Comparative genomics of the nonlegume Parasponia reveals insights into evolution of nitrogen-fixing rhizobium symbioses.

Nodules harboring nitrogen-fixing rhizobia are a well-known trait of legumes, but nodules also occur in other plant lineages, with rhizobia or the actinomycete Frankia as microsymbiont. It is generally assumed that nodulation evolved independently multiple times. However, molecular-genetic support for this hypothesis is lacking, as the genetic changes underlying nodule evolution remain elusive. We conducted genetic and comparative genomics studies by using Parasponia species (Cannabaceae), the only nonlegumes that can establish nitrogen-fixing nodules with rhizobium. Intergeneric crosses between Parasponia andersonii and its nonnodulating relative Trema tomentosa demonstrated that nodule organogenesis, but not intracellular infection, is a dominant genetic trait. Comparative transcriptomics of P. andersonii and the legume Medicago truncatula revealed utilization of at least 290 orthologous symbiosis genes in nodules. Among these are key genes that, in legumes, are essential for nodulation, including NODULE INCEPTION (NIN) and RHIZOBIUM-DIRECTED POLAR GROWTH (RPG). Comparative analysis of genomes from three Parasponia species and related nonnodulating plant species show evidence of parallel loss in nonnodulating species of putative orthologs of NIN, RPG, and NOD FACTOR PERCEPTION Parallel loss of these symbiosis genes indicates that these nonnodulating lineages lost the potential to nodulate. Taken together, our results challenge the view that nodulation evolved in parallel and raises the possibility that nodulation originated ∼100 Mya in a common ancestor of all nodulating plant species, but was subsequently lost in many descendant lineages. This will have profound implications for translational approaches aimed at engineering nitrogen-fixing nodules in crop plants.

Title: Enhanced salt tolerance and photosynthetic performance: Implication of ɤ-amino butyric acid application in salt-exposed lettuce (Lactuca sativa L.) plants.

Gamma-Amino Butyric Acid (GABA) is a substantial component of the free amino acid pool with low concentration in plant tissues. Enhanced GABA content occurs during plant growth and developmental processes like seed germination. GABA level, basically, alters in response to many endogenous and exogenous stimuli. In the current study, GABA effects were studied on germination, photosynthetic performance and oxidative damages in salt-exposed lettuce plants. Three NaCl (0, 40 and 80 mM) and two GABA (0 and 25 µM) concentrations were applied on lettuce during two different developmental (seed germination and seedlings growth) stages. Negative effects of salinity on germination and plant growth were removed by GABA application. GABA significantly reduced mean germination time (MGT) in salt-exposed lettuce seeds. Although, salinity caused a significant decline in maximum quantum yield of photosystem II (Fv/Fm) during distinct steps of plant growth, GABA application improved Fv/Fm particularly on high salinity level. GABA decreased specific energy fluxes per reaction center (RC) for energy absorption and dissipation, while enhanced-electron transport flux in photosynthetic apparatus of lettuce plants was observed in GABA-supplemented plants. Moreover, decline in non-photochemical quenching (NPQ) and quenching coefficients (qP, qL, qN) by salt stress were recovered by GABA application. Elevated electrolyte leakage considerably decreased by GABA exposure on salt-treated plants. Although, proline level increased by NaCl treatments in a concentration dependent manner, combined application of salt with GABA caused a significant reduction in proline content. Catalase; EC 1.11.1.6 (CAT), l-ascorbate peroxidase; EC 1.11.1.11 (APX), and superoxide dismutase; EC 1.15.1.1 (SOD) activities were increased by GABA exposure in salt-supplemented plants that resulted in regulated hydrogen peroxide level. In conclusion, a multifaceted role for GABA is suggested for minimizing detrimental effects of salinity on lettuce through improvement of photosynthetic functionality and regulation of oxidative stress.