Evolutionary history and ecology shape the diversity and abundance of phytochemical arsenals across monkeyflowers.

We examine the extent to which phylogenetic effects and ecology are associated with macroevolutionary patterns of phytochemical defence production across the Mimulus phylogeny. We grew plants from 21 species representing the five major sections of the Mimulus phylogeny in a common garden to assess how the arsenals (NMDS groupings) and abundances (concentrations) of a phytochemical defence, phenylpropanoid glycosides (PPGs), vary across the phylogeny. Very few PPGs are widespread across the genus, but many are common to multiple sections of the genus. Phytochemical arsenals cluster among sections in an NMDS and are not associated with total concentration of PPGs. There is a strong phylogenetic signal for phytochemical arsenal composition across the Mimulus genus, whereas ecological variables such as growing season length, latitude, and elevation do not significantly influence arsenal. In contrast, there is little phylogenetic signal for total PPG concentration, and this trait is significantly influenced by several ecological factors. Phytochemical arsenals and abundances are influenced by plant life history form. Both phylogenetic effects and ecology are related to phytochemical patterns across species, albeit in different ways. The independence of phytochemical defence concentrations from arsenal compositions indicates that these aspects of defence may continue to evolve independently of one another.

Molecular and phytochemical variability among genus Albizia: a phylogenetic prospect for future breeding.

Fabaceae, the third-largest Angiosperm family, exhibits great morphological diversity with significantly high species diversification rate. Albizia, one of the largest genera of the legume family, possesses high ecological, economical and medicinal application prospects and displays a global distribution. The taxonomic classification among Albizia remains, however, unclear and has been subjected to changes. The resolution of phylogenetic relationships among members of genus Albizia is a priority. Nine Albizia species cultivated in Egypt; Albizia lebbeck, A. julibrissin, A. odoratissima, A. procera, A. anthelmintica, A. guachapele, A. myriophylla, A. richardiana and A. lucida were subjected to molecular classification via DNA fingerprinting techniques viz. Inter Simple Sequence Repeat (ISSR) and Start Codon Targeted polymorphism (SCoT) using ten primers, five for each technique. The total number of bands produced by ISSR and SCoT primers was 28 and 40, respectively. The percentage of polymorphism varied from 64.28% in ISSR to 67.50% in SCoT analysis. Additionally, chemotaxonomic analysis was implemented based on UV spectroscopic profiling and total phenolic content coupled to unsupervised chemometric tools; Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA). Interspecific relationships were confirmed via molecular and phytochemical analyses between A. procera and A. guachapele; A. lebbeck and A. odoratissima; and A. julibrissin and A. lucida. The study reveals that chemotaxonomic data can reflect phylogenetic relationships among examined Albizia species and provides insights to the significance of utilizing the strengths of both molecular taxonomy and chemotaxonomy to resolve phylogenetic relationship among this genus which offers baseline for breeding programs. Future strategies to enrich taxonomic classification among Albizia includes extensive morphological characterization, DNA barcoding techniques and metabolomic profiling.

Variation in Phytochemical, Morphological, and Ploidy Levels of Iranian Thymus Species.

Thymus is one of the most important genera of the Lamiaceae family. This work was performed to assess inter and intra species variation, which is an indispensable prerequisite for the selection and the exploitation of the germplasm, using yield, secondary metabolites, and ploidy level criteria. Nineteen Iranian populations belonging to 11 Thymus species which includes T. vulgaris were used in this study. The results of cytological observations on the 19 populations revealed the three root-tip chromosome numbers of 2n=2x=30, 2n=4x=56 or 60 (diploid and tetraploid). This study also presents the results of a two-year field experiment that evaluates the agronomic and morphology of the 19 populations of Thymus spp. Cluster analysis grouped the populations into six groups and explained the relationships among ploidy levels, morphological traits, and essential oils (EOs). In general, diploid species belonged to the thymol chemotype, whilst carvacrol chemotype consistently dependent on the gene-dosage effect. Thymus migricus, T. daenensis-2, T. serpyllum, and T. trautvetteri populations with diverse thymol background were the best selection as the parents to improve thymol in a breeding program. Moreover, dry and fresh weight criteria can be used to improve EO content in thyme. Achieving this goal would be expected by crossing T. migricus and T. daenensis-2. Finally, providing relevant information on the ploidy level of Thymus species, with emphasis on morphology and EO components variations, may be recommended for the selection of populations or species to improve bioactive components as well as morphological traits in future breeding programs.

