Metabolic engineering-induced transcriptome reprogramming of lipid biosynthesis enhances oil composition in oat.

The endeavour to elevate the nutritional value of oat (Avena sativa) by altering the oil composition and content positions it as an optimal crop for fostering human health and animal feed. However, optimization of oil traits on oat through conventional breeding is challenging due to its quantitative nature and complexity of the oat genome. We introduced two constructs containing three key genes integral to lipid biosynthesis and/or regulatory pathways from Arabidopsis (AtWRI1 and AtDGAT1) and Sesame (SiOLEOSIN) into the oat cultivar 'Park' to modify the fatty acid composition. Four homozygous transgenic lines were generated with a transformation frequency of 7%. The expression of these introduced genes initiated a comprehensive transcriptional reprogramming in oat grains and leaves. Notably, endogenous DGAT, WRI1 and OLEOSIN genes experienced upregulation, while genes associated with fatty acid biosynthesis, such as KASII, SACPD and FAD2, displayed antagonistic expression patterns between oat grains and leaves. Transcriptomic analyses highlighted significant differential gene expression, particularly enriched in lipid metabolism. Comparing the transgenic oat plants with the wild type, we observed a remarkable increase of up to 34% in oleic acid content in oat grains. Furthermore, there were marked improvements in the total oil content in oat leaves, as well as primary metabolites changes in both oat grains and leaves, while maintaining homeostasis in the transgenic oat plants. These findings underscore the effectiveness of genetic engineering in manipulating oat oil composition and content, offering promising implications for human consumption and animal feeding through oat crop improvement programmes.

Germicidal efficacy of continuous and pulsed ultraviolet-C radiation on pathogen models and SARS-CoV-2.

Ultraviolet radiation's germicidal efficacy depends on several parameters, including wavelength, radiant exposure, microbial physiology, biological matrices, and surfaces. In this work, several ultraviolet radiation sources (a low-pressure mercury lamp, a KrCl excimer, and four UV LEDs) emitting continuous or pulsed irradiation were compared. The greatest log reductions in E. coli cells and B. subtilis endospores were 4.1 ± 0.2 (18 mJ cm-2) and 4.5 ± 0.1 (42 mJ cm-2) with continuous 222 nm, respectively. The highest MS2 log reduction observed was 2.7 ± 0.1 (277 nm at 3809 mJ cm-2). Log reductions of SARS-CoV-2 with continuous 222 nm and 277 nm were ≥ 3.4 ± 0.7, with 13.3 mJ cm-2 and 60 mJ cm-2, respectively. There was no statistical difference between continuous and pulsed irradiation (0.83-16.7% [222 nm and 277 nm] or 0.83-20% [280 nm] duty rates) on E. coli inactivation. Pulsed 260 nm radiation (0.5% duty rate) at 260 nm yielded significantly greater log reduction for both bacteria than continuous 260 nm radiation. There was no statistical difference in SARS-CoV-2 inactivation between continuous and pulsed 222 nm UV-C radiation and pulsed 277 nm radiation demonstrated greater germicidal efficacy than continuous 277 nm radiation. Greater radiant exposure for all radiation sources was required to inactivate MS2 bacteriophage. Findings demonstrate that pulsed irradiation could be more useful than continuous UV radiation in human-occupied spaces, but threshold limit values should be respected. Pathogen-specific sensitivities, experimental setup, and quantification methods for determining germicidal efficacy remain important factors when optimizing ultraviolet radiation for surface decontamination or other applications.

Cold Ethanol Extraction of Cannabinoids and Terpenes from Cannabis Using Response Surface Methodology: Optimization and Comparative Study.

Efficient cannabis biomass extraction can increase yield while reducing costs and minimizing waste. Cold ethanol extraction was evaluated to maximize yield and concentrations of cannabinoids and terpenes at different temperatures. Central composite rotatable design was used to optimize two independent factors: sample-to-solvent ratio (1:2.9 to 1:17.1) and extraction time (5.7 min-34.1 min). With response surface methodology, predicted optimal conditions at different extraction temperatures were a cannabis-to-ethanol ratio of 1:15 and a 10 min extraction time. With these conditions, yields (g 100 g dry matter-1) were 18.2, 19.7, and 18.5 for -20 °C, -40 °C and room temperature, respectively. Compared to the reference ground sample, tetrahydrocannabinolic acid changed from 17.9 (g 100 g dry matter-1) to 15, 17.5, and 18.3 with an extraction efficiency of 83.6%, 97.7%, 102.1% for -20 °C, -40 °C, and room temperature, respectively. Terpene content decreased by 54.1% and 32.2% for extraction at -20 °C and room temperature, respectively, compared to extraction at -40 °C. Principal component analysis showed that principal component 1 and principal component 2 account for 88% and 7.31% of total variance, respectively, although no significant differences in cold ethanol extraction at different temperatures were observed.

Microwave- and Ultrasound-Assisted Extraction of Cannabinoids and Terpenes from Cannabis Using Response Surface Methodology.

Limited studies have explored different extraction techniques that improve cannabis extraction with scale-up potential. Ultrasound-assisted and microwave-assisted extraction were evaluated to maximize the yield and concentration of cannabinoids and terpenes. A central composite rotatable design was used to optimize independent factors (sample-to-solvent ratio, extraction time, extraction temperature, and duty cycle). The optimal conditions for ultrasound- and microwave-assisted extraction were the sample-to-solvent ratios of 1:15 and 1:14.4, respectively, for 30 min at 60 °C. Ultrasound-assisted extraction yielded 14.4% and 14.2% more oil and terpenes, respectively, compared with microwave-assisted extracts. Ultrