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This study investigates the impact of sweet potato plant sanitation on the yield and external and internal quality root storage exploring the nutritional content affected by various cooking methods (raw, boiled, and oven-cooked). The presence of viruses, and concretely of the sweet potato leaf curl virus (SPLCV), in sweet potato propagation material is shown to significantly reduce yield and modify storage root quality. Notably, the research reveals a substantial improvement in crop yield and external quality, reinforcing the efficacy of plant sanitation methods, specifically apical meristem culture, in preserving the overall productivity of sweet potato crops. Furthermore, the investigation identifies a noteworthy decrease in starch content, suggesting a dynamic interaction between plant sanitation and starch metabolism in response to viral diseases. The study also delves into the alteration of mineral absorption patterns, shedding light on how plant sanitation influences the uptake of essential minerals in sweet potato storage roots. While the health status of the plants only slightly affected magnesium (Mg) and manganese (Mn) accumulation, indicating a potential resilience of mineral balance under virus-infected conditions. Moreover, the research identifies significant modifications in antioxidant levels, emphasizing the role of plant sanitation in enhancing the nutritional quality of sweet potatoes. Heat-treated storage roots, subjected to various cooking methods such as boiling and oven-cooking, exhibit notable differences in internal quality parameters. These differences include increased concentrations of total soluble solids (SS) and heightened levels of antioxidant compounds, particularly phenolic and flavonoid compounds. The observed increase in antioxidant capacity underscores the potential health-promoting benefits associated with plant sanitation practices. Overall, the study underscores the critical importance of plant sanitation in enhancing sweet potato production sustainability, contributing to food security, and supporting local agricultural economies. The results emphasize the need for further research to optimize plant sanitation methods and promote their widespread adoption globally, providing valuable insights into the complex relationships in food quality.
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High salinity decreases the productivity of crops worldwide. Pepper is particularly sensitive to high salt concentrations. Herein, we subjected three tolerant pepper accessions (C12, B14 and A25) to high sodium chloride concentration (70 mM NaCl). The aerial and root biomass, leaf and root osmotic potential (Ψπ ), Na+ , Cl- , K+ and proline concentrations and the relative expression of the putative genes CaSOS1, CaHKT1, three CaNHXs and CaP5CS were measured. Different salinity tolerance strategies depending on the pepper accession were identified. In C12, tolerance was attributed to the accumulation of Na+ in vacuoles and endosomes by the activation of vacuolar CaNHXs genes and the reduction in Ψπ ; additionally, the activation of CaHKT1 and CaSOS1 in leaves and roots moved and accumulated Na+ ions in the xylem and xylem parenchyma cells (XPC) as well as expulsed it out of the root cells. A25 accession, on the contrary, was specialized in compartmentalizing Na+ ions in root and leaf vacuoles and root XPC by the up-regulation of CaNHXs and CaHKT1, respectively, avoiding a toxic accumulation in leaves. Finally, B14 accession moved and accumulated Na+ in xylem and XPC, reducing its concentration in roots by the activation of CaSOS1 and CaHKT1. This study shade light on different tolerance mechanisms of pepper plants to overcome salt stress.
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Salinidad , Sodio , Sodio/metabolismo , Ósmosis , Iones/metabolismo , Expresión Génica , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismoRESUMEN
Phytohormones play an important role in regulating the plant behavior to drought. In previous studies, NIBER® pepper rootstock showed tolerance to drought in terms of production and fruit quality compared to ungrafted plants. In this study, our hypothesis was that short-term exposure to water stress in young, grafted pepper plants would shed light on tolerance to drought in terms of modulation of the hormonal balance. To validate this hypothesis, fresh weight, water use efficiency (WUE) and the main hormone classes were analyzed in self-grafted pepper plants (variety onto variety, V/V) and variety grafted onto NIBER® (V/N) at 4, 24, and 48h after severe water stress was induced by PEG addition. After 48h, WUE in V/N was higher than in V/V, due to major stomata closure to maintain water retention in the leaves. This can be explained by the higher abscisic acid (ABA) levels observed in the leaves of V/N plants. Despite the interaction between ABA and the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), in relation to stomata closure is controversial, we observed an important increase of ACC at the end of the experiment in V/N plants coinciding with an important rise of the WUE and ABA. The maximum concentration of jasmonic acid and salicylic acid after 48h was found in the leaves of V/N, associated with their role in abiotic stress signaling and tolerance. Respect to auxins and cytokinins, the highest concentrations were linked to water stress and NIBER®, but this effect did not occur for gibberellins. These results show that hormone balance was affected by water stress and rootstock genotype, where NIBER® rootstock displayed a better ability to overcome short-term water stress.
