Variation and correlation of properties in different grades of maple syrup.

Thirty five commercial maple syrups from twelve producers in Southern Ontario were evaluated for properties including light transmittance, autofluorescence, density, pH, total soluble solids (TSS), glucose and fructose content, total phenol content, antioxidant potential and mineral content (Mg, Mn, P, Zn, Ca, K, Fe and Pb). A high degree of variability was found in many characteristics, often exceeding an order of magnitude. Syrups were categorized based on light transmission at 560 nm into amber (12), dark (13) and very dark (10) using International Maple Syrup Institute (IMSI) guidelines. No statistical differences were found among grades of syrup for density, pH, TSS, glucose, fructose, total reducing sugars, glucose:fructose ratio, magnesium, manganese or potassium. Darker syrups showed significantly higher autofluorescence, total phenol content, antioxidant potential, phosphorous, calcium and total mineral content. Significant negative correlations of percent transmission with total phenol content, antioxidant potential and total mineral content are reported. Significant positive correlations among total phenol content, antioxidant potential and total mineral content are also described. The results from this study suggest that darker syrups tend to contain more beneficial traits and may be applied in developing functional foods and value added products.

Foliar Symptomology, Nutrient Content, Yield, and Secondary Metabolite Variability of Cannabis Grown Hydroponically with Different Single-Element Nutrient Deficiencies.

In controlled environment production systems, Cannabis sativa (hereafter cannabis) is a commodity with high nutrient demands due to prolific growth under optimized environmental conditions. Since nutrient deficiencies can reduce yield and quality, cultivators need tools to rapidly detect and evaluate deficiency symptoms so corrective actions can be taken quickly to minimize losses. We grew cannabis plants in solution culture with different individual nutrient elements withheld from the solutions to identify deficiency symptoms. Control plants received a complete nutrient recipe, whereas the following single elements were withheld from the respective nutrient deficiency treatments: N, P, K, Ca, Mg, S, Fe, and Mn. The nutrient treatments began when the photoperiod was switched to a 12/12 h (light/dark), and plants were grown to commercial maturity. Plants were monitored daily, and the development of visual deficiency symptoms were recorded. Photographs of each plant were taken weekly. Upon the onset of visual deficiency symptoms, both upper- and lower-canopy foliage were analyzed for nutrient element concentrations. At harvest, plants were evaluated for biomass partitioning, and the cannabinoid composition of inflorescence tissues. This manuscript describes the onset and progression of nutrient deficiency symptoms (with pictures), relates symptomology to foliar nutrient analyses, and contextualizes the relationships between nutrient deficiencies and cannabis growth, yield, and quality. Aboveground vegetative fresh weights were reduced by 73% in the -N treatment and 59% in the -P treatment, compared with the control. All deficiency treatments except for -Fe and -Mn had floral yields reduced by between 33% to 72%, compared with the control. Overall, deficiencies of individual nutrients can substantially reduce vegetative growth and inflorescence yield, although only minor effects were observed in secondary metabolite composition. The onset of individual deficiency symptoms did not always correspond with elemental analysis of foliar tissues. Cultivators should take an integrated approach in diagnosing nutrient deficiencies and take timely corrective actions to optimize productivity and minimize losses to yield and quality.

Understanding and Rebalancing: A Rapid Scoping Review of Cannabis Research Among Indigenous People.

