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Methionine (Met) is an essential sulfur-containing amino acid in animals. Cereal and legume crops with limiting levels of Met represent the major food and feed sources for animals. In plants, cystathionine gamma-synthase (CGS), methionine methyltransferase (MMT) and homocysteine methyltransferase (HMT) are committing enzymes synergistically synthesizing Met through the aspartate (Asp) family pathway and the S-methylmethionine (SMM) cycle. The biological functions of  CGS ,  MMT  and  HMT  genes have been respectively studied, whereas their evolution patterns and their contribution to the evolution of Met biosynthetic pathway in plants are unknown. In the present study, to reveal their evolution patterns and contribution, the evolutionary relationship of  CGS ,  MMT  and  HMT  gene families were reconstructed. The results showed that  MMT s began in the ancestor of the land plants and kept conserved during evolution, while the  CGS s and  HMT s had diverged. The  CGS  genes were divided into two branches in the angiosperms, Class 1 and Class 2, of which Class 2 only contained the grasses. However, the  HMT  genes diverged into Class 1 and Class 2 in all of the seed plants. Further, the gene structure analysis revealed that the  CGS s,  MMT s and  HMT s were relatively conserved except for the  CGS s in Class 2. According to the expression of  CGS ,  HMT  and  MMT  genes in soybeans, as well as in the database of soybean, rice and  Arabidopsis , the expression patterns of the  MMT s were shown to be consistently higher in leaves than in seeds. However, the expression of  CGS s and  HMT s had diverged, either expressed higher in leaves or seeds, or showing fluctuated expression. Additionally, the functions of  HMT  genes had diverged into the repair of  S -adenosylmethionine and SMM catabolism during the evolution. The results indicated that the  CGS  and  HMT  genes have experienced partial subfunctionalization. Finally, given the evolution and expression of the  CGS ,  HMT  and  MMT  gene families, we built the evolutionary model of the Met biosynthetic pathways in plants. The model proposed that the Asp family pathway existed in all the plant lineages, while the SMM cycle began in the ancestor of land plants and then began to diverge in the ancestor of seed plants. The model suggested that the evolution of Met biosynthetic pathway is basically consistent with that of plants, which might be vital to the growth and development of different botanical lineages during evolution.

Molecular evolution and expression divergence of three key Met biosynthetic genes in plants: CGS, HMT and MMT