Chemical Profile and Biological Activity of Cherimoya (Annona cherimola Mill.) and Atemoya (Annona atemoya) Leaves.

Annona cherimola (Cherimoya) and Annona atemoya (Atemoya) are tropical plants known for their edible fruit. Scientific data suggest that their leaves, used in traditional medicine in the form of teas or infusions without evidence of toxicity, contain several bioactive compounds. However, only Annona muricata among all the Annona species is currently used in the nutraceutical field, and its dried leaves are marketed for tea preparation. In this work, we explored the nutraceutical potential of Atemoya and Cherimoya leaves, by evaluating their chemical profile and functional properties. Phytochemical analyses showed large amounts of phenolic compounds, in particular proanthocyanidins, and identified 18 compounds, either flavonoids or alkaloids. Concerning biological activity, we found antioxidative properties correlated with polyphenols, and antiproliferative activity against HeLa and HepG2 cell lines correlated with alkaloids. The obtained results demonstrate the potential use of Annona cherimola leaves for the preparation of dietary supplements aimed to promote the physiological redox balance. Moreover, the varietal comparison suggests that two commercial cultivars (Campas and White) and the local Torre 1, better suit this purpose. On the other hand, among the studied cultivars, Campas and Torre 1 are also the richest in alkaloids and, in consideration of the anti-proliferative properties of their extracts, dietary supplements based on these cultivars might also have chemo-preventive effects.

Reduction of the geomagnetic field delays Arabidopsis thaliana flowering time through downregulation of flowering-related genes.

Variations in magnetic field (MF) intensity are known to induce plant morphological and gene expression changes. In Arabidopsis thaliana Col-0, near-null magnetic field (NNMF, i.e., <100 nT MF) causes a delay in the transition to flowering, but the expression of genes involved in this response has been poorly studied. Here, we showed a time-course quantitative analysis of the expression of both leaf (including clock genes, photoperiod pathway, GA20ox, SVP, and vernalization pathway) and floral meristem (including GA2ox, SOC1, AGL24, LFY, AP1, FD, and FLC) genes involved in the transition to flowering in A. thaliana under NNMF. NNMF induced a delayed flowering time and a significant reduction of leaf area index and flowering stem length, with respect to controls under geomagnetic field. Generation experiments (F1 - and F2 -NNMF) showed retention of flowering delay. The quantitative expression (qPCR) of some A. thaliana genes expressed in leaves and floral meristem was studied during transition to flowering. In leaves and flowering meristem, NNMF caused an early downregulation of clock, photoperiod, gibberellin, and vernalization pathways and a later downregulation of TSF, AP1, and FLC. In the floral meristem, the downregulation of AP1, AGL24, FT, and FLC in early phases of floral development was accompanied by a downregulation of the gibberellin pathway. The progressive upregulation of AGL24 and AP1 was also correlated to the delayed flowering by NNMF. The flowering delay is associated with the strong downregulation of FT, FLC, and GA20ox in the floral meristem and FT, TSF, FLC, and GA20ox in leaves. Bioelectromagnetics. 39:361-374, 2018. © 2018 The Authors. Bioelectromagnetics Published by Wiley Periodicals, Inc.

Origanum vulgare Terpenoids Induce Oxidative Stress and Reduce the Feeding Activity of Spodoptera littoralis.

Terpenoids are toxic compounds produced by plants as a defense strategy against insect herbivores. We tested the effect of Origanum vulgare terpenoids on the generalist herbivore Spodoptera littoralis and the response of the plant to herbivory. Terpenoids were analyzed by GC-FID and GC-MS and quantitative gene expression (qPCR) was evaluated on selected plant genes involved in both terpene biosynthesis. The insect detoxification response to terpenes was evaluated by monitoring antioxidant enzymes activity and expression of insect genes involved in terpene detoxification. O. vulgare terpenoid biosynthesis and gene expression was modulated by S. littoralis feeding. The herbivore-induced increased level of terpenoids (particularly carvacrol and p-cymene) interacted with the herbivore by decreasing larval survival and growth rate. The assimilation by S. littoralis of more than 50% of ingested terpenes correlated with the possible toxic effects of O. vulgare terpenoids. In choice test experiments, carvacrol and γ-terpinene mediated the larval feeding preferences, wherease the prolonged feeding on O. vulgare terpenoids (particularly on γ-terpinene) exerted relevant antinutritional effects on larvae. S. littoralis was found to react to O. vulgare terpenoids by increasing its antioxidant enzymes activities and gene expression, although this was not sufficient to sustain the toxicity of O. vulgare terpenoids.

