Recombinant Protein L

Saline conditions can significantly affect plant growth and development, leading to massive reduction in crop yield. Herein, we show that a protein repairing enzyme PROTEIN L-ISOASPARTYL METHYLTRANSFERASE imparts salinity stress tolerance in Arabidopsis thaliana by repairing deleterious isoAsp accumulation during salinity stress. We demonstrate that salinity stress accelerates isoAsp accumulation in proteins and also induces PIMT activity in Arabidopsis. Transcript analysis indicates that both PIMT1 and PIMT2 are upregulated in response to salinity stress. Subsequent functional analysis reveals that PIMT1 and PIMT2 overexpression lines are tolerant, while RNAi lines are hyper sensitive to salinity stress in comparison to wild type (WT). Biochemical analyses of thesePIMT transgenic lines also reveals that compromised salinity tolerance of RNAi lines are linked to increased isoAsp accumulation, while improved tolerance of overexpression lines is associated with reduced isoAsp accumulation in proteins. Histochemical and biochemical studies further confirm lower accumulation of ROS and reduced lipid peroxidation in PIMT overexpression lines, while increased ROS accumulation and increased lipid peroxidation in RNAi lines as compared to WT under salinity stress. Interestingly, PIMToverexpression lines exhibit improved antioxidant enzyme efficiency, while RNAi lines display compromised antioxidant enzyme efficacy as compared to WT type plants. Our study suggests that PIMT improves salinity stress tolerance by restricting salt induced-excess ROS accumulation possibly by repairing isoAsp mediated protein damage of antioxidant enzymes. Our study can be utilized for enhancing salinity stress tolerance of economically important crops.

To facilitate selective breeding of polyphenol-rich peanuts, we looked for mass spectrometry-based proteomic evidence, investigating a subset of recombinant inbred lines (RILs) developed by the Australian peanut breeding program. To do this, we used label-free shotgun proteomics for protein and peptide quantitation, statistically analyzed normalized spectral abundance factors using R-package, as well as assayed important antioxidants. Results revealed statistically significant protein expression changes in 82 proteins classified between high or low polyphenols expressing RILs. Metabolic changes in polyphenol-rich RIL p27-362 point towards increased en-zymatic breakdown of sugars and phenylalanine biosynthesis. The study revealed phenylpropanoid pathway overexpression resulting in increased polyphenols biosynthesis. Overexpression of antioxidant enzymes such as catalase, by 73.4 fold was also observed. A strong metabolic correlation exists with the observed phenotypic traits. Peanut RIL p27-362 presents a superior nutritional composition with antioxidant-rich peanut phenotype and could yield commercial profits. Data are available via ProteomeXchange with identifier PXD015493.