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Soil salinity is one of the major threats that pose challenges to global cereal productivity and food security. Cereals have evolved sophisticated mechanisms to circumvent stress at morpho-physiological, biochemical, and molecular levels. Salt stress cues are perceived by the roots, which trigger the underlying signaling pathways that involve phytohormones. Each phytohormone triggers a specific signaling pathway integrated in a complex manner to produce antagonistic, synergistic, and additive responses. Phytohormones induce salt-responsive signaling pathways to modulate various physiological and anatomical mechanisms, including cell wall repair, apoplastic pH regulation, ion homeostasis, root hair formation, chlorophyll content, and leaf morphology. Exogenous applications of phytohormones moderate the adverse effects of salinity and improve growth. Understanding the complex hormonal crosstalk in cereals under salt stress will advance the knowledge about cooperation or antagonistic mechanisms among hormones and their role in developing salt-tolerant cereals to enhance the productivity of saline agricultural land. In this context, the present review focuses on the mechanisms of hormonal crosstalk that mediate the salt stress response and adaptation in graminaceous crops.

Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) is a deadly infection, and increasing resistance worsens an already bad scenario. In this work, a new nanomedicine antibacterial agent, based on dihydroartemisinin (DHA) and chitosan (CS), has been successfully developed to overcome MTB’s drug-resistant. To enhance DHA’s solubility, we have prepared nanoparticles of DHA loaded CS by an ionic crosslinking method with sodium tripolyphosphate (STPP) as the crosslinking agent. The DHA-CS nanoparticles (DHA-CS NPs) have been fully characterized by scanning electron microscopy, Fourier transforms infrared spectroscopy, dynamic light scattering, and ultraviolet spectrophotometry. DHA-CS NPs show an excellent antibacterial effect on the rifampicin (RFP)-resistant strain (ATCC 35838) and, at a concentration of 8.0 μg/ml, the antibacterial impact reaches up to 61.0 ± 2.13% (n = 3). The results of Gram staining, acid-fast staining, auramine “O” staining and electron microscopy show that the cell wall of RFP-resistant strains is destroyed by DHA-CS NPs (n = 3), and it is further verified by gas chromatography-mass spectrometry. Since all the metabolites identified in DHA-CS NPs treated RFP-resistant strains indicate an increase in fatty acid synthesis and cell wall repair, it can be concluded that DHA-CS NPs act by disrupting the cell wall. In addition, the resistance of 12 strains is effectively reduced by 8.0 μg/ml DHA-CS NPs combined with RFP, with an effective rate of 66.0%. The obtained results indicate that DHA-CS NPs combined with RFP may have potential use for TB treatment.

Актуальность исследования и наличие пробелов в существующем знании на тему. Мониторинговые исследования биологической безопасности пищевого сырья по микробиологическим показателям – актуальная проблема, в связи с увеличением числа пищевых инфекций во всем мире. Цель работы – изучить особенности формирования биопленок и некультивируемых микроорганизмов при различных условиях культивирования Методы. Морфометрические и денситометрические показатели биопленок и некультивируемых микроорганизмов исследовали при различных условиях культивирования. Для изучения роста и развития популяций микроорганизмов использовали среды, содержащие ростовые факторы для репарации клеточной стенки и реверсии L-форм микроорганизмов. Результаты и их обсуждение. При микробиологическом контроле критических точек технологии животноводства и пищевых производств изучены видовой состав и этиологическая значимость факторов вирулентности штаммов, продуцирующих адгезивные антигены, бактериоцины, гемолизины, токсины, β-лактамазы расширенного спектра, обусловливающие тенденцию роста множественной лекарственной устойчивости. Изучены морфофункциональные признаки биопленок, представляющих собой сообщества микроорганизмов, секретирующих полимерный матрикс и адгезированных к тканям восприимчивых видов животных и абиотическим поверхностям животноводческих помещений и пищевых производств. Установлены прямые коррелятивные зависимости между филаментацией, дисперсией поливидовых биопленок микроорганизмов и развитием дистрофических и некротические процессов в тканях и органах млекопитающих и птиц. Для оптимизации схемы микробиологической диагностики инфекционной патологии апробированы и подобраны эффективные способы детекции гетероморфных биоплёнок и некультивируемых микроорганизмов. Для предотвращения формирования биопленок патогенных микроорганизмов перспективными являются препараты, снижающие уровень микробиологических показателей первичной контаминации; минимизации адгезивных свойств, а также биоцидов разрушающих межклеточный матрикс. Выводы. Способность формирования биопленок, вариабельность фенотипических признаков, множественность факторов вирулентности, возникновение устойчивых форм бактерий за счет синтеза экзополисахаридов, значительно снижают эффективность противоэпизоотических и диагностических мероприятий. Разработка ускоренных методов детекции биопленок и дифференциации некультивируемых микроорганизмов позволит научно обосновать и разработать комплекс мероприятий, направленных на предупреждение заболеваний животных и получение безопасных продуктов животноводства, с целью профилактики заболеваний человека.