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In the current climate change scenario, water stress is a serious threat to limit crop growth and yields. It is necessary to develop tolerant plants that cope with water stress and, for this purpose, tolerance mechanisms should be studied. NIBER® is a proven water stress- and salt-tolerant pepper hybrid rootstock (Gisbert-Mullor et al., 2020; López-Serrano et al., 2020), but tolerance mechanisms remain unclear. In this experiment, NIBER® and A10 (a sensitive pepper accession (Penella et al., 2014)) response to short-term water stress at 5 h and 24 h was studied in terms of gene expression and metabolites content in roots. GO terms and gene expression analyses evidenced constitutive differences in the transcriptomic profile of NIBER® and A10, associated with detoxification systems of reactive oxygen species (ROS). Upon water stress, transcription factors like DREBs and MYC are upregulated and the levels of auxins, abscisic acid and jasmonic acid are increased in NIBER®. NIBER® tolerance mechanisms involve an increase in osmoprotectant sugars (i.e., trehalose, raffinose) and in antioxidants (spermidine), but lower contents of oxidized glutathione compared to A10, which indicates less oxidative damage. Moreover, the gene expression for aquaporins and chaperones is enhanced. These results show the main NIBER® strategies to overcome water stress.
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Capsicum , Estrés Fisiológico , Estrés Fisiológico/genética , Transcriptoma , Deshidratación , Perfilación de la Expresión Génica , Capsicum/genéticaRESUMEN
Interest in the cultivation of lettuce landraces is increasing because native varieties, as high-quality products, are particularly attractive to consumers. Lettuce is a popular leafy vegetable worldwide, and interest in the consumption of first leaves (microgreens) and seedlings (baby leaves) has grown due to the general belief that young plants offer higher nutritional value. The content of some bioactive compounds and antioxidants (chlorophylls, carotenoids, anthocyanins, ascorbic acid, phenols, antioxidant activity) was monitored in six lettuce landraces and five commercial varieties, and compared across three development stages: microgreen, baby, and adult. Ascorbic acid and phenolic contents were 42% and 79% higher, respectively, in the early stages than in adult lettuces, and red-leaf varieties (CL4 and L11) stood out. This finding agrees with lettuce's marked antioxidant capacity and correlates with its pigment contents, especially anthocyanins. The nutritional value of adult lettuce is conditioned by its size, shape, and head structure as phytochemical concentrations are regulated by light. The low content of ascorbic acid, phenolics, and anthocyanins in crisphead lettuce (CL5) is a clear example (49, 67%, and 27% lower, respectively, than the adult mean). Our results indicate the wide variability of lettuces' nutritional characteristics and emphasize that traditional varieties are a helpful source of agricultural biodiversity.
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Given the little variability among commercialised eggplants mainly in developed countries, exploring, and structuring of traditional varieties germplasm collections have become a key element for extending ecotypes and promoting biodiversity preservation and consumption. Thirty-one eggplant landraces from Spain were characterised with 22 quantitative and 14 qualitative conventional morphological descriptors. Landraces were grouped based on their fruit skin colour (black-purple, striped, white, and reddish). Landraces B7, B20, and B24 were left out for their distinctive fruit characteristics. Wide variation for plant, leaf, flower, and fruit phenology traits was observed across the local landraces, and fruit descriptors were considered the most important ones. In a second experiment, landraces, B14, B16, and B17 were selected to determine fruit quality. By contemplating the benefits provided by antioxidants and sugars for human health, pulp antioxidant capacity, total phenolic, ascorbic acid, carotenoid, flavonoid, and total sugar content were determined. Significant differences were observed across these three landraces, and B14 was highlighted for its antioxidant properties, while B17 stood out for its high sugar content. B16 did not stand out for any traits. The results indicate the wide variability in eggplants for their phenotypic and nutritional characteristics, which emphasises the importance of traditional varieties as the main source of agricultural biodiversity.
