Monitoring Plant Height and Spatial Distribution of Biometrics with a Low-Cost Proximal Platform.

Measuring canopy height is important for phenotyping as it has been identified as the most relevant parameter for the fast determination of plant mass and carbon stock, as well as crop responses and their spatial variability. In this work, we develop a low-cost tool for measuring plant height proximally based on an ultrasound sensor for flexible use in static or on-the-go mode. The tool was lab-tested and field-tested on crop systems of different geometry and spacings: in a static setting on faba bean (Vicia faba L.) and in an on-the-go setting on chia (Salvia hispanica L.), alfalfa (Medicago sativa L.), and wheat (Triticum durum Desf.). Cross-correlation (CC) or a dynamic time-warping algorithm (DTW) was used to analyze and correct shifts between manual and sensor data in chia. Sensor data were able to reproduce with minor shifts in canopy profile and plant status indicators in the field when plant heights varied gradually in narrow-spaced chia (R2 = 0.98), faba bean (R2 = 0.96), and wheat (R2 = up to 0.99). Abrupt height changes resulted in systematic errors in height estimation, and short-scale variations were not well reproduced (e.g., R2 in widely spaced chia was 0.57 to 0.66 after shifting based on CC or DTW, respectively)). In alfalfa, ultrasound data were a better predictor than NDVI (Normalized Difference Vegetation Index) for Leaf Area Index and biomass (R2 from 0.81 to 0.84). Maps of ultrasound-determined height showed that clusters were useful for spatial management. The good performance of the tool both in a static setting and in the on-the-go setting provides flexibility for the determination of plant height and spatial variation of plant responses in different conditions from natural to managed systems.

Wastewater irrigation and Trichoderma colonization in tomato plants: effects on plant traits, antioxidant activity, and performance of the insect pest Macrosiphum euphorbiae.

The scarcity of freshwater for agriculture in many regions has led to the application of sewage and saline water for irrigation. Irrigation with non-conventional water sources could become a non-harmful process for plant cultivation, and the effects of their use on crops should be monitored in order to develop optimal management strategies. One possibility to overcome potential barriers is to use biostimulants such as Trichoderma spp. fungi. Tomato is a crop of great economic importance in the world. This study investigated the joint effects of Trichoderma afroharzianum T-22 on tomato plants irrigated with simulated unconventional waters. The experiment consisted of a control and three water treatments. In the control, the plants were watered with distilled water. The three water treatments were obtained by using an irrigation water added with nitrogen, a wastewater effluent, and a mixed groundwater-wastewater effluents. Potted tomato plants (variety Bobcat) were grown in a controlled growth chamber. Antioxidant activity, susceptibility to the aphids Macrosiphum euphorbiae, and tomato plant growth parameters were estimated. Trichoderma afroharzianum T-22 had a positive effect on plant growth and antioxidant defenses when plants were irrigated with distilled water. Instead, no significant morphological effects induced by T. afroharzianum T-22 on plants were observed when unconventional water was used for irrigation. However, inoculation with T. afroharzianum T-22 activated a stress response that made the colonized plants more susceptible to aphid development and increased their fecundity and longevity. Thanks to this study, it may be possible for the first time to open a new discussion on the practical possibility of using reclaimed wastewater for crop irrigation with the addition of a growth-promoting fungal symbiont.

Enhancement of Yield, Phytochemical Content and Biological Activity of a Leafy Vegetable (Beta vulgaris L. var. cycla) by Using Organic Amendments as an Alternative to Chemical Fertilizer.

