Chlorella vulgaris and Tetradesmus obliquus Protect Spinach (Spinacia oleracea L.) against Fusarium oxysporum.

Chlorella vulgaris and Tetradesmus obliquus were tested as biocontrol agents against the phytopathogenic fungus Fusarium oxysporum. This evaluation was conducted through in vitro and in vivo trials with spinach (Spinacia oleracea L.). The in vitro trials showed that C. vulgaris and T. obliquus were able to inhibit the phytopathogen, showing a similar inhibitory effect to that of the positive controls (Rovral, BASF® and Biocontrol T34, Biocontrol Technologies® S.L.). C. vulgaris aqueous suspensions at 3.0 g L-1 led to a hyphal growth of 0.55 cm, each corresponding to a reduction of 63% of fungal growth. With T. obliquus, the hyphal growth was 0.53 cm when applied at a concentration of 0.75 g L-1, having an inhibition of fungus growth of 64%. Thereafter, these results were validated in an in vivo trial on spinach using the same controls. The results revealed a lower severity and disease incidence and a reduction in the disease's AUDPC (area under the disease progress curve) when spinach was treated with the microalgae suspensions. Overall, these findings highlight the potential of C. vulgaris and T. obliquus suspensions as promising biocontrol agents against F. oxysporum in spinach when applied through irrigation.

Nutritional Performance of Five Citrus Rootstocks under Different Fe Levels.

Iron is an essential micronutrient for citrus, playing an important role in photosynthesis and yield. The aim of this paper was to evaluate the tolerance to Fe deficiency of five citrus rootstocks: sour orange (S), Carrizo citrange (C), Citrus macrophylla (M), Troyer citrange (T), and Volkamer lemon (V). Plants were grown for 5 weeks in nutrient solution that contained the following Fe concentrations (in µM): 0, 5, 10, 15, and 20. At the end of the experiment, biomass (dry weight-DW), leaf area, total leaf chlorophyll (CHL), and the activity of root chelate reductase (FCR) were recorded. Additionally, the mineral composition of roots (R) and shoots (S) was evaluated. Principal component analysis was used to study the relationships between all parameters and, subsequently, the relations between rootstocks. In the first component, N-S, P-S, Ca-S, Cu-S, Zn-S, Mn-S, Zn-R, and Mn-R concentrations were related to leaf CHL and FCR. Increases in leaf CHL, Mg-R, and DW (shoots and roots) were inversely related to Cu-R, which was shown in the second component. The values obtained were consistent for V10, C15, and C20, but in contrast for S0 and S5. In conclusion, micronutrient homeostasis in roots and shoots of all rootstocks were affected by Fe stress conditions. The Fe/Cu ratio was significantly related to CHL, which may be used to assist rootstock performance.

Effects of foliar application of organic acids on strawberry plants.

The large economic costs and environmental impacts of iron-chelate treatments has led to the search for alternative methods and compounds to control iron (Fe) deficiency chlorosis. Strawberry plants (Fragaria x ananassa) were grown in Hoagland's nutrient solution in a greenhouse with two levels of Fe: 0 and 10 µM Fe(III)-EDDHA. After 20 days, plants growing without Fe showed typical symptoms of Fe deficiency chlorosis in young leaves. Then, the adaxial and abaxial sides of one mature or one young leaf in each plant were brushed with 10 mM malic (MA), citric (CA) or succinic (SA) acids. Eight applications were done over a two-week period. At the end of the experiment, the newly emerged (therefore untreated), young and mature leaves were sampled for nutritional and metabolomic analysis, to assess the effectiveness of treatments. Leaf regreening was monitored using a SPAD-502 apparatus, and the activity of the ferric chelate-reductase activity (FCR) was measured using root tips. Iron deficiency negatively affected biomass and leaf chlorophyll but did not increase FCR activity. Application of succinic acid alleviated the decrease in chlorophyll observed in other treatments, and the overall nutritional balance in the plant was also changed. The concentrations of two quinic acid derivatives increased under Fe deficiency and decreased in plants treated with succinic acid, and thus they are proposed as Fe stress markers. Data suggest that foliage treatments with carboxylates may be, in some cases, environmentally friendly alternatives to Fe(III)-chelates. The importance of Fe mobilization pathways in the formulation of new fertilizers is also discussed.

Algae as Food in Europe: An Overview of Species Diversity and Their Application.

Algae have been consumed for millennia in several parts of the world as food, food supplements, and additives, due to their unique organoleptic properties and nutritional and health benefits. Algae are sustainable sources of proteins, minerals, and fiber, with well-balanced essential amino acids, pigments, and fatty acids, among other relevant metabolites for human nutrition. This review covers the historical consumption of algae in Europe, developments in the current European market, challenges when introducing new species to the market, bottlenecks in production technology, consumer acceptance, and legislation. The current algae species that are consumed and commercialized in Europe were investigated, according to their status under the European Union (EU) Novel Food legislation, along with the market perspectives in terms of the current research and development initiatives, while evaluating the interest and potential in the European market. The regular consumption of more than 150 algae species was identified, of which only 20% are approved under the EU Novel Food legislation, which demonstrates that the current legislation is not broad enough and requires an urgent update. Finally, the potential of the European algae market growth was indicated by the analysis of the trends in research, technological advances, and market initiatives to promote algae commercialization and consumption.