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BACKGROUND: Pepper is one of the most cultivated crops worldwide, but is sensitive to salinity. This sensitivity is dependent on varieties and our knowledge about how they can face such stress is limited, mainly according to a molecular point of view. This is the main reason why we decided to develop this transcriptomic analysis. Tolerant and sensitive accessions, respectively called A25 and A6, were grown for 14 days under control conditions and irrigated with 70 mM of NaCl. Biomass, different physiological parameters and differentially expressed genes were analysed to give response to differential salinity mechanisms between both accessions. RESULTS: The genetic changes found between the accessions under both control and stress conditions could explain the physiological behaviour in A25 by the decrease of osmotic potential that could be due mainly to an increase in potassium and proline accumulation, improved growth (e.g. expansins), more efficient starch accumulation (e.g. BAM1), ion homeostasis (e.g. CBL9, HAI3, BASS1), photosynthetic protection (e.g. FIB1A, TIL, JAR1) and antioxidant activity (e.g. PSDS3, SnRK2.10). In addition, misregulation of ABA signalling (e.g. HAB1, ERD4, HAI3) and other stress signalling genes (e.g. JAR1) would appear crucial to explain the different sensitivity to NaCl in both accessions. CONCLUSIONS: After analysing the physiological behaviour and transcriptomic results, we have concluded that A25 accession utilizes different strategies to cope better salt stress, being ABA-signalling a pivotal point of regulation. However, other strategies, such as the decrease in osmotic potential to preserve water status in leaves seem to be important to explain the defence response to salinity in pepper A25 plants.
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Capsicum/fisiología , Tolerancia a la Sal/genética , Transcriptoma/fisiología , Capsicum/genéticaRESUMEN
Currently, limited water supply is a major problem in many parts of the world. Grafting peppers onto adequate rootstocks is a sustainable technique used to cope with water scarcity in plants. For 1 month, this work compared grafted peppers by employing two rootstocks (H92 and H90), with different sensitivities to water stress, and ungrafted plants in biomass, photosynthesis, and antioxidant response terms to identify physiological-antioxidant pathways of water stress tolerance. Water stress significantly stunted growth in all the plant types, although tolerant grafted plants (variety grafted onto H92, Var/H92) had higher leaf area and fresh weight values. Var/H92 showed photosynthesis and stomata conductance maintenance, compared to sensitive grafted plants (Var/H90) and ungrafted plants under water stress, linked with greater instantaneous water use efficiency. The antioxidant system was effective in removing reactive oxygen species (ROS) that could damage photosynthesis; a significant positive and negative linear correlation was observed between the rate of CO2 uptake and ascorbic acid (AsA)/total AsA (AsAt) and proline, respectively. Moreover, in Var/H92 under water stress, both higher proline and ascorbate concentration were observed. Consequently, less membrane lipid peroxidation was quantified in Var/H92.
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Sweet pepper is one of the most important economic fruits with nutritional attributes. In this sense, the nutraceutical value of consumed products is a major concern nowadays so the content of some bioactive compounds and antioxidants (phenols, ascorbic acid, lycopene, carotenoids, chlorophylls, and antioxidant activity) was monitored in 18 sweet pepper landraces at two maturity stages (green and red). All the traits except chlorophylls significantly increased in red fruits (between 1.5- and 2.3-fold for phenols, ascorbic acid, and 2-2-diphenyl-1-picrylhydrazyl (DPPH) inhibition activity, 4.8-fold for carotenoid and 27.4-fold for lycopene content), which suggests that ripening is key for obtaining desired fruit quality. Among landraces, P-44 in green fruits is highlighted for its content in carotenoids, chlorophylls, phenols, and ascorbic acid, and P-46 for its antioxidant capacity and lycopene content. Upon maturity, P-48, P-44, and P-41 presented higher levels of phenols and lycopene, and P-39 of phenols, carotenoid, and DPPH. This work reflects a wide variability in the 18 pepper landraces at bioactive compounds concentration and in relation to fruit ripeness. The importance of traditional landraces in terms of organoleptic properties is emphasized as they are the main source of agricultural biodiversity today and could be helpful for breeders to develop new functional pepper varieties.
