Enhancement of bioactive compounds in baby leaf Amaranthus tricolor L. using short-term application of UV-B irradiation.

Baby-leaf vegetables are a trade name for leafy vegetables sold as leaves with petioles at the seedling stage. Amaranth (Amaranthus tricolor L.) is a nutritious baby-leaf vegetable containing many bioactive compounds. The effects of short-term ultraviolet B (UV-B) treatments on the growth and quality of baby leaf amaranth were studied, including the conditions of a 24-h recovery period after irradiation, and different irradiation intensities (3.0-9.0 W m-2), irradiation periods (4-16 h), and cumulative energies (130-170 kJ m-2). A recovery period experiment was conducted to observe the changes in the growth and quality of leaves at 0 and 24 h after UV-B irradiation. The results showed that the concentrations of phenolic compounds, flavonoids, anthocyanin, and ascorbic acid in the leaves, as well as the leaf antioxidant capacity increased 24 h after UV-B irradiation. Increases in target compound concentrations and antioxidant capacity without negative growth and appearance effects were observed in leaves irradiated with UV-B at 3, 6, and 9 W m-2 for irradiation periods of 12 and 16, 8 and 12, and 4 h, respectively. The highest bioactive compound concentration was found in leaves irradiated with UV-B at 6 W m-2 for 7 h (cumulative energy: 150 kJ m-2). It was concluded that UV-B irradiation at 6 W m-2 with a cumulative energy of 150 kJ m-2 and a 24 h post-irradiation recovery period could be an appropriate treatment to increase bioactive compounds in baby leaf amaranth without causing appearance abnormalities.

Speed breeding short-day crops by LED-controlled light schemes.

KEY MESSAGE: A simple and rapid speed breeding system was developed for short-day crops that enables up to five generations per year using LED lighting systems that allow very specific adjustments regarding light intensity and quality. Plant breeding is a key element for future agricultural production that needs to cope with a growing human population and climate change. However, the process of developing suitable cultivars is time-consuming, not least because of the long generation times of crops. Recently, speed breeding has been introduced for long-day crops, but a similar protocol for short-day crops is lacking to date. In this study, we present a speed breeding protocol based on light-emitting diodes (LEDs) that allow to modify light quality, and exemplarily demonstrate its effectiveness for the short-day crops soybean (Glycine max), rice (Oryza sativa) and amaranth (Amaranthus spp.). Adjusting the photoperiod to 10 h and using a blue-light enriched, far-red-deprived light spectrum facilitated the growth of short and sturdy soybean plants that flowered ~ 23 days after sowing and matured within 77 days, thus allowing up to five generations per year. In rice and amaranth, flowering was achieved ~ 60 and ~ 35 days after sowing, respectively. Interestingly, the use of far-red light advanced flowering by 10 and 20 days in some amaranth and rice genotypes, respectively, but had no impact on flowering in soybeans, highlighting the importance of light quality for speed breeding protocols. Taken together, our short-day crops' speed breeding protocol enables several generations per year using crop-specific LED-based lighting regimes, without the need of tissue culture tools such as embryo rescue. Moreover, this approach can be readily applied to a multi-storey 96-cell tray-based system to integrate speed breeding with genomics, toward a higher improvement rate in breeding.

Reactive oxygen species trigger the fast action of glufosinate.

MAIN CONCLUSION: Glufosinate is primarily toxic to plants due to a massive light-dependent generation of reactive oxygen species rather than ammonia accumulation or carbon assimilation inhibition. Glutamine synthetase (GS) plays a key role in plant nitrogen metabolism and photorespiration. Glufosinate (C5H12NO4P) targets GS and causes catastrophic consequences leading to rapid plant cell death, and the causes for phytoxicity have been attributed to ammonia accumulation and carbon assimilation restriction. This study aimed to examine the biochemical and physiological consequences of GS inhibition to identify the actual cause for rapid phytotoxicity. Monocotyledonous and dicotyledonous species with different forms of carbon assimilation (C3 versus C4) were selected as model plants. Glufosinate sensitivity was proportional to the uptake of herbicide between species. Herbicide uptake also correlated with the level of GS inhibition and ammonia accumulation in planta even with all species having the same levels of enzyme sensitivity in vitro. Depletion of both glutamine and glutamate occurred in glufosinate-treated leaves; however, amino acid starvation would be expected to cause a slow plant response. Ammonia accumulation in response to GS inhibition, often reported as the driver of glufosinate phytotoxicity, occurred in all species, but did not correlate with either reductions in carbon assimilation or cell death. This is supported by the fact that plants can accumulate high levels of ammonia but show low inhibition of carbon assimilation and absence of phytotoxicity. Glufosinate-treated plants showed a massive light-dependent generation of reactive oxygen species, followed by malondialdehyde accumulation. Consequently, we propose that glufosinate is toxic to plants not because of ammonia accumulation nor carbon assimilation inhibition, but the production of reactive oxygen species driving the catastrophic lipid peroxidation of the cell membranes and rapid cell death.

