Rhus coriaria induces senescence and autophagic cell death in breast cancer cells through a mechanism involving p38 and ERK1/2 activation

Here, we investigated the anticancer effect of Rhus coriaria on three breast cancer cell lines. We demonstrated that Rhus coriaria ethanolic extract (RCE) inhibits the proliferation of these cell lines in a time- and concentration-dependent manner. RCE induced senescence and cell cycle arrest at G1 phase. These changes were concomitant with upregulation of p21, downregulation of cyclin D1, p27, PCNA, c-myc, phospho-RB and expression of senescence-associated β-galactosidase activity. No proliferative recovery was detected after RCE removal. Annexin V staining and PARP cleavage analysis revealed a minimal induction of apoptosis in MDA-MB-231 cells. Electron microscopy revealed the presence of autophagic vacuoles in RCE-treated cells. Interestingly, blocking autophagy by 3-methyladenine (3-MA) or chloroquine (CQ) reduced RCE-induced cell death and senescence. RCE was also found to activate p38 and ERK1/2 signaling pathways which coincided with induction of autophagy. Furthermore, we found that while both autophagy inhibitors abolished p38 phosphorylation, only CQ led to significant decrease in pERK1/2. Finally, RCE induced DNA damage and reduced mutant p53, two events that preceded autophagy. Our findings provide strong evidence that R. coriaria possesses strong anti-breast cancer activity through induction of senescence and autophagic cell death, making it a promising alternative or adjunct therapeutic candidate against breast cancer.UAEU Program for Advanced Research (Grant 31S111-UPAR) and by the Zayed Center for Health Sciences (ZCHS) research grant (grant 31R021) and College of Science Individual Research Grant (grant 31S123) to Rabah Iratni

The 1-aminocyclopropane-1-carboxylic acid deaminase-producing Streptomyces violaceoruber UAE1 can provide protection from sudden decline syndrome on date palm

In the United Arab Emirates (UAE), sudden decline syndrome (SDS) is one of the major fungal diseases caused by Fusarium solani affecting date palm plantations. To minimize the impact of the causal agent of SDS on date palm, native actinobacterial strains isolated from rhizosphere soils of healthy date palm plants were characterized according to their antifungal activities against F. solani DSM 106836 (Fs). Based on their in vitro abilities, two promising biocontrol agents (BCAs), namely Streptomyces tendae UAE1 (St) andStreptomyces violaceoruber UAE1 (Sv), were selected for the production of antifungal compounds and cell wall degrading enzymes (CWDEs), albeit their variations in synthesizing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase (ACCD). Although both isolates showed antagonism when applied 7 days before the pathogen in the greenhouse experiments, the ACCD-producing Sv was relatively superior in its efficacy against SDS over the non-ACCD-producing St. This was evident from the symptoms of SDS in diseased date palm seedlings which were greatly reduced by Sv compared to St. On a scale of 5.0, the estimated disease severity indices in Fs-diseased seedlings were significantly (P < 0.05) reduced from 4.8 to 1.5 and 0.5 by St and Sv, respectively. Thus, the number of conidia of Fs recovered from plants pre-treated with both BCAs was comparable, but significantly (P < 0.05) reduced compared to plants without any BCA treatment. In addition, a significant (P < 0.05) decrease in ACC levels of both the root and shoot tissues was detected inSv + Fs seedlings to almost similar levels of healthy seedlings. However, in planta ACC levels highly increased in seedlings grown in soils infested with the pathogen alone or amended with St prior to F. solani infestation (St + Fs). This suggests a major role of ACCD production in relieving the stress of date palm seedlings infected with F. solani, thus supporting the integrated preventive disease management programs against this pathogen. This is the first report of effective rhizosphere actinobacterial BCAs to provide protection against SDS on date palm, and to help increase agricultural productivity in a more sustainable manner in the UAE and the other arid regions

Impact of Folic Acid in Modulating Antioxidant Activity, Osmoprotectants, Anatomical Responses, and Photosynthetic Efficiency of Plectranthus amboinicus Under Salinity Conditions