Effect of Methyl Jasmonate on Thymol, Carvacrol, Phytochemical Accumulation, and Expression of Key Genes Involved in Thymol/Carvacrol Biosynthetic Pathway in Some Iranian Thyme Species.

Thyme species are a good source of thymol and carvacrol, which play a key role in controlling diseases. For the first time, the expression patterns of γ-terpinene synthase (TPS2), CYP71D178, and CYP71D180 genes and the amount of phenolics compounds were evaluated in T. migricus and T. daenensis after different methyl jasmonate (MeJA) treatments. The highest thymol and carvacrol contents were observed in T. migricus (86.27%) and T. daenensis (17.87%) at MeJA 100 µM, which was consistent with the expression patterns of the three investigated genes. All species treated showed high total phenolic and flavonoid content compared to control plants for which the highest amounts were observed in T. vulgaris treated with 100 µM and 10 µM MeJA. Furthermore, in the 100 µM MeJA treatment, the relative expression of TPS2 and CYP71D178 in T. migricus increased 7.47 and 9.86-fold compared with the control, respectively. The highest level of CYP71D180 transcripts (5.15-fold) was also observed for T. daenensis treated. This finding highlights the notion that thymol was known as the dominant component of the essential oil rather than carvacrol in diffident thyme species. This implies that MeJA at different concentrations influenced metabolic pathways and induced expression changes, resulting in a rise in essential oil levels.

Analyses of chemosensory genes provide insight into the evolution of behavioral differences to phytochemicals in Bactrocera species.

Insect olfactory systems have evolved to recognize phytochemicals and respond to olfactory-triggered cues that vary depending on needs. Several Bactrocera species are attracted explicitly to particular phytochemicals, including methyl eugenol (ME) and cue lure/raspberry ketone (CL/RK). The attraction of Bactrocerans to ME/RK is primarily driven by olfaction. Therefore, the divergent behavioral phenotypes are due to the differences in genes expressed in antennae. High quality transcriptomes were generated with mRNA from dissected antennae, to analyze the differences in olfaction-related genes of three ME-responders B. dorsalis, B. papayae and B. correcta and two RK-responders B. cucurbitae and B. tau. Many distinct quantitative and qualitative differences were identified in their respective chemosensory repertoires. Tissue- and sex-specific expression analyses identified antennae-predominant and sex-biased chemosensory genes. Sequence comparison revealed variations among family members of odorant-binding proteins and odorant-receptors between ME-responders and RK-responders. Differences in composition, expression levels, and sequence of proteins encoded by olfactory-related genes were identified between ME-responders and RK-responders. Some of the differences might contribute to the divergence in response to plant-derived odorants. Taken together, our results provide insights into the evolution of an olfactory system at molecular level in Bactrocera ME- and CL/RK-responding species.

Synthetic biology strategies toward heterologous phytochemical production.

Covering: 2006 to 2018 Phytochemicals are important sources for the discovery and development of agricultural and pharmaceutical compounds, such as pesticides and medicines. However, these compounds are typically present in low abundance in nature, and the biosynthetic pathways for most phytochemicals are not fully elucidated. Heterologous production of phytochemicals in plant, bacterial, and yeast hosts has been pursued as a potential approach to address sourcing issues associated with many valuable phytochemicals, and more recently has been utilized as a tool to aid in the elucidation of plant biosynthetic pathways. Due to the structural complexity of certain phytochemicals and the associated biosynthetic pathways, reconstitution of plant pathways in heterologous hosts can encounter numerous challenges. Synthetic biology approaches have been developed to address these challenges in areas such as precise control over heterologous gene expression, improving functional expression of heterologous enzymes, and modifying central metabolism to increase the supply of precursor compounds into the pathway. These strategies have been applied to advance plant pathway reconstitution and phytochemical production in a wide variety of heterologous hosts. Here, we review synthetic biology strategies that have been recently applied to advance complex phytochemical production in heterologous hosts.