Nutritional and Functional Evaluation of Inula crithmoides and Mesembryanthemum nodiflorum Grown in Different Salinities for Human Consumption.

The nutritional composition and productivity of halophytes is strongly related to the biotic/abiotic stress to which these extremophile salt tolerant plants are subjected during their cultivation cycle. In this study, two commercial halophyte species (Inula crithmoides and Mesembryanthemum nodiflorum) were cultivated at six levels of salinity using a soilless cultivation system. In this way, it was possible to understand the response mechanisms of these halophytes to salt stress. The relative productivity decreased from the salinities of 110 and 200 mmol L-1 upwards for I. crithmoides and M. nodiflorum, respectively. Nonetheless, the nutritional profile for human consumption remained balanced. In general, I. crithmoides vitamin (B1 and B6) contents were significantly higher than those of M. nodiflorum. For both species, ß-carotene and lutein were induced by salinity, possibly as a response to oxidative stress. Phenolic compounds were more abundant in plants cultivated at lower salinities, while the antioxidant activity increased as a response to salt stress. Sensory characteristics were evaluated by a panel of culinary chefs showing a preference for plants grown at the salt concentration of 350 mmol L-1. In summary, salinity stress was effective in boosting important nutritional components in these species, and the soilless system promotes the sustainable and safe production of halophyte plants for human consumption.

Wild vs cultivated halophytes: Nutritional and functional differences.

Some halophyte plants are currently used in gourmet cuisine due to their unique organoleptic properties. Moreover, they exhibit excellent nutritional and functional properties, being rich in polyphenolics and vitamins. These compounds are associated to strong antioxidant activity and enhanced health benefits. This work compared the nutritional properties and antioxidant potential of three species (Mesembryanthemum nodiflorum, Suaeda maritima and Sarcocornia fruticosa) collected in saltmarshes from Portugal and Spain with those of cultivated plants. The latter were generally more succulent and had higher contents of minerals than plants obtained from the wild and contained less fibre. All species assayed are a good source of proteins, fibres and minerals. Additionally, they are good sources of carotenoids and vitamins A, C and B6 and showed good antioxidant potential particularly S. maritima. Chromatographic analysis of the phenolic profile revealed that ferulic and caffeic acids as the most relevant phenolic compounds detected in the halophytes tested.

Influence of cultivation salinity in the nutritional composition, antioxidant capacity and microbial quality of Salicornia ramosissima commercially produced in soilless systems.

The consumption of halophytes as healthy gourmet food has increased considerably in the past few years. However, knowledge on the nutritional profile of domesticated halophytes is scarce and little is known on which cultivation conditions can produce plants with the best nutritional and functional properties. In this context, Salicornia ramosissima J. Woods was cultivated in six different salt concentrations, ranging from 35 to 465 mM of NaCl. Both the nutritional profile, the antioxidant capacity, and microbial quality of the produced plants were evaluated including minerals and vitamins. Salt has a marked effect on growth, which decreases for salinities higher than 110 mM. Nonetheless, plants cultivated with intermediate levels of salinity (110 and 200 mM) revealed better antioxidant status with higher amounts of phenolic compounds. Overall, results from this paper indicated that soilless culture systems using low-intermediate salinities produces S. ramosissima plants fit for commercialization and human consumption.

Silencing of the FRO1 gene and its effects on iron partition in Nicotiana benthamiana.

To evaluate the dynamic role of the ferric-chelate reductase enzyme (FCR) and to identify possible pathways of regulation of its activity in different plant organs an investigation was conducted by virus-induced gene silencing (VIGS) using tobacco rattle virus (TRV) to silence the ferric reductase oxidase gene (FRO1) that encodes the FCR enzyme. Half of Nicotiana benthamiana plants received the VIGS vector and the rest remained as control. Four treatments were imposed: two levels of Fe in the nutrient solution (0 or 2.5 µM of Fe), each one with silenced or non-silenced (VIGS-0; VIGS-2.5) plants. Plants grown without iron (0; VIGS-0) developed typical symptoms of iron deficiency in the youngest leaves. To prove that FRO1 silencing had occurred, resupply of Fe (R) was done by adding 2.5 µM of Fe to the nutrient solution in a subset of chlorotic plants (0-R; VIGS-R). Twelve days after resupply, 0-R plants had recovered from Fe deficiency while plants containing the VIGS vector (VIGS-R) remained chlorotic and both FRO1 gene expression and FCR activity were considerably reduced, consequently preventing Fe uptake. With the VIGS technique we were able to silence the FRO1 gene in N. benthamiana and point out its importance in chlorophyll synthesis and Fe partition.

