Journal: Plant Physiology

Article Title: An intrinsically disordered region-containing protein mitigates the drought–growth trade-off to boost yields

doi: 10.1093/plphys/kiad074

Figure Lengend Snippet: Analysis of the relationship between SGI and CATs as well as other drought-responsive factors. A) Pull down assays of the interaction between SGI and CATs using GST-SGI recombinant protein as bait. Anti-MPB or -GST tag antibodies was employed for detecting the protein of interest. The pull down experiments were repeated 4 times with similar results. B) Similar as in A) except for MBP-CAT recombinant protein being used as bait. C) Assays of recombinant protein catalase activity of CATs in the absence or presence of SGI. All activities were calculated relative to that of SGI. Data represent means ± SD of 4 replicate experiments. D) Analysis of the accumulation of CATs in the rapeseed SGI mutant and OE lines under drought stress. Relative protein abundance of each sample is given below, and the level of proteins from each genotype without drought treatment was designated as 100%. The western blot experiments were repeated 3 times with similar results. E) Y2H assays of the interactions between SGI and drought-responsive factors. Shown above are dilution series for the co-transformed yeast vectors. Both positive and negative controls were employed during the assay. The Y2H experiments were repeated 3 times with similar results. F) Pull-down assays of the interactions between SGI and drought-responsive factors. Anti-His or -GST tag antibodies was employed for detecting the protein of interest. # denote degradation and/or unspecific bands. The pull-down experiments were repeated 3 times with similar results.

Article Snippet: Primary antibodies for CAT proteins were purchased from the PhytoAB company.

Techniques: Recombinant, Activity Assay, Mutagenesis, Quantitative Proteomics, Western Blot, Transformation Assay

Journal: Plant Physiology

Article Title: An intrinsically disordered region-containing protein mitigates the drought–growth trade-off to boost yields

doi: 10.1093/plphys/kiad074

Figure Lengend Snippet: Schematic model for how SGI protein factor alleviates drought-growth trade-off in plants. In SGI -OE lines, the induced expression of SGI in response to drought facilitates the interactions of SGI protein with dehydrins (LEA3 and LEA4-5), catalase isoforms (CAT1, CAT2, and CAT3), stress regulators (LSD1, NCA1, and VOC) and itself. It is also known that NCA1 interacts with CAT2 and that LSD1 interacts with all 3 CAT isoforms to positively regulate stress responses via guarding ROS homeostasis. It would be stimulating to test whether SGI-associated protein complexes might potentially be formed. The interactions can stabilize CAT proteins during drought stress and promote the catalase activity, to maintain basal cellular ROS levels and photosynthetic efficiency, conferring plant tolerance to drought with improved growth, and eventually increased yield. In contrast, because of the absence of these SGI-mediated interactions, SGI -CRISPR mutants overaccumulate ROS, causing plant susceptible to drought with retarded growth, and hence reduced yield. It should be noted that, based on our present results and the previous reports, no interactions have been demonstrated between LEA3 and LEA4-5 or between LEAs and any other factors depicted in the model. Likewise, no public data available indicate that VOC interacts with any of these protein factors. SGI, Stress and Growth Interconnector; CAT, catalase; LSD1, Lesion Simulating Disease1; NCA1, No Catalase Activity1; VOC, Vicinal Oxygen Chelate; LEA, Late Embryogenesis Abundant proteins; ROS, reactive oxygen species.

Article Snippet: Primary antibodies for CAT proteins were purchased from the PhytoAB company.

Techniques: Expressing, Activity Assay, CRISPR

Journal: Plants

Article Title: Modulation of Photorespiratory Enzymes by Oxidative and Photo-Oxidative Stress Induced by Menadione in Leaves of Pea ( Pisum sativum )

doi: 10.3390/plants10050987

Figure Lengend Snippet: The transcript levels of glycolate oxidase (GO) ( a ), catalase (CAT) ( b ), hydroxypyruvate reductase (HPR) ( c ), glycerate kinase (GK), ( d ), and phosphoglycolate phosphatase (PGLP) ( e ), in extracts from leaf discs on exposure to MD and/or high light (HL, 1200 µmol m −2 s −1 ). The expression of genes was represented as fold-change after normalization with actin 2. Each data point is the average of three replicates, and error bars represent ±SE. Asterisks indicate the significance of MD effect compared to the respective control. * p < 0.05; ** p < 0.001.

Article Snippet: Primary antibodies for GO, CAT, HPR, GR, and SOD (CuZn-SOD and Fe-SOD) and secondary antibody goat anti-rabbit IgG conjugated with alkaline phosphatase were from Agrisera AB (Vännäs, Sweden).

Techniques: Expressing, Control

Journal: Plants

Article Title: Modulation of Photorespiratory Enzymes by Oxidative and Photo-Oxidative Stress Induced by Menadione in Leaves of Pea ( Pisum sativum )

doi: 10.3390/plants10050987

Figure Lengend Snippet: The levels of enzyme proteins: glycolate oxidase (GO) ( a ), catalase (CAT) ( b ), and hydroxypyruvate reductase (HPR) ( c ), in extracts from leaf discs on treatment with MD either normal or high light (1200 µmol m −2 s −1 ). The ratios of each protein (such as GO, CAT, and HPR) to Rubisco large subunits (loading control) were calculated based on ImageJ software. Each data point is the average of three replicates, and error bars represent ±SE. Asterisks indicate the significance of MD effect compared to the respective control. * p < 0.05; ** p < 0.001.

Article Snippet: Primary antibodies for GO, CAT, HPR, GR, and SOD (CuZn-SOD and Fe-SOD) and secondary antibody goat anti-rabbit IgG conjugated with alkaline phosphatase were from Agrisera AB (Vännäs, Sweden).

Techniques: Control, Software