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Plant-made biopharmaceuticals are biotechnological valuable tools in aquaculture. Herein, the immunogenic potential of an oral corn smut (‘huitlacoche’)-made antigen (CS) was assessed in Pacific red snapper. The B subunit of the cholera toxin (CTB) was used as an immunostimulant antigen model due to its well-proven mucosal immunostimulatory activity. Test fish received corn smut-based CTB (CS-CTB) or wild-type (CS-WT) as control on day 1 and 7. Changes in humoral and cellular innate immune parameters were evaluated at 7 and 14 dpi (days post-first immunostimulation). Interestingly, total IgM and anti-CTB IgM levels in skin mucus increased in all the assessed time points in the group treated with CS-CTB. Peroxidase activity significantly increased in both serum and skin mucus from immunostimulated fish. The SOD and CAT activities were significantly increased in mucus from stimulated fish with the CS-CTB. The test fish that received CS-CTB showed enhanced phagocytic ability and radical production in head-kidney leukocytes compared with the control group. In addition, oral administration of CS-WT significantly increased the expression levels of pro-inflammatory cytokines. These findings support the use of the corn smut as a biofactory and oral delivery vehicle of novel immunostimulants, such as CTB, for fish aquaculture.

Plants have developed mechanisms that allow them to tolerate different abiotic stresses. Among these mechanisms, the accumulation of specific proteins such as dehydrins (DHNs) and aquaporins (AQPs) can protect other proteins from damage during dehydration and may allow the control of water loss, respectively. Although both types of proteins are involved in plant protection against dehydration stress, a direct interaction between them has not been explored. A previous screen to identify potential OpsDHN1 protein interactions revealed an aquaporin as a possible candidate. Here, we used the Bimolecular Fluorescence Complementation (BiFC) approach to investigate the direct interaction of the cactus OpsDHN1 protein with the Arabidopsis plasma membrane PIP family aquaporin AtPIP2B (At2G37170). Since AtPIP2B is a membrane protein and OpsDHN1 is a cytosolic protein that may be peripherally associated with membranes, we propose that OpsDHN1/AtPIP2B interaction takes place at cellular membranes. Furthermore, we also demonstrate the interaction of AtPIP2B with the three Arabidopsis dehydrins COR47 (AT1G20440), ERD10 (At1g20450), and RAB18 (At5g66400).

Ustilago maydis is a dimorphic fungus that has emerged as a model organism for the study of fungal phytopathogenicity and RNA biology. In a previous study, we isolated the U. maydis UmRrm75 gene. The deletion of the UmRrm75 gene affected morphogenesis and pathogenicity. UmRrm75 gene encodes a protein containing three RNA recognition motifs. Here we determined that UmRrm75 has chaperone activity in Escherichia coli using the transcription anti-termination assay. Subsequently, we analyzed the growth of ΔUmRrm75 mutants at 15 °C and 37 °C, observing that mutant strains had reduced growth in comparison to parental strains. UmRrm75 gene expression was induced under these non-optimal temperatures. ΔUmRrm75 mutant colonies displayed a dark-brown color at 28 °C, which was confirmed to be melanin based on spectroscopic analysis and spectrometric data. Furthermore, ΔUmRrm75 mutant strains showed the presence of peroxisomes, and increased H2O2 levels, even at 28 °C. The ΔUmRrm75 mutant strains displayed a higher expression of redox-sensor UmYap1 gene and increased catalase activity than the parental strains. Our data show that deletion of the UmRrm75 gene results in higher levels of H2O2, increased melanin content, and abiotic stress sensitivity.

