The Karrikin Receptor Karrikin Insensitive2 Positively Regulates Heat Stress Tolerance in Arabidopsis thaliana.

In this study, we investigated the potential role of the karrikin receptor KARRIKIN INSENSITIVE2 (KAI2) in the response of Arabidopsis seedlings to high-temperature stress. We performed phenotypic, physiological and transcriptome analyses of Arabidopsis kai2 mutants and wild-type (WT) plants under control (kai2_C and WT_C, respectively) and 6- and 24-h heat stress conditions (kai2_H6, kai2_H24, WT_H6 and WT_H24, respectively) to understand the basis for KAI2-regulated heat stress tolerance. We discovered that the kai2 mutants exhibited hypersensitivity to high-temperature stress relative to WT plants, which might be associated with a more highly increased leaf surface temperature and cell membrane damage in kai2 mutant plants. Next, we performed comparative transcriptome analysis of kai2_C, kai2_H6, kai2_H24, WT_C, WT_H6 and WT_H24 to identify transcriptome differences between WT and kai2 mutants in response to heat stress. K-mean clustering of normalized gene expression separated the investigated genotypes into three clusters based on heat-treated and non-treated control conditions. Within each cluster, the kai2 mutants were separated from WT plants, implying that kai2 mutants exhibited distinct transcriptome profiles relative to WT plants. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed a repression in 'misfolded protein binding', 'heat shock protein binding', 'unfolded protein binding' and 'protein processing in endoplasmic reticulum' pathways, which was consistent with the downregulation of several genes encoding heat shock proteins and heat shock transcription factors in the kai2 mutant versus WT plants under control and heat stress conditions. Our findings suggest that chemical or genetic manipulation of KAI2 signaling may provide a novel way to improve heat tolerance in plants.

Bioremoval capacity of phenol by some selected endophytic fungi isolated from Hibiscus sabdariffa and batch biodegradation of phenol in paper and pulp effluents.

BACKGROUND AND OBJECTIVES: The use of endophytic fungi for management of phenol residue in paper and pulp industries has been shown as cost-effective and eco-friendly approach. In this study, isolation of endophytic fungi from roots, stems, and leaves of Hibiscus sabdariffa was conducted. Additionally, the isolated fungi were examined for their ability to degrade phenol and its derivatives in paper and pulp industrial samples, using different growth conditions. MATERIALS AND METHODS: Out of 35 isolated endophyitc fungi, 31 were examined for their phenol biodegradation capacity using Czapek Dox broth medium containing Catechol and Resorcinol as a sole carbon source at final concentrations of 0.4, 0.6 and 0.8%. RESULTS: A total of 35 fungal species belonging to 18 fungal genera were isolated and identified from different parts of H. sabdariffa plants. All strains have the capability for degrading phenol and their derivatives with variable extents. The optimum condition of degrading phenol in paper and pulp effluent samples by Fusarium poae11r7 were at pH 3-5, temperature at 28-35°C, good agitation speed at no agitation and 100 rpm. CONCLUSION: All endophytic fungal species can utilize phenol and its derivatives as a carbon source and be the potential to degrade phenol in industrial contaminants.

Heat Sensing and Lipid Reprograming as a Signaling Switch for Heat Stress Responses in Wheat.

Temperature is an essential physical factor that affects the plant life cycle. Almost all plant species have evolved a robust signal transduction system that enables them to sense changes in the surrounding temperature, relay this message and accordingly adjust their metabolism and cellular functions to avoid heat stress-related damage. Wheat (Triticum aestivum), being a cool-season crop, is very sensitive to heat stress. Any increase in the ambient temperature, especially at the reproductive and grain-filling stages, can cause a drastic loss in wheat yield. Heat stress causes lipid peroxidation due to oxidative stress, resulting in the damage of thylakoid membranes and the disruption of their function, which ultimately decreases photosynthesis and crop yield. The cell membrane/plasma membrane plays prominent roles as an interface system that perceives and translates the changes in environmental signals into intracellular responses. Thus, membrane lipid composition is a critical factor in heat stress tolerance or susceptibility in wheat. In this review, we elucidate the possible involvement of calcium influx as an early heat stress-responsive mechanism in wheat plants. In addition, the physiological implications underlying the changes in lipid metabolism under high-temperature stress in wheat and other plant species will be discussed. In-depth knowledge about wheat lipid reprograming can help develop heat-tolerant wheat varieties and provide approaches to solve the impact of global climate change.

