Project description:Metaproteomics can be used to study functionally active biofilm-based bacterial populations in reclaimed water distribution systems, which in turn result in bacterial regrowth that impacts the water quality. However, existing protein extraction methods have differences in their protein recovery and have not been evaluated for their efficacies in reclaimed water biofilm samples. In this study, we first evaluated six different protein extraction methods with diverse chemical and physical properties on a mixture of bacterial cell culture. Based on a weighting scores-based evaluation, the extraction protocols in order of decreasing performance are listed as B-PER > RIPA > PreOmics > SDS > AllPrep > Urea. The highest four optimal methods on cell culture were further tested against treated wastewater non-chlorinated and chlorinated effluent biofilms. In terms of protein yield, our findings showed that RIPA performed the best; however, the highest number of proteins were extracted from SDS and PreOmics. Furthermore, SDS and PreOmics worked best to rupture gram-positive and gram-negative bacterial cell walls. Considering the five evaluation factors, PreOmics obtained highest weighted score, indicating its potential effectiveness in extracting proteins from biofilms. This study provides the first insight into evaluating protein extraction methods to facilitate metaproteomics for complex reclaimed water matrices.
2023-08-04 | PXD042292 | Pride
Project description:Dynamics of microbial communities on spinach irrigated by ground water, reclaimed water and roof-harvest water
Project description:Quinoa is an Andean crop whose cultivation has been extended to many different parts of the world in the last decade. It shows a great capacity for adaptation to diverse climate conditions, including environmental stressors, and moreover, the seeds are very nutritious in part due to their high protein content which is rich in essential amino acids. They also contain good amounts of other nutrients such as unsaturated fatty acids, vitamins, or minerals and are gluten-free seeds. Also, the use of quinoa hydrolysates and peptides has been linked to numerous health benefits. Altogether, these aspects have situated quinoa as a crop able to contribute to food security worldwide. Aiming to deepen our understanding of the protein quality and function of quinoa seeds and how they can vary when this crop is subjected to water-limiting conditions, a shotgun proteomics analysis was performed to obtain the proteomes of quinoa seeds harvested from two different water regimes in field: rainfed and irrigated conditions. Also, a quantitative analysis was developed to determine the total number of proteins identified in both samples, which of them are present in both conditions, and also, which ones were present exclusively in seeds under rainfed or irrigated conditions.
Project description:A field experiment was conducted at the Federal University of Alagoas in Maceio, Brazil. varieties RB86-7515, RB85-5536 and RB92-579 were distributed in a randomized block design with four replicates in two conditions: irrigated (control) and non-irrigated (treatment).
Project description:The response to moderate and heavy drought of two Solanum tuberosum ssp. Andigena varieties, Sullu (NP 03.03) and SA 2563 (NP 03.01), planted in rain- and soil water protected fields in the Peruvian highlands are compared. Previous experiments indicate that Sullu has a greater capacity for yield maintenance under drought than SA 2563. Both clones have similar morphological properties, vegetative periods and rooting depths, so it can be assumed that the cause for increased drought tolerance of clone NP 03.03 is rather due to physiological or biochemical mechanisms, than to drought escape by deep rooting or earliness. Sullu and SA 2563 were planted in a random block design with 5 plants per bloc and 7 repetitions per treatment. Treatments: (1) drought stress, (2) irrigated control The plants were drip-irrigated in both treatments until tuberization (until day 84 after planting). Subsequently, the irrigation was stopped in the drought field, but continued in the control field. The soil moisture content in the control field was kept near field capacity. Planting date: October 05 2004 Start of drought treatment (during tuberization, 84 days after planting): December 28 2004 First sampling (soil water potential: -0.3 mPa 114 days after planting): January 27 2005 Second sampling (soil water potential –0.6 MPa, 134 days after planting): February 15 2005 Harvest: March 19 2005 (165 days after planting) The experimental design includes gene expression analysis in leaves, roots and stolons at two time points, when soil water potential reaches -0.3 and –0.6 MPa. Gene expression changes will be set in relation with physiological and agronomical data obtained in the same experiment. Keywords: Direct comparison