Salinity is a major threat to the sustainability of agricultural production systems. Salt stress has unfavorable implications on various plant physio-morphological and biochemical reactions, causing osmotic and ionic stress. Exogenously applied folic acid (FA) may at least provide one mechanism to evade the injurious stress effects of saline irrigation water on Plectranthus amboinicus. In this regard, two pot trials were performed during the 2018–2019 and 2019–2020 seasons in an open greenhouse of an experimental farm (29°17'N; 30°53'E) in Fayoum, Egypt. We tested four levels of saline irrigation water (SW): 34, 68, and 102 mM NaCl, plus tap water as the control = 0), combined with FA at three concentrations (25 and 50 μM, plus spray with distilled water as the control = 0). The growth parameters, biochemistry, physiology, elemental leaf status, essential oil content, and anatomical responses were assessed. Salt markedly reduced photosynthetic productivity [Fv/Fm and performance index (PI)], total chlorophyll [soil plant analysis development (SPAD)], and leaf osmoprotectant compounds, i.e., total soluble sugars (TSS), free amino acids, proline, and total phenolics, thus hampering P. amboinicus growth and essential oil yield. However, the addition of FA as a foliar spray to P. amboinicus irrigated with saline water induced increases in Fv/Fm, SPAD, and PI. These were linked with enriched stem anatomical structures, leaf osmoprotectant compounds, and enhanced leaf enzymatic activity, e.g., superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, glutathione, ascorbic acid, and antioxidant content. Under salt stress, supplementation of 25 and 50 μM FA increased the growth and production of essential oil by 27.8 and 55.6%, respectively, compared with no applied FA. The highest growth characteristics and elemental leaf contents were obtained when P. amboinicus was irrigated with 0 mM saline water and treated foliarly with 50 μM of FA compared with non-treated plants. Overall, these data showed that foliar spraying with FA reduces the impact of salt stress on P. amboinicus irrigated with saline water

Soil application of effective microorganisms and nitrogen alleviates salt stress in hot pepper (Capsicum annum L.) plants

The application of effective microorganisms (EMs) and/or nitrogen (N) have a stimulating effect on plants against abiotic stress conditions. The aim of the present study was to determine the impact of the co-application of EMs and N on growth, physio-biochemical attributes, anatomical structures, nutrients acquisition, capsaicin, protein, and osmoprotectant contents, as well as the antioxidative defense system of hot pepper (Capsicum annum L.) plants. In the field trials, EMs were not applied (EMs-) or applied (EMs+) along with three N rates of 120, 150, and 180 kg unit N ha-1 (designated as N120, N150, and N180, respectively) to hot pepper plants grown in saline soils (9.6 dS m-1). The application of EMs and/or high N levels attenuated the salt-induced damages to hot pepper growth and yield. The application of EMs+ with either N150 or N180 increased the number, average weight and yield of fruits by 14.4 or 17.0%, 20.8 or 20.8% and 28.4 or 27.5%, respectively, compared to hot pepper plants treated with the recommended dose (EMs- × N150). When EMs+ was individually applied or combined with either N150 or N180, increased accumulation of capsaicin were observed by 16.7 or 20.8%, protein by 12.5 or 16.7%, proline by 19.0 or 14.3%, and total soluble sugars by 3.7 or 7.4%, respectively, in comparison with those treated with the integrative EMs- × N150. In addition, the non-enzymatic contents (ascorbate, and glutathione) and enzymatic activities (catalase, superoxide dismutase, and glutathione reductase) of the antioxidant defense systems significantly increased in hot pepper plants treated with EMs+ alone or combined with N150 or N180 under salt stress conditions. Higher accumulation of nutrients (N, P, K+, and Ca2+) along with reduced Na+ acquisition was also evidenced in response to EMs+ or/and high N levels. Most anatomical features of stems and leaves recovered in hot pepper plants grown in saline soils and supplied with EMs+ and N. The application of EMs and N is undoubtedly opening new sustainable approaches toward enhancing abiotic stress tolerance in crops (e.g. hot pepper)

