Terpene Profiles Research Articles

Insights into terpenes profiling and transcriptional analyses during flowering of different Cannabis sativa L. chemotypes

Terpenes, volatile compounds known for their aromatic and therapeutic properties, play a pivotal role in shaping the overall chemical profile of Cannabis sativa L. Their biosynthesis in planta occurs in trichomes and involves the 2-C-methyl-D-erythritol 4-phosphate (MEP) and the mevalonic acid (MVA) pathways, responsible for producing the substrates utilized by a family of enzymes, the terpene synthases (TPS), for terpene production. In this work, a comprehensive approach combining chemical analyses of the volatile compounds characterizing the aroma of the inflorescences three C. sativa genotypes collected at three stages of maturity and the transcriptional analyses of key genes involved in the terpene biosynthesis was adopted to study this pathway. The results revealed different terpene profiles among genotypes, which were characterized by peculiar compounds belonging to the sesqui- (CINBOL and Fibrante) or monoterpene (Ermo) categories. Both structural and putative regulatory genes were analysed by RT-qPCR, revealing distinct transcriptional profiles of Terpene Synthases, contributing to the diversity of mono and sesquiterpenes synthesized. Furthermore, the research delved into potential regulatory genes associated with trichome formation, a crucial factor influencing terpene accumulation. This integrated approach highlighted complex mechanisms governing terpene accumulation in cannabis, while also offering potential regulators putatively involved in this pathway.

Physical and chemical characterization of wild tomato Solanum habrochaites accessions for resistance to the sweetpotato whitefly

Sweetpotato whitefly Bemisia tabaci is a significant worldwide insect pest affecting tomato production, causing direct damage during feeding and transmitting multiple plant viruses to this crop. Continuing research to improve integrated pest management (IPM) of sweetpotato whitefly is required to maintain profitable tomato production for growers. This study evaluated the resistance of eight wild tomato accessions of Solanum habrochaites and four commercial tomato cultivars of Solanum lycopersicum to sweetpotato whitefly infestation. Field experiments were conducted at the Wiregrass Research and Extension Center at Auburn University, where whitefly populations were reported in vegetable crops. Accessions and cultivars were evaluated for resistance to whiteflies through the ability to reduce the insect population, analysis of the presence of glandular trichomes, and the profile and quantification of terpenes of each tomato species. Results indicated significant variations in whitefly population dynamics among the wild tomato accessions and commercial tomato cultivars. Wild tomato accessions of S. habrochaites had higher numbers of glandular trichomes and distinct terpene profiles, particularly sesquiterpenes, known for their pest-repellent properties. Multivariate analysis identified three clusters based on terpene profiles and trichome types correlating with whitefly resistance. Accessions of S. habrochaites PI127826, PI134418, and G29258 had prominent results. They significantly reduced whitefly nymph development compared to cultivars of S. lycopersicum. The GC-MS analysis of leaflet extracts revealed that accessions of S. habrochaites had higher levels of sesquiterpenes, including α-zingiberene, β-caryophyllene, and α-humulene, with PI127826 uniquely producing Ar-curcumene and 9-hydroxy-zingiberene. Results highlight the potential of wild tomato accessions of S. habrochaites in breeding programs to enhance whitefly resistance in tomatoes, which can support a sustainable IPM program.

Development of GC–MS coupled to GC–FID method for the quantification of cannabis terpenes and terpenoids: Application to the analysis of five commercial varieties of medicinal cannabis

