Bioremediation on anthropogenic affected areas: Ectomycorrhizal and plant growth bacteria as promoters of pine establishment

The recovery of damaged areas due to inadequate farming policies and increased industrial sediment deposition, have contaminated not only soil and surrounding areas but also other natural resources. The potential use of disturbed sites for agriculture and forestry is jeopardised and their remediation is critical and expensive. The utilization of biotechnological tools, such as plant growth promoting bacteria (PGPB) and ectomycorrhizal fungi (ECM) could help remediation of such soils as they can be used as plant facilitators for land recovery. The aim of this study was to assess the potential of PGPB and ECM to enhance the growth of Pinus pinaster in antropogenic sediments and forest soil. Pine seedlings were inoculated with Suillus bovinus, Pisolithus tinctorius and Paxillus involutus, and co-inocualted with Bacillus spp. and Mesorhizobium spp. Plants were harvested after 6 month growth and parametric and nutritional data determined. Results show that P. involutus increased seedling growth(height) in industrial sediments soil, whereas in forest soil, plant performance was higher with S. bovinus. The effect of inoculation on the fungal communityin seedling roots and bacterial rhizosphere was also analysed by PCR-DGGE and differences arose between inoculated and uninoculated soil, indicating that PGPB and ECM may significantly influence the plant growth performance over a period of time. The study shows that PGPB and ECM fungi may be used as a biotechnology tool contributing to the successful plant establishment in disturbed environments

Silk fibroin-spider silk-like protein biomaterials for preventing microbial infections

Publicado em "Frontiers in Bioengineering and Biotechnology. Conference Abstract: 10th World Biomaterials Congress"Introduction: Microbial contamination of medical devices, such as sutures, are one of the major causes of hospital infections despite improvements in medical healthcare[1]. Thus, it is important to explore new biomaterials with antimicrobial properties in order to overcome microbial colonisation and biofilm formation. Spider silk has been considered an excellent biomaterial because of its toughness, strength and outstanding elasticity[2]. Also, through recombinant DNA technology, we can bioengineer and functionalize spider silk-based materials with antimicrobial peptides (AMP), thus evolving a new type of biomaterials[3]. Herein, the aim of this study was to develop silk-based fibers with antimicrobial properties by combining silk fibroin (SF) with recombinant spider silk proteins functionalized with AMP. Materials and Methods: Silk-based sutures were produced by combining spider silk chimeric proteins SSP (6mer and 6mer-HNP1) with different ratios of SF extracted from Bombyx mori through wet-spinning. The structure and topography of the sutures were characterized by scanning electron microscopy (SEM) and the tensile properties and knot strength of the sutures was assessed using an INSTRON 5540 Universal Machine. The formation of biofilm on the fibers was assessed and hemolytic effects of the materials were evaluated. Results and Discussion: Silk fibers containing spider silk 6mer or 6mer-HNP1 showed improved tensile behaviour when compared to silk fibers without SPP (Figure 1). The results suggest that the combination of spider silk chimeric proteins with SF increased the tensile stress of the fibers, when compared to those without spider silk chimeric proteins. No bacterial biofilm was observed on the fibers containing SSP functionalized with AMP, suggesting that the presence of the 6mer-HNP1 prevented the formation of biofilm. Conclusions: The outcomes suggest that silk-based fibers functionalized with AMP showed better mechanical properties when compared to silk fibers alone. The data also demonstrate the positive effect of the presence of AMP in preventing biofilm formation on the fibers, suggesting that the functionalized silk fibers could be used as a new strategy to produce sutures capable of preventing microbial proliferation, while retaining useful mechanical properties. Further investigation is still need in order to understand the in vivo performance of sutures and their physical changes during wound healing.Portuguese Foundation for Science and Technology under the scope of the project PTDC/BBB­BIO/0827/201

Management of nursery practices for efficient ectomycorrhizal fungi application in the production of Quercus ilex

The application of ectomycorrhizal (ECM) fungi on forest nursery production is regarded as part of good management practice. However, before employing large scale inoculations in a nursery the interaction between ECM symbionts, growth substrate and fertilisation input should be studied to select the most suitable nursery practices for promoting plant growth and ECM colonisation. In this study, seedlings of Quercus ilex were inoculated with Paxillus involutus, Hebeloma mesophaeum or Cenococcum geophilum and grown in three different substrates commonly used in forest nurseries: peat-based compost, forest soil or composted pine bark. The effect of various fertilisation regimes was also studied. The choice of substrate had a significant effect on plant growth and ECM colonisation. The most appropriate combination of substrate and ECM fungus for Q. ilex growth and nutrition was peat and H. mesophaeum. Plants grown on a peat-based compost and inoculated with H. mesophaeum had a significantly greater biomass and leaf phosphorus concentration without fertilisation. Composted pine bark was found not to be suitable for growth or for mycorrhization. If the appropriate growth substrate is selected, it is possible to replace the use of chemical fertilisers by inoculation with selected ECM fungi. This results in a significant increase in plant development, and thus ECM fungi can be recommended as a more environmental friendly biotechnological approach to plant management in the nursery.info:eu-repo/semantics/publishedVersio