This study evaluates the effect of a chemical fertilizer (ammonium nitrate), a compost (vermicompost from cattle manure) and two biochars (from vine prunings and wood chips, respectively), applied to the soil alone or in mixture, on the yield, phytochemical content and biological activity of Beta vulgaris L. var. cycla (Swiss chard). The respective treatments, each replicated four times, were arranged according to a completely randomized block design. Results showed that vermicompost, both alone and in mixture with vine pruning biochar, significantly increased yield parameters (plant height and leaf area) and yield over the untreated soil and the biochars alone, similar to ammonium nitrate. Moreover, vermicompost, both alone and in mixture, respectively, with the two biochars, determined lower total N and NO3- contents than ammonium nitrate, both alone and in mixture, respectively, with the two biochars. In particular, NO3- content was within the safe thresholds fixed for leafy vegetables by the European Commission to prevent any adverse implication on human health from dietary NO3- exposure. The biochars alone resulted in very low yield and leaf total N content, likely due to a limited release of N for plant uptake, also evidenced by the undetectable NO3- leaf content, similarly shown by plants grown in untreated soil. Vermicompost, alone or in mixture, respectively, with the two biochars, increased the content of specialized metabolites, with a positive effect on antioxidant activity. The organic amendments, particularly compost, could be an alternative to chemical fertilizers to reach a trade-off between yield, nutritional and health qualities in Swiss chard, meeting the needs of farmers and consumers as well as the targets for sustainable food production.

A Complete Survey of Glycoalkaloids Using LC-FTICR-MS and IRMPD in a Commercial Variety and a Local Landrace of Eggplant (Solanum melongena L.) and their Anticholinesterase and Antioxidant Activities.

Eggplant contains glycoalkaloids (GAs), a class of nitrogen-containing secondary metabolites of great structural variety that may have both adverse and beneficial biological effects. In this study, we performed a complete survey of GAs and their malonylated form, in two genotypes of eggplants: A commercial cultivated type, Mirabella (Mir), with purple peel and bitter taste and a local landrace, named Melanzana Bianca di Senise (Sen), characterized by white peel with purple strip and a typical sweet aroma. Besides the analysis of their morphological traits, nineteen glycoalkaloids were tentatively identified in eggplant berry extracts based upon LC-ESI-FTICR-MS analysis using retention times, elution orders, high-resolution mass spectra, as well as high-resolution fragmentation by IRMPD. The relative signal intensities (i.e., ion counts) of the GAs identified in Mir and Sen pulp extracts showed as solamargine, and its isomers are the most abundant. In addition, anticholinesterase and antioxidant activities of the extracts were evaluated. Pulp tissue was found to be more active in inhibiting acetylcholinesterase enzyme than peel showing an inhibitory effect higher than 20% for Mir pulp. The identification of new malonylated GAs in eggplant is proposed.

Glucosinolate profile and distribution among plant tissues and phenological stages of field-grown horseradish.

Profile and distribution of glucosinolates (GLS) were detected in plant tissues of horseradish at different developmental stages: beginning of vegetative re-growth, flowering and silique formation. The GLS profile varied widely in the different tissues: we identified 17 GLS in roots and sprouts, one of which was not previously characterized in horseradish, i.e. the 2(S)-hydroxy-2-phenylethyl-GLS (glucobarbarin) and/or 2(R)-hydroxy-2-phenylethyl-GLS (epiglucobarbarin), 11 already found in the roots, including the putative 2-methylsulfonyl-oxo-ethyl-GLS, and 5 previously recognized only in the sprouts. Fifteen of those GLS were also identified in young and cauline leaves, 12 in the mature leaves and 13 in the inflorescences. No difference in GLS profile was observed in plant among the phenological stages. Differences in concentrations of GLS, quantified as desulfated, were found in plant. At the beginning of vegetative re-growth, sprouts while showing the same profile of the roots were much richer in GLS having the highest total GLS concentrations (117.5 and 7.7µmolg(-1) dry weight in sprouts and roots, respectively). During flowering and silique forming stages, the roots still maintained lower amount of total GLS (7.4µmolg(-1) of dry weight, on average) with respect to the epigeous tissues, in which mature and young leaves showed the highest total concentrations (70.5 and 73.8µmolg(-1) of dry weight on average, respectively). Regardless of the phenological stages, the aliphatic GLS were always predominant in all tissues (95%) followed by indolic (2.6%) and benzenic (2.4%) GLS. Sinigrin contributed more than 90% of the total GLS concentration. Aliphatic GLS concentrations were much higher in the epigeous tissues, particularly in the mature and young leaves, while benzenic and indolic GLS concentrations were higher in the roots. Through the phenological stages, GLS concentration increased in young and mature leaves and decreased in cauline leaves and inflorescences, while it remained constant over time in roots.

