C1 metabolism and chlorophyll synthesis: the Mg-protoporphyrin IX methyltransferase activity is dependent on the folate status.

* Tetrahydrofolate derivatives are central cofactors of C1 metabolism. Using methotrexate as a specific inhibitor of folate biosynthesis, we altered the folate status in 10-d-old etiolated pea (Pisum sativum) leaves and followed the rate of chlorophyll synthesis upon illumination. * In our conditions, the folate concentration decreased only from 5.7 to 4.2 nmol g(-1) FW, but the amount of chlorophyll after 24 h of illumination was reduced 2.5 times. Folate status and rate of chlorophyll synthesis were apparently correlated through the methyl cycle. * Indeed, we observed that methyl-tetrahydrofolate was the folate derivative most affected by the treatment; the decrease of methyl-tetrahydrofolate was associated with a sharp rise in homocysteine and S-adenosylhomocysteine concentrations, which are normally maintained at very low values, shifting the methylation index (S-adenosylmethionine/S-adenosylhomocysteine ratio) from 7 to 1; the decrease of the methylation index reduced by a factor of 3 the activity of the Mg-protoporphyrin IX methyltransferase (CHLM), an essential enzyme for chlorophyll synthesis. CHLM gene expression and protein concentration remained unchanged, suggesting that this inhibition relied essentially on metabolic regulation. * These results point out that an even moderate change in the folate status may affect plant development and adaptation.