RESUMO
Faced with a global pandemic such as the one triggered by the SARS-CoV-2 virus, the medical supply chain has been highly demanded. An item in which this manifested itself more clearly, are the N95 masks, designed to be disposable items, in many cases they have had to be reused. In these emergency conditions, it was necessary to apply an effective and safe method that can be used locally. Here a device for disinfection by ultraviolet C light was developed that allows irradiating N95 masks with a known and reproducible dose. Thus being able to apply a safe and effective disinfection method according to existing information. The use of a common model of UV-C lamps and the simple construction of the device allows it to be built at low cost and with widely available materials. The effectiveness of the device was demonstrated against an enveloped RNA virus, characteristics shared with the virus that causes COVID19, being capable of reducing the viral load by 4 orders of magnitude.
RESUMO
The moss Physcomitrella patens can withstand extreme environmental conditions including drought and salt stress. Tolerance to dehydration in mosses is thought to rely on efficient limitation of stress-induced cell damage and repair of cell injury upon stress relief. Dehydrin proteins (DHNs) are part of a conserved cell protecting mechanism in plants although their role in stress tolerance is not well understood. Four DHNs and two DHN-like proteins were identified in the predicted proteome of P. patens. Expression of PpDHNA and PpDHNB was induced by salt and osmotic stress and controlled by abscisic acid. Subcellular localization of the encoded proteins suggested that these dehydrins are localized in cytosol and accumulate near membranes during stress. Comparative analysis of dhnA and dhnB targeted knockout mutants of P. patens revealed that both genes play a role in cellular protection during salt and osmotic stress, although PpDHNA has a higher contribution to stress tolerance. Overexpression of PpDHNA and PpDHNB genes in transgenic Arabidopsis improved rosette and root growth in stress conditions, although PpDHNA was more efficient in this role. These results suggest that specific DHNs contribute considerably to the high stress tolerance of mosses and offer novel tools for genetic engineering stress tolerance of higher plants.
Assuntos
Adaptação Fisiológica/genética , Bryopsida/genética , Genes de Plantas/genética , Osmose/fisiologia , Proteínas de Plantas/genética , Cloreto de Sódio/farmacologia , Estresse Fisiológico/genética , Adaptação Fisiológica/efeitos dos fármacos , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Bryopsida/efeitos dos fármacos , Bryopsida/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Técnicas de Inativação de Genes , Mutação/genética , Osmose/efeitos dos fármacos , Filogenia , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , Transporte Proteico/efeitos dos fármacos , Proteínas Recombinantes de Fusão/metabolismo , Reprodutibilidade dos Testes , Estresse Fisiológico/efeitos dos fármacos , Frações Subcelulares/efeitos dos fármacos , Frações Subcelulares/metabolismoRESUMO
BACKGROUND: Vascular plants respond to pathogens by activating a diverse array of defense mechanisms. Studies with these plants have provided a wealth of information on pathogen recognition, signal transduction and the activation of defense responses. However, very little is known about the infection and defense responses of the bryophyte, Physcomitrella patens, to well-studied phytopathogens. The purpose of this study was to determine: i) whether two representative broad host range pathogens, Erwinia carotovora ssp. carotovora (E.c. carotovora) and Botrytis cinerea (B. cinerea), could infect Physcomitrella, and ii) whether B. cinerea, elicitors of a harpin (HrpN) producing E.c. carotovora strain (SCC1) or a HrpN-negative strain (SCC3193), could cause disease symptoms and induce defense responses in Physcomitrella. RESULTS: B. cinerea and E.c. carotovora were found to readily infect Physcomitrella gametophytic tissues and cause disease symptoms. Treatments with B. cinerea spores or cell-free culture filtrates from E.c. carotovoraSCC1 (CF(SCC1)), resulted in disease development with severe maceration of Physcomitrella tissues, while CF(SCC3193) produced only mild maceration. Although increased cell death was observed with either the CFs or B. cinerea, the occurrence of cytoplasmic shrinkage was only visible in Evans blue stained protonemal cells treated with CF(SCC1) or inoculated with B. cinerea. Most cells showing cytoplasmic shrinkage accumulated autofluorescent compounds and brown chloroplasts were evident in a high proportion of these cells. CF treatments and B. cinerea inoculation induced the expression of the defense-related genes: PR-1, PAL, CHS and LOX. CONCLUSION: B. cinerea and E.c. carotovora elicitors induce a defense response in Physcomitrella, as evidenced by enhanced expression of conserved plant defense-related genes. Since cytoplasmic shrinkage is the most common morphological change observed in plant PCD, and that harpins and B. cinerea induce this type of cell death in vascular plants, our results suggest that E.c. carotovora CFSCC1 containing HrpN and B. cinerea could also induce this type of cell death in Physcomitrella. Our studies thus establish Physcomitrella as an experimental host for investigation of plant-pathogen interactions and B. cinerea and elicitors of E.c. carotovora as promising tools for understanding the mechanisms involved in defense responses and in pathogen-mediated cell death in this simple land plant.