Biofortificação de pera Rocha em cálcio: itinerário técnico para a produção e caracterização química

A biofortificação visa o aumento dos teores de um dado nutriente-alvo nas partes edíveis das plantas, para colmatar défices nutricionais na dieta do Homem que podem resultar no desenvolvimento de patologias. Por sua vez, a pêra Rocha destaca-se como um produto tipicamente português cuja produção média anual atinge as 173000 ton. Neste enquadramento, o objetivo deste estudo consistiu no desenvolvimento de um itinerário técnico de biofortificação agronómica com recurso a aplicações foliares, visando a obtenção de pêra Rocha biofortificada em cálcio, cujo processamento, possibilitará a obtenção de alimentos funcionais. O ensaio decorreu no campo experimental da HBio Lda. (coordenadas GPS 39º 23’ 29,675’’N; 9º 4’ 52,396’’O), localizado na região Oeste em Portugal. Efetuaram-se 7 aplicações foliares entre Maio e Agosto de 2018: as duas primeiras aplicações com recurso a Ca(NO3)2 e CaCl2 (nas seguintes concentrações: 0 kg ha-1, 0,1 kg ha-1, 0,3 kg ha-1, 0,6 kg ha-1 e 0 kg ha-1, 0,4 kg ha-1, 0,8 kg ha-1, 1,6 kg ha-1 respetivamente), a partir da terceira aplicação aplicou-se apenas CaCl2 em todas as árvores, primeiro com 4 kg ha-1 e as restantes quatro aplicações com 8 kg ha-1. Inicialmente, procedeu-se à caracterização do campo experimental com análises ao solo e qualidade da água. Posteriormente realizaram-se análises aos frutos e folhas, obtidos nas 5 recolhas realizadas, nomeadamente: (1) no caso das folhas: parâmetros colorimétricos e quantificação de elementos minerais por Fluorescência de Raios-X; (2) no caso dos frutos: aspetos relacionados com a sua morfometria e colorimetria, quantificação de elementos minerais, e características físicas, químicas e organolépticas da polpa de pêra de cada tratamento. Relativamente aos aspectos edafoclimáticos, apesar do ensaio experimental ter decorrido num ano de clima atípico, o solo e água do campo experimental evidenciaram-se compatíveis com a prática agrícola. A ausência de aspectos fitotóxicos nas folhas, indicam a ausência de danos resultantes da biofortificação na cultura das pereiras, não condicionado a absorção de Ca pela via foliar. Face ao itinerário de biofortificação aplicado, constataram-se teores superiores de Ca com acumulação heterogénea nas peras sujeitas a biofortificação, oscilando globalmente entre 3,6 e 14,3 %, prevalecendo no plano equatorial do fruto, teores que oscilam entre 33 e 67 %. A morfometria e colorimetria dos frutos, e por conseguinte, as propriedades físicas, químicas e organolépticas das polpas de pêra Rocha não foram afetadas pelo processo de biofortificação. O processo de biofortificação agronómica foi bem-sucedido, pois não alterou os parâmetros de qualidades da pêra Rocha, solidificando o seu potencial com o aumento efetivo dos teores de Ca no fruto

Mineral Monitorization in Different Tissues of Solanum tuberosum L. during Calcium Biofortification Process

Calcium is one of the 16 essential elements for plants, being required as Ca2+ and being involved in several fundamental processes (namely, in the stability and integrity of the cell wall, the development of plant tissue, cell division, and in stress responses). Moreover, Ca plays an important role in potato production. In this context, this study aimed to monitor the culture development (in situ and using an unmanned aerial vehicle (UAV)) and the mineral content of four essential elements (Ca, P, K, and S) in different organs of Solanum tuberosum L. (roots, stems, leaves, and tubers) during a calcium biofortification process, carried out with two types of solutions (CaCl2 and Ca- EDTA) with two concentrations (12 and 24 kg ha1). The calcium content generally increased in the S. tuberosum L. organs of both varieties and showed, after the last foliar application, an increase in Ca content that varied between 5.7–95.6% and 20.7–33%, for the Picasso and Agria varieties, respectively. The patterns of accumulation in both varieties during the biofortification process were different between the variety and mineral element. Regarding the quality analysis carried out during the development of the tubers, only the Agria variety was suitable for industrial processing after the last foliar applicationinfo:eu-repo/semantics/publishedVersio

