Unraveling microRNA Mediated Gene Regulation in Centella asiatica (L.) Urb. by High-throughput Sequencing based Small-RNA Profiling.

Centella asiatica is a widely spread herb mostly found in the tropics having extensive medicinal values. Here, we report for the first time, transcriptome-wide characterization of miRNA profile from the leaves of C. asiatica using high-throughput Illumina sequencing. We identified 227 conserved and 109 putative novel miRNAs. Computational screening revealed potential mRNA targets for both the conserved and novel miRNAs encoding diverse transcription factors and enzymes involved in plant development, disease resistance, metabolic and signaling pathways. Gene ontology annotation and KEGG analysis revealed the miRNA targets to be involved in a wide range of metabolomic and regulatory pathways. The differential expression of the miRNA encoding genes in diverse tissues was determined by real-time PCR analysis. We also found that gene expression levels of miR156, 159 and 1171 was reduced in salicylic acid treated axenic shoot cultures of C. asiatica compared to its control. Furthermore, RLM-RACE experiments mapped miRNA-mediated cleavage at two of the mRNA targets. The present study represents the large-scale identification of microRNAs from C. asiatica and contributes to the base for the up-coming studies on miRNA-mediated gene regulation of plant secondary metabolite pathways in particular.

Microbes in the Datura Rootzone Contribute to an Antioxidant Support System of Flavonoids and Other Aromatic Compounds

The purpose of this paper is to elucidate the roles that microbes may be playing in the rootzone of the medicinal plant Datura inoxia. We hypothesized that rhizospheric and endophytic microbes would be found that were capable of performing the same secondary metabolic functions of the plant rootzone they inhabited. We also hypothesized that the microbial functions would be co-operative with and supportive to plant secondary metabolite production, for example, by providing precursors to important plant bioactive molecules. The methods employed were mi-crobial barcoding, tests of essential oils against antibiotic resistant bacteria and other soil bacterial isolates, 16S Next Generation Sequencing (NGS) metabarcoding, and Whole Genome Shotgun (WGS) taxonomic and functional. A few of the main bacterial genera of interest that were dis-covered in the Datura root microbiome were Flavobacterium, Chitinophaga, Pseudomonas, Strepto-myces, Rhizobium, and Bacillus. In the context of known interactions, and current results, plants and microbes influence the flavonoid biosynthetic pathways of one other, in terms of the regulation of the phenylpropanoid pathway. This is important because these compounds are phyto-protective antioxidants and are precursors to many aromatic bioactive compounds that are relevant to human health. There was strong evidence to support the notion that synergistic production of plant de-rived secondary metabolites by microbes occurred, as well as the ability for the compounds to enter plant cells. There are possible biopharmaceutical and agricultural applications of the natural interplay that was discovered during this study of the Datura inoxia rhizosphere.

Plant secondary metabolite diversity reflects both phylogeny and ecological adaptation

A phylogenetic framework explaining plant secondary metabolite diversity is lacking, but metabolite classes could represent adaptations to habitat resource availability. We test the hypothesis that primary adaptive strategies (competitors, C; stress-tolerators, S; ruderals, R) are associated, respectively, with nitrogenous metabolites synthesized in persistent organs (alkaloids), nitrogen-lacking aromatic terpenes and phenolics, and nitrogenous compounds prevalent in reproductive tissues (cyanogenic glucosides and glucosinolates). A matrix was compiled of 1019 species for which secondary metabolite pathways and CSR strategies are known. Accounting for phylogenetic relatedness and native biomes, we found that most phytochemical pathways did not correlate with strategy axes, but certain key associations were evident. C-selection was positively associated with amino acid-derived phenylpropanoids (low phylogenetic relatedness; λ<0.5) and pyrrolizidine alkaloids and galloyl derivatives (high λ), and negatively with N-lacking linear monoterpenes (low λ). Nitrogenous cyanogenic glucosides positively correlated with R-selection (low λ). Terpenoids were widely distributed, but correlated positively with S- and negatively with R-selection (low λ). Twenty-six correlations between phytochemicals and biomes (low λ) were evident. Most secondary metabolite synthesis pathways are widespread, reflecting common roles and obligate defence, and strong phylogenetic effects are often evident. However, the character of phytochemical/adaptive strategy associations agrees with ecological theory and thus reflects adaptation.

Plant Secondary Metabolite Transporters: Diversity, Functionality, and Their Modulation

Secondary metabolites (SMs) play crucial roles in the vital functioning of plants such as growth, development, defense, and survival via their transportation and accumulation at the required site. However, unlike primary metabolites, the transport mechanisms of SMs are not yet well explored. There exists a huge gap between the abundant presence of SM transporters, their identification, and functional characterization. A better understanding of plant SM transporters will surely be a step forward to fulfill the steeply increasing demand for bioactive compounds for the formulation of herbal medicines. Thus, the engineering of transporters by modulating their expression is emerging as the most viable option to achieve the long-term goal of systemic metabolic engineering for enhanced metabolite production at minimum cost. In this review article, we are updating the understanding of recent advancements in the field of plant SM transporters, particularly those discovered in the past two decades. Herein, we provide notable insights about various types of fully or partially characterized transporters from the ABC, MATE, PUP, and NPF families including their diverse functionalities, structural information, potential approaches for their identification and characterization, several regulatory parameters, and their modulation. A novel perspective to the concept of “Transporter Engineering” has also been unveiled by highlighting its potential applications particularly in plant stress (biotic and abiotic) tolerance, SM accumulation, and removal of anti-nutritional compounds, which will be of great value for the crop improvement program. The present study creates a roadmap for easy identification and a better understanding of various transporters, which can be utilized as suitable targets for transporter engineering in future research.