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Antioxidantes/farmacología , Capsicum/química , Ecotipo , Fitoquímicos/farmacología , Ácido Ascórbico/análisis , Clorofila/análisis , Suplementos Dietéticos/análisis , Frutas/química , Licopeno/análisis , Fenoles/análisis , Pigmentación , Análisis de Componente PrincipalRESUMEN
Grafting is a widely utilized agronomical technique to improve yield, disease resistance, and quality of fruit and vegetables. This work aims to assess the effect of grafting and fruit ripening on the production, physico-chemical characteristics, and nutritional quality of fruit from Spanish local pepper landraces. Landraces "Cuerno," "Sueca," and "Valencia" were used as scions, and "NIBER®" as the rootstock. Two ripening stages of the fruits were sampled: green and red. Grafting improved the yield and marketable quality and did not negatively influence the physico-chemical and nutritional characteristics of the fruit. It was noteworthy that the bioactive compound contents and antioxidant capacity were more related to maturity stage and genotype, and red fruit had a higher antioxidant capacity than green fruit. However, in all the scions, grafting significantly enhanced lycopene content in both red and green fruit. Another important effect of grafting was the volatile compound composition evidenced by discriminant analyses, which was characterized for the first time in the fruit of these landraces. The rootstock and scion combination could be a way to improve not only the production, but also the fruit quality of peppers.
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In pepper crops, rootstocks that tolerate salt stress are not used because available commercial rootstocks offer limited profits. In this context, we obtained the hybrid NIBER®, a new salinity-tolerant rootstock that has been tested under real salinity field conditions for 3 years with 32%-80% higher yields than ungrafted pepper plants. This study aimed to set up the initial mechanisms involved in the salinity tolerance of grafted pepper plants using NIBER® as a rootstock to study root-shoot behavior, a basic requirement to develop efficient rootstocks. Gas exchange, Na+/K+, antioxidant capacity, nitrate reductase activity, ABA, proline, H2O2, phenols, MDA concentration and biomass were measured in ungrafted plants of cultivar Adige (A), self-grafted (A/A), grafted onto NIBER® (A/N) and reciprocal grafted plants (N/A), all exposed to 0 mM and 70 mM NaCl over a 10-day period. Salinity significantly and quickly decreased photosynthesis, stomatal conductance and nitrate reductase activity, but to lower extent in A/N plants compared to A, A/A and N/A. A/N plants showed decreases in the Na+/K+ ratio, ABA content and lipid peroxidation activity. This oxidative damage alleviation in A/N was probably due to an enhanced H2O2 level that activates antioxidant capacity to cope salinity stress, and acts as a signal molecule rather than a damaging one by contributing a major increase in phenols and, to a lesser extent, in proline concentration. These traits led to a minor impact on biomass in A/N plants under salinity conditions. Only the plants with the NIBER® rootstock controlled the scion by modulating responses to salinity.
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Raíces de Plantas , Salinidad , Estrés Fisiológico , Hibridación Genética , Raíces de Plantas/fisiología , Tolerancia a la Sal/genética , Estrés Fisiológico/genéticaRESUMEN
In vegetables, tolerance to drought can be improved by grafting commercial varieties onto drought tolerant rootstocks. Grafting has emerged as a tool that copes with drought stress. In previous results, the A25 pepper rootstock accession showed good tolerance to drought in fruit production terms compared with non-grafted plants and other rootstocks. The aim of this work was to study if short-term exposure to drought in grafted plants using A25 as a rootstock would show tolerance to drought now. To fulfill this objective, some physiological processes involved in roots (rootstock) and leaves (scion) of grafted pepper plants were analyzed. Pepper plants not grafted (A), self-grafted (A/A), and grafted onto a tolerant pepper rootstock A25 (A/A25) were grown under severe water stress induced by PEG addition (-0.55 MPa) or under control conditions for 7 days in hydroponic pure solution. According to our results, water stress severity was alleviated by using the A25 rootstock in grafted plants (A/A25), which indicated that mechanisms stimulated by roots are essential to withstand stress. A/A25 had a bigger root biomass compared with plants A and A/A that resulted in better water absorption, water retention capacity and a sustained CO2 assimilation rate. Consequently, plants A/A25 had a better carbon balance, supported by greater nitrate reductase activity located mainly in leaves. In the non-grafted and self-grafted plants, the photosynthesis rate lowered due to stomatal closure, which limited transpiration. Consequently, part of NO3 - uptake was reduced in roots. This condition limited water uptake and CO2 fixation in plants A and A/A under drought stress, and accelerated oxidative damage by producing reactive oxygen species (ROS) and H2O2, which were highest in their leaves, indicating great sensitivity to drought stress and induced membrane lipid peroxidation. However, drought deleterious effects were slightly marked in plants A compared to A/A. To conclude, the A25 rootstock protects the scion against oxidative stress, which is provoked by drought, and shows better C and N balances that enabled the biomass to be maintained under water stress for short-term exposure, with higher yields in the field.