Evidence-based perspectives on and patterns of cannabis use are vital to addressing ethical, legal, and regulatory controversies, but have not yet been mapped for Indigenous people. We searched five databases and used a rapid scoping review methodology to analyze empirical studies with a primary focus on cannabis and Indigenous peoples. Studies were examined for year of publication, origin of study and author groups, methods, and thematic foci. We analyzed 68 studies with publication dates between1983 and 2022. Approximately 90% of articles were written by authors in the same geographic location as the study population. Seventy-one percent (71%) of the articles were written by authors of multiple articles. Four articles acknowledged author Indigeneity. None contained author positionality statements. The majority of studies utilized mixed methods that integrated both qualitative and quantitative components. Two major categories of focus that emerged from the analysis are substance use disorders and prevalence rates (n=35) and predictors of and motivators for use (n=27), together representing the majority of articles (n=52/68). Impact on mental health (n=6), treatment, and management of cannabis use disorder (CUD) (n=3), legalization and criminalization (n=2), genomic heritability and dependence (n=2), and economics of cannabis use (n=1) were the focus of the remaining articles in the sample. Mixed methods empirical research largely focuses on risks of cannabis use among Indigenous people worldwide. The small, repeating pool of senior authors represents an opportunity for capacity building. A lack of transparency about author positionality and absence of empirical studies that explore the lived experiences of Indigenous peoples and cannabis are significant gaps poised to be filled for future research and regulation.

Inhibition of phenylpropanoid biosynthesis increases cell wall digestibility, protoplast isolation, and facilitates sustained cell division in American elm (Ulmus americana).

BACKGROUND: Protoplast technologies offer unique opportunities for fundamental research and to develop novel germplasm through somatic hybridization, organelle transfer, protoclonal variation, and direct insertion of DNA. Applying protoplast technologies to develop Dutch elm disease resistant American elms (Ulmus americana L.) was proposed over 30 years ago, but has not been achieved. A primary factor restricting protoplast technology to American elm is the resistance of the cell walls to enzymatic degradation and a long lag phase prior to cell wall re-synthesis and cell division. RESULTS: This study suggests that resistance to enzymatic degradation in American elm was due to water soluble phenylpropanoids. Incubating tobacco (Nicotiana tabacum L.) leaf tissue, an easily digestible species, in aqueous elm extract inhibits cell wall digestion in a dose dependent manner. This can be mimicked by p-coumaric or ferulic acid, phenylpropanoids known to re-enforce cell walls. Culturing American elm tissue in the presence of 2-aminoindane-2-phosphonic acid (AIP; 10-150 µM), an inhibitor of phenylalanine ammonia lyase (PAL), reduced flavonoid content, decreased tissue browning, and increased isolation rates significantly from 11.8% (±3.27) in controls to 65.3% (±4.60). Protoplasts isolated from callus grown in 100 µM AIP developed cell walls by day 2, had a division rate of 28.5% (±3.59) by day 6, and proliferated into callus by day 14. Heterokaryons were successfully produced using electrofusion and fused protoplasts remained viable when embedded in agarose. CONCLUSIONS: This study describes a novel approach of modifying phenylpropanoid biosynthesis to facilitate efficient protoplast isolation which has historically been problematic for American elm. This isolation system has facilitated recovery of viable protoplasts capable of rapid cell wall re-synthesis and sustained cell division to form callus. Further, isolated protoplasts survived electrofusion and viable heterokaryons were produced. Together, these results provide the first evidence of sustained cell division, callus regeneration, and potential application of somatic cell fusion in American elm, suggesting that this source of protoplasts may be ideal for genetic manipulation of this species. The technological advance made with American elm in this study has potential implications in other woody species for fundamental and applied research which require availability of viable protoplasts.

Mutualism breakdown in breadfruit domestication.

During the process of plant domestication, below-ground communities are rarely considered. Some studies have attempted to understand the changes in root symbionts owing to domestication, but little is known about how it influences mycorrhizal response in domesticated crops. We hypothesized that selection for above-ground traits may also result in decreased mycorrhizal abundance in roots. Breadfruit (Artocarpus sp.) has a long domestication history, with a strong geographical movement of cultivars from west to east across the Melanesian and Polynesian islands. Our results clearly show a decrease in arbuscular mycorrhizas (AMs) along a domestication gradient from wild to recently derived cultivars. We showed that the vesicular and arbuscular colonization rate decreased significantly in more recently derived breadfruit cultivars. In addition, molecular analyses of breadfruit roots indicated that AM fungal species richness also responded along the domestication gradient. These results suggest that human-driven selection for plant cultivars can have unintended effects on below-ground mutualists, with potential impacts on the stress tolerance of crops and long-term food security.