Three new polyacetylene glycosides (PAGs) from the aerial part of Launaea capitata (Asteraceae) with anti-biofilm activity against Staphylococcus aureus.

Four polyacetylenic glycosides, three of which are new, together with two known flavonoids were isolated from the methanol extract of the aerial parts of Launaea capitate, designated bidensyneoside A1 (1), 6´-O-acetyl-bidensyneoside A1 (2), bidensyneoside E (3), bidensyneoside F (4), luteolin (5) and luteolin-7-glucoside (6) also known as cynaroside. Their structures were elucidated by comprehensive analysis of 1D, 2D-NMR and HR-MS data. The absolute configuration of the bidensyneosides was determined by Mosher ester analysis and the optical rotation values. The isolated compounds were tested against biofilm formation of Staphylococcus aureus as well as against several pathogens including Gram-positive bacteria, Gram-negative bacteria, fungi and yeasts. Furthermore, they were tested for their cytotoxicity against two cancer cell lines L929 and KB-3-1. Compound 4 showed moderate inhibition of S. aureus biofilm formation with 30% and 25% at 256 and 128 µg/mL, respectively, while compounds 1 and 5 showed weak inhibition with 20% at 256 µg/mL. Compound 5 showed moderate cytotoxicity against both cell lines L929 and KB-3-1, with IC50 values of 18 µg/mL.

The role of the endophytic fungus, Thermomyces lanuginosus, on mitigation of heat stress to its host desert plant Cullen plicata.

INTRODUCTION: Endophytic fungi associated with desert plants have a crucial role to enable these plants to tolerate abiotic stress, such as heat and drought. METHODS: In this study, a thermophilic fungal endophyte was isolated from a hot desert-adapted plant, Cullen plicata Delile. The endophytic fungus was (molecularly) identified as Thermomyces lanuginosus, and inoculated plants were coded as E+ and the control as E-. RESULTS: This fungus had an effective growth-promoting activity on its host plant and increased the plant resistance to heat stress as well. DISCUSSION: Our findings demonstrate that thermophilic fungal endophytes can enhance drought and heat stress tolerance in desert plants by ecophysiological mechanisms and improve growth of its host plants.

Metabolomics and Transcriptomics in Legumes Under Phosphate Deficiency in Relation to Nitrogen Fixation by Root Nodules.

Phosphate (Pi) deficiency is a critical environmental constraint that affects the growth and development of several legume crops that are usually cultivated in semi-arid regions and marginal areas. Pi deficiency is known to be a significant limitation for symbiotic nitrogen (N2) fixation (SNF), and variability in SNF is strongly interlinked with the concentrations of Pi in the nodules. To deal with Pi deficiency, plants trigger various adaptive responses, including the induction and secretion of acid phosphatases, maintenance of Pi homeostasis in nodules and other organs, and improvement of oxygen (O2) consumption per unit of nodule mass. These molecular and physiological responses can be observed in terms of changes in growth, photosynthesis, and respiration. In this mini review, we provide a brief introduction to the problem of Pi deficiency in legume crops. We then summarize the current understanding of how Pi deficiency is regulated in legumes by changes in the transcriptomes and metabolomes found in different plant organs. Finally, we will provide perspectives on future directions for research in this field.

Correction: RNA-sequencing-based transcriptome and biochemical analyses of steroidal saponin pathway in a complete set of Allium fistulosum-A. cepa monosomic addition lines.

[This corrects the article DOI: 10.1371/journal.pone.0181784.].

Bacillus methylotrophicus ASWU-C2, a strain inhabiting hot desert soil, a new source for antibacterial bacillopyrone, pyrophen, and cyclopeptides.

A strain of Bacillus methylotrophicus was isolated from a soil sample collected in Aswan eastern desert, which is known for its extremely arid climate. After fermentation of the strain in liquid culture and subsequent extraction, a bioassay-guided isolation procedure yielded five compounds: 2-benzyl-4H-pyran-4-one, named bacillopyrone (1), pyrophen (2), macrolactin A (3) and the cyclopeptides malformin A1 (4), and bacillopeptin A (5). The structures were determined by interpretation of nuclear magnetic resonance (NMR) spectroscopy and high resolution mass spectrometry (HR-MS) data. This is the first report on the isolation of compounds 1 and 2 from Bacillus species; compound 1 was reported previously as synthetic product. Bacillopyrone (1) exhibited moderate activity against the Gram-negative Chromobacterium violaceum with minimum inhibitory concentration 266.6 µg/mL, while macrolactin A (3) and malformin A1 (4) inhibited Staphylococcus aureus (minimum inhibitory concentrations 13.3 and 133.3 µg/mL, respectively).