Comprehensive Analysis of Phytochemical Constituents and Ethnopharmacological Investigation of Genus Datura.

The genus Datura comprises wild shrub plants that belong to the Solanaceae family. Naturally, they possess both medicinal and poisonous properties due to the presence of many biologically active phytochemical constituents. Traditionally, Datura had been used for mystic and religious purposes, as a natural drug to treat asthma, pain, gout, boils, abscesses, and wounds, and as psychoactive infusions and fumitories. Different Datura species exhibit diverse ethnopharmacological activities against different diseases, and many ancient and traditional cultures have used various forms of Datura to treat ailments and to prevent many diseases. In this article, we comprehensively summarize various phytochemical constituents isolated from Datura, their pharmacological properties against different diseases, parts of the plants used as traditional therapeutic agents, regions where they are located, and botanical descriptions of different Datura species. The ethnopharmacological properties of Datura may provide new insights for discovery and development of natural drugs. Further research is needed for the investigation of mechanisms of action and to develop safety profiles of the phytochemical constituents isolated from Datura species.

A Phytochemical-Based Copolymer Derived from Coriolus versicolor Polysaccharopeptides for Gene Delivery.

Coriolus versicolor is an herb widely used for cancer treatment in traditional Chinese medicine. Its active ingredients, polysaccharopeptides (PSP), have been used for adjuvant therapies in cancer treatment. This study conjugates Coriolus versicolor PSP with poly(ethylenimine) (PEI) to generate a PSP-PEI copolymer for gene transfer. After PEI conjugation, both the pH buffering capacity and DNA compaction ability of PSP are significantly increased. Compared with that of PSP, the transfection efficiency of PSP-PEI is 10 to 20-fold higher in vitro. This is a proof-of-concept study reporting the direct use of bioactive phytochemicals from traditional Chinese medicine for gene vector development. The promising performance of PSP-PEI raises the possibility that bioactive herbal ingredients can be further developed as a multi-therapeutic gene carrier for tackling cancers.

Bioactive Phytochemicals and Antioxidant Properties of the Grains and Sprouts of Colored Wheat Genotypes.

The grains and sprouts of colored wheat genotypes (having blue, purple and yellow colored grains) contain specific anthocyanidins, such as pelargonidin and cyanidin derivatives, that produce beneficial health effects. The objective of the presented study is to compare the antioxidant capacity and contents of bioactive phytochemicals in grains and sprouts of wheat genotypes that differ in grain color. The methods α, α-diphenyl-ß-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activities, together with spectrophotometrical and high-performance thin-layer chromatography (HPTLC) methods, were used to study the presence of total phenolics, flavonoids, anthocyanins and anthocyanidins (pelargonidin, peonidin, cyanidin, delphinidin) content. It was predicted that the sprouts of all colored wheat genotypes would have significantly higher total flavonoids, total phenolics, anthocyanidin levels and antioxidant activity than the grains. The correlation results between antioxidant activity and contents of bioactive phytochemicals in grains and sprouts of colored wheat genotypes have shown a high correlation for cyanidin and pelargonidin, especially in grains, as well as quercetin in sprouts. It was found that total anthocyanin, quercetin and pelargonidin contents were significantly higher in the sprouts of the purple wheat genotypes than in the blue or yellow wheat genotypes. Delphinidin was detected at a higher level in the grains than in the sprouts of the blue wheat genotypes. Peonidin was present at very low quantities in the grains of all colored wheat genotypes. The sprouts of the purple wheat genotypes, among the colored wheat genotypes, had the highest pelargonidin, cyanidin and quercetin contents and, therefore, can be a promising source for functional food use.

Influence of Genotype, Environment, and Gypsy Moth Herbivory on Local and Systemic Chemical Defenses in Trembling Aspen (Populus tremuloides).