Indoor grown cannabis yield increased proportionally with light intensity, but ultraviolet radiation did not affect yield or cannabinoid content.

Cannabis (Cannabis sativa) flourishes under high light intensities (LI); making it an expensive commodity to grow in controlled environments, despite its high market value. It is commonly believed that cannabis secondary metabolite levels may be enhanced both by increasing LI and exposure to ultraviolet radiation (UV). However, the sparse scientific evidence is insufficient to guide cultivators for optimizing their lighting protocols. We explored the effects of LI and UV exposure on yield and secondary metabolite composition of a high Δ9-tetrahydrocannabinol (THC) cannabis cultivar 'Meridian'. Plants were grown under short day conditions for 45 days under average canopy photosynthetic photon flux densities (PPFD, 400-700 nm) of 600, 800, and 1,000 µmol m-2 s-1, provided by light emitting diodes (LEDs). Plants exposed to UV had PPFD of 600 µmol m-2 s-1 plus either (1) UVA; 50 µmol m-2 s-1 of UVA (315-400 nm) from 385 nm peak LEDs from 06:30 to 18:30 HR for 45 days or (2) UVA + UVB; a photon flux ratio of ≈1:1 of UVA and UVB (280-315 nm) from a fluorescent source at a photon flux density of 3.0 µmol m-2 s-1, provided daily from 13:30 to 18:30 HR during the last 20 days of the trial. All aboveground biomass metrics were 1.3-1.5 times higher in the highest vs. lowest PPFD treatments, except inflorescence dry weight - the most economically relevant parameter - which was 1.6 times higher. Plants in the highest vs. lowest PPFD treatment also allocated relatively more biomass to inflorescence tissues with a 7% higher harvest index. There were no UV treatment effects on aboveground biomass metrics. There were also no intensity or UV treatment effects on inflorescence cannabinoid concentrations. Sugar leaves (i.e., small leaves associated with inflorescences) of plants in the UVA + UVB treatment had ≈30% higher THC concentrations; however, UV did not have any effect on the total THC in thesefoliar tissues. Overall, high PPFD levels can substantially increase cannabis yield, but we found no commercially relevant benefits of adding UV to indoor cannabis production.

Improved in vitro rooting in liquid culture using a two piece scaffold system.

Plant tissue culture techniques have been used to propagate horticultural crops at a commercial scale for more than three decades. However, due to the high cost it is generally only used for high value crops. To increase production efficiency and make micropropagation viable for a wider range of species, new approaches to address key steps of the process with high labor inputs need to be evaluated. For this study, a two-piece scaffold system was designed, prototyped using 3D printing, and tested to physically hold plants upright thereby facilitating liquid based rooting. This system was evaluated with Malus domestica, Betula lenta, and Musa sp. using static liquid culture as well as rocker based temporary immersion system and compared to rooting in semi-solid based medium as is commonly practiced. Significantly, earlier rooting was observed in all three species in liquid when compared to semi-solid culture system, and plants cultured in liquid on the rocker generally performed better than those in static liquid. In addition to quicker, more uniform rooting, reducing labor requirements, and preventing root damage. This newly designed system is simple, easy to use, will help to improve efficiency, and reduce the cost of micropropagation.

Isolation and identification of mosquito biting deterrents from the North American mosquito repelling folk remedy plant, Matricaria discoidea DC.