RNA-sequencing-based transcriptome and biochemical analyses of steroidal saponin pathway in a complete set of Allium fistulosum-A. cepa monosomic addition lines.

The genus Allium is a rich source of steroidal saponins, and its medicinal properties have been attributed to these bioactive compounds. The saponin compounds with diverse structures play a pivotal role in Allium's defense mechanism. Despite numerous studies on the occurrence and chemical structure of steroidal saponins, their biosynthetic pathway in Allium species is poorly understood. The monosomic addition lines (MALs) of the Japanese bunching onion (A. fistulosum, FF) with an extra chromosome from the shallot (A. cepa Aggregatum group, AA) are powerful genetic resources that enable us to understand many physiological traits of Allium. In the present study, we were able to isolate and identify Alliospiroside A saponin compound in A. fistulosum with extra chromosome 2A from shallot (FF2A) and its role in the defense mechanism against Fusarium pathogens. Furthermore, to gain molecular insight into the Allium saponin biosynthesis pathway, high-throughput RNA-Seq of the root, bulb, and leaf of AA, MALs, and FF was carried out using Illumina's HiSeq 2500 platform. An open access Allium Transcript Database (Allium TDB, http://alliumtdb.kazusa.or.jp) was generated based on RNA-Seq data. The resulting assembled transcripts were functionally annotated, revealing 50 unigenes involved in saponin biosynthesis. Differential gene expression (DGE) analyses of AA and MALs as compared with FF (as a control) revealed a strong up-regulation of the saponin downstream pathway, including cytochrome P450, glycosyltransferase, and beta-glucosidase in chromosome 2A. An understanding of the saponin compounds and biosynthesis-related genes would facilitate the development of plants with unique saponin content and, subsequently, improved disease resistance.

Isolation and characterization of Cepa2, a natural alliospiroside A, from shallot (Allium cepa L. Aggregatum group) with anticancer activity.

Exploration of new and promising anticancer compounds continues to be one of the main tasks of cancer research because of the drug resistance, high cytotoxicity and limitations of tumor selectivity. Natural products represent a better choice for cancer treatment in comparison with synthetic compounds because of their pharmacokinetic properties and lower side effects. In the current study, we isolated a steroidal saponin, named Cepa2, from the dry roots of shallot (Allium cepa L. Aggregatum group), and determined its structure by using two-dimensional nuclear manganic resonance (2D NMR). The 1H NMR and 13C NMR data revealed that the newly isolated Cepa2 compound is identical to alliospiroside A (C38H60O12) [(25S)-3ß-hydroxyspirost-5-en-1ß-yl-2-O-(6-deoxy-α-L-mannopyranosyl)-α-L-arabinopyranoside], whose anticancer activity remains elusive. Our in vitro examination of the cytotoxic activity of the identified Cepa2 against P3U1 myeloma cancer cell line showed its high efficiency as an anticancer with 91.13% reduction in P3U1 cell viability 12 h post-treatment. The reduction of cell viability was correlated with the increase in reactive oxygen species levels in Cepa2-treated P3U1 cells, as compared with untreated cells. Moreover, scanning electron microscope results demonstrated apoptosis of the Cepa2-treated P3U1 cells in a time course-dependent manner. The results of our study provide evidence for the anticancer properties of the natural Cepa2/alliospiroside A extracted from shallot plants, and a strong foundation for in-depth investigations to build theoretical bases for cell apoptosis and development of novel anticancer drugs.

The "STAY-GREEN" trait and phytohormone signaling networks in plants under heat stress.

The increasing demand for food and the heavy yield losses in primary crops due to global warming mean that there is an urgent need to improve food security. Therefore, understanding how plants respond to heat stress and its consequences, such as drought and increased soil salinity, has received much attention in plant science community. Plants exhibit stress tolerance, escape or avoidance via adaptation and acclimatization mechanisms. These mechanisms rely on a high degree of plasticity in their cellular metabolism, in which phytohormones play an important role. "STAY-GREEN" is a crucial trait for genetic improvement of several crops, which allows plants to keep their leaves on the active photosynthetic level under stress conditions. Understanding the physiological and molecular mechanisms concomitant with "STAY-GREEN" trait or delayed leaf senescence, as well as those regulating photosynthetic capability of plants under heat stress, with a certain focus on the hormonal pathways, may be a key to break the plateau of productivity associated with adaptation to high temperature. This review will discuss the recent findings that advance our understanding of the mechanisms controlling leaf senescence and hormone signaling cascades under heat stress.