Investigation of glucosinolate profile and qualitative aspects in sprouts and roots of horseradish (Armoracia rusticana) using LC-ESI-hybrid linear ion trap with Fourier transform ion cyclotron resonance mass spectrometry and infrared multiphoton dissociation.

Within the family of Brassicaceae, an important source of glucosinolates (GLSs) is represented by horseradish ( Armoracia rusticana P. Gaertner, B. Meyer & Scherbius), cultivated for its roots, which are grated fresh or processed into a sauce and used as a condiment. The characteristic pungent flavor of the root depends on the abundance of the bioactive GLS molecules. In crude plant extracts (sprouts and roots) of an accession of horseradish largely diffused in the Basilicata region (southern Italy), which develops many sprouts and produces white, fiery, and sharp-flavored marketable roots, we characterized the GLS profile by LC-ESI-LTQ-FTICR-MS and IRMPD. In sprouts and roots we identified 16 and 11 GLSs, respectively. We confirmed the presence of sinigrin, 4-hydroxyglucobrassicin, glucobrassicin, gluconasturtin, and 4-methoxyglucobrassicin and identified glucoiberin, gluconapin, glucocochlearin, glucoconringianin, glucosativin, glucoibarin, 5-hydroxyglucobrassicin, glucocapparilinearisin or glucobrassicanapin, glucotropaeolin, and glucoarabishirsutain, not previously characterized in horseradish. Of particular note was the presence of the putative 2-methylsulfonyl-oxo-ethyl-GLS.

Accumulation of cadmium, zinc, and copper by Helianthus annuus L.: impact on plant growth and uptake of nutritional elements.

We investigated the effects on physiological response, trace elements and nutrients accumulation of sunflower plants grown in soil contaminated with: 5 mg kg(-1) of Cd; 5 and 300 mg kg(-1) of Cd and Zn, respectively; 5, 300, and 400 mg kg(-1) of Cd, Zn, and Cu, respectively. Contaminants applied did not produce large effects on growth, except in Cd-Zn-Cu treatment in which leaf area and total dry matter were reduced, by 15%. The contamination with Cd alone did not affect neither growth nor physiological parameters, despite considerable amounts of Cd accumulated in roots and older leaves, with a high bioconcentration factor from soil to plant. By adding Zn and then Cu to Cd in soil, significant were the toxic effects on chlorophyll content and water relations due to greater accumulation of trace elements in tissues, with imbalances in nutrients uptake. Highly significant was the interaction between shoot elements concentration (Cd, Zn, Cu, Fe, Mg, K, Ca) and treatments. Heavy metals concentrations in roots always exceeded those in stem and leaves, with a lower translocation from roots to shoots, suggesting a strategy of sunflower to compartmentalise the potentially toxic elements in physiologically less active parts in order to preserve younger tissues.

Immunological evaluation of the alcohol-soluble protein fraction from gluten-free grains in relation to celiac disease.

Celiac disease (CD) is a gluten-sensitive enteropathy with an immune basis. We established the immune reactivity of the alcohol-soluble fraction from two minor cereals (tef and millet) and two pseudocereals (amaranth and quinoa) which are believed to be nontoxic based on taxonomy. Grains were examined in intestinal T-cell lines (iTCLs), cultures of duodenal explants from HLA-DQ2(+) CD patients and HLA-DQ8 transgenic mice for signs of activation. Our data indicated that tef, millet, amaranth, and quinoa did not show any immune cross-reactivity toward wheat gliadin, and therefore confirming their safety in the diet of CD patients.

Factors affecting CO2 assimilation, leaf injury and growth in salt-stressed durum wheat.