Cannabis terpenes and terpenoids are among the major classes of pharmacologically active secondary metabolites of therapeutic interest. Indeed, these hydrocarbon molecules, responsible for the characteristic aroma of cannabis flowers, are thought to be involved in a synergistic effect known as the “entourage effect”, together with cannabinoids. Numerous analytical studies have been carried out to characterize the terpene and terpenoid contents of some cannabis varieties, but they have not proposed any real quantification or have described a limited number of analytical standards or average response factors, which may have led to over- or underestimation of the real content of the cannabis flowers. Real and reliable quantification is necessary to justify the entourage effect. Here, we report a rigorous and precise GC–FID and GC–MS method for the identification and quantification of cannabis terpenes and terpenoids. This method is distinguished by the use of a high number of analytical standards, the determination of retention indices for all compounds studied, an exhaustive comparison of databases and scientific literature, the use of relevant response factors, and internal calibration for reliable results. It was applied to the study of terpenic compounds in five commercial varieties of medicinal cannabis produced by Bedrocan International: a CBD-rich (Bedrolite®), a THC/CBD balanced (Bediol®), and three THC-dominant (Bedrocan®, Bedica® and Bedrobinol®). Two extraction solvents are described (ethanol and hexane) to compare their selectivity towards target molecules, and to describe as exhaustively as possible the terpenic profile of the five pharmaceutical-grade varieties. Twenty-three standards were used for accurate dosages. This work highlights that the choice of solvent and the analysis method reliability are critical for the study of these terpenic compounds, regarding their contribution to the entourage effect.

Profile of Polyphenols, Fatty Acids, and Terpenes in Henola Hemp Seeds Depending on the Method of Fertilization.

Botanical varieties of hemp differ in chemical composition, plant morphology, agronomy, and industrial suitability. Hemp is popular for cultivation for the production of cannabinoid oil, fiber production, biomass, etc. The fertilization process is one of the most important factors affecting the plant, both its condition and chemical composition. So far, research has been carried out proving that hemp is a valuable source of, among others: fatty acids, amino acids, acids, vitamins, numerous micro- and macroelements, and antioxidant compounds. In this experiment, it was decided to check the possibility of harvesting hemp panicles twice in one year. The purpose of this treatment is to use one plant to produce cannabidiol oil and grain. The main aim of the research was to determine bioactive compounds in hemp seeds and to determine whether the cultivation method affects their content and quantity. Based on the research conducted, it was observed that hemp can be grown in two directions at the same time and harvested twice because its health-promoting properties do not lose their value. It was found that regardless of whether hemp is grown solely for seeds or to obtain essential oils and then seeds, the type of fertilization does not affect the content of phenolic acids (e.g., syringic acid: 69.69-75.14 μg/100 g, vanillic acid: 1.47-1.63 μg/100 g). Based on the conducted research, it was found that essential oils can be obtained from one plant in the summer and seeds from Henola hemp cultivation in the autumn, because such a treatment does not affect the content of the discussed compounds.

Volatile Terpene Profiles of Needle and Phloem Tissues of Healthy and Tomicus destruens-Infested Pinus brutia Trees.

Coniferous trees produce secondary or defense chemicals, such as terpenes, against pest insects. Terpenes could serve as constitutive or induced defensive mechanisms, defending the tree from invasive herbivores. The Mediterranean pine shoot beetle Tomicus destruens colonizes stems and branches of Pinus brutia trees and even can kill mature trees during periodic outbreaks. We investigated whether terpene profiles of needle and stem of P. brutia trees differ between health and those infested by T. destruens. We selected 20 healthy and T. destruens-infested trees and analyzed the monoterpenes and sesquiterpenes of their needles and phloem. We found higher concentrations of tricyclene, camphene and p-cymene in the phloem of infested trees. Similarly, the needles of infested trees had higher concentrations of α-pinene, β-pinene, myrcene, limonene, trans-β-caryophyllene and α-humulene than healthy trees. These results show that the monoterpene and sesquiterpene profiles of P. brutia trees differed between healthy and infested trees, suggesting that volatile terpenes may be an important part of plant-induced responses against T. destruens.

Classification of Cannabis Strains Based on their Chemical Fingerprint-A Broad Analysis of Chemovars in the German Market.

