45 research outputs found
The chloroplast protein HCF164 is predicted to be associated with Coffea SH 9 resistance factor against Hemileia vastatrix
This work was funded by Portuguese national funds through FCT—Fundação para a CiĂŞncia e Tecnologia, I.P., under the Projects: UID/AGR/04129/2020 of LEAF; UIDP/04378/2020 and UIDB/04378/2020 of UCIBIO; and LA/P/0140/2020 of i4HB and FCT and FEDER funds through PORNorte under the projects: HDT-Coffee (PTDC/ASP-PLA/32429/2017) and CoffeeRES (PTDC/ASP-PLA/29779/2017). H.A. was supported by Portuguese national funds through FCT within the scope of the Stimulus of Scientific Employment—Individual Support (CEECIND/00399/2017/CP1423/CT0004). A.O. was supported at the University of Bristol by Oracle for Research and the Biological and Biotechnological Sciences Research Council ([BB/X009831/1] and [BBW003449/1]). All molecular modelling work was carried out using the computational facilities of the Advanced Computing Research Centre, University of Bristol (http://www.bris.ac.uk/acrc).To explore the connection between chloroplast and coffee resistance factors, designated as SH1 to SH9, whole genomic DNA of 42 coffee genotypes was sequenced, and entire chloroplast genomes were de novo assembled. The chloroplast phylogenetic haplotype network clustered individuals per species instead of SH factors. However, for the first time, it allowed the molecular validation of Coffea arabica as the maternal parent of the spontaneous hybrid “HĂbrido de Timor”. Individual reads were also aligned on the C. arabica reference genome to relate SH factors with chloroplast metabolism, and an in-silico analysis of selected nuclear-encoded chloroplast proteins (132 proteins) was performed. The nuclear-encoded thioredoxin-like membrane protein HCF164 enabled the discrimination of individuals with and without the SH9 factor, due to specific DNA variants linked to chromosome 7c (from C. canephora-derived sub-genome). The absence of both the thioredoxin domain and redox-active disulphide center in the HCF164 protein, observed in SH9 individuals, raises the possibility of potential implications on redox regulation. For the first time, the identification of specific DNA variants of chloroplast proteins allows discriminating individuals according to the SH profile. This study introduces an unexplored strategy for identifying protein/genes associated with SH factors and candidate targets of H. vastatrix effectors, thereby creating new perspectives for coffee breeding programs.CoffeeRES
CEECIND/00399/2017/CP1423/CT0004, PTDC/ASP-PLA/29779/2017University of Bristol by Oracle for ResearchBiotechnology and Biological Sciences Research Council
BB/X009831/1, BBW003449/1 BBSRCUniversity of BristolFundação para a Ciência e a Tecnologia
LA/P/0140/2020, UID/AGR/04129/2020, UIDB/04378/2020, UIDP/04378/2020 FCTEuropean Regional Development Fund
PTDC/ASP-PLA/32429/2017 ERD
Designing and implementing a distance education plan: a case study at a school in Portugal in pandemic context, Part I of the Research
Considering the epidemiological situation of the new coronavirus - COVID 19 in Portugal, the XXII Constitutional Government decided, in March 2020, to suspend all classroom teaching and non-teaching activities in all educational establishments in the national territory. Consequently, all Portuguese educational institutions had to adapt to the new educational reality, where technologies play a leading role in ensuring communication, interaction and collaboration between all stakeholders in the teaching-learning process. Naturally, this adaptation took time and was accompanied by several opinions and guidelines issued by the Ministry of Education and other responsible entities. In general, these guidelines aimed to ensure that all students continue to learn in a pandemic context, regardless of geographical and temporal barriers. One of the guidelines issued was the design, development and implementation of a Distance Learning Plan in each of the Portuguese schools. Thus, based on the case study methodology, we intend to demonstrate the process of designing and implementing a distance education plan in a school in Portugal. The plan was categorized into three main areas - technology, pedagogy, and content - and arise from the theoretical model Technological, Pedagogical and Content Knowledge (TPACK). This plan was developed based on the consultation of all educational agents in that school, but, in a special way, of the students and teachers who were the protagonists in the educational process. Participant observation, recorded in the researcher's diary, was also another instrument used that contributed to the design of the distance education plan. The results of the study suggest that the plan designed, despite the improvements that it naturally had to have during its implementation, was adequate for the realization of the process of teaching and learning at a distance in times of pandemic.info:eu-repo/semantics/publishedVersio
EPPN2020/FLOWERINGUNDERSTRESS – sensor and deep physiological phenotyping of terminal drought and heat-effects on chickpea flowering, seed yield and quality
Chickpeas (Cicer arietinum L.) are included in the Zero Hunger program, are the world’s
second most cultivated food legume, and have one of the best nutritional compositions among
the grain legumes. Drought typically reduces grain yield, namely when occurring during pod
set and seed filling, which is further reduced in combination with elevated temperatures. With
this EPPN2020 project, we focused the phenotyping during the critical flowering and seed filling
stages. Phenological responses to terminal drought and heat were evaluated by non-invasive
thermo-, multireflectance- and multifluorescence imaging in the Phenolab at the University of
Copenhagen (DK) to relate physiological responses at plant level with the activity signatures of
key enzymes of carbohydrate and antioxidant metabolism and its final impact on seed yield
and quality. Two chickpea genotypes with high production potential were evaluated: Elvar,
adapted to dry conditions; Electra, selected due to large seed size. The impact of high
temperature was combined with a 40% or 10% watering regime and compared with plants
kept at 40% watering regime and ambient temperature. The sensor based phenotyping in the
Phenolab was complemented with seed yield and quality analysis. Both seed weight and seed
number were negatively affected by high temperatures irrespectively of the watering regime.