Differential sensitivity of spinach and amaranthus to enhanced UV-B at varying soil nutrient levels: association with gas exchange, UV-B-absorbing compounds and membrane damage.

The metabolic reasons associated with differential sensitivity of C3 and C4 plant species to enhanced UV-B under varying soil nutrient levels are not well understood. In the present study, spinach (Spinacia oleracea L. var All Green), a C3 and amaranthus (Amaranthus tricolor L. var Pusa Badi Chaulai), a C4 plant were subjected to enhanced UV-B (280-315 nm; 7.2 kJ m(-2) day(-1)) over ambient under varying soil nutrient levels. The nutrient amendments were recommended Nitrogen (N), Phosphorus (P), Potassium (K), 1.5× recommended NPK, 1.5× recommended N and 1.5× recommended K. Enhanced UV-B negatively affected both the species at all nutrient levels, but the reductions varied with nutrient concentration and combinations. Reductions in photosynthetic rate, stomatal conductance and chlorophyll content were significantly more in spinach compared with amaranthus. The reduction in photosynthetic rate was maximum at 1.5× recommended K and minimum in 1.5× NPK amended plants. The oxidative damage to membranes measured in terms of malondialdehyde content was significantly higher in spinach compared with amaranthus. Enhanced UV-B reduced SOD activity in both the plants except in amaranthus at 1.5× recommended K. POX activity increased under enhanced UV-B at all nutrient levels in amaranthus, but only at 1.5× K in spinach. Amaranthus had significantly higher UV-B-absorbing compounds than spinach even under UV-B stress. Lowest reductions in yield and total biomass under enhanced UV-B compared with ambient were observed in amaranthus grown at 1.5× recommended NPK. Enhanced UV-B did not significantly change the nitrogen use efficiency in amaranthus at all NPK levels, but reduced in spinach except at 1.5× K. These findings suggest that the differential sensitivity of the test species under enhanced UV-B at varying nutrient levels is due to varying antioxidative and UV-B screening capacity, and their ability to utilize nutrients. Amaranthus tolerated enhanced UV-B stress more than spinach at all nutrient levels and 1.5× recommended NPK lowered the sensitivity maximally to enhanced UV-B with respect to photosynthesis, biomass and yield. PCA score has also confirmed the lower sensitivity of amaranthus compared with spinach with respect to the measured physiological and biochemical parameters.

[Effect of decimeter polarized electromagnetic radiation on germinating capacity of seeds].

The effect of a polarization structure of electromagnetic radiation on the germinating capacity of seeds of such weeds as Green foxtail (Setaria viridis) and Green amaranth (Amaranthus retroflexus) has been studied. Seeds have been exposed to impulse electromagnetic radiation in a frequency of 896 MHz with linear, elliptical right-handed and elliptical left-handed polarizations at different power flux density levels. It is determined that the effect of the right-handed polarized electromagnetic radiation increases and the influence of the left-handed polarized one reduces the germinating capacity of seeds compared to the effect of the linearly polarized electromagnetic radiation. It is shown that the seeds have an amplitude polarization selectivity as evinced by the major effect of the right-handed polarized radiation on seeds. An electrodynamic model as the right-handed elliptically polarized antenna with the given quantity of the ellipticity of polarization is suggested to use in description of this selectivity.

The effects of host defence elicitors on betacyanin accumulation in Amaranthus mangostanus seedlings.

The effect of elicitors associated with host defence on betacyanin accumulation in Amaranthus mangostanus seedlings was investigated. Under the conditions of the experiments, betacyanin accumulation was generally enhanced by light. Methyl jasmonate (MeJA) treatment increased betacyanin synthesis in a concentration-dependent response. Seedlings treated with ethylene as 5mM Ethephon also had elevated levels of betacyanin. In contrast, salicylic acid (SA) and H(2)O(2) treatments had no influence on betacyanin contents in light or dark. Combined MeJA with Ethephon or H(2)O(2) had an additive effect on betacyanin accumulation in dark-grown seedlings. However, a decline was recorded in light-grown seedlings. Moreover, an antagonistic effect on betacyanin synthesis was found when MeJA and SA were added simultaneously. Our results indicate that betacyanin content in A. mangostanus seedlings can be upregulated by MeJA and ethylene. Both additive and antagonistic effects in regulating betacyanin synthesis in A. mangostanus seedlings were observed between MeJA and other elicitors.

