F3). Col-0 seeds and seeds
of 18 T-DNA mutants with Col-0 background were purchased from NASC/ABRC
[21]. Segregating lines were genotyped using specific primers and
multiplied for T2 generations. The non-expression of the genes was
validated through qPCR (Dataset_SD1).