Seeds from both genotypes obtained under high temperature conditions showed increased
protein content and lower starch. Protein content was further affected by the watering regime.
Alterations in protein-to-starch indicates modifications in sink-source relationships. The
differential effects were mirrored in distinct sensor based and biochemical biosignatures. This
study contributes to the mechanistic knowledge on how combined effects of severe terminal
drought and high temperature modulate sink capacity and chickpea productivity (yield and
quality) and identifies predictive biomarkers for the breeding.info:eu-repo/semantics/publishedVersio
An in‑planta comparative study of Plasmopara viticola proteome reveals diferent infection strategies towards susceptible and Rpv3‑mediated resistance hosts
Plasmopara viticola, an obligate biotrophic oomycete, is the causal agent of one of the most harmful grapevine diseases, downy mildew. Within this pathosystem, much information is gathered on the host, as characterization of pathogenicity and infection strategy of a biotrophic pathogen is quite challenging. Molecular insights into P. viticola development and pathogenicity are just beginning to be uncovered, mainly by transcriptomic studies. Plasmopara viticola proteome and secretome were only predicted based on transcriptome data. In this study, we have identified the in-planta proteome of P. viticola during infection of a susceptible ('Trincadeira') and a Rpv3-mediated resistance ('Regent') grapevine cultivar. Four hundred and twenty P. viticola proteins were identified on a label-free mass spectrometry-based approach of the apoplastic fluid of grapevine leaves. Overall, our study suggests that, in the compatible interaction, P. viticola manipulates salicylic-acid pathway and isoprenoid biosynthesis to enhance plant colonization. Furthermore, during the incompatible interaction, development-associated proteins increased while oxidoreductases protect P. viticola from ROS-associated plant defence mechanism. Up to our knowledge this is the first in-planta proteome characterization of this biotrophic pathogen, thus this study will open new insights into our understanding of this pathogen colonization strategy of both susceptible and Rpv3-mediated resistance grapevine genotypes.info:eu-repo/semantics/publishedVersio
Dietary antioxidants in coffee leaves: impact of botanical origin and maturity on chlorogenic acids and xanthones
Natural polyphenols are important dietary antioxidants that significantly benefit human
health. Coffee and tea have been shown to largely contribute to the dietary intake of these antioxidants
in several populations. More recently, the use of coffee leaves to produce tea has become a potential
commercial target, therefore prompting studies on the quantification of polyphenols in coffee leaves.
In this study a variety of coffee leaf species, at different development stages, were analyzed using
ultra-high pressure liquid chromatography. The results demonstrate that both the botanical origin of
the samples and their maturity influence significantly the concentration of the antioxidants; for total
chlorogenic acids a two-fold difference was found between different species and up to a three-fold
variation was observed between young and mature leaves. Furthermore, the range of concentrations
of chlorogenic acids in young leaves (35.7–80.8 mg/g of dry matter) were found to be comparable to
the one reported for green coffee beans. The results provide important data from which potential new
commercial products can be developedinfo:eu-repo/semantics/publishedVersio
Evaluation of virus-induced gene silencing (VIGS) in coffee plants
29th Conference of Association for the Science and Information on Coffee, 11 Sept. - 14 Sept. 2023 Hanoi, Vietnaminfo:eu-repo/semantics/publishedVersio
Flowering under stress: phenomics and proteomics approaches to chickpea grain yield and quality.