Methicillin-resistant Staphylococcus aureus (MRSA) is a multidrug-resistant pathogen of acute clinical importance. Combination treatment with an FtsZ inhibitor potentiates the activity of penicillin binding protein (PBP)-targeting β-lactam antibiotics against MRSA. To explore the mechanism underlying this synergistic behavior, we examined the impact of treatment with the FtsZ inhibitor TXA707 on the spatial localization of the five PBP proteins expressed in MRSA. In the absence of drug treatment, PBP1, PBP2, PBP3, and PBP4 colocalize with FtsZ at the septum, contributing to new cell wall formation. By contrast, PBP2a localizes to distinct foci along the cell periphery. Upon treatment with TXA707, septum formation becomes disrupted and FtsZ relocalizes away from mid-cell. PBP1 and PBP3 remain significantly colocalized with FtsZ, while PBP2, PBP4, and PBP2a localize away from FtsZ to specific sites along the periphery of the enlarged cells. We also examined the impact on PBP2a and PBP2 localization of treatment with β-lactam antibiotic oxacillin alone and in synergistic combination with TXA707. Significantly, PBP2a localizes to the septum in approximately 15% of the oxacillin-treated cells, a behavior that likely contributes to the β-lactam resistance of MRSA. Combination treatment with TXA707 causes both PBP2a and PBP2 to localize in malformed septal-like structures. Our collective results suggest that PBP2, PBP4, and PBP2a may function collaboratively in peripheral cell wall repair and maintenance in response to FtsZ inhibition by TXA707. Cotreatment with oxacillin, appears to reduce the availability of PBP2a to assist in this repair, thereby rendering the MRSA cells more susceptible to the β-lactam. IMPORTANCE MRSA is a multidrug-resistant bacterial pathogen of acute clinical importance, infecting many thousands of individuals globally each year. The essential cell division protein FtsZ has been identified as an appealing target for the development of new drugs to combat MRSA infections. Through synergistic actions, FtsZ-targeting agents can sensitize MRSA to antibiotics like the β-lactams that would otherwise be ineffective. This study provides key insights into the mechanism underlying this synergistic behavior as well as MRSA resistance to β-lactam drugs. The results of this work will help guide the identification and optimization of combination drug regimens that can effectively treat MRSA infections and reduce the potential for future resistance.