Enrichment of Grapes with Zinc-Efficiency of Foliar Fertilization with ZnSO4 and ZnO and Implications on Winemaking

Grapes and wine are widely consumed in the world, yet their mineral content can be influenced by many factors such as the mineral composition of soils, viticulture practices and environmental conditions. In this context, considering the importance of Zn in the human physiology, the enrichment of Moscatel and Castelão grapes (white and red variety, respectively) with this nutrient prompted this study; further assessment of tissue deposition and some implications for wine production. Using two foliar fertilizers (ZnO or ZnSO4, at 150, 450 and 900 g ha1), decreases in net photosynthesis and stomatal conductance occurred in both varieties, suggesting that the physiological threshold of Zn toxicity was reached without visible symptoms. Following foliar spraying with both fertilizers, the content of Zn in leaves of the Castelão and Moscatel varieties showed higher values in all treatments relative to the control. Moreover, in grapes this tendency occurred only in Castelão. Concerning Cu, Fe, Ca, K, S and P, some significant differences also happened in leaves and grapes among treatments. At harvest, the indexes of Zn enrichment in grapes increased between 2.14- and 8.38-fold and between 1.02- and 1.44-fold in Castelão and Moscatel varieties, respectively. Zinc in the dried skin of Castelão only increased with ZnO and ZnSO4 sprayed at 900 g ha1 (ca. 2.71- and 1.5-fold relative to the control, respectively), but in Moscatel a clear accumulation trend could not be found. The dry weight of grapes ranged (in %) between 16 and 23 (but did not vary significantly among treatments of each variety or in each treatment between varieties), and total soluble solids (e.g., mainly soluble sugars and proteins) and color parameters showed some significant variations. Through winemaking, the contents of Zn increased in both varieties (1.34- and 3.57-fold, in Castelão and Moscatel, respectively) and in all treatments, although non-significantly in Castelão. It is concluded that, to increase the contents of Zn in grapes without reaching the threshold of toxicity ZnO or ZnSO4 can be used for foliar spraying of Castelão and Moscatel varieties until 900 g ha1 and that winemaking augments the level of this nutrientinfo:eu-repo/semantics/publishedVersio

Implications on Mineral Elements, Sugars and Fatty Acids Accumulation in Tissues

Funding Information: This work received funding from PDR2020-101-030734 and Fundação para a Ciência e a Tecnologia, I.P. (FCT), Portugal, through the research units UIDB/04035/2020 (GeoBioTec), UIDB/00239/2020 (CEF) and UID/FIS/04559/2020 (LIBPhys) from the FCT/MCTES/PIDDAC. This work was further supported by the grant of Fundação para a Ciência e Tecnologia (FCT) UI/BD/150718/2020. Publisher Copyright: © 2022 by the authors.Following an agronomic approach for the Ca enrichment of Rocha pears, this study aimed to assess the interactions between mineral nutrients in fruit tissues at harvest and after storage for 5 months and to characterize the implications on the profile of sugars and fatty acids (FA). A total of seven foliar sprays (with concentrations of 0.1–0.6 kg·ha−1 Ca(NO3)2 and 0.8–8 kg·ha−1 CaCl2) were applied to pear trees. After harvest, the fruits were stored for 5 months, in environmentally controlled chambers, and the mineral contents in five regions (on the equatorial section) of the fruits were assessed, while the sugar and FA content were quantified. For both dates, all foliar sprayed treatments, at different extends, increased Ca content in the center and near the epidermis of Rocha pear fruits and the levels of K, Mn, Fe, Zn and Cu also varied. At harvest, the Ca treatments did not affect the levels of sucrose, glucose, fructose and sorbitol and, after storage, their concentrations remained higher in Ca-treated fruits. Additionally, the tendency of the relative proportions of FA was C18:2 > C18:1 > C16:0 > C18:3 > C18:0 > chains inferior to 16 C ( C16:0 > C18:3 > C18:0 > C18:1 > chains inferior to 16 C (<16:0). It is concluded that the heterogeneous distribution of Ca in the tissues of Rocha pear fruits results from its absorption in the peel after Ca(NO3)2 and CaCl2 sprays and from the xylemic flux in the core prior to maturity. Additionally, the hydrolysis of complex polysaccharides affects the contents of simpler sugars during maturation, ripening and senescence, while storage decreases the amount of total fatty acids (TFA), but the double bond index (DBI) indicate that cell membrane fluidity remains unaffected.publishersversionpublishe