Numerous studies have explored the impacts of intraspecific genetic variation and environment on the induction of plant chemical defenses by herbivory. Relatively few, however, have considered how those factors affect within-plant distribution of induced defenses. This work examined the impacts of plant genotype and soil nutrients on the local and systemic phytochemical responses of trembling aspen (Populus tremuloides) to defoliation by gypsy moth (Lymantria dispar). We deployed larvae onto foliage on individual tree branches for 15 days and then measured chemistry in leaves from: 1) branches receiving damage, 2) undamaged branches of insect-damaged trees, and 3) branches of undamaged control trees. The relationship between post-herbivory phytochemical variation and insect performance also was examined. Plant genotype, soil nutrients, and damage all influenced phytochemistry, with genotype and soil nutrients being stronger determinants than damage. Generally, insect damage decreased foliar nitrogen, increased levels of salicinoids and condensed tannins, but had little effect on levels of a Kunitz trypsin inhibitor, TI3. The largest damage-mediated tannin increases occurred in leaves on branches receiving damage, whereas the largest salicinoid increases occurred in leaves of adjacent, undamaged branches. Foliar nitrogen and the salicinoid tremulacin had the strongest positive and negative relationships, respectively, with insect growth. Overall, plant genetics and environment concomitantly influenced both local and systemic phytochemical responses to herbivory. These findings suggest that herbivory can contribute to phytochemical heterogeneity in aspen foliage, which may in turn influence future patterns of herbivory and nutrient cycling over larger spatial scales.

Immunostimulatory properties of heat-resistant RNA in a decoction of Glycyrrhizae Radix.

Research on the bioactive components of herbal medicines have been conducted mainly on the secondary metabolites of herbal plants. Accordingly, limited information is available on primary metabolites (carbohydrates, amino acids, lipids, and nucleic acids) and their biological effects. Here, we focused on the heat-resistant RNA of a decoction of Glycyrrhizae Radix and showed its immunostimulatory effects. The RNA activated NF-κB/AP-1 and induced TNF-α production in murine macrophages. Further analysis revealed that the RNA was around 90 nucleotides long. RNA sequencing (RNA-Seq) by next generation sequencing (NGS) showed that approximately 30% of the NGS reads were mapped to the genome of Glycyrrhiza uralensis, which is plant material of Glycyrrhizae Radix. Further analysis of the other 70% of reads indicated that the RNA contained RNA sequences that could be mapped to various microorganisms. Together, these results propose nucleic acids as a new research field in the bioactive components of herbal medicines.

Effect of Polyploidy Induction on Natural Metabolite Production in Medicinal Plants.

Polyploidy plays an important role in plant diversification and speciation. The ploidy level of plants is associated with morphological and biochemical characteristics, and its modification has been used as a strategy to alter the quantitative and qualitative patterns of secondary metabolite production in different medicinal plants. Polyploidization can be induced by many anti-mitotic agents, among which colchicine, oryzalin, and trifluralin are the most common. Other variables involved in the induction process include the culture media, explant types, and exposure times. Due to the effects of polyploidization on plant growth and development, chromosome doubling has been applied in plant breeding to increase the levels of target compounds and improve morphological characteristics. Prompted by the importance of herbal medicines and the increasing demand for drugs based on plant secondary metabolites, this review presents an overview of how polyploidy can be used to enhance metabolite production in medicinal plants.

Noncoding RNAs in Medicinal Plants and their Regulatory Roles in Bioactive Compound Production.

BACKGROUND: Noncoding RNAs (ncRNAs), such as microRNAs (miRNAs), small interfering RNAs (siRNAs) and long noncoding RNAs (lncRNAs), play significant regulatory roles in plant development and secondary metabolism and are involved in plant response to biotic and abiotic stresses. They have been intensively studied in model systems and crops for approximately two decades and massive amount of information have been obtained. However, for medicinal plants, ncRNAs, particularly their regulatory roles in bioactive compound biosynthesis, are just emerging as a hot research field. OBJECTIVE: This review aims to summarize current knowledge on herbal ncRNAs and their regulatory roles in bioactive compound production. RESULTS: So far, scientists have identified thousands of miRNA candidates from over 50 medicinal plant species and 11794 lncRNAs from Salvia miltiorrhiza, Panax ginseng, and Digitalis purpurea. Among them, more than 30 miRNAs and five lncRNAs have been predicted to regulate bioactive compound production. CONCLUSION: The regulation may achieve through various regulatory modules and pathways, such as the miR397-LAC module, the miR12112-PPO module, the miR156-SPL module, the miR828-MYB module, the miR858-MYB module, and other siRNA and lncRNA regulatory pathways. Further functional analysis of herbal ncRNAs will provide useful information for quality and quantity improvement of medicinal plants.

