Increased ascorbate biosynthesis does not improve nitrogen fixation nor alleviate the effect of drought stress in nodulated Medicago truncatula plants

Legume plants are able to establish nitrogen-fixing symbiotic relations with Rhizobium bacteria. This symbiosis is, however, affected by a number of abiotic constraints, particularly drought. One of the consequences of drought stress is the overproduction of reactive oxygen (ROS) and nitrogen species (RNS), leading to cellular damage and, ultimately, cell death. Ascorbic acid (AsA), also known as vitamin C, is one of the antioxidant compounds that plants synthesize to counteract this oxidative damage. One promising strategy for the improvement of plant growth and symbiotic performance under drought stress is the overproduction of AsA via the overexpression of enzymes in the Smirnoff-Wheeler biosynthesis pathway. In the current work, we generated Medicago truncatula plants with increased AsA biosynthesis by overexpressing MtVTC2, a gene coding for GDP-L-galactose phosphorylase. We characterized the growth and physiological responses of symbiotic plants both under well-watered conditions and during a progressive water deficit. Results show that increased AsA availability did not provide an advantage in terms of plant growth or symbiotic performance either under well-watered conditions or in response to drought.This work had been funded by the Spanish Ministry of Science and Innovation-European Regional Development Fund (grant RTI2018-094623-B-C22) and the Government of Navarra (project PC112-113 LEGUSI). EL was a Ramón y Cajal fellow (RYC2018-023867-I) and LC-P and MR were Formación de Personal Investigador fellows from the Spanish Ministry of Economy and Competitiveness (BES-2015-074411 and BES-2012-059972, respectively)