Matricaria discoidea DC. (Asteraceae) has documented use as an insect repellent by Blackfoot Indians and other indigenous groups of North America. This investigation was conducted to evaluate this practice and systematically identify chemical constituents responsible for any insect repelling effect by utilizing a mosquito (Aedes aegypti (Linnaeus)) biting deterrent bioactivity-directed purification approach. Hydrodistilled oil from dried aerial parts of M. discoidea was the most bioactive crude extract generated and was as. Fractionation of this extract, followed by re-evaluation for mosquito biting deterrence using the K & D (Klun and Debboun) bioassay, produced many active fractions that were subsequently evaluated by spectroscopic techniques and the most active compounds were determined to be α-terpineol, spathulenol, and neryl isovalerate. A & K (Ali and Khan) large cage in vitro evaluation of pure compounds isolated from M. discoidea indicated α-terpineol to be the most active compound providing complete protection at 25 µg/cm2. This is the first report on the mosquito repellency of neryl isovalerate and scientific evidence reported here validates the traditional use of M. discoidea as a biting-insect deterrent.

Application of 3D printing to prototype and develop novel plant tissue culture systems.

BACKGROUND: Due to the complex process of designing and manufacturing new plant tissue culture vessels through conventional means there have been limited efforts to innovate improved designs. Further, development and availability of low cost, energy efficient LEDs of various spectra has made it a promising light source for plant growth in controlled environments. However, direct replacement of conventional lighting sources with LEDs does not address problems with uniformity, spectral control, or the challenges in conducting statistically valid experiments to assess the effects of light. Prototyping using 3D printing and LED based light sources could help overcome these limitations and lead to improved culture systems. RESULTS: A modular culture vessel design in which the fluence rate and spectrum of light are independently controlled was designed, prototyped using 3D printing, and evaluated for plant growth. This design is compatible with semi-solid and liquid based culture systems. Observations on morphology, chlorophyll content, and chlorophyll fluorescence based stress parameters from in vitro plants cultured under different light spectra with similar overall fluence rate indicated different responses in Nicotiana tabacum and Artemisia annua plantlets. This experiment validates the utility of 3D printing to design and test functional vessels and demonstrated that optimal light spectra for in vitro plant growth is species-specific. CONCLUSIONS: 3D printing was successfully used to prototype novel culture vessels with independently controlled variable fluence rate/spectra LED lighting. This system addresses several limitations associated with current lighting systems, providing more uniform lighting and allowing proper replication/randomization for experimental plant biology while increasing energy efficiency. A complete procedure including the design and prototyping of a culture vessel using 3D printing, commercial scale injection molding of the prototype, and conducting a properly replicated experiment are discussed. This open source design has the scope for further improvement and adaptation and demonstrates the power of 3D printing to improve the design of culture systems.

Isolation and Identification of Mosquito (Aedes aegypti) Biting-Deterrent Compounds from the Native American Ethnobotanical Remedy Plant Hierochloë odorata (Sweetgrass).

Hierochloë odorata (L.) P. Beauv. (Poaceae), commonly known as sweetgrass, has documented use as an insect repellent by the Flatheads of Montana and Blackfoot of Alberta. Both the Flatheads of Montana and Blackfoot of Alberta would use braided plant material in a sachet in clothing or burn them from one end as incense, air/clothing freshener, and insect repellent. This study evaluated the insect-repellent properties of this plant using an in vitro mosquito Aedes aegypti feeding bioassay-directed approach to identify the compound(s) responsible for the observed activities. Evaluation of crude extracts produced from H. odorata revealed that the hydrodistillate had the highest level of mosquito biting deterrence. Fractionation of this extract, followed by re-evaluation for mosquito biting deterrence, produced many active fractions, which were evaluated by spectroscopic techniques and determined to contain phytol, coumarin, and 2-methoxy-4-vinylphenol. Phytol and coumarin were both determined to be responsible for the Ae. aegypti biting deterrency. Scientific evidence reported here validates its traditional use as a biting-insect deterrent.

Quantification of pyrrolizidine alkaloids in North American plants and honey by LC-MS: single laboratory validation.

