Maping the thm1 mutation via bulked segregation analysis (BSA)
To map the thm1 mutation via bulked segregation analysis (BSA), the thm1 mutant was backcrossed once with WT to generate the BC1F1 population. Phenotypes in the second filial generation (BC1F2) progeny were analyzed for their ability to trigger HSCE in response to heat shock. Three pools, each consisting of 25 plants showing thm1 like, 25 plants showing WT like, and 18 WT reference individuals. Genomic DNA was extracted from rosette leaves of the bulks and WT using a 2 % cetyltrimethylammonium bromide (CTAB) method. The WT DNA was used as an internal reference. The whole genome sequencing and analysis were carried out by Novogene.