In one carbon reaction, tetrahydrofolate works as cofactor for the synthesis of glycine, serine, methionine, purines and thymidylate. Folate or vitamin B9 is not synthesized in humans therefore plants are the main source of this essential vitamin. Deficiency of vitamin B9 may cause severe health consequences like birth defects, megaloblastic anemia, cardiovascular disorders, cancers, etc. To recover folate intake worldwide in the daily diet it is essential to engineer genetically universal food plants with intrinsic capacity to synthesize folate. In this review, we have precised recent advances in determination, biosynthesis enzymes, transport and compartmentation of folate.