Pyrrolizidine alkaloids (PAs) are a class of naturally occurring compounds produced by many flowering plants around the World. Their presence as contaminants in food systems has become a significant concern in recent years. For example, PAs are often found as contaminants in honey through pollen transfer. A validated method was developed for the quantification of four pyrrolizidine alkaloids and one pyrrolizidine alkaloid N-oxide in plants and honey grown and produced in British Columbia. The method was optimised for extraction efficiency from the plant materials and then subjected to a single-laboratory validation to assess repeatability, accuracy, selectivity, LOD, LOQ and method linearity. The PA content in plants ranged from1.0 to 307.8 µg/g with repeatability precision between 3.8 and 20.8% RSD. HorRat values were within acceptable limits and ranged from 0.62 to 1.63 for plant material and 0.56-1.82 for honey samples. Method accuracy was determined through spike studies with recoveries ranging from 84.6 to 108.2% from the raw material negative control and from 82.1-106.0 % for the pyrrolizidine alkaloids in corn syrup. Based on the findings in this single-laboratory validation, this method is suitable for the quantitation of lycopsamine, senecionine, senecionine N-oxide, heliosupine and echimidine in common comfrey (Symphytum officinale), tansy ragwort (Senecio jacobaea), blueweed (Echium vulgare) and hound's tongue (Cynoglossum officinale) and for PA quantitation in honey and found that PA contaminants were present at low levels in BC honey.

Galanthamine, an anticholinesterase drug, effects plant growth and development in Artemisia tridentate Nutt. via modulation of auxin and neutrotransmitter signaling.

Galanthamine is a naturally occurring acetylcholinesterase (AchE) inhibitor that has been well established as a drug for treatment of mild to moderate Alzheimer disease, but the role of the compound in plant metabolism is not known. The current study was designed to investigate whether galanthamine could redirect morphogenesis of Artemisia tridentata Nutt. cultures by altering concentration of endogenous neurosignaling molecules acetylcholine (Ach), auxin (IAA), melatonin (Mel), and serotonin (5HT). Exposure of axenic A. tridentata cultures to 10 µM galanthamine decreased the concentration of endogenous Ach, IAA, MEL, and AchE, and altered plant growth in a manner reminiscent of 2-4D toxicity. Galanthamine itself demonstrated IAA activity in an oat coleotile elongation bioassay, 20 µM galanthamine showed no significant difference compared with 5 µM IAA or 5 µM 1-Naphthaleneacetic acid (NAA). Metabolomic analysis detected between 20,921 to 27,891 compounds in A. tridentata plantlets and showed greater commonality between control and 5 µM treatments. Furthermore, metabolomic analysis putatively identified coumarins scopoletin/isoscopoletin, and scopolin in A. tridentata leaf extracts and these metabolites linearly increased in response to galanthamine treatments. Overall, these data indicate that galanthamine is an allelopathic phytochemical and support the hypothesis that neurologically active compounds in plants help ensure plant survival and adaptation to environmental challenges.

Isolation and identification of mosquito (Aedes aegypti ) biting deterrent fatty acids from male inflorescences of breadfruit (Artocarpus altilis (Parkinson) Fosberg).

Dried male inflorescences of breadfruit ( Artocarpus altilis , Moraceae) are burned in communities throughout Oceania to repel flying insects, including mosquitoes. This study was conducted to identify chemicals responsible for mosquito deterrence. Various crude extracts were evaluated, and the most active, the hydrodistillate, was used for bioassay-guided fractionation. The hydrodistillate and all fractions displayed significant deterrent activity. Exploratory GC-MS analysis revealed more than 100 distinctive peaks, and more than 30 compounds were putatively identified, including a mixture of terpenes, aldehydes, fatty acids, and aromatics. A systematic bioassay-directed study using adult Aedes aegypti females identified capric, undecanoic, and lauric acid as primary deterrent constituents. A synthetic mixture of fatty acids present in the most active fraction and individual fatty acids were all significantly more active than N,N-diethyl-m-toluamide (DEET). These results provide support for this traditional practice and indicate the potential of male breadfruit flowers and fatty acids as mosquito repellents.