*4.4. Chromosome Location and Covariance Analysis*

Chromosome lengths and gene positions were obtained from *M. sativa* genome annotation files, and MG2C v.2 (http://mg2c.iask.in/mg2c\_v2.0/) (accessed on 8 August 2022) was used to visualize gene positions on chromosomes. *M. sativa* protein sequences were aligned to each other or to those of *A. thaliana*, *G. max*, *Z. mays*, or *M. truncatula* using TBtools software. MCScanX was used along with default parameters to identify homozygous relationships between gene replication events and HSF proteins, and results were visualized using Circos and Dual Synteny Plot in TBtools.