Microarray analysis of Arabidopsis WRKY33 mutants in response to the necrotrophic fungus Botrytis cinerea

The WRKY33 transcription factor was reported for resistance to the necrotrophic fungus Botrytis cinerea. Using microarray-based analysis, we compared Arabidopsis WRKY33 overexpressing lines and wrky33 mutant that showed altered susceptibility to B. cinerea with their corresponding wild-type plants. In the wild-type, about 1660 genes (7% of the transcriptome) were induced and 1054 genes (5% of the transcriptome) were repressed at least twofold at early stages of inoculation with B. cinerea, confirming previous data of the contribution of these genes in B. cinerea resistance. In Arabidopsis wild-type plant infected with B. cinerea, the expressions of the differentially expressed genes encoding for proteins and metabolites involved in pathogen defense and non-defense responses, seem to be dependent on a functional WRKY33 gene. The expression profile of 12-oxo-phytodienoic acid- and phytoprostane A1-treated Arabidopsis plants in response to B. cinerea revealed that cyclopentenones can also modulate WRKY33 regulation upon inoculation with B. cinerea. These results support the role of electrophilic oxylipins in mediating plant responses to B. cinerea infection through the TGA transcription factor. Future directions toward the identification of the molecular components in cyclopentenone signaling will elucidate the novel oxylipin signal transduction pathways in plant defense

Omics and plant responses to Botrytis cinerea

Botrytis cinerea is a dangerous plant pathogenic fungus with wide host ranges. This aggressive pathogen uses multiple weapons to invade and cause serious damages on its host plants. The continuing efforts of how to solve the “puzzle” of the multigenic nature of B. cinerea’s pathogenesis and plant defense mechanisms against the disease caused by this mold, the integration of omic approaches, including genomics, transcriptomics, proteomics and metabolomics, along with functional analysis could be a potential solution. Omic studies will provide a foundation for development of genetic manipulation and breeding programs that will eventually lead to crop improvement and protection. In this mini-review, we will highlight the current progresses in research in plant stress responses to B. cinerea using high-throughput omic technologies. We also discuss the opportunities that omic technologies can provide to research on B. cinerea-plant interactions as an example showing the impacts of omics on agricultural research

The Arabidopsis Mitochondria-Localized Pentatricopeptide Repeat Protein PGN Functions in Defense against Necrotrophic Fungi and Abiotic Stress Tolerance1[C][W][OA]

Pentatricopeptide repeat (PPR) proteins (PPRPs) are encoded by a large gene family in Arabidopsis (Arabidopsis thaliana), and their functions are largely unknown. The few studied PPRPs are implicated in different developmental processes through their function in RNA metabolism and posttranscriptional regulation in plant organelles. Here, we studied the functions of Arabidopsis PENTATRICOPEPTIDE REPEAT PROTEIN FOR GERMINATION ON NaCl (PGN) in plant defense and abiotic stress responses. Inactivation of PGN results in susceptibility to necrotrophic fungal pathogens as well as hypersensitivity to abscisic acid (ABA), glucose, and salinity. Interestingly, ectopic expression of PGN results in the same phenotypes as the pgn null allele, indicating that a tight regulation of the PGN transcript is required for normal function. Loss of PGN function dramatically enhanced reactive oxygen species accumulation in seedlings in response to salt stress. Inhibition of ABA synthesis and signaling partially alleviates the glucose sensitivity of pgn, suggesting that the mutant accumulates high endogenous ABA. Accordingly, induction of NCED3, encoding the rate-limiting enzyme in stress-induced ABA biosynthesis, is significantly higher in pgn, and the mutant has higher basal ABA levels, which may underlie its phenotypes. The pgn mutant has altered expression of other ABA-related genes as well as mitochondria-associated transcripts, most notably elevated levels of ABI4 and ALTERNATIVE OXIDASE1a, which are known for their roles in retrograde signaling induced by changes in or inhibition of mitochondrial function. These data, coupled with its mitochondrial localization, suggest that PGN functions in regulation of reactive oxygen species homeostasis in mitochondria during abiotic and biotic stress responses, likely through involvement in retrograde signaling