Introduction: Cannabis cultivars were usually categorized based on their genetic profile as sativa, indica, or hybrid types. However, these three criteria do not allow sufficient differentiation between the numerous varieties of cannabis strains. Furthermore, this classification is based on morphological and bio-geographical properties of the plants and does not represent the chemical composition of different cultivars. The concentration of cannabinoids and terpenes are crucial for the pharmacological effect, not only because of the known entourage effect, and therefore needs to be considered by categorization. Materials and Methods: A total of 140 medicinal cannabis flowers available on the German market were analyzed regarding their individual terpene profile using GC-MS analysis. Statistical evaluation was performed to investigate correlations and data relations as well as for clustering. Results: Multivariate analysis showed correlations between individual terpenes. However, there was no statistical correlation between terpene profiles and their respective genetic profile. Terpene profiles of sativa, indica, and hybrid strains are quite heterogenous and clearly showed that there is no relation between terpenes and the estimated pharmacological effect. As a result, we suggest a new classification system based on individual terpene profiles to faster a comprehensive understanding of the expected medical effect. Discussion: Considering main terpenes, we established a concept of six clusters with various terpene profiles being attributed to different medicinal applications. We excluded tetrahydrocannabinol (THC) and cannabidiol (CBD) content from clustering as most of the strains were THC dominant and therefore distort the results. Our pattern of strains with similar terpene profiles might refine the existing classes of chemotypes with different THC:CBD content. Conclusion: The categorization of cannabis strains based on their terpene profiles allows a clearer, finer and, above all, more meaningful classification than the existing sativa/indica classification. Due to the entourage effect and the interactions between cannabinoids and terpenes, this group of substances is also given the necessary consideration when selecting the right medicine for the individual. Within the next steps, further studies are needed with the aim of mapping clinical validated effects to our chemovars. If it is possible to correlate therapy of symptoms to specific chemical profiles personalized cannabinoid therapy will be possible.

Effect of Hemp Extraction Procedures on Cannabinoid and Terpenoid Composition.

A variety of techniques have been developed to extract hemp phytochemicals for research and consumption. Some of the most common processes in the industry include supercritical CO2 extraction, hydrodistillation, and solvent-based (ethanol) extractions. Each of these processes has the potential to differentially extract various phytochemicals, which would impact their efficacy, tolerability, and safety. However, despite these differences, there has been no direct comparison of the methods and the resulting phytochemical composition. This work aimed to compare cannabinoid and terpene profiles using the three primary commercial procedures, using hemp inflorescence from a CBD/CBG dominant Cannabis sativa L. cultivar. Extracts were then evaluated for their terpene and cannabinoid content using GC-MS and LC-MS/MS, respectively. Hydrodistilled extracts contained the most variety and abundance of terpenes with β-caryophyllene to be the most concentrated terpene (25-42 mg/g). Supercritical CO2 extracts displayed a minimal variety of terpenes, but the most variety and abundance of cannabinoids with CBD ranging from 12.8-20.6 mg/g. Ethanol extracts contained the most acidic cannabinoids with 3.2-4.1 mg/g of CBDA along with minor terpene levels. The resulting extracts demonstrated substantially different chemical profiles and highlight how the process used to extract hemp can play a large role in product composition and potential biological effects.

Evaluation of Terpene Decomposition in Kaffir Lime Juice during Storage Using Gas Chromatography-Mass Spectrometry and Proton Transfer Reaction-Mass Spectrometry.

Kaffir lime juice, often treated as production waste, can be a good source of terpenes. These compounds undergo various decomposition processes under the influence of external factors, especially during transportation and storage. In this paper, it was possible to monitor changes in the terpene profile of kaffir lime juice under different storage conditions, namely, 4 °C, 20 °C, and 35 °C. The identification of key decomposition products was achieved using gas chromatography-mass spectrometry (GC-MS) and a data mining protocol. It was followed by tracing those products in different storage conditions using a high-throughput proton transfer reaction mass spectrometry (PTR-MS) approach. Based on our findings, degradation pathways were presented, showing that the main products resulting from storage are p-cymene, p-cymenene, terpinene-4-ol, and α-terpineol. It was shown that conversion to p-cymenene occurs after 5 days of storage. Terpinene-4-ol and α-terpineol were found to be the final products of the conversion at all temperatures. Changes in the composition of terpenes are important from the point of view of their bioactive properties.