Sonocatalytic degradation of amaranth catalyzed by La3+ doped TiO2 under ultrasonic irradiation.

The relationship between the physicochemical properties and the activity of sonocatalysts is investigated and elucidated in the sonodegradation of amaranth. The sonocatalyts are composed of La(3+) doped TiO(2) synthesized via a sol-gel process. The sonocatalysts are characterized by X-ray diffraction, scanning electron microscopy, energy dispersive X-ray spectroscopic analysis, UV-Vis absorption spectroscopy, and X-ray photoelectron emission spectroscopy. The sonodegradation products of amaranth are analyzed by UV-Vis absorption spectroscopy. The presence of the La(3+) doped TiO(2) catalysts substantially enhances the sonocatalytic degradation of amaranth in aqueous suspensions. The possible sonocatalytic mechanisms for such are discussed.

Interplay of light and temperature during the in planta modulation of C4 phosphoenolpyruvate carboxylase from the leaves of Amaranthus hypochondriacus L.: diurnal and seasonal effects manifested at molecular levels.

The interactive effects of light and temperature on C(4) phosphoenolpyruvate carboxylase (PEPC) were examined both in vivo and in situ using the leaves of Amaranthus hypochondriacus collected at different times during a day and in each month during the year. The maximum activity of PEPC, least inhibition by malate, and highest activation by glucose-6-phosphate were at 15.00 h during a typical day, in all the months. This peak was preceded by maximum ambient light but coincided with high temperature in the field. The highest magnitude in such responses was in the summer (e.g. May) and least in the winter (e.g. December). Light appeared to dominate in modulating the PEPC catalytic activity, whereas temperature had a strong influence on the regulatory properties, suggesting interesting molecular interactions. The molecular mechanisms involved in such interactive effects were determined by examining the PEPC protein/phosphorylation/mRNA levels. A marked diurnal rhythm could be seen in the PEPC protein levels and phosphorylation status during May (summer month). In contrast, only the phosphorylation status increased during the day in December (winter month). The mRNA peaks were not as strong as those of phosphorylation. Thus, the phosphorylation status and the protein levels of PEPC were crucial in modulating the daily and seasonal patterns in C(4) leaves in situ. This is the first detailed study on the diurnal as well as seasonal patterns in PEPC activity, its regulatory properties, protein levels, phosphorylation status, and mRNA levels, in relation to light and temperature intensities in the field.

Co-regulation of dark and light reactions in three biochemical subtypes of C(4) species.

Regulation of light harvesting in response to changes in light intensity, CO(2) and O(2) concentration was studied in C(4) species representing three different metabolic subtypes: Sorghum bicolor (NADP-malic enzyme), Amaranthus edulis (NAD-malic enzyme), and Panicum texanum (PEP-carboxykinase). Several photosynthetic parameters were measured on the intact leaf level including CO(2) assimilation rates, O(2) evolution, photosystem II activities, thylakoid proton circuit and dissipation of excitation energy. Gross rates of O(2) evolution (J(O)2'), measured by analysis of chlorophyll fluorescence), net rates of O(2) evolution and CO(2) assimilation responded in parallel to changes in light and CO(2) levels. The C(4) subtypes had similar energy requirements for photosynthesis since there were no significant differences in maximal quantum efficiencies for gross rates of O(2) evolution (average value = 0.072 O(2)/quanta absorbed, approximately 14 quanta per O(2) evolved). At saturating actinic light intensities, when photosynthesis was suppressed by decreasing CO(2), ATP synthase proton conductivity (g (H) (+)) responded strongly to changes in electron flow, decreasing linearly with J(O)2', which was previously observed in C(3) plants. It is proposed that g (H) (+) is controlled at the substrate level by inorganic phosphate availability. The results suggest development of nonphotochemical quenching in C(4) plants is controlled by a decrease in g (H) (+), which causes an increase in proton motive force by restricting proton efflux from the lumen, rather than by cyclic or pseudocyclic electron flow.

Improved discrimination between monocotyledonous and dicotyledonous plants for weed control based on the blue-green region of ultraviolet-induced fluorescence spectra.