Alkaline soil primes the recovery from drought in Populus nigra plants through physiological and chemical adjustments.

Perennial plants are frequently exposed to severe and prolonged drought, and when the balance between water transport and transpirational demand is compromised trees are in danger of embolism formation. To maintain the physiological balance, plants can rely on mechanisms to quickly recover the lost xylem hydraulic capacity and reduce the prolonged impact on photosynthetic activity upon rehydration. Among factors helpful for plants to sustain acclimation and adaptation responses to drought and promote recovery, maintaining an optimal nutritional status is crucial for plant survival. This study aimed to investigate the physiological and biochemical responses under drought and recovery of Populus nigra plants grown in soil with impaired nutrient bioavailability obtained by adding calcium oxide (CaO) to the substrate. Although the CaO treatment did not affect plant growth, in well-watered conditions, treated poplars displayed an impaired inorganic ions profile in tissues. Under drought, although CaO-treated and untreated plants showed similar physiological responses, the former closed the stomata earlier. During water stress relief, the CaO-treated poplars exhibited a faster stomatal opening and a higher capacity to restore xylem hydraulic conductivity compared to not-treated plants, probably due to the higher osmolyte accumulation during drought. The content of some inorganic ions (e.g, Ca2+ and Cl-) was also higher in the xylem sap collected from stressed CaO-treated plants, thus contributing to increase the osmotic gradient necessary for the recovery. Taken together, our results suggest that CaO treatment promotes a faster and more efficient plant recovery after drought due to a modulation of ions homeostasis.

A new protein hydrolysate-based biostimulant applied by fertigation promotes relief from drought stress in Capsicum annuum L.

Recently, biostimulants have been used in sustainable agriculture as priming agents able to increase crop tolerance to abiotic stressors. Here, a soil application of GHI_16_VHL, a plant protein hydrolysate-based biostimulant, was tested for its capability to mitigate severe water stress effects on Capsicum annuum at flowering time. The biostimulant influence on plant physiological status was monitored upon stress and its relief, by measuring chlorophyll levels, stomatal density, stem water potential, leaf gas exchanges and plant growth. Moreover, leaf osmoregulation and oxidative stress levels were also evaluated by quantifying free proline, total non-structural carbohydrates (NSC), ROS-scavenging activity and H2O2 level. Although biostimulant-primed plants showed a quicker decrease of stem water potential with respect to untreated plants upon drought imposition, they recovered faster probably due to the higher leaf osmolyte accumulation, namely NSC during drought. Moreover, leaf gas exchange recovery was prompted in biostimulant-treated plants, which showed an incremented stomatal density and the same chlorophyll level of well-watered plants at the end of the recovery phase. Hydrogen peroxide level was significantly lower during stress and early recovery in biostimulant primed plants, probably due to the higher catalase activity in treated plants before drought or to the higher level of non-enzymatic antioxidant scavengers in primed stressed plants. Finally, the biostimulant priming increased aboveground relative growth rate and final fruit yield of stressed plants. Taken together, our data suggest that the biostimulant priming treatment promotes a faster and more efficient plant recovery after drought.

Transcriptome Analyses and Antioxidant Activity Profiling Reveal the Role of a Lignin-Derived Biostimulant Seed Treatment in Enhancing Heat Stress Tolerance in Soybean.

Soybean (Glycine max Merr.) is a worldwide important legume crop, whose growth and yield are negatively affected by heat stress at germination time. Here, we tested the role of a biostimulant based on lignin derivatives, plant-derived amino acids, and molybdenum in enhancing soybean heat stress tolerance when applied on seeds. After treatment with the biostimulant at 35 °C, the seed biometric parameters were positively influenced after 24 h, meanwhile, germination percentage was increased after 72 h (+10%). RNA-Seq analyses revealed a modulation of 879 genes (51 upregulated and 828 downregulated) in biostimulant-treated seeds as compared with the control, at 24 h after incubation at 35 °C. Surprisingly, more than 33% of upregulated genes encoded for ribosomal RNA (rRNA) methyltransferases and proteins involved in the ribosome assembly, acting in a specific protein network. Conversely, the downregulated genes were involved in stress response, hormone signaling, and primary metabolism. Finally, from a biochemical point of view, the dramatic H2O2 reduction 40%) correlated to a strong increase in non-protein thiols (+150%), suggested a lower oxidative stress level in biostimulant-treated seeds, at 24 h after incubation at 35 °C. Our results provide insights on the biostimulant mechanism of action and on its application for seed treatments to improve heat stress tolerance during germination.