Morpho- and Chemotyping of Holopelagic Sargassum Species Causing Massive Strandings in the Caribbean Region

The specific identification of three major morphotypes of the tropical holopelagic Sargassum species causing massive strandings on the African and Caribbean coastlines was attempted by morphological characterisation as well as quantitative and qualitative analyses of several metabolites. Of the 25 morphological variables studied on 208 samples from the North Atlantic Ocean, 22 were used to establish a dichotomous identification key, allowing without any doubt the identification of each morphotype based on their morphological criteria alone. We also attempted to differentiate morphotypes using chemical fingerprintings (HR-MAS NMR) and markers by analysing pigment level and composition using High Pressure Liquid Chromatography, terpene profiles by Thin Layer Chromatography, phenolic compound levels by the Folin-Ciocalteu assay and structures by 2D Nuclear Magnetic Resonance spectroscopy, and fatty acid composition by Gas Chromatography. While pigment level and composition, terpene profiles, and phenolic contents were not discriminating, quantification of eight fatty acids enabled the differentiation of the three morphotypes. Furthermore, phlorotannin purification permitted their structural characterisation allowing discrimination between the three morphotypes. Our study highlights the potential of the free fatty acid profile and phlorotannin structure as good chemomarkers in order to discriminate between the three morphotypes of holopelagic Sargassum.

Optimization of Trimming Techniques for Enhancing Cannabinoid and Terpene Content in Medical Cannabis Inflorescences.

Cannabis sativa L. inflorescences are widely used in the medicinal field as treatments for a variety of symptoms and illnesses due to their unique phytochemicals such as cannabinoids and terpenes. Common postharvest procedures for cannabis inflorescence include trimming, followed by drying, curing, and subsequent storage. The postharvest trimming step, particularly its timing (pre- or post-drying) and the extent of trimming, is not optimally refined in terms of its impact on the cannabinoid and terpene content. In this study, our objective was to identify the optimal trimming conditions for a commercially available medicinal cannabis hybrid chemovar, with the goal of maximizing its cannabinoid and terpene content. To achieve this, we investigated the effects of pre- versus post-drying trimming and evaluated the impact of mild versus aggressive trimming prior to drying on the cannabinoid and terpene profiles using liquid and gas chromatography. Our results indicated that pre-drying mild trimming yielded the highest cannabinoid concentration, possibly due to optimal balance between stress signals and precursor influx from the sugar leaves to the inflorescence. On the other, post-drying trimming yielded the highest terpene content. Identifying the optimal trimming conditions that maximize both cannabinoid and terpene levels in cannabis is challenging. Therefore, growers face a decision in their trimming practices: to prioritize either enhanced cannabinoid content or increased aromatic terpene concentrations, as optimizing for both simultaneously appears to be difficult.

An improved genome assembly of Chrysanthemum nankingense reveals expansion and functional diversification of terpene synthase gene family

BackgroundTerpenes are important components of plant aromas, and terpene synthases (TPSs) are the key enzymes driving terpene diversification. In this study, we characterized the volatile terpenes in five different Chrysanthemum nankingense tissues. In addition, genome-wide identification and expression analysis of TPS genes was conducted utilizing an improved chromosome-scale genome assembly and tissue-specific transcriptomes. The biochemical functions of three representative TPSs were also investigated.ResultsWe identified tissue-specific volatile organic compound (VOC) and volatile terpene profiles. The improved Chrysanthemum nankingense genome assembly was high-quality, including a larger assembled size (3.26 Gb) and a better contig N50 length (3.18 Mb) compared to the old version. A total of 140 CnTPS genes were identified, with the majority representing the TPS-a and TPS-b subfamilies. The chromosomal distribution of these TPS genes was uneven, and 26 genes were included in biosynthetic gene clusters. Closely-related Chrysanthemum taxa were also found to contain diverse TPS genes, and the expression profiles of most CnTPSs were tissue-specific. The three investigated CnTPS enzymes exhibited versatile activities, suggesting multifunctionality.ConclusionsWe systematically characterized the structure and diversity of TPS genes across the Chrysanthemum nankingense genome, as well as the potential biochemical functions of representative genes. Our results provide a basis for future studies of terpene biosynthesis in chrysanthemums, as well as for the breeding of improved chrysanthemum varieties.