Dissection of Trichoderma longibrachiatum-induced defense in onion (Allium cepa L.) against Fusarium oxysporum f. sp. cepa by target metabolite profiling.

Trichoderma spp. are versatile opportunistic plant symbionts that can cause substantial changes in the metabolism of host plants, thereby increasing plant growth and activating plant defense to various diseases. Target metabolite profiling approach was selected to demonstrate that Trichoderma longibrachiatum isolated from desert soil can confer beneficial agronomic traits to onion and induce defense mechanism against Fusarium oxysporum f. sp. cepa (FOC), through triggering a number of primary and secondary metabolite pathways. Onion seeds primed with Trichoderma T1 strain displayed early seedling emergence and enhanced growth compared with Trichoderma T2-treatment and untreated control. Therefore, T1 was selected for further investigations under greenhouse conditions, which revealed remarkable improvement in the onion bulb growth parameters and resistance against FOC. The metabolite platform of T1-primed onion (T1) and T1-primed onion challenged with FOC (T1+FOC) displayed significant accumulation of 25 abiotic and biotic stress-responsive metabolites, representing carbohydrate, phenylpropanoid and sulfur assimilation metabolic pathways. In addition, T1- and T1+FOC-treated onion plants showed discrete antioxidant capacity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) compared with control. Our findings demonstrated the contribution of T. longibrachiatum to the accumulation of key metabolites, which subsequently leads to the improvement of onion growth, as well as its resistance to oxidative stress and FOC.

Biodegradation of poly (ε-caprolactone) (PCL) film and foam plastic by Pseudozyma japonica sp. nov., a novel cutinolytic ustilaginomycetous yeast species.

Aliphatic polyesters poly (ε-caprolactone) (PCL) and foam plastic have been shown to be biodegradable by microorganisms, which possess cutinolytic enzymes. Pseudozyma japonica-Y7-09, showed both high growth and enzyme activity on Yeast malt (YM) medium fed with PCL film than on YM medium. The hydrolytic enzyme activity of the culture on p-nitrophenyl butyrate indicated the occurrence of cutinase enzyme. This activity was confirmed by the degradation of PCL film which reached to the maximum (93.33 %) at 15 days and the degradation of foam plastic which reached 43.2 % at 30 days. These results suggest that the extracellular cutinase enzyme of Pseudozyma japonica-Y7-09 may be useful for the biological degradation of plastic wastes.

Identification and biological activity of antifungal saponins from shallot ( Allium cepa L. Aggregatum group).

The n-butanol extract of shallot basal plates and roots showed antifungal activity against plant pathogenic fungi. The purified compounds from the extract were examined for antifungal activity to determine the predominant antifungal compounds in the extract. Two major antifungal compounds purified were determined to be alliospiroside A (ALA) and alliospiroside B. ALA had prominent antifungal activity against a wide range of fungi. The products of acid hydrolysis of ALA showed a reduced antifungal activity, suggesting that the compound's sugar chain is essential for its antifungal activity. Fungal cells treated with ALA showed rapid production of reactive oxygen species. The fungicidal action of ALA was partially inhibited by a superoxide scavenger, Tiron, suggesting that superoxide anion generation in the fungal cells may be related to the compound's action. Inoculation experiments showed that ALA protected strawberry plants against Colletotrichum gloeosporioides , indicating that ALA has the potential to control anthracnose of the plant.

Discovery of a new source of resistance to Fusarium oxysporum, cause of Fusarium wilt in Allium fistulosum, located on chromosome 2 of Allium cepa Aggregatum group.