Unraveling MicroRNA Mediated Gene Regulation in Centella Asiatica (L.) Urb. by High-Throughput Sequencing based Small-RNA Profiling.

Centella asiatica is a widely spread herb mostly found in the tropics having extensive medicinal values. Here, we report for the first time, transcriptome-wide characterization of miRNA profile from the leaves of C. asiatica using high-throughput Illumina sequencing. We identified 227 conserved and 109 putative novel miRNAs. Computational screening revealed potential mRNA targets for both the conserved and novel miRNAs encoding diverse transcription factors and enzymes involved in plant development, disease resistance, metabolic and signaling pathways. Gene ontology annotation and KEGG analysis revealed the miRNA targets to be involved in a wide range of metabolomic and regulatory pathways. The differential expression of the miRNA encoding genes in diverse tissues was determined by real-time PCR analysis. We also found that gene expression levels of miR156, 159 and 1171 was reduced in salicylic acid treated axenic shoot cultures of C. asiatica compared to its control. Furthermore, RLM-RACE experiments mapped miRNA-mediated cleavage at two of the mRNA targets. The present study represents the large-scale identification of microRNAs from C. asiatica and contributes to the base for the up-coming studies on miRNA-mediated gene regulation of plant secondary metabolite pathways in particular.

Improvement of the quality in hydroponically grown fresh aromatic herbs by inducing mild salinity stress is species-specific

Profitable hydroponic production requires high quality fresh water, which is often not available for agricultural use, while desalinisation of salty water is an expensive and unsustainable technology. In the present study, we assessed the effect of mild salinity stress during the soilless cultivation of fresh peppermint and spearmint in the floating system on biomass yield, produce quality and plant secondary metabolite content. Peppermint and spearmint plants were grown for 25 days on a nutrient solution (NS) supplemented with three different NaCl concentrations (0 mM, 10 mM or 20 mM NaCl). The plant height, root length, fresh and dry weight were recorded and composition was determined on fresh tissue. The composition of essential oil was determined upon hydrodistillation and that of polyphenolic compounds by targeted ultra-performance liquid chromatography coupled with mass spectrometer (UPLC-MS/MS). Plant growth was not suspended by the addition of NaCl in the NS, except for the plant height at the highest salinity level. In peppermint, the nutritional composition was not affected by the salinity, whereas it was significantly improved in spearmint as confirmed by the nitrate content decrease and the total antioxidant capacity, total soluble phenol, total carotenoid and essential oil content increases. Simultaneously, no effect of the salinity on essential oil or polyphenolic composition in both plants was induced. In conclusion, peppermint and spearmint production is feasible in the floating system even under mild salinity conditions, without negatively affecting either the crop yield or the plant's essential oil or phenolic composition. Indeed, low salinity levels improved the nutritional composition of spearmint plants.

KCNQ Potassium Channels as Targets of Botanical Folk Medicines

Since prehistory, human species have depended on plants for both food and medicine. Even in countries with ready access to modern medicines, alternative treatments are still highly regarded and commonly used. Unlike modern pharmaceuticals, many botanical medicines are in widespread use despite a lack of safety and efficacy data derived from controlled clinical trials and often unclear mechanisms of action. Contributing to this are the complex and undefined composition and likely multifactorial mechanisms of action and multiple targets of many botanical medicines. Here, we review the newfound importance of the ubiquitous KCNQ subfamily of voltage-gated potassium channels as targets for botanical medicines, including basil, capers, cilantro, lavender, fennel, chamomile, ginger, and Camellia, Sophora, and Mallotus species. We discuss the implications for the traditional use of these plants for disorders such as seizures, hypertension, and diabetes and the molecular mechanisms of plant secondary metabolite effects on KCNQ channels. Expected final online publication date for the Annual Review of Pharmacology and Toxicology, Volume 62 is January 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

The Opportunity of Spent Bleaching Earth (Bentonite) and Silica Solubilizing Bacteria as Silica Source for Induction of Secondary Metabolites Production in Plants

Background: CPO refining which produces solid waste namely spent bleaching earth (SBE) in large quantities can pollute the environment. SBE from bentonite ores contains large amounts of silica, so it can be an alternative source of silica minerals. Silica plays an important role in increasing plant resistance and bioactive plant compound products. Methods: The application of Si in plants can increase secondary metabolites such as phenolic and anti-fungal compounds in response to disease pathogens. However, the low solubility of silica makes silica not sufficiently available for plants. Using microorganisms as silica solubilizing bacteria helps increasing solubility of silica in the soil. Bacteria dissolve silica by removing organic acids and producing indole acetic acid (AAI), which stimulates root hairs. Results: This review presents the results of a study on the utilization of silica-rich SBE waste as a source of available silica for plants with solubilizing method using bacteria to increase plant growth and resistance, as well as increase plant secondary metabolite compounds. Conclusions: The application of silica solubilizing bacteria has been known to play an important role in providing silica for plants, through enzymatic mechanisms, namely the production of organic acids and extracellular polysaccharides.