Simultaneous quantification of terpenes and cannabinoids by reversed-phase LC-APCI-MS/MS in Cannabis sativa L. samples combined with a subsequent chemometric analysis

Cannabis sativa L. has been the most discussed medicinal plant in recent years. In particular, the dynamic shift from a formerly illicit and tightly controlled substance to a plant recognized for both medicinal and recreational purposes has brought C. sativa into the global spotlight. Due to the ongoing international legalization processes, fast and convenient analytical methods for the quality control of C. sativa flowers for medicinal and recreational purposes are of tremendous interest. In this study, we report the development and validation of a liquid chromatography-tandem mass spectrometry (LC–MS/MS)-based method applying atmospheric pressure chemical ionization (APCI) to fully quantify 16 terpenes and 7 cannabinoids including their acidic forms by a single chromatographic method. The method presented here is unique and simple, as it eliminates the need for derivatization reactions and includes the unconventional analysis of volatile compounds by liquid chromatography. Samples were prepared by a simple and fast ethanolic extraction. Separation was accomplished within 25 min on a reversed-phase C18 column. Method validation was conducted according to international guidelines regarding selectivity, accuracy, precision, robustness, and linearity. Detection was done in multiple reaction monitoring, which allowed the simultaneous quantification of co-eluting analytes applying two selective mass transitions. In addition, due to reproducible in-source decarboxylation, the acidic forms of cannabinoids were reliably quantified using mass transitions of the neutral forms. The accuracy given as the bias was below 15% for all analytes. Matrix effects for cannabinoids were studied by spiking Humulus lupulus extracts with the analytes at varying concentrations. APCI did not show susceptibility toward ion suppression or enhancement. In addition, the recovery effect after spiking was between 80 and 120% for terpenes. Further, 55 authentic C. sativa extracts were fully quantified, and the obtained results for the terpene profiles were compared to state-of-the-art gas chromatography coupled to flame ionization detection. Comparable results were achieved, emphasizing the method’s applicability for cannabinoids and terpenes. Further, acquired metabolite patterns for C. sativa samples were studied, identifying a relationship between cannabinoid and terpene patterns, as well as the abundance of myrcene in CBD-dominant C. sativa strains.Graphical

A multimodal analytical approach is important in accurately assessing terpene composition in edible essential oils

Terpenes in essential oils (EOs) have recently received significant attention due to their potential to improve brain and whole-body health. A deeper understanding of the terpene composition of edible EOs is important for fully exploring their possible applications. In our study, we employed a comprehensive study using four different methods to analyze EO samples, including GC–MS with solid phase microextraction (SPME), liquid injection (LI), derivatization to trimethylsilyl ethers (TMSE), and LC-MS. Our findings revealed that relying on a single analytical method may be insufficient for detecting all terpenes in EOs. Despite identifying a total of 156 terpenes in the samples, only 58 were detectable across all 4 methods. To obtain a more accurate terpene profile of EOs, we advocate for the combined use of LI-GC and TMSE-GC. The terpenes detected by these two methods are complementary, enabling the detection of all terpenes with high VIP in the samples.

Monitoring the Phenolic and Terpenic Profile of Olives, Olive Oils and By-Products throughout the Production Process.