Precision weeding by spot spraying in real time requires sensors to discriminate between weeds and crop without contact. Among the optical based solutions, the ultraviolet (UV) induced fluorescence of the plants appears as a promising alternative. In a first paper, the feasibility of discriminating between corn hybrids, monocotyledonous, and dicotyledonous weeds was demonstrated on the basis of the complete spectra. Some considerations about the different sources of fluorescence oriented the focus to the blue-green fluorescence (BGF) part, ignoring the chlorophyll fluorescence that is inherently more variable in time. This paper investigates the potential of performing weed/crop discrimination on the basis of several large spectral bands in the BGF area. A partial least squares discriminant analysis (PLS-DA) was performed on a set of 1908 spectra of corn and weed plants over 3 years and various growing conditions. The discrimination between monocotyledonous and dicotyledonous plants based on the blue-green fluorescence yielded robust models (classification error between 1.3 and 4.6% for between-year validation). On the basis of the analysis of the PLS-DA model, two large bands were chosen in the blue-green fluorescence zone (400-425 nm and 425-490 nm). A linear discriminant analysis based on the signal from these two bands also provided very robust inter-year results (classification error from 1.5% to 5.2%). The same selection process was applied to discriminate between monocotyledonous weeds and maize but yielded no robust models (up to 50% inter-year error). Further work will be required to solve this problem and provide a complete UV fluorescence based sensor for weed-maize discrimination.

Relationships between quantum yield for CO2 assimilation, activity of key enzymes and CO2 leakiness in Amaranthus cruentus, a C4 dicot, grown in high or low light.

In C(4) photosynthesis, a part of CO(2) fixed by phosphoenolpyruvate carboxylase (PEPC) leaks from the bundle-sheath cells. Because the CO(2) leak wastes ATP consumed in the C(4) cycle, the leak may decrease the efficiency of CO(2) assimilation. To examine this possibility, we studied the light dependence of CO(2) leakiness (phi), estimated by the concurrent measurements of gas exchange and carbon isotope discrimination, initial activities of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and pyruvate, orthophosphate dikinase (PPDK), the phosphorylation state of PEPC and the CO(2) assimilation rate using leaves of Amaranthus cruentus (NAD-malic enzyme subtype, dicot) plants grown in high light (HL) and low light (LL). phi was constant at photon flux densities (PFDs) >200 micromol m(-2) s(-1) and was around 0.3. At PFDs <150 micromol m(-2) s(-1), phi increased markedly as PFD decreased. At 40 micromol m(-2) s(-1), phi was 0.76 in HL and 0.55 in LL leaves, indicating that the efficiency of CO(2) assimilation at low PFD was greater in LL leaves. The activities of Rubisco and PPDK, and the phosphorylated state of PEPC all decreased as PFD decreased. Theoretical calculations with a mathematical model clearly showed that the increase in phi with decreasing PFD contributed to the decrease in the CO(2) assimilation rate. It was also shown that the 'conventional' quantum yield of photosynthesis obtained by fitting the straight line to the light response curve of the CO(2) assimilation rate at the low PFD region is seriously overestimated. Ecological implications of the increase in phi in LL are discussed.

Mutual stimulation of temperature and light effects on C(4) phosphoenolpyruvate carboxylase in leaf discs and leaves of Amaranthus hypochondriacus.

This article reports marked modulation of the activity and regulatory properties of phosphoenolpyruvate carboxylase (PEPC) by temperature and light in leaf discs as well as leaves of Amaranthus hypochondriacus. The activity of PEPC increased by 1.7-fold at 45 degrees C over 25 degrees C. Warm temperature also stimulated the photoactivation of PEPC. The activation by light of PEPC was 1.9-fold at 25 degrees C and increased to 2.2-fold at 45 degrees C. The sensitivity of PEPC to its inhibitor malate was less and the activation by glucose-6-phosphate (G-6-P) or inorganic phosphate (Pi) was more at 45 degrees C than that at 25 degrees C. These effects of temperature were quite pronounced in light. Similar responses were observed when detached leaves were exposed to varying ambient temperature (dry heat). The activity of PEPC increased by 1.6-fold at 45 degrees C over 25 degrees C in the dark. The activation of PEPC by light was 2.1-fold at 25 degrees C and increased to 2.6-fold at 45 degrees C. Inhibition by malate was less and activation by G-6-P or Pi was more at 45 degrees C than that at 25 degrees C. Thus, there was a marked modulation of not only the activity but also the regulatory properties of the enzyme by temperature and light, independently as well as cooperatively with each other. Further experiments suggested that PEPC was able to memorize to a significant extent the changes induced by warm temperature and that these changes were complemented by subsequent illumination. These effects were not due to changes in PEPC protein levels. We conclude that temperature and light can modulate PEPC activity and regulatory properties not only individually but also in a significantly cooperative manner with each other. As significant increases in temperature are common during daytime in tropical or subtropical conditions, we suggest that the synergistic effects of temperature and light are quite relevant in optimizing the activity of PEPC in leaves of C(4) plants.