Phosphate Nutrition and Defoliation Effects on Growth and Root Physiology of Alfalfa

Phosphorus (P) deficiency reduces forage yield and stand persistence of alfalfa (Medicago sativa L.). The objective of this study was to determine the influence of P nutrition and defoliation on alfalfa shoot growth, root carbohydrate and protein metabolism, and steady-state mRNA levels for high-affinity P transporters. In a greenhouse study, P-deprived plants were provided with 0, 0.25, 2, and 6 mM P beginning 7 d before shoot removal. Plants were sampled immediately (day -7) on days -5, -2, 0 (day of shoot removal), and on days 1, 2, 6, and 9 post-shoot removal. Addition of P to P-deficient plants stimulated growth of shoots but not roots. Taproot bark sugar concentrations were reduced significantly in cut plants at any rate of P, whereas only the 6 mM P treatment reduced taproot wood sugar concentrations in uncut plants. There was a significant defoliation-induced decline in both wood and bark sugar and amino-acid concentration that was enhanced at high P rates. Low P reduced utilization of starch and protein reserves in taproots. Transcripts for a high-affinity P transporter were not detected in any root or shoot tissue assayed, irrespective of defoliation or P treatment. The uncertain relationship between P availability and P-transporter transcript abundance in our greenhouse-grown plants requires additional investigation

Phosphate nutrition effects on growth, phosphate transporter transcript levels and physiology of alfalfa cells

Phosphorus deficiency reduces forage yield and stand persistence of alfalfa (Medicago sativa L.). Our objectives were to isolate and characterize a high-affinity phosphate-transporter (P-transporter) from alfalfa roots (Medicago sativa L.); determine how phosphorus (P) nutrition impacts P-uptake, growth, and carbohydrate and protein metabolism of alfalfa cells; and learn how expression of the P-transporter is influenced by P nutrition. An 1087-base pair (bp) sequence was isolated using RT-PCR that possessed high nucleotide and amino acid sequence similarity to high-affinity P-transporters. Cultured cells were sampled at 3-day intervals for 9 days while growing in media containing P concentrations ranging from 0 to 10 mM. Media P concentrations declined rapidly in all P treatments by day 6. Low media P concentrations (0, 0.1 and 0.5 mM) reduced cell growth rates compared to higher media P levels (2.5, 5 and 10 mM). Suspension cell cultures supplied 0.5, 2.5, 5, and 10 mM P had lower starch concentrations by day 3 compared to cells cultured in media containing 0 and 0.1 mM P. Steady-state transcript levels for the high-affinity P- transporter were high in P-deprived cells, but declined within 1 day when cells were provided 10 mM P

Differential Degradation and Detoxification of an Aromatic Pollutant by Two Different Peroxidases

Enzymatic degradation of organic pollutants is a new and promising remediation approach. Peroxidases are one of the most commonly used classes of enzymes to degrade organic pollutants. However, it is generally assumed that all peroxidases behave similarly and produce similar degradation products. In this study, we conducted detailed studies of the degradation of a model aromatic pollutant, Sulforhodamine B dye (SRB dye), using two peroxidases—soybean peroxidase (SBP) and chloroperoxidase (CPO). Our results show that these two related enzymes had different optimum conditions (pH, temperature, H2O2 concentration, etc.) for efficiently degrading SRB dye. High-performance liquid chromatography and liquid chromatography –mass spectrometry analyses confirmed that both SBP and CPO transformed the SRB dye into low molecular weight intermediates. While most of the intermediates produced by the two enzymes were the same, the CPO treatment produced at least one different intermediate. Furthermore, toxicological evaluation using lettuce (Lactuca sativa) seeds demonstrated that the SBP-based treatment was able to eliminate the phytotoxicity of SRB dye, but the CPO-based treatment did not. Our results show, for the first time, that while both of these related enzymes can be used to efficiently degrade organic pollutants, they have different optimum reaction conditions and may not be equally efficient in detoxification of organic pollutants