Chickpea (Cicer arietinum) is a major player in the FAO Zero Hunger program “toolkit”. It is the second most cultivated food legume, a source of sustainable protein (and other nutrients) and contributes to improved soil health and lower fertilization input. Grain yield, seed protein content and nutritional quality were found to be largely affected by Genotype x Environment. In the Mediterranean regions, higher frequency, intensity and duration of droughts, as well as hotter droughts are being registered, thus interfering with agroecosystems’ structure, composition, and functions. A controlled conditions assay (non-invasive phenotyping@PhenoLab) was undertaken in two genotypes from the Portuguese chickpea breeding program (Elvas, INIAV). Results revealed that high temperature (32º C during the reproductive phase) and water regime (40% vs 10% soil water content) play a significant role in chickpea development, seed production and composition, and protein content. As seed biochemical signatures allow to discriminate between genotypes, the power protein-based methodologies in genotype assessment are highlighted. Because some proteins can resist gastrointestinal digestion and influence human health, the processed seed proteome (i.e. following soaking, boiling and in-vitro digestion) in three consecutive years was further analysed in field-grown chickpeas (Elvas, INIAV). The seed proteome was found to be highly conserved, with minor changes being attributed to the seed development conditions. Furthermore, in vitro digestion efficiently removed many anti-nutritional proteins. Combined phenotyping and omic´s approaches contribute to the mechanistic knowledge of how severe terminal drought and high temperature modulate sink capacity and productivity (yield and quality). The integrated use of phenomics and omics methodologies has significant potential to increase our understanding of plant growth and development and, thus, an efficient, knowledge-based management of crops and resources.Work supported by EMPHASIS-GO (HORIZON-INFRA-2021-DEV-02 contract n. 101079772), by the European Plant Phenotyping Network 2020 (EXCELLENT SCIENCE - Research Infrastructures contract n. 731013) and by national funds from FCT - Fundação para a Ciência e a Tecnologia, I.P. namelly: UCIBIO – Research Unit on Applied Molecular Biosciences (FCT UIDP/04378/2020; FCT UIDB/04378/ 2020); 4HB - Associate Laboratory Institute for Health and Bioeconomy (FCT LA/P/0140/2020); LEAF – Linking Landscape, Environment, Agriculture and Food (FCT UID/AGR/04129/2020); PhD fellowship SFRH/BD/70345/2010.info:eu-repo/semantics/publishedVersio
Metabolicoffee - How primary metabolism modulates coffea spp. - Hemileia Vastatrix interactions?
Coffee leaf rust (Hemileia vastatrix) is the most important disease of Coffea arabica. This
project aims to identify coffee’s primary metabolic components essential for growth and
development that are simultaneously involved in plant defense responses. We studied the
interaction of two contrasting coffee genotypes [C. arabica S4 Agaro and Kawisari hybrid
(C. arabica Ă— C. liberica)] inoculated with different H. vastatrix races to establish compatible
(susceptible - S) and incompatible (resistant - R) interactions. Mock-inoculated leaves were
used as control (C). Leaves were collected during the infection process, and the fungal
colonization was cytologically evaluated. Inoculated and mock-inoculated coffee leaves were
prepared and, metabolites, proteins, and starch were extracted from a single sample using a
fractionated extraction method. At IPK, metabolite analysis by GC-TOF was undertaken while
using the Golm Metabolome Database (GMD). The microscopic evaluation of H. vastatrix pre-penetration stages revealed a high rate of urediniospore germination and appressoria
differentiation over the stomata which ensured the success of the infection. For both
genotypes, starch content increased along the infection process. A preliminary Principal
Component Analysis (PCA) was performed on GC-TOF data showing a clear separation of the
two coffee genotypes. In the Kawisari genotype, the PCA analysis of sugar-related features
revealed an evident split between samples C, R and S, at each time point. The role of these
metabolic features in the coffee defense responses will be discussed.info:eu-repo/semantics/publishedVersio
A chromosome-level genome resource for studying virulence mechanisms and evolution of the coffee rust pathogen Hemileia vastatrix
29th Conference of Association for the Science and Information on Coffee, 11 Sept. - 14 Sept. 2023 Hanoi, Vietnaminfo:eu-repo/semantics/publishedVersio