Effects of growth light and nitrogen nutrition on the organization of the photosynthetic apparatus in leaves of a C4 plant, Amaranthus cruentus.

Properties of C4 photosynthesis were examined in Amaranthus cruentus L. (NAD-malic enzyme (ME) subtype, dicot) grown under different light and nitrogen (N) conditions, from the viewpoint of N investment into their photosynthetic components. In low-light (LL) leaves, chlorophyll content per leaf area was greater and chlorophyll alb ratio was lower than in high-light (HL) leaves. These indicate that LL leaves invest more N into their light-harvesting systems. However, this N investment did not contribute to the increase in the quantum yield of photosynthesis on the incident photon flux density (PFD) basis (Qi) in LL leaves. N allocation to ribulose 1,5-bisphosphate carboxylasel oxygenase (Rubisco) was significantly higher in HL-high N (HN) leaves than in other leaves. On the other hand, N allocation to C4 enzymes [phosphoenolpyruvate carboxylase (PEPC) and pyruvate Pi dikinase (PPDK)] was unaffected by the growth conditions. Maximum photosynthetic rates (Pmax) per Rubisco content were similar irrespective of the growth light treatments. Carbon isotope ratios (delta13 C) in the leaf dry matter were more negative in LL leaves than in HL leaves (LL = -19.3% per hundred, HL = -16.0% per hundred) and independent of leaf N. Vein density was highest in HL-HN leaves, and leaf thickness was unaffected by the growth light treatments. From these results, we conclude that A. cruentus leaves would not acclimate efficiently to low growth light.

Effect of rare earth element europium on amaranthin synthesis in Amarathus caudatus seedlings.

Considering the resemblances between Eu3+ and Ca2+ in their atomic radius and structures of the valence electron, the effects of Eu3+ on amaramthin synthesis in Amarathus caudatus seedling were studied. Eu3+ had both promoting and inhibiting effects on amaramthin synthesis. The optimum promoting concentration and half inhibiting concentration of Eu3+ to synthesis of amaranthin were 0.4 mmol/L and 2.5 mmol/L, respectively. In the dark, A23187 (ions carrier) could carry Eu3+ into cells through the Ca2+ channel. When Ca2+ was chelated with EGTA, the synthesis of amaranthin could be partly retrieved by Eu3+. Eu3+ treatment could also activate Ca2+- ATPase on plasma membrane. Moreover, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of total proteins from the plants treated by Eu3+ and Ca2+ were similar but slightly different in the contents. It suggested that the effects of Eu3+ and Ca2+ on amaranthin synthesis were similar. After being treated by Eu3+ or Ca2+, the outside Ca2+ could enter into cells to promote synthesis of amaranthin. The results above indicated that Eu3+ might replace Ca2+ in the calcium/calmidulindependent phytochrome signal transduction system and play important roles in plant development by promoting calcium transportation across plasma membrane.

Decoupling of light intensity effects on the growth and development of C3 and C4 weed species through sucrose supplementation.

Light availability has a profound effect on plant growth and development. One of the ways to study the effects of light intensity on plant growth and development without the confounding problem of photosynthate availability is sucrose injection/supplementation. A greenhouse experiment was conducted to evaluate the effects of light levels (0% and 75% shade) and sucrose injection (distilled water or 150 g sucrose l(-1)) on three weed species: redroot pigweed (Amaranthus retroflexus L., C4), lambsquarters (Chenopodium album L., C3) and velvetleaf (Abutilon theophrasti Medic., C3). The average total sucrose uptake was 7.6 and 5.9 g per plant for 0% and 75% shading, respectively, representing 47% of the average total weed dry weight. Plants injected with sucrose had greater dry weights and shoot-to-root ratios under both light levels. In spite of sucrose supplementation the reduction in dry matter due to shading was greater for roots and reproductive structures than vegetative shoot tissues, indicating light level regulation of morphological changes resulting in changed C allocation that are independent of photosynthate availability. Dry weights of plants injected with sucrose under 75% shading were not different from distilled water-injected unshaded plants. However, both sucrose-injected and control plants, regardless of their photosynthetic pathways, underwent similar changes in allocation of dry matter and morphology due to shading, suggesting that these effects are strictly due to light intensity and not related to photosynthate availability.