Olive oil is a food of great importance in the Mediterranean diet and culture. However, during its production, the olive oil industry generates a large amount of waste by-products that can be an important source of bioactive compounds, such as phenolic compounds and terpenes, revalorizing them in the context of the circular economy. Therefore, it is of great interest to study the distribution and abundance of these bioactive compounds in the different by-products. This research is a screening focused on phytochemical analysis, with particular emphasis on the identification and quantification of the phenolic and terpenic fractions. Both the main products of the olive industry (olives, olive paste and produced oil) and the by-products generated throughout the oil production process (leaf, "alpeorujo", liquid and solid residues generated during decanting commonly named "borras" and washing water) were analyzed. For this purpose, different optimized extraction procedures were performed for each matrix, followed by high-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry (HPLC-ESI-TOF/MS) analysis. Although no phenolic alcohols were quantified in the leaf and the presence of secoiridoids was low, this by-product was notable for its flavonoid (720 ± 20 µg/g) and terpene (5000 ± 300 µg/g) contents. "Alpeorujo" presented a complete profile of compounds of interest, being abundant in phenolic alcohols (900 ± 100 µg/g), secoiridoids (4500 ± 500 µg/g) and terpenes (1200 ± 100 µg/g), among others. On the other hand, while the solid residue of the borras was the most abundant in phenolic alcohols (3700 ± 200 µg/g) and secoiridoids (680 ± 20 µg/g), the liquid fraction of this waste was notable for its content of elenolic acid derivatives (1700 ± 100 µg/mL) and phenolic alcohols (3000 ± 300 µg/mL). Furthermore, to our knowledge, this is the first time that the terpene content of this by-product has been monitored, demonstrating that it is an important source of these compounds, especially maslinic acid (120 ± 20 µg/g). Finally, the phytochemical content in wash water was lower than expected, and only elenolic acid derivatives were detected (6 ± 1 µg/mL). The results highlighted the potential of the olive by-products as possible alternative sources of a wide variety of olive bioactive compounds for their revalorization into value-added products.

Biological activity of essential oils of Calocedrus decurrens and Cupressus arizonica on Lymantria dispar larvae and Phytophthora root pathogens

Composition of Calocedrus decurrens and Cupressus arizonica essential oils (CDEO and CAEO, respectively), their antifeedant activity on spongy moth (Lymantria dispar) larvae, as well as their antimicrobial activity on three Phytophthora species were investigated. In leaves of these two conifers from the Cupressaceae family, monoterpenes strongly dominated (90.5 and 85.0%, respectively), but their terpene profiles were different. The dominant compound of CDEO was δ-3-carene (49.5%), followed by myrcene (9.4%), terpinolene (8.6%) and α-pinene (7.0%). In CAEO, the most dominant compounds were limonene (23.3%) and α-pinene (20.5%), followed by umbellulone (12.0%) and terpinen-4-ol (9.5%). CDEO showed a strong antifeedant activity, while leaf discs treated with CAEO had phagostimulatory effect on spongy moth larvae. CAEO had no significant effect on relative consumption and larval growth rate, while CDEO reduced consumption and stimulated growth at the lowest EO concentration (0.05%). Both EOs significantly affected the growth of colonies of the Phytophthora species tested. At concentrations of 0.5% and 1%, a 100% inhibition was observed in almost all experimental groups, with the exception of P. plurivora on 0.5% CDEO. At a concentration of 0.1% inhibition rate varied from 15% in P. ×cambivora treated with CDEO to 90% in P. quercina treated with CAEO. These findings indicate that these two EOs could be used to control tree pests.