Functional relationship of vegetable colors and bioactive compounds: Implications in human health.

Vegetables are essential protective diet ingredients that supply ample amounts of minerals, vitamins, carbohydrates, proteins, dietary fiber, and various nutraceutical compounds for protection against various disease conditions. Color is the most important quality parameter for the farmers to access the harvest maturity while for the consumer's reliable indices to define acceptability or rejection. The colored vegetables contain functional compounds like chlorophylls, carotenoids, betalains, anthocyanins, etc. well recognized for their antioxidant, antimicrobial, hypolipidemic, neuroprotective, antiaging, diuretic, and antidiabetic properties. Recently, there has been a shift in food consumption patterns from processed to semi-processed or fresh fruits and vegetables to ensure a healthy disease-free life. This shifted the focus of agriculture scientists and food processors from food security to nutrition security. This has resulted in recent improvements to existing crops like blue tomato, orange cauliflower, colored and/or black carrots, with improved color, and thus enriched bioactive compounds. Exhaustive laboratory trials though are required to document and establish their minimum effective concentrations, bioavailability, and specific health benefits. Efforts should also be directed to breed color-rich cultivars or to improve the existing varieties through conventional and molecular breeding approaches. The present review has been devoted to a better understanding of vegetable colors with specific health benefits and to provide in-hand information about the effect of specific pigment on body organs, the effect of processing on their bioavailability, and recent improvements in colors to ensure a healthy lifestyle.

Study of the effects of Lemna minor extracts on human immune cell populations.

OBJECTIVE: Lemna minor is a plant with a huge repertoire of secondary metabolites. The literature indicates that extracts of Lemna minor have antioxidant, antiradical, immunomodulatory and anti-inflammatory properties. The objective of the present study was to find a suitable technique to extract active compounds from this plant and verify whether these extracts have immunomodulatory activity. MATERIALS AND METHODS: We grew L. minor on a standard medium with Gamborg B5 and vitamins. We extracted compounds from the plant by maceration and decoction. The phytochemical profile of the extracts was characterized by chromatography, spectrophotometry, and spectroscopy. The extracts were tested on cultures of mononuclear cells from four human subjects. These cells were pulsed with carboxyfluorescein succinimidyl ester, grown in triplicate in standard culture medium without (control) and with increasing concentrations of Lemna extracts. Flow cytometry was used to evaluate cell death and proliferation of the total mononuclear cell population and of CD4+, CD8+, B cell and monocyte populations. RESULTS: The Lemna extracts were not cytotoxic and did not cause cell necrosis or apoptosis in immune cells. At low concentrations, they induced very limited proliferation of CD4+ cells within 48 hours. At high concentrations, they induced proliferation of CD8+ cells and B lymphocytes within 48 hours. CONCLUSIONS: Unfortunately, we failed to confirm any immunomodulatory activity of Lemna extracts. Growth and death rates of human immune cells were not significantly affected by adding Lemna extracts to the culture medium.

Deciphering the multi-scale mechanisms of Tephrosia purpurea against polycystic ovarian syndrome (PCOS) and its major psychiatric comorbidities: Studies from network pharmacological perspective.