Ustilago maydis is a biotrophic plant pathogenic fungus that leads to tumor development in the aerial tissues of its host, Zea mays. These tumors are the result of cell hypertrophy and hyperplasia, and are accompanied by the reprograming of primary and secondary metabolism of infected plants. Up to now, little is known regarding key plant actors and their role in tumor development during the interaction with U. maydis. Polyamines are small aliphatic amines that regulate plant growth, development and stress responses. In a previous study, we found substantial increases of polyamine levels in tumors. In the present work, we describe the maize polyamine oxidase (PAO) gene family, its contribution to hydrogen peroxide (H2O2) production and its possible role in tumor development induced by U. maydis. Histochemical analysis revealed that chlorotic lesions and maize tumors induced by U. maydis accumulate H2O2 to significant levels. Maize plants inoculated with U. maydis and treated with the PAO inhibitor 1,8-diaminooctane exhibit a notable reduction of H2O2 accumulation in infected tissues and a significant drop in PAO activity. This treatment also reduced disease symptoms in infected plants. Finally, among six maize PAO genes only the ZmPAO1, which encodes an extracellular enzyme, is up-regulated in tumors. Our data suggest that H2O2 produced through PA catabolism by ZmPAO1 plays an important role in tumor development during the maize-U. maydis interaction. Fil: Jasso Robles, Francisco Ignacio. Universidad Autónoma de San Luis Potosí. Facultad de Ciencias; México Fil: Jiménez Bremont, Juan Francisco. Instituto Potosino de Investigación Científica y Tecnológica; México Fil: Becerra Flora, Alicia. Instituto Potosino de Investigación Científica y Tecnológica; México Fil: Juárez Montiel, Margarita. Instituto Potosino de Investigación Científica y Tecnológica; México Fil: Gonzalez, Maria Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Pieckenstain, Fernando Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: García de la Cruz, Ramón Fernando. Universidad Autónoma de San Luis Potosí. Facultad de Ciencias Químicas; México Fil: Rodríguez Kessler, Margarita. Universidad Autónoma de San Luis Potosí. Facultad de Ciencias; México

Arabidopsis amiR:ADC-L2 is a non-lethal line with several developmental defects, it is characterized by a drastic reduction in free polyamine content. Herein, we found that catalase application had growth-promoting effects in amiR:ADC-L2 and parental Ws seedlings. Differences in ROS content between amiR:ADC-L2 and Ws seedlings were detected. Increased H2O2 levels were found in the amiR:ADC-L2, as well as low AtCAT2 gene expression and reduced catalase activity. Estimation of polyamine oxidase activity in amiR:ADC-L2 line indicated that the over-accumulation of H2O2 is independent of polyamine catabolism. However, increments in NADPH oxidase activity and O2•− content could be associated to the higher H2O2 levels in the amiR:ADC-L2 line. Our data suggest that low polyamine levels in Arabidopsis seedlings are responsible for the accumulation of ROS, by altering the activities of enzymes involved in ROS production and detoxification.

Dehydrins (DHNs) are intrinsically disordered proteins expressed under cellular dehydration-related stresses. In this study, we identified potential proteolytic PEST sequences located at the central and C-terminal regions from the Opuntia streptacantha OpsDHN1 protein. In order to evaluate these PEST sequences as proteolytic tags, we generated a translational fusion with the GUS reporter protein and OpsDHN1 coding sequence. We found a GUS degradation effect in tobacco agro-infiltrated leaves and Arabidopsis transgenic lines that expressed the fusion GUS::OpsDHN1 full-length. Also, two additional translational fusions between OpsDHN1 protein fragments that include the central (GUS::PEST-1) or the C-terminal (GUS::PEST-2) PEST sequences were able to decrease the GUS activity, with PEST-2 showing the greatest reduction in GUS activity. GUS signal was abated when the OpsDHN1 fragment that includes both PEST sequences (GUS::PEST-1-2) were fused to GUS. Treatment with the MG132 proteasome inhibitor attenuated the PEST-mediated GUS degradation. Point mutations of phosphorylatable residues in PEST sequences reestablished GUS signal, hence these sequences are important during protein degradation. Finally, in silico analysis identified potential PEST sequences in other plant DHNs. This is the first study reporting presence of PEST motifs in dehydrins.

Proteins with glycine-rich repeats have been identified in plants, mammalians, fungi, and bacteria. Plant glycine-rich proteins have been associated to stress response. Previously, we reported that the Arabidopsis thaliana AtGRDP2 gene, which encodes a protein with a glycine-rich domain, plays a role in growth and development of A. thaliana and Lactuca sativa. In this study, we generated composite Phaseolus vulgaris plants that overexpress the AtGRDP2 gene in hairy roots generated by Agrobacterium rhizogenes. We observed that hairy roots harboring the AtGRDP2 gene developed more abundant and faster-growing roots than control hairy roots generated with the wild type A. rhizogenes. In addition, composite common bean plants overexpressing the AtGRDP2 gene in roots were more tolerant to salt stress showing increments in their fresh and dry weight. Our data further support the role of plant GRDP genes in development and stress response.