The memory of iron stress in strawberry plants.

To provide information towards optimization of strategies to treat Fe deficiency, experiments were conducted to study the responses of Fe-deficient plants to the resupply of Fe. Strawberry (Fragaria × ananassa Duch.) was used as model plant. Bare-root transplants of strawberry (cv. 'Diamante') were grown for 42 days in Hoagland's nutrient solutions without Fe (Fe0) and containing 10 µM of Fe as Fe-EDDHA (control, Fe10). For plants under Fe0 the total chlorophyll concentration of young leaves decreased progressively on time, showing the typical symptoms of iron chlorosis. After 35 days the Fe concentration was 6% of that observed for plants growing under Fe10. Half of plants growing under Fe0 were then Fe-resupplied by adding 10 µM of Fe to the Fe0 nutrient solution (FeR). Full Chlorophyll recovery of young leaves took place within 12 days. Root ferric chelate-reductase activity (FCR) and succinic and citric acid concentrations increased in FeR plants. Fe partition revealed that FeR plants expressively accumulated this nutrient in the crown and flowers. This observation can be due to a passive deactivation mechanism of the FCR activity, associated with continuous synthesis of succinic and citric acids at root level, and consequent greater uptake of Fe.

Changes in the concentration of organic acids in roots and leaves of carob-tree under Fe deficiency.

Several fruit trees are able to cope with iron (Fe) deficiency when grown in calcareous soils in the Mediterranean region, although information regarding well adapted slow-growing species is scarce, and the mechanisms activated by these species are not described in the literature. A crucial issue related to tolerance is the need to transport Fe over relatively long distances inside the plant. To evaluate the possible role of organic acids in the movement of Fe in tolerant plants, we studied the concentration of low molecular weight organic acids in several organs of 1-year old carob plants grown for 55 days in nutrient solutions without Fe (0µM Fe) or with 1µM Fe and 10µM Fe. Roots, stems and leaves were harvested, and the biomass, Fe and organic acid contents quantified. Total leaf chlorophyll (Chl) was evaluated in young leaves over the experimental period and the activity of root ferric chelate-reductase (FC-R; EC 1.16.1.17) was determined after 35 days, when deficiency symptoms appeared. Iron chlorosis was observed only at the end of the experiment in plants grown in the absence of Fe, and these plants had a smaller DW of leaves and also significant greater activity of root FC-R. Iron deficiency (Fe0 and Fe1 treatments) induced significant changes in the concentrations of succinic, malic, citric and fumaric acids, which increased in roots, or in basal, middle and apical leaves. There were significant correlations between most organic acids (with the exceptions of 2-oxoglutaric and tartaric acids) and leaf Chl. Analysis of each type of leaf showed that more succinic and malic acids were present in young chlorotic leaves while the reverse was true for quinic acid. These changes in organic acids followed a root-to-foliage pathway that was similar in all leaf types and particularly evident in young chlorotic leaves. We hypothesised that it was associated with Fe transport from roots to aboveground tissues, as there were significant differences in Fe contents between treatments with and without Fe.

Development and recovery of iron deficiency by iron resupply to roots or leaves of strawberry plants.

Bare-root transplants of strawberry (Fragaria ananassa Duch. cv. 'Selva') were transferred to nutrient solutions with or without iron (Fe). After six weeks of growth, plants grown in solution lacking Fe were chlorotic and showed morphological changes in roots typical of Fe deficiency. Subsequently, four treatments were applied for nine days: plants grown in continued absence of Fe (Fe0); plants grown in continued presence of 10 µM Fe (Fe10); foliar application of ferrous sulphate every two days to chlorotic plants (Fe-leaves); and growth of chlorotic plants in solution with ferrous sulphate (Fe-solution). After six days, the chlorophyll (Chl) content in leaves of Fe-solution plants was similar to that in Fe10 plants. Under the Fe-leaves treatment, a slight regreening of new leaves was observed only by the end of the experiment. After nine days, ferric chelate reductase (FC-R) activity was unchanged in Fe10 but increased in Fe0 plants. The FC-R activity of Fe-solution plants was similar to the initial value for chlorotic plants, whereas it was reduced drastically under the Fe-leaves treatment. The Fe concentration in leaves of Fe0 and Fe10 was similar, whereas the Fe-solution and Fe-leaves treatments enhanced leaf Fe concentration. In contrast to the Fe-solution treatment, foliar application of Fe did not increase the Fe concentration in roots. Under our experimental conditions, FC-R activity in strawberry appeared to be deactivated rapidly by pulses of Fe applied by foliar sprays. Deactivation was slower if Fe was applied directly to roots, which suggested that the plants had greater opportunity to take Fe.