Sesquiterpenes of the ectomycorrhizal fungus Pisolithus microcarpus alter root growth and promote host colonization

Trees form symbioses with ectomycorrhizal (ECM) fungi, maintained in part through mutual benefit to both organisms. Our understanding of the signaling events leading to the successful interaction between the two partners requires further study. This is especially true for understanding the role of volatile signals produced by ECM fungi. Terpenoids are a predominant class of volatiles produced by ECM fungi. While several ECM genomes are enriched in the enzymes responsible for the production of these volatiles (i.e., terpene synthases (TPSs)) when compared to other fungi, we have limited understanding of the biochemical products associated with each enzyme and the physiological impact of specific terpenes on plant growth. Using a combination of phylogenetic analyses, RNA sequencing, and functional characterization of five TPSs from two distantly related ECM fungi (Laccaria bicolor and Pisolithus microcarpus), we investigated the role of these secondary metabolites during the establishment of symbiosis. We found that despite phylogenetic divergence, these TPSs produced very similar terpene profiles. We focused on the role of P. microcarpus terpenes and found that the fungus expressed a diverse array of mono-, di-, and sesquiterpenes prior to contact with the host. However, these metabolites were repressed following physical contact with the host Eucalyptus grandis. Exposure of E. grandis to heterologously produced terpenes (enriched primarily in γ\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\gamma$$\\end{document}-cadinene) led to a reduction in the root growth rate and an increase in P. microcarpus–colonized root tips. These results support a very early putative role of fungal-produced terpenes in the establishment of symbiosis between mycorrhizal fungi and their hosts.

DoAP2/ERF89 activated the terpene synthase gene DoPAES in Dendrobium officinale and participated in the synthesis of β-patchoulene.

Dendrobium officinale Kimura et Migo is a tonic plant that has both ornamental and medicinal properties. Terpenoids are significant and diverse secondary metabolites in plants, and are one of the important natural active ingredients in D. officinale. The AP2/ERF gene family plays a major role in primary and secondary metabolism. However, the AP2/ERF transcription factor family has not been identified in D. officinale, and it is unclear if it is involved in the regulation of terpenoid biosynthesis. This study identified a sesquiterpene synthetase-β-patchoulene synthase (DoPAES) using transcriptome and terpenic metabolic profile analyses. A total of 111 members of the AP2/ERF family were identified through the whole genome of D. officinale. The tissue-specific expression and gene co-expression pattern of the DoAP2/ERF family members were analyzed. The results showed that the expression of DoPAES was highly correlated with the expression of DoAP2/ERF89 and DoAP2/ERF47. The yeast one-hybrid (Y1H) assays and dual-luciferase experiments demonstrated that DoAP2/ERF89 and DoAP2/ERF47 could regulate the expression of DoPAES. The transcriptional regulatory effects were examined using homologous transient expression of DoAP2/ERF89 in protocorms of D. officinale. DoAP2/ERF89 positively regulated the biosynthesis of β-patchoulene. This study showed that DoAP2/ERF89 can bind to the promoter region of DoPAES to control its expression and further regulate the biosynthesis of β-patchoulene in D. officinale. These results provide new insights on the regulation of terpenoid biosynthesis.

Exploring the versatility of sesquiterpene biosynthesis in guava plants: a comparative genome-wide analysis of two cultivars

Psidium guajava L., a fruit crop belonging to the Myrtaceae family, is highly valued for its nutritional and medicinal properties. The family exhibits a diverse chemical profile of essential oils and serves as a valuable resource due to its ecological interactions, adaptability, and dispersal capacity. The Myrtaceae family has been extensively studied for its terpenoids. Genetic studies have focused on foliar terpene yield in species from the Eucalypteae and Melaleucaceae tribes. To understand the evolutionary trends in guava breeding, this study predicted terpene synthase genes (TPS) from different cultivars. Through this analysis, 43 full-length TPS genes were identified, and approximately 77% of them exhibited relative expression in at least one of the five investigated plant tissues (root, leaf, bud, flower, and fruit) of two guava cultivars. We identified intra-species variation in the terpene profile and single nucleotide polymorphisms (SNPs) in twelve TPS genes, resulting in the clustering of 62 genotypes according to their essential oil chemotypes. The high concentration of sesquiterpenes is supported by the higher number of TPS-a genes and their expression. The expansion for TPS sub-families in P. guajava occurred after the expansion of other rosids species. Providing insight into the origin of structural diversification and expansion in each clade of the TPS gene family within Myrtaceae. This study can provide insights into the diversity of genes for specialized metabolites such as terpenes, and their regulation, which can lead to a diverse chemotype of essential oil in different tissues and genotypes. This suggests a mode of enzymatic evolution that could lead to high sesquiterpene production, act as a chemical defense and contribute to the adaptive capacity of this species to different habitats.