Tephrosia purpurea (T. purpurea), a plant belonging to Fabaceae (pea) family, is a well-known Ayurvedic herb and commonly known as Sarapunkha in traditional Indian medicinal system. Described as "Sarwa wranvishapaka", i.e. having a capability to heal all types of wounds, it is particularly recognized for its usage in splenomegaly. Towards exploring the comprehensive effects of T. purpurea against polycystic ovarian syndrome (PCOS) and three comorbid neuropsychiatric diseases (anxiety, depression, and bipolar disorder), its constituent phytochemicals (PCs) were extensively reviewed and their network pharmacology evaluation was carried out in this study. The complex regulatory potential of its 76 PCs against PCOS is enquired by developing and analyzing high confidence tripartite networks of protein targets of each phytochemical at both pathway and disease association scales. We also developed a high-confidence human Protein-Protein Interaction (PPI) sub-network specific to PCOS, explored its modular architecture, and probed 30 drug-like phytochemicals (DPCs) having multi-module regulatory potential. The phytochemicals showing good binding affinity towards their protein targets were also evaluated for similarity against currently available approved drugs present in DrugBank. Multi-targeting and synergistic capacities of 12 DPCs against 10 protein targets were identified and evaluated using molecular docking and interaction analyses. Eight DPCs as a potential source of PCOS and its comorbidity regulators are reported in T. purpurea. The results of network-pharmacology study highlight the therapeutic relevance of T. purpurea as PCOS-regulator and demonstrate the effectiveness of the approach in revealing action-mechanism of Ayurvedic herbs from holistic perspective.

Genetic analysis of phenylpropanoids and antioxidant capacity in strawberry fruit reveals mQTL hotspots and candidate genes.

Phenylpropanoids are a large class of plant secondary metabolites, which play essential roles in human health mainly associated with their antioxidant activity. Strawberry (Fragaria × ananassa) is a rich source of phytonutrients, including phenylpropanoids, which have been shown to have beneficial effects on human health. In this study, using the F. × ananassa '232' × '1392' F1 segregating population, we analyzed the genetic control of individual phenylpropanoid metabolites, total polyphenol content (TPC) and antioxidant capacity (TEAC) in strawberry fruit over two seasons. We have identified a total of 7, 9, and 309 quantitative trait loci (QTL) for TPC, TEAC and for 77 polar secondary metabolites, respectively. Hotspots of stable QTL for health-related antioxidant compounds were detected on linkage groups LG IV-3, LG V-2 and V-4, and LG VI-1 and VI-2, where associated markers represent useful targets for marker-assisted selection of new varieties with increased levels of antioxidant secondary compounds. Moreover, differential expression of candidate genes for major and stable mQTLs was studied in fruits of contrasting lines in important flavonoids. Our results indicate that higher expression of FaF3'H, which encodes the flavonoid 3'-hydroxylase, is associated with increased content of these important flavonoids.

Transcriptome sequencing of the apricot (Prunus armeniaca L.) and identification of differentially expressed genes involved in drought stress.

Apricot (Prunus armeniaca L.) is an important fruit crop that is widely planted throughout the world. But drought affects both yield and quality of apricot. In order to study the effects of long-term drought on the molecular and physiological mechanisms of apricot, we used transcriptome sequencing and measured physiological indices. First, 322 million high-quality clean reads were obtained, and 74,892 unigenes were generated for the transcriptome. Among the assembled unigenes, 18,671 simple sequence repeats (SSRs) and 5581 differentially expressed genes (DEGs) were identified. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the DEGs revealed that starch and sucrose metabolism, plant-pathogen interaction and plant hormone signal transduction pathways are enriched. Additionally, we used quantitative real-time PCR (qRT-PCR) to confirm the RNA-seq results with 11 drought-related DEGs. Second, through the physiological analysis of apricot leaves under constant drought stress, and the results show the internal microstructure of apricot leaves changed to withstand drought stress. At the same time, plants exposed to long-term drought stress showed higher degree of membrane damage, which reduced photosynthesis in the damaged leaves. Our findings enrich the genome resources for apricot and refine our understanding of the molecular and physiological mechanisms of drought response in this fruit crop, providing insights into drought adaptation of the apricot.

Tailoring of microbes for the production of high value plant-derived compounds: From pathway engineering to fermentative production.

Plant natural products have been an attracting platform for the isolation of various active drugs and other bioactives. However large-scale extraction of these compounds is affected by the difficulty in mass cultivation of these plants and absence of strategies for successful extraction. Even though, synthesis by chemical method is an alternative method; it is less efficient as their chemical structure is highly complex which involve enantio-selectivity. Thus an alternate bio-system for heterologous production of plant natural products using microbes has emerged. Advent of various omics, synthetic and metabolic engineering strategies revolutionised the field of heterologous plant metabolite production. In this context, various engineering methods taken to synthesise plant natural products are described with an additional focus to fermentation strategies.