Mineral Monitorization in Different Tissues of Solanum tuberosum L. during Calcium Biofortification Process

Funding Information: This work received funding from PDR2020-101-030719 and the Fundação para a Ciência e a Tecnologia, I.P. (FCT), Portugal, through the research units UIDP/04035/2020 (GeoBioTec), UIDB/00239/2020 (CEF), and UID/FIS/04559/2013 (LIBPhys). This work was further supported by the grant of the Fundação para a Ciência e Tecnologia (FCT) UI/BD/150806/2020. Publisher Copyright: © 2022 by the authors.Calcium is one of the 16 essential elements for plants, being required as Ca2+ and being involved in several fundamental processes (namely, in the stability and integrity of the cell wall, the development of plant tissue, cell division, and in stress responses). Moreover, Ca plays an important role in potato production. In this context, this study aimed to monitor the culture development (in situ and using an unmanned aerial vehicle (UAV)) and the mineral content of four essential elements (Ca, P, K, and S) in different organs of Solanum tuberosum L. (roots, stems, leaves, and tubers) during a calcium biofortification process, carried out with two types of solutions (CaCl2 and Ca-EDTA) with two concentrations (12 and 24 kg∙ha−1). The calcium content generally increased in the S. tuberosum L. organs of both varieties and showed, after the last foliar application, an increase in Ca content that varied between 5.7–95.6% and 20.7–33%, for the Picasso and Agria varieties, respectively. The patterns of accumulation in both varieties during the biofortification process were different between the variety and mineral element. Regarding the quality analysis carried out during the development of the tubers, only the Agria variety was suitable for industrial processing after the last foliar application.publishersversionpublishe

Macaronesia Acts as a Museum of Genetic Diversity of Relict Ferns: The Case of Diplazium caudatum (Athyriaceae)

Macaronesia has been considered a refuge region of the formerly widespread subtropical lauroid flora that lived in Southern Europe during the Tertiary. The study of relict angiosperms has shown that Macaronesian relict taxa preserve genetic variation and revealed general patterns of colonization and dispersal. However, information on the conservation of genetic diversity and range dynamics rapidly diminishes when referring to pteridophytes, despite their dominance of the herbaceous stratum in the European tropical palaeoflora. Here we aim to elucidate the pattern of genetic diversity and phylogeography of Diplazium caudatum, a hypothesized species of the Tertiary Palaeotropical flora and currently with its populations restricted across Macaronesia and disjunctly in the Sierras de Algeciras (Andalusia, southern Iberian Peninsula). We analysed 12 populations across the species range using eight microsatellite loci, sequences of a region of plastid DNA, and carry out species-distribution modelling analyses. Our dating results confirm the Tertiary origin of this species. The Macaronesian archipelagos served as a refuge during at least the Quaternary glacial cycles, where populations of D. caudatum preserved higher levels of genetic variation than mainland populations. Our data suggest the disappearance of the species in the continent and the subsequent recolonization from Macaronesia. The results of the AMOVA analysis and the indices of clonal diversity and linkage disequilibrium suggest that D. caudatum is a species in which inter-gametophytic outcrossing predominates, and that in the Andalusian populations there was a shift in mating system toward increased inbreeding and/or clonality. The model that best explains the genetic diversity distribution pattern observed in Macaronesia is, the initial and recurrent colonization between islands and archipelagos and the relatively recent diversification of restricted area lineages, probably due to the decrease of favorable habitats and competition with lineages previously established. This study extends to ferns the concept of Macaronesia archipelagos as refugia for genetic variation

Mineral Interaction in Biofortified Tomatoes (Lycopersicum esculentum L.) with Magnesium

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Zinc Biofortification in Vitis vinifera: Implications for Quality and Wine Production