Underutilized Canadian wild berries as potential sources of lipophilic bioactive compounds with antihypertensive properties.

Traditional berries are small fruits and are widely distributed in the Canadian prairies. The current study investigates the lipophilic bioactive compounds such as fatty acids, phytosterols, and terpenes, and their bioactivities, such as lipid peroxidation, as well as the antihypertensive activities of fourteen underutilized Canadian wild berries. These berries include Saskatoon berries (Amelanchier alnifolia), gooseberries (Ribes hirtellum), wild grapes (Vitis riparia), blackcurrants (Ribes nigrum), redcurrants (Ribes rubrum), haskap berries (Lonicera caerulea), wild raspberries (Rubus idaeus), wild blueberries (Vaccinium angustifolium), chokeberries (Aronia melanocarpa), buckthorn (Rhamnus cathartica), highbush cranberries (Viburnum trilobum), chokecherries (Prunus virginiana), nannyberries (Viburnum lentago) and snowberries (Symphoricarpos albus). The fatty acids, phytosterols, and terpenes were identified using Gas Chromatography-Mass Spectrometry (GC-MS). Lipid peroxidation and the antihypertensive activity assessed by measuring the berries' angiotensin converting enzyme 1 (ACE1) inhibitory activity were determined using in vitro methods. Notably, wild grapes exhibited the highest (p < 0.05) total fat content (7659 ± 312 μg per g DW), followed by haskap berries (4650 ± 184 μg per g DW). Polyunsaturated fatty acids (PUFAs) were highest (p < 0.05) in wild grapes (74%). Predominant phytosterols and terpenes identified in Canadian wild berries included β-sitosterol, isofucosterol, phytol, and α-amyrin. Saskatoon berries and gooseberries showed a distinct phytosterol and terpene profile compared to the other wild berries. Snowberries demonstrated the highest (p < 0.05) lipid peroxidation and the lowest (p < 0.05) angiotensin converting enzyme (ACE1) activity. This research provides valuable insights into the lipophilic bioactive compounds and their potential activities in vitro of the Canadian wild berries, offering a foundation for further exploration and potential applications in the context of nutraceuticals and functional foods.

Influence of prolonged maceration on phenolic compounds, volatile profile and sensory properties of wines from Minutolo and Verdeca, two Apulian white grape varieties

Prolonged maceration was applied to two Apulian white grape cultivars, Minutolo and Verdeca. The impact of prolonged maceration (up to six months) on the quality of the wines was evaluated through characterization of phenolic, volatile and sensory profiles. Macerated wines presented a peak of extraction after 4 months, followed by a decrease probably due to precipitation/reabsorption. Extraction of seed phenols occurred during longer maceration times. Volatile profile was also modified, showing a different behavior at different maceration times. Oxidation and aging compounds appeared during maceration, that are unusual for traditional white wines. Furthermore, maceration successfully enhanced varietal identity of Minutolo. The terpene profile of Minutolo wines changed during maceration, with a maximum of terpenols after 1 month of maceration, while longer maceration added a pattern of oxidized terpenes. Sensory analysis highlighted clearly distinct profiles of wines, according to both cultivar and maceration time. A good attitude was observed of both varieties to undergo maceration. In particular, Verdeca appeared suitable for longer maceration times, with flowery, citrus and apricot notes after 4 months, while enhanced apricot notes were perceived after 6 months. On the other hand, Minutolo wine after 6 months of maceration was characterized by a marked astringency and peach flavor.