Reduction of geomagnetic field (GMF) to near null magnetic field (NNMF) affects some Arabidopsis thaliana clock genes amplitude in a light independent manner.

Plant endogenous clock consists of self-sustained interlocked transcriptional/translational feedback loops whose oscillation regulates many circadian processes, including gene expression. Its free running rhythm can be entrained by external cues, which can influence all clock parameters. Among external cues, the geomagnetic field (GMF) has been demonstrated to influence plant growth and development. We evaluated the quantitative expression (qRT-PCR) of three clock genes (LHY, GI and PRR7) in time-course experiments under either continuous darkness (CD) or long days (LD) conditions in Arabidopsis thaliana seedlings exposed to GMF (∼40 µT) and Near Null Magnetic Field (NNMF; ∼40 nT) conditions. Under both LD and CD conditions, reduction of GMF to NNMF prompted a significant increase of the gene expression of LHY and PRR7, whereas an opposite trend was found for GI gene expression. Exposure of Arabidopsis to NNMF altered clock gene amplitude, regardless the presence of light, by reinforcing the morning loop. Our data are consistent with the existence of a plant magnetoreceptor that affects the Arabidopsis endogenous clock.

Geomagnetic field impacts on cryptochrome and phytochrome signaling.

The geomagnetic field (GMF) is an environmental element whose instability affects plant growth and development. Despite known plant responses to GMF direction and intensity, the mechanism of magnetoreception in plants is still not known. Magnetic field variations affect many light-dependent plant processes, suggesting that the magnetoreception could require light. The objective of this work was to comprehensively investigate the influence of GMF on Arabidopsis thaliana (Col-0) photoreceptor signaling. Wild-type Arabidopsis seedlings and photoreceptor-deficient mutants (cry1cry2, phot1, phyA and phyAphyB) were exposed to near null magnetic field (NNMF, ≤40 nT) and GMF (~43 µT) under darkness and different light wavelengths. The GMF did not alter skotomorphogenic or photomorphogenic seedling development but had a significant impact on gene expression pathways downstream of cryptochrome and phytochrome photoactivation. GMF-induced changes in gene expression observed under blue light were partially associated with an alteration of cryptochrome activation. GMF impacts on phytochrome-regulated gene expression could be attributed to alterations in phytochrome protein abundance that were also dependent on the presence of cry1, cry2 and phot1. Moreover, the GMF was found to impact photomorphogenic-promoting gene expression in etiolated seedlings, indicating the existence of a light-independent magnetoreception mechanism. In conclusion, our data shows that magnetoreception alters photoreceptor signaling in Arabidopsis, but it does not necessarily depend on light.

Geomagnetic Field (Gmf) and Plant Evolution: Investigating the Effects of Gmf Reversal on Arabidopsis thaliana Development and Gene Expression.

One of the most stimulating observations in plant evolution is a correlation between the occurrence of geomagnetic field (GMF) reversals (or excursions) and the moment of the radiation of Angiosperms. This led to the hypothesis that alterations in GMF polarity may play a role in plant evolution. Here, we describe a method to test this hypothesis by exposing Arabidopsis thaliana to artificially reversed GMF conditions. We used a three-axis magnetometer and the collected data were used to calculate the magnitude of the GMF. Three DC power supplies were connected to three Helmholtz coil pairs and were controlled by a computer to alter the GMF conditions. Plants grown in Petri plates were exposed to both normal and reversed GMF conditions. Sham exposure experiments were also performed. Exposed plants were photographed during the experiment and images were analyzed to calculate root length and leaf areas. Arabidopsis total RNA was extracted and Quantitative Real Time-PCR (qPCR) analyses were performed on gene expression of CRUCIFERIN 3 (CRU3), copper transport protein1 (COTP1), Redox Responsive Transcription Factor1 (RRTF1), Fe Superoxide Dismutase 1, (FSD1), Catalase3 (CAT3), Thylakoidal Ascorbate Peroxidase (TAPX), a cytosolic Ascorbate Peroxidase1 (APX1), and NADPH/respiratory burst oxidase protein D (RbohD). Four different reference genes were analysed to normalize the results of the qPCR. The best of the four genes was selected and the most stable gene for normalization was used. Our data show for the first time that reversing the GMF polarity using triaxial coils has significant effects on plant growth and gene expression. This supports the hypothesis that GMF reversal contributes to inducing changes in plant development that might justify a higher selective pressure, eventually leading to plant evolution.