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Exogenous application of biochemicals has been found to improve water stress tolerance in herbaceous crops but there are limited studies on deciduous fruit trees. The goal of this research was to study if ascorbic acid applications could improve physiological mechanisms associated with water stress tolerance in young fruit trees. Ascorbic acid was foliarly applied at a concentration of 250 ppm to water-stressed and well-watered peach trees (control) of two cultivars ('Scarletprince' and 'CaroTiger'). Trees received either one or two applications, and 1 week after the second application all trees were rewatered to field capacity. Upon rewatering, CO2 assimilation and stomatal conductance of water-stressed 'Scarletprince' trees sprayed with ascorbic acid (one or two applications) were similar to those of well-irrigated trees, but water-stressed trees that had not received ascorbic acid did not recover photosynthetical functions. Also, water status in sprayed water-stressed 'Scarletprince' trees was improved to values similar to control trees. On the other hand, water-stressed 'CaroTiger' trees needed two applications of ascorbic acid to reach values of CO2 assimilation similar to control trees but these applications did not improve their water status. In general terms, different response mechanisms to cope with water stress in presence of ascorbic acid were found in each cultivar, with 'Scarletprince' trees preferentially using proline as compatible solute and 'CaroTiger' trees relying on stomatal regulation. The application of ascorbic acid reduced cell membrane damage and increased catalase activity in water-stressed trees of both cultivars. These results suggest that foliar applications of ascorbic acid could be used as a management practice for improving water stress tolerance of young trees under suboptimal water regimes.
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Using chlorophyll (Chl) a fluorescence many aspects of the photosynthetic apparatus can be studied, both in vitro and, noninvasively, in vivo. Complementary techniques can help to interpret changes in the Chl a fluorescence kinetics. Kalaji et al. (Photosynth Res 122:121-158, 2014a) addressed several questions about instruments, methods and applications based on Chl a fluorescence. Here, additional Chl a fluorescence-related topics are discussed again in a question and answer format. Examples are the effect of connectivity on photochemical quenching, the correction of F V /F M values for PSI fluorescence, the energy partitioning concept, the interpretation of the complementary area, probing the donor side of PSII, the assignment of bands of 77 K fluorescence emission spectra to fluorescence emitters, the relationship between prompt and delayed fluorescence, potential problems when sampling tree canopies, the use of fluorescence parameters in QTL studies, the use of Chl a fluorescence in biosensor applications and the application of neural network approaches for the analysis of fluorescence measurements. The answers draw on knowledge from different Chl a fluorescence analysis domains, yielding in several cases new insights.
