A rare non-canonical splice site in Trema orientalis SYMRK does not affect its dual symbiotic functioning in endomycorrhiza and rhizobium nodulation.

BACKGROUND: Nitrogen-fixing nodules occur in ten related taxonomic lineages interspersed with lineages of non-nodulating plant species. Nodules result from an endosymbiosis between plants and diazotrophic bacteria; rhizobia in the case of legumes and Parasponia and Frankia in the case of actinorhizal species. Nodulating plants share a conserved set of symbiosis genes, whereas related non-nodulating sister species show pseudogenization of several key nodulation-specific genes. Signalling and cellular mechanisms critical for nodulation have been co-opted from the more ancient plant-fungal arbuscular endomycorrhizal symbiosis. Studies in legumes and actinorhizal plants uncovered a key component in symbiotic signalling, the LRR-type SYMBIOSIS RECEPTOR KINASE (SYMRK). SYMRK is essential for nodulation and arbuscular endomycorrhizal symbiosis. To our surprise, however, despite its arbuscular endomycorrhizal symbiosis capacities, we observed a seemingly critical mutation in a donor splice site in the SYMRK gene of Trema orientalis, the non-nodulating sister species of Parasponia. This led us to investigate the symbiotic functioning of SYMRK in the Trema-Parasponia lineage and to address the question of to what extent a single nucleotide polymorphism in a donor splice site affects the symbiotic functioning of SYMRK. RESULTS: We show that SYMRK is essential for nodulation and endomycorrhization in Parasponia andersonii. Subsequently, it is revealed that the 5'-intron donor splice site of SYMRK intron 12 is variable and, in most dicotyledon species, doesn't contain the canonical dinucleotide 'GT' signature but the much less common motif 'GC'. Strikingly, in T. orientalis, this motif is converted into a rare non-canonical 5'-intron donor splice site 'GA'. This SYMRK allele, however, is fully functional and spreads in the T. orientalis population of Malaysian Borneo. A further investigation into the occurrence of the non-canonical GA-AG splice sites confirmed that these are extremely rare. CONCLUSION: SYMRK functioning is highly conserved in legumes, actinorhizal plants, and Parasponia. The gene possesses a non-common 5'-intron GC donor splice site in intron 12, which is converted into a GA in T. orientalis accessions of Malaysian Borneo. The discovery of this functional GA-AG splice site in SYMRK highlights a gap in our understanding of splice donor sites.

Alleviation of Herbicide Toxicity in Solanum lycopersicum L.-An Antioxidant Stimulation Approach.

Application of the herbicide glyphosate in crops is a common practice among farmers around the world. Tomato is one of the crops that are treated with glyphosate to fight weed growth and loss of crop. However, tomato plants often show phytotoxic effects from glyphosate. In this study, the ability of pongamia oil derived from Pongamia pinnata (known also as Millettia pinnata) tree to alleviate the herbicide glyphosate toxicity effects in tomato (S.lycopersicum L. cv. Micro-tom) plants was tested. Tomato plants were treated with a mixture of a dose of (GLY) glyphosate (10 mg kg−1) and different doses of pongamia oil (PO) foliar spray (5, 10, 50, and 100 mM) and compared with the herbicide or oil control (glyphosate 10 mg kg−1 or pongamia oil PO 50 mM). Some morphological features, non-enzymatic and enzymatic antioxidants, and gene expression were observed. Glyphosate-treated plants sprayed with PO 50 mM (GLY + PO 50) showed increased root biomass (0.28 g-p ≤ 0.001), shoot biomass (1.2 g-p ≤ 0.01), H2O2 (68 nmol/g), and the activities of superoxide dismutase (SOD; 40 mg-p ≤ 0.001), catalase (CAT; 81.21 mg-p ≤ 0.05), ascorbate peroxidase (APX; 80 mg-p ≤ 0.01) and glutathione reductase (GR; 53 min/mg-F4,20 = 15.88, p ≤ 0.05). In contrast, these plants showed reduced contents of Malondialdehyde (MDA; 30 nmol/g-F4,20 = 18.55, p ≤ 0.01), O2 (0.6 Abs/g), Prolne (Pro; 345 µg/g), Glutathine (GSH; 341 nmol/mg-p ≤ 0.001), ascorbate (AsA; 1.8 µmol/gm), ascorbic acid (AA; 1.62 mg-p ≤ 0.05) and dehydroascorbate (DHAR; 0.32 mg p ≤ 0.05). The gene expression analysis was conducted for seven oxidative stress related genes besides the house-keeping gene Actin as a reference. The gene CYP1A1450 showed the highest mRNA expression level (6.8 fold ± 0.4) in GLY-treated tomato plants, whereas GLY-treated plants + PO 50 showed 2.9 fold. The study concluded that foliar spray of 50 mM pongamia oil alleviated the toxic effects of glyphosate on tomato plants in the form of increased root and shoot biomass, SOD, CAT, APX, and GR activity, while reduced MDA, O2, Pro, GSH, AsA, AA, DHAR, and gene CYP1A1450 expression.