Salinity is among the major abiotic stresses negatively affecting the growth and productivity of crop plants. Sodium nitroprusside (SNP) -an external nitric oxide (NO) donor- has been found effective to impart salinity tolerance to plants. Soybean (Glycine max L.) is widely cultivated around the world; however, salinity stress hampers its growth and productivity. Therefore, the current study evaluated the role of SNP in improving morphological, physiological and biochemical attributes of soybean under salinity stress. Data relating to biomass, chlorophyll and malondialdehyde (MDA) contents, activities of various antioxidant enzymes, ion content and ultrastructural analysis were collected. The SNP application ameliorated the negative effects of salinity stress to significant extent by regulating antioxidant mechanism. Root and shoot length, fresh and dry weight, chlorophyll contents, activities of various antioxidant enzymes, i.e., catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) were improved with SNP application under salinity stress compared to control treatment. Similarly, plants treated with SNP observed less damage to cell organelles of roots and leaves under salinity stress. The results revealed pivotal functions of SNP in salinity tolerance of soybean, including cell wall repair, sequestration of sodium ion in the vacuole and maintenance of normal chloroplasts with no swelling of thylakoids. Minor distortions of cell membrane and large number of starch grains indicates an increase in the photosynthetic activity. Therefore, SNP can be used as a regulator to improve the salinity tolerance of soybean in salt affected soils.

International audience; Cell shape and cell-envelope integrity of bacteria are determined by the peptidoglycan cell wall. In rod-shaped Escherichia coli, two conserved sets of machinery are essential for cell-wall insertion in the cylindrical part of the cell: the Rod complex and the class-A penicillin-binding proteins (aPBPs). While the Rod complex governs rod-like cell shape, aPBP function is less well understood. aPBPs were previously hypothesized to either work in concert with the Rod complex or to independently repair cell-wall defects. First, we demonstrate through modulation of enzyme levels that aPBPs do not contribute to rod-like cell shape but are required for mechanical stability, supporting their independent activity. By combining measurements of cell-wall stiffness, cell-wall insertion, and PBP1b motion at the single-molecule level, we then present evidence that PBP1b, the major aPBP, contributes to cell-wall integrity by repairing cell wall defects.

Nanotechnology is currently undergoing rapid development partly due to the increasing use of carbon-based nanoparticles, such as Few Layer Graphene (FLG). Owing to its numerous applications, its industrial production is likely to lead to environmental release, including into aquatic ecosystems. In this study, a transcriptomic approach was used to assess the effect of FLG at low (0.1 mg L-1) and high (50 mg L-1) concentrations on the benthic freshwater diatom Nitzschia palea after 48 h of exposure. Direct contact with FLG and induced shading were distinguished to compare the transcriptomic responses. Genes that were differentially expressed after exposure compared with control conditions were identified, and their functional descriptions are discussed. A slight transcriptomic response related to cell wall repair was observed in diatoms exposed to the low FLG concentration. Exposure to the high FLG concentration induced a strong response involving 1907 transcripts. Notably, 16 transcripts involved in the chlorophyll biosynthesis process were under-expressed (log2 fold change between -3 and -1.2), suggesting a down-regulation of the photosynthetic metabolism. Diatoms exposed to the high FLG concentration over-expressed about 13 transcripts encoding for extracellular proteins that play a role in cellular adhesion, and two transcripts involved in cell wall repair were highly up-regulated. Light deprivation caused by shading induced a downregulation of genes involved in the energetic metabolism of N. palea. Overall, these results revealed that metabolic pathways impacted by FLG exposure are concentration and contact dependent. Moreover, this study suggests that a low FLG concentration, close to that in environmental conditions, will have a minor impact on diatom biofilms whereas a high FLG concentration, mimicking accidental release, might be critical for ecosystems.

Considering the disadvantageous of first line anti-tuberculosis (TB) drugs, including poor patient adherence, drug side effects, the long treatment duration and rapidly increasing microbe resistance, alternative treatment strategies are needed. Colistin sulfate (CS), a polymyxin antibiotic considered a last-resort antibiotics for treating multidrug-resistant Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter, has antimicrobial activity towards mycobacteria, and could serve as a possible anti-TB drug. Using GCxGC-TOFMS metabolomics, we compared the metabolic profiles of Mycobacterium tuberculosis (Mtb) cultured in the presence and absence of CS, to elucidate the mechanisms by which this drug may exert its antimicrobial effects. The principal component analysis of the metabolite data indicated significant variation in the underlying metabolite profiles of the groups. Those metabolites best explaining this differentiation, were acetic acid, and cell wall associated methylated and unmethylated fatty acids, and their alcohol and alkane derivatives. The elevated glucose levels, and various glyoxylate and glycerolipid metabolic intermediates, indicates an elevated flux in these metabolic pathways. Since all the metabolites identified in the colistin treated Mtb indicates an increase in fatty acid synthesis and cell wall repair, it can be concluded that CS acts by disrupting the cell wall in Mtb, confirming a similar drug action to other organisms.