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Clorofila/química , Clorofila/metabolismo , Fluorescencia , Técnicas Biosensibles , Clorofila A , Productos Agrícolas , Complejo de Citocromo b6f/metabolismo , Citocromos b6/metabolismo , Transporte de Electrón , Herbicidas/toxicidad , Luz , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Estrés Fisiológico , Temperatura , ÁrbolesRESUMEN
The performance of a salt-tolerant pepper (Capsicum annuum L.) accession (A25) utilized as a rootstock was assessed in two experiments. In a first field experiment under natural salinity conditions, we observed a larger amount of marketable fruit (+75%) and lower Blossom-end Root incidence (-31%) in commercial pepper cultivar Adige (A) grafted onto A25 (A/A25) when compared with ungrafted plants. In order to understand this behavior a second greenhouse experiment was conducted to determine growth, mineral partitioning, gas exchange and chlorophyll a fluorescence parameters, antioxidant systems and proline content in A and A/A25 plants under salinity conditions (80 mM NaCl for 14 days). Salt stress induced significantly stunted growth of A plants (-40.6% of leaf dry weight) compared to the control conditions, while no alterations were observed in A/A25 at the end of the experiment. Accumulation of Na(+) and Cl(-) in leaves and roots was similar in either grafted or ungrafted plants. Despite the activation of protective mechanisms (increment of superoxide dismutase, catalase, ascorbate peroxidase activity and non-photochemical quenching), A plants showed severely reduced photosynthetic CO2 assimilation (-45.6% of AN390) and substantial buildup of malondialdehyde (MDA) by-product, suggesting the inability to counteract salt-triggered damage. In contrast, A/A25 plants, which had a constitutive enhanced root apparatus, were able to maintain the shoot and root growth under salinity conditions by supporting the maintained photosynthetic performance. No increases in catalase and ascorbate peroxidase activities were observed in response to salinity, and MDA levels increased only slightly; indicating that alleviation of oxidative stress did not occur in A/A25 plants. In these plants the increased proline levels could protect enzymatic stability from salt-triggered damage, preserving the photosynthetic performance. The results could indicate that salt stress was vanished by the lack of negative effects on photosynthesis that support the maintained plant growth and increased marketable yield of the grafted plants.
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Capsicum/fisiología , Fotosíntesis/fisiología , Cloruro de Sodio/farmacología , Antioxidantes/metabolismo , Capsicum/efectos de los fármacos , Catalasa/metabolismo , Clorofila/metabolismo , Clorofila A , Malondialdehído/metabolismo , Estrés Oxidativo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/fisiología , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/fisiología , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/fisiología , Transpiración de Plantas/fisiología , Prolina/metabolismo , Salinidad , Tolerancia a la Sal , Estrés Fisiológico , Superóxido Dismutasa/metabolismoRESUMEN
Grafting has been proposed as an interesting strategy that improves the responses of crops under salinity. In pepper, we reported increased fruit yield of the commercial 'Adige' cultivar under salinity when grafted onto accessions Capsicum chinense Jacq. 'ECU-973' (12) and Capsicum baccatum L. var. pendulum 'BOL-58' (14), whereas no effect was observed when grafted onto accession Capsicum annuum L var. 'Serrano' (5). We also analysed the physiological and biochemical mechanisms related to the tolerance conferred by these rootstocks. Responses to salinity (40 mM NaCl) were studied in the different plant combinations for 30 days by determining water relations, mineral content, proline accumulation, photosynthetic parameters, nitrate reductase activity and antioxidant capacity. Higher salt tolerance was achieved when the 'Adige' cultivar was grafted onto the 12 genotype, which allowed not only lower Na(+) and Cl(-) accumulation in the scion, but also ion selectivity maintenance, particularly Na(+)/K(+) discrimination. These traits led to a minor negative impact on photosynthesis, nitrate reductase activity and lipid peroxidation in grafted scion leaves. This work suggests that using tolerant pepper rootstocks that maintain the scion's ion homeostasis is a promising strategy to provide salinity tolerance and can consequently improve crop yield.
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Capsicum/fisiología , Transporte Iónico , Tolerancia a la Sal , Capsicum/crecimiento & desarrollo , Capsicum/metabolismo , Peroxidación de Lípido , Nitrato-Reductasa/metabolismo , Presión Osmótica , Fotosíntesis , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/fisiología , Prolina/metabolismo , Salinidad , Estrés FisiológicoRESUMEN
The aim of this educational review is to provide practical information on the hardware, methodology, and the hands on application of chlorophyll (Chl) a fluorescence technology. We present the paper in a question and answer format like frequently asked questions. Although nearly all information on the application of Chl a fluorescence can be found in the literature, it is not always easily accessible. This paper is primarily aimed at scientists who have some experience with the application of Chl a fluorescence but are still in the process of discovering what it all means and how it can be used. Topics discussed are (among other things) the kind of information that can be obtained using different fluorescence techniques, the interpretation of Chl a fluorescence signals, specific applications of these techniques, and practical advice on different subjects, such as on the length of dark adaptation before measurement of the Chl a fluorescence transient. The paper also provides the physiological background for some of the applied procedures. It also serves as a source of reference for experienced scientists.