To examine the factors that affect tolerance to high internal salt concentrations, two tetraploid wheat genotypes that differ in the degree of salt-induced leaf injury (Wollaroi and Line 455) were grown in 150 mM NaCl for 4 weeks. Shoot biomass of both genotypes was substantially reduced by salinity, but genotypic differences appeared only after 3 weeks, when durum cultivar Wollaroi showed greater leaf injury and a greater reduction in biomass than Line 455. Ion accumulation, water relations, chlorophyll fluorescence and gas exchange were followed on one leaf (leaf 3) throughout its life. Salinity caused a large decrease in stomatal conductance (gs) of both genotypes. This was not due to poor water relations, as leaf turgor of both genotypes was higher in the salt treatment than in controls, so chemical signals were likely to have caused the decrease in gs. Reductions in assimilation rate were initially due to gs and, with time, were due to a combination of stomatal and non-stomatal limitations. The non-stomatal limitations were associated with a build up of Na+ above 250 mM. The efficiency of PSII photochemistry in Line 455 was unaffected throughout. However, in Wollaroi, the potential and actual quantum yield of PSII photochemistry began to decline as the leaf aged and the thermal energy dissipation of excess light energy (NPQ) increased. This coincided with high Na+ and Cl- concentrations in the leaf and with chlorophyll degradation, indicating that these later reductions in CO2 assimilation in Wollaroi were a consequence of a direct toxic ion effect. The earlier reduction in CO2 assimilation and greater leaf injury explain why growth of Wollaroi was less than Line 455. The most sensitive indicator of salinity stress was gs, followed by CO2 assimilation, with fluorescence parameters other than NPQ being no more sensitive than chlorophyll itself.

Effects of salinity on gas exchange, water relations and growth of sunflower (Helianthus annuus).

The aim of this study was to determine the response of sunflower (Helianthus annuus L. cv. Romsum HS90) to salinity in terms of gas exchange, ionic and water relations, and growth. Experiments were carried out in the glasshouse, where sunflower plants were exposed to increasing salinity levels using water with a wide range of electrical conductivity (0.39-20 dS m-1) to provide different degrees of salt stress. The CO2 assimilation rate (A), stomatal conductance and plant aboveground dry weight (DW) significantly decreased as electrical conductivity of the soil increased. The decline in photosynthesis measured in response to salt stress was proportionally greater than the decline in transpiration, resulting in a reduction of water use efficiency, at both the leaf and whole-plant levels. Among the factors inhibiting photosynthetic activity, those of a non-stomatal nature had a greater effect. In particular, an analysis of photosynthetic CO2 assimilation rate vs intercellular CO2 concentration (A vs Ci curves) indicated a reduction in activity of Rubisco (EC 4.1.1.39) as salinity levels increased. Under severe salt-stress conditions, chlorophyll fluorescence showed a slowing of electron transport at the PSII level. Salt accumulation in the rhizosphere caused a reduction in tissue water status that was partly associated with a decline in osmotic potential (Ψπ). Leaf ionic concentration was clearly correlated with values of leaf Ψπ. However, leaf ionic concentration showed discontinuous distribution between younger and older leaves, reflecting a strategy of plants to preserve younger and more metabolically-active leaves from accumulating salt to toxic levels.

Effect of salinity on water relations and growth of wheat genotypes with contrasting sodium uptake.

Four wheat genotypes with contrasting degrees of Na+ exclusion were selected to see if low Na+ uptake had an adverse effect on water relations or growth rates during exposure to saline conditions. Plants were grown in supported hydroponics with and without 150 mM NaCl, and sampled for measurements of water relations, biomass, stomatal conductance, and ion accumulation. After 4 weeks exposure to salt, biomass was reduced in all genotypes to a similar extent (about 50%), with the effect of salinity on relative growth rate confined largely to the first week. There was little difference between genotypes in the effect of salinity on water relations, as indicated by their relative water content and estimated turgor. Osmotic adjustment occurred in all genotypes, with one of the low-Na+ genotypes having the greatest adjustment. In the low-Na+ genotypes, osmotic adjustment depended on higher K+ and high organic solute accumulation. Stomatal conductance of all genotypes was reduced by saline conditions, but the reduction was greater in the low-Na+ genotypes. These genotypes also showed a larger fall in the value of carbon isotope discrimination measured in expanding leaves, indicating a greater transpiration efficiency when exposed to saline conditions. Chlorophyll fluorescence measurements failed to indicate damage to photochemical pathways in either high- or low-Na+ genotypes. These data indicate that selecting lines with low Na+ accumulation for the purpose of improving salt tolerance is unlikely to introduce limitations for osmotic adjustment.