This study was carried out to evaluate the antifungal effect of Allium cepa Aggregatum group (shallot) metabolites on Fusarium oxysporum and to determine the shallot chromosome(s) related to Fusarium wilt resistance using a complete set of eight Allium fistulosum - shallot monosomic addition lines. The antifungal effects of hexane, butanol, and water extraction fractions from bulbs of shallot on 35 isolates of F. oxysporum were examined using the disc diffusion method. Only hexane and butanol fractions showed high antifungal activity. Shallot showed no symptom of disease after inoculation with F. oxysporum f. sp. cepae. The phenolic content of the roots and the saponin content of root exudates of inoculated shallot increased to much higher levels than those of the control at 3 days after inoculation. Application of freeze-dried shallot root exudates to seeds of A. fistulosum soaked in a spore suspension of F. oxysporum resulted in protection of seedlings against infection. Among eight monosomic addition lines and A. fistulosum, FF+2A showed the highest resistance to Fusarium wilt. This monosomic addition line also showed a specific saponin band derived from shallot on the thin layer chromatography profile of saponins in the eight monosomic addition lines. The chromosome 2A of shallot might possess some of the genes related to Fusarium wilt resistance.

Triterpenes from Euphorbia rigida.

Phytochemical studies of the aerial parts of Euphorbia rigida afforded three triterpenes: betulin (1), cycloart-23Z-ene-3, 25-diol (2) and cycloartan-3, 24, 25-triol (3), firstly isolated from this plant. The structures and relative stereochemistry were determined on the basis of extensive spectroscopic analyses, including 1D and 2D NMR experiments (1H NMR, 13C NMR, COSY, NOESY, HMQC and HMBC).

Cytotoxicity of 3-O-(beta-D-glucopyranosyl) etioline, a steroidal alkaloid from Solanum diphyllum L.

In continuation of our interest in phytochemical screening of the Egyptian flora for potential drugs, the reinvestigation of the methanolic extract of the roots of Solanum diphyllum, which grows naturally in the south of Egypt and is recorded as new to the Egyptian flora, afforded an interesting, highly cytotoxic compound, named 3-O-(beta-D-glucopyranosyl) etioline [(25S)-22,26-epimino-3beta-(beta-D-glucopyranosyloxy) cholesta-5,22(N)-dien-16alpha-ol]. The chemical structure of this compound was determined by comprehensive NMR studies, including DEPT, COSY, HMQC, and MS. The compound exhibited high cytotoxic effects against the cervical cancer cell line, Hela cells, with an IC50 value of 150 microg/mL.

alpha-Tomatine, the major saponin in tomato, induces programmed cell death mediated by reactive oxygen species in the fungal pathogen Fusarium oxysporum.

The tomato saponin alpha-tomatine has been proposed to kill sensitive cells by binding to cell membranes followed by leakage of cell components. However, details of the modes of action of the compound on fungal cells are poorly understood. In the present study, mechanisms involved in alpha-tomatine-induced cell death of fungi were examined using a filamentous pathogenic fungus Fusarium oxysporum. alpha-Tomatine-induced cell death of F. oxysporum (TICDF) occurred only under aerobic conditions and was blocked by the mitochondrial F(0)F(1)-ATPase inhibitor oligomycin, the caspase inhibitor D-VAD-fmk, and protein synthesis inhibitor cycloheximide. Fungal cells exposed to alpha-tomatine showed TUNEL-positive nuclei, depolarization of transmembrane potential of mitochondria, and reactive oxygen species (ROS) accumulation. These results suggest that TICDF occurs through a programmed cell death process in which mitochondria play a pivotal role. Pharmacological studies using inhibitors suggest that alpha-tomatine activates phosphotyrosine kinase and monomeric G-protein signaling pathways leading to Ca(2+) elevation and ROS burst in F. oxysporum cells.

Methyljasmonate accelerates catabolism of monoterpenoid indole alkaloids in Catharanthus roseus during leaf processing.

Variations in alkaloid pattern during drying of leaves (leaf processing) showed that treatment with methyljasmonate can induce formation of bisindole alkaloids as a result of catabolism of the monomeric alkaloids catharanthine and vindoline. A two-fold increase in 3',4'-anhydrovinblastine was shown in treated leaves especially from day 8 until day 21. Serpentine also increased in the same period under the treatment as a catabolic product of ajmalicine. Basic peroxidases that are responsible for the formation of anhydrovinblastine and serpentine showed high activity at days 8 and 21 in treated leaves, causing the increase in anhydrovinblastine and serpentine.

Alkaloid accumulation in Catharanthus roseus cell suspension cultures fed with stemmadenine.

Feeding stemmadenine to Catharanthus roseus cell suspension culture resulted in the accumulation of catharanthine, tabersonine and condylocarpine. Condylocarpine is not an intermediate in the pathway to catharanthine or tabersonine when it is fed to the cultures. The results support the hypothesis that stemmadenine is an intermediate in the pathway to catharanthine and tabersonine.