Project description:The response to moderate and heavy drought of two Solanum tuberosum ssp. Andigena varieties, Sullu (NP 03.03) and SA 2563 (NP 03.01), planted in rain- and soil water protected fields in the Peruvian highlands are compared. Previous experiments indicate that Sullu has a greater capacity for yield maintenance under drought than SA 2563. Both clones have similar morphological properties, vegetative periods and rooting depths, so it can be assumed that the cause for increased drought tolerance of clone NP 03.03 is rather due to physiological or biochemical mechanisms, than to drought escape by deep rooting or earliness. Sullu and SA 2563 were planted in a random block design with 5 plants per bloc and 7 repetitions per treatment. Treatments: (1) drought stress, (2) irrigated control The plants were drip-irrigated in both treatments until tuberization (until day 84 after planting). Subsequently, the irrigation was stopped in the drought field, but continued in the control field. The soil moisture content in the control field was kept near field capacity. Planting date: October 05 2004 Start of drought treatment (during tuberization, 84 days after planting): December 28 2004 First sampling (soil water potential: -0.3 mPa 114 days after planting): January 27 2005 Second sampling (soil water potential –0.6 MPa, 134 days after planting): February 15 2005 Harvest: March 19 2005 (165 days after planting) The experimental design includes gene expression analysis in leaves, roots and stolons at two time points, when soil water potential reaches -0.3 and –0.6 MPa. Gene expression changes will be set in relation with physiological and agronomical data obtained in the same experiment. Keywords: Direct comparison 19 hybs total
Project description:The aim of this study was to evaluate the physiological response and the expression analysis of `Dusa´ avocado rootstocks subjected to two different levels of water stress, and their subsequent recovery. At the beginning of the experiment, avocado plants were divided in Control plants (watered to field capacity (Fc) throughout the experiment) and stressed plants that were subjected to controlled substrate drying-up until they reached 50% of Fc (mild-WS) and 25% of Fc (severe-WS), respectively. Afterwards, plants were fully irrigated to assess drought recovery response. A set of physiological measurement were taken at leaf and whole-plant levels to assess avocado response to each level of water stress and rewatering. Root samples were collected in mild-WS and severe-WS and gene expression analysis was carried out using a targeted cDNA avocado stress microarray containing transcripts from de novo sequencing of 'Dusa' in response to biotic and abiotic stress. Avocado gene expression profiles under different levels of water stress are discussed in order to shed light on the molecular mechanisms associated with water deprivation in Dusa avocado rootstocks.
Project description:Maize transgenic event MON810, grown and commercialised worldwide, is the only cultivated GM event in EU. Maize MON810, variety DKC6575, and the corresponding near-isogenic Tietar were studied in different growing conditions, to assess their behaviour in response to drought. Profiling gene expression in water deficit regimes and in generalised water stress showed an up-regulation of different stress- responsive genes. A greater number of differentially expressed genes was observed in Tietar rather than in DKC6575, with genes belonging to transcription factor families and genes encoding HSPs, LEAs and detoxification enzymes. Since these genes have been from literature, indicated as typical of stress responses, their activation in Tietar rather than in DKC6575 may be reminiscent of a more efficient water stress response. DKC6575 was also analysed for the expression of the transgene CryIAb (encoding for the delta-endotoxin insecticidal protein) in water limiting conditions. In all the experiments the CryIAb transcript was not influenced by water stress, but expressed at a constant level. This suggests that though a different pattern of sensitivity to stress, the transgenic variety maintains the same expression level for the transgene. The Maize Oligonucleotide microarray was used to measure differences in gene expression levels under drought stress in DKC6575 and in the near-isogenic variety Tietar growing in the field. The gene expression profile of each variety was evaluated at T1 and at T2 for samples fully irrigated and at T2 for samples with irrigation deficit.