An overview of the antimicrobial properties of lignocellulosic materials

Pathogenic microbes are a major source of health and environmental problems, mostly due to their easy proliferation on most surfaces. Currently, new classes of antimicrobial agents are under development to prevent microbial adhesion and biofilm formation. However, they are mostly from synthetic origin and present several disadvantages. The use of natural biopolymers such as cellulose, hemicellulose, and lignin, derived from lignocellulosic materials as antimicrobial agents has a promising potential. Lignocellulosic materials are one of the most abundant natural materials from renewable sources, and they present attractive characteristics, such as low density and biodegradability, are low-cost, high availability, and environmentally friendly. This review aims to provide new insights into the current usage and potential of lignocellulosic materials (biopolymer and fibers) as antimicrobial materials, highlighting their future application as a novel drug-free antimicrobial polymer.This research was partially funded through the project TERM RES Hub—Infraestrutura Científica para a Engenharia de Tecidos e Medicina Regenerativa, Refª Norte-01-0145-FEDER02219015, co-financed by the European Regional Development Fund (FEDER) through the North Regional Operational Programme (NORTE2020) and national funds, by the Portuguese Foundation for Science and Technology (FCT)

Succession dynamics of ectomycorrhizal fungi in inoculated Quercus rubra seedlings – a field study

Selected ectomycorrhizal (ECM) fungi may have a beneficial effect on the establishment of nursery grown seedlings and may influence their survival on the first years. By increasing seedling capacity to capture nutrients, ECM fungi often contribute to a successful plant establishment. We investigated the performance of pre-inoculated Quercus rubra seedlings on a reforestation site and monitored ECM fungal persistence and succession dynamics between selected ECM fungi and native fungal community. Nursery grown Q. rubra seedlings inoculated with a mixture of ECM fungi (Cenococcum geophilum, Hebeloma crustuliniforme, H. mesophaeum, H. velutipes, Paxillus involutus, Scleroderma citrinum) and non-inoculated control oak seedlings, were transplanted to Serra da Cabreira in Northern Portugal. Two years after planting, the subsistence of the inoculated ECM fungi was assessed using ITS-DGGE. Results demonstrated that inoculation with selected ECM fungi at nursery stage promoted the establishment and improved the growth performance of out planted oak seedlings. C. geophilum was significantly associated with inoculated saplings but other ECM fungi species were also found. Moreover, ECM fungal communities of inoculated and non-inoculated samples are significantly different. Further monitoring is required to increase knowledge on the persistence of ECM fungal communities and their succession dynamics

Study of symptoms and gene expression in four Pinus species after pinewood nematode infection

Pine wilt disease, caused by the pinewood nematode Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle, is originating severe infections in pine trees. The disease is detected when external symptoms appear (e.g. needle chlorosis), but trees could remain asymptomatic for long periods and serve as a long-term host. The primary goal of this study was to assess the effect of inoculation with an avirulent isolate of B. xylophilus (C14-5) on different Pinus spp. seedlings (P. sylvestris, P. nigra, P. pinea and P. pinaster). At the same time, seedlings were also inoculated with a virulent strain, HF, in order to compare the phenotypic and genomic results of the two types of inoculations. The effect of inoculation was determined in terms of expression of various Pinus genes potentially involved in the response to the disease.The results suggest that P. pinea and P. nigra are more resistant to infection by the nematode than P. sylvestris and P. pinaster. The phenotypic and genetic differences were more marked among P. pinea and P. pinaster.info:eu-repo/semantics/acceptedVersio

Reforestation of burned stands: the effect of ectomycorrhizal fungi on Pinus pinaster establishment

The area occupied by Pinus pinaster in Portugal is rapidly diminishing because of forest fires. Ectomycorrhizal fungi form obligate, mutually beneficial associations with P. pinaster which improve plant growth and resistance to adverse conditions. The aim of this work was to assess whether native ectomycorrhizal fungi could be a useful tool in the reforestation of burned areas. The work was conducted in a forest nursery greenhouse, where P. pinaster seedlings were inoculated with compatible ectomycorrhizal fungal isolates: Suillus bovinus, Pisolithus tinctorius, Rhizopogon roseolus, and a mixture of the three fungi, using burned and unburned forest soil as substrate. Inoculation significantly enhanced the growth of P. pinaster, with R. roseolus proving to be the most effective in burned soil, with an 8-fold increase in plant fresh weight. Overall, inoculation stimulated growth most in burned than in unburned soil. This study suggests that inoculation with selected ectomycorrhizal fungi in containerised nurseries can be an advantageous approach for the successful establishment of P. pinaster in burned soil. The obtained results point out to the interest of extending these studies into fire-impacted areas, using ectomycorrhizal fungi as a biological tool.info:eu-repo/semantics/acceptedVersio