Nowadays, there is a growing concern about micronutrient deficits in food products, with agronomic biofortification being considered a mitigation strategy. In this context, as Zn is essential for growth and maintenance of human health, a workflow for the biofortification of grapes from the Vitis vinifera variety Fernão Pires, which contains this nutrient, was carried out considering the soil properties of the vineyard. Additionally, Zn accumulation in the tissues of the grapes and the implications for some quality parameters and on winemaking were assessed. Vines were sprayed three times with ZnO and ZnSO4 at concentrations of 150, 450, and 900 g ha1 during the production cycle. Physiological data were obtained through chlorophyll a fluorescence data, to access the potential symptoms of toxicity. At harvest, treated grapes revealed significant increases of Zn concentration relative to the control, being more pronounced for ZnO and ZnSO4 in the skin and seeds, respectively. After winemaking, an increase was also found regarding the control (i.e., 1.59-fold with ZnSO4-450 g ha1). The contents of the sugars and fatty acids, as well as the colorimetric analyses, were also assessed, but significant variations were not found among treatments. In general, Zn biofortification increased with ZnO and ZnSO4, without significantly affecting the physicochemical characteristics of grapesinfo:eu-repo/semantics/publishedVersio

Magnesium Accumulation in Two Contrasting Varieties of Lycopersicum esculentum L. Fruits: Interaction with Calcium at Tissue Level and Implications on Quality

As the productivity and quality of tomato fruits are responsive to Mg applications, without surpassing the threshold of toxicity, the assessment of potential levels of Mg accumulation in tissues, as well as the interactions with Ca and physicochemical properties, prompt this study. An agronomic workflow for Mg enrichment, consisting of six foliar applications of MgSO4 with four concentrations (0%, 0.25%, 1% and 4%), equivalent to 0, 43.9, 175.5 and 702 g ha-1 , was applied on two tomato (Lycopersicum esculentum L.) genotypes (Heinz1534 and Heinz9205). During fruit development, leaf gas exchange was screened, with only minor physiological deviations being found. At harvest, Mg contents among tissues and the interactions with Ca were analyzed, and it was found that in both varieties a higher Mg/Ca ratio prevailed in the most external part of the fruit sprayed with 4% MgSO4. However, Mg distribution prevailed relatively near the epidermis in H1534, while in H9205 the higher contents of this nutrient occurred in the core of the fruit, which indicated a decrease of the relative proportion of Ca. The morphologic (height and diameter), physical (dry weight and density) and colorimetric parameters, and the total soluble solids of fruits, did not reveal significant changes in both tomato varieties. It was further concluded that foliar application until 4% MgSO4 does not have physiological impacts in the fruit’s quality of both varieties, but in spite of the different patterns of Mg accumulation in tissues, if the mean value in the whole fruit is considered, this nutrient prevails in H1534. This study thus suggests that variety H1534 can be used to attain tomato fruits with added value, providing an option of further processing to achieve food products with functional properties, ultimately proving a beneficial option to producers, the food processing industry and consumers. Moreover, the study reinforces the importance of variety choice when designing enrichment workflowsinfo:eu-repo/semantics/publishedVersio

Lipophosphoglycans from \u3cem\u3eLeishmania amazonensis\u3c/em\u3e Strains Display Immunomodulatory Properties via TLR4 and Do Not Affect Sand Fly Infection

The immunomodulatory properties of lipophosphoglycans (LPG) from New World species of Leishmania have been assessed in Leishmania infantum and Leishmania braziliensis, the causative agents of visceral and cutaneous leishmaniasis, respectively. This glycoconjugate is highly polymorphic among species with variation in sugars that branch off the conserved Gal(β1,4)Man(α1)-PO4 backbone of repeat units. Here, the immunomodulatory activity of LPGs from Leishmania amazonensis, the causative agent of diffuse cutaneous leishmaniasis, was evaluated in two strains from Brazil. One strain (PH8) was originally isolated from the sand fly and the other (Josefa) was isolated from a human case. The ability of purified LPGs from both strains was investigated during in vitro interaction with peritoneal murine macrophages and CHO cells and in vivo infection with Lutzomyia migonei. In peritoneal murine macrophages, the LPGs from both strains activated TLR4. Both LPGs equally activate MAPKs and the NF-κB inhibitor p-IκBα, but were not able to translocate NF-κB. In vivo experiments with sand flies showed that both stains were able to sustain infection in L. migonei. A preliminary biochemical analysis indicates intraspecies variation in the LPG sugar moieties. However, they did not result in different activation profiles of the innate immune system. Also those polymorphisms did not affect infectivity to the sand fly