Despite the widespread use of monochloramine in drinking water treatment, there is surprisingly little information about its mode of action. In this study, DNA microarrays were used to investigate the global gene expression of Escherichia coli cells exposed to sub-lethal concentrations of monochloramine, with a focus on temporal dynamics. Genes induced by monochloramine were associated with several stress response functions, including oxidative stress, DNA repair, multidrug efflux, biofilm formation, antibiotic resistance, and cell wall repair. The diversity of functional associations supports a model of monochloramine action involving multiple cellular targets including cell membranes, nucleic acids, and proteins. These data suggest that E. coli responds to monochloramine exposure by activating diverse defense responses rather than a single antioxidant system and the exposure may also induce biofilm formation. The induction of multidrug efflux pumps and specific antibiotic resistance genes further suggests that exposure to monochloramine may contribute to reduced susceptibility to some antibiotics.

Zds1/Zds2–PP2ACdc55 forms a complex with Rho1 GTPase and specifies Rho1 signaling outcome by regulating Rho1 GAPs in budding yeast., Budding yeast Rho1 guanosine triphosphatase (GTPase) plays an essential role in polarized cell growth by regulating cell wall glucan synthesis and actin organization. Upon cell wall damage, Rho1 blocks polarized cell growth and repairs the wounds by activating the cell wall integrity (CWI) Pkc1–mitogen-activated protein kinase (MAPK) pathway. A fundamental question is how active Rho1 promotes distinct signaling outputs under different conditions. Here we identified the Zds1/Zds2–protein phosphatase 2ACdc55 (PP2ACdc55) complex as a novel Rho1 effector that regulates Rho1 signaling specificity. Zds1/Zds2–PP2ACdc55 promotes polarized growth and cell wall synthesis by inhibiting Rho1 GTPase-activating protein (GAP) Lrg1 but inhibits CWI pathway by stabilizing another Rho1 GAP, Sac7, suggesting that active Rho1 is biased toward cell growth over stress response. Conversely, upon cell wall damage, Pkc1–Mpk1 activity inhibits cortical PP2ACdc55, ensuring that Rho1 preferentially activates the CWI pathway for cell wall repair. We propose that PP2ACdc55 specifies Rho1 signaling output and that reciprocal antagonism between Rho1–PP2ACdc55 and Rho1–Pkc1 explains how only one signaling pathway is robustly activated at a time.

The yeast cell wall contains beta1,3-glucanase-extractable and beta1,3-glucanase-resistant mannoproteins. The beta1,3-glucanase-extractable proteins are retained in the cell wall by attachment to a beta1,6-glucan moiety, which in its turn is linked to beta1,3-glucan (J. C. Kapteyn, R. C. Montijn, E. Vink, J. De La Cruz, A. Llobell, J. E. Douwes, H. Shimoi, P. N. Lipke, and F. M. Klis, Glycobiology 6:337-345, 1996). The beta1,3-glucanase-resistant protein fraction could be largely released by exochitinase treatment and contained the same set of beta1,6-glucosylated proteins, including Cwp1p, as the B1,3-glucanase-extractable fraction. Chitin was linked to the proteins in the beta1,3-glucanase-resistant fraction through a beta1,6-glucan moiety. In wild-type cell walls, the beta1,3-glucanase-resistant protein fraction represented only 1 to 2% of the covalently linked cell wall proteins, whereas in cell walls of fks1 and gas1 deletion strains, which contain much less beta1,3-glucan but more chitin, beta1,3-glucanase-resistant proteins represented about 40% of the total. We propose that the increased cross-linking of cell wall proteins via beta1,6-glucan to chitin represents a cell wall repair mechanism in yeast, which is activated in response to cell wall weakening.

Background Exophiala dermatitidis causes a variety of illnesses in humans which are always refractory to available treatment modalities. Hsp90 governs crucial stress responses, cell wall repair mechanisms and antifungal resistance in pathogenic fungi. Thus, targeting Hsp90 with specific inhibitors holds considerable promise as combination strategy. Objectives To investigate the antifungal effect of 17-AAG alone or combined with azoles against E. dermatitidis. Methods In vitro interactions of 17-AAG, a Hsp90 inhibitor, and azoles including itraconazole, voriconazole and posaconazole against E. dermatitidis were evaluated via broth microdilution chequerboard technique, adapted from the CLSI M38-A2 method. A total of 18 clinical strains were studied. Candida parapsilosis (ATCC22019) was included to ensure quality control. Results and conclusions 17-AAG alone exhibited minimal antifungal activity against all tested isolates. However, synergistic effects between 17-AAG and posaconazole, itraconazole or voriconazole were observed against 15 (83.3%), 12 (66.7%) and 1 (5.6%) isolates of E. dermatitidis, respectively. The effective working ranges of 17-AAG in synergistic combinations were mostly within 2-8 μg/mL. No antagonism was observed. In conclusion, harnessing fungal Hsp90 with 17-AAG might prove a potential antifungal regimen for E. dermatitidis infections. However, due to the host toxicity of 17-AAG, more efforts are needed to develop fungal specific Hsp90 inhibitors.

Storage roots of cassava (Manihot esculenta Crantz) exhibit a rapid post-harvest physiological deterioration (PPD) response that can occur within 24–72 h of harvest. PPD is an enzymatically mediated oxidative process with parallels to plant wound, senescence and defence responses. To characterise those genes that show significant change in expression during the PPD response we have used cDNA microarray technology to carry out a large-scale analysis of the cassava root transcriptome during the post-harvest period. We identified 72 non-redundant expressed sequence tags which showed altered regulation during the post-harvest period. Of these 63 were induced, whilst 9 were down-regulated. RNA blot analysis of selected genes was used to verify and extend the microarray data. Additional microarray hybridisation experiments allowed the identification of 21 root-specific and 24 root-wounding-specific sequences. Many of the up-regulated and PPD-specific expressed sequence tags were predicted to play a role in cellular processes including reactive oxygen species turnover, cell wall repair, programmed cell death, ion, water or metabolite transport, signal transduction or perception, stress response, metabolism and biosynthesis, and activation of protein synthesis.

Our knowledge of the yeast cell wall has increased rapidly in the past few years, allowing for the first time a description of its structure in molecular terms. Two types of cell wall proteins (CWPs) have been identified that are covalently linked to beta-glucan, namely GPI-CWPs and Pir-CWPs. Both define a characteristic supramolecular complex or structural unit. The GPI building block has the core structure GPI-CWP-->beta1,6-glucan-->beta1,3-glucan, which may become extended with one or more chitin chains. The Pir building block is less well characterized, but preliminary evidence points to the structure, Pir-CWP-->beta1,3-glucan, which probably also may become extended with one or more chitin chains. The molecular architecture of the cell wall is not fixed. The cell can make considerable adjustments to the composition and structure of its wall, for example, during the cell cycle or in response to environmental conditions such as nutrient and oxygen availability, temperature, and pH. When the cell wall is defective, dramatic changes can occur in its molecular architecture, pointing to the existence of cell wall repair mechanisms that compensate for cell damage. Finally, evidence is emerging that at least to a considerable extent the cell wall of Saccharomyces cerevisiae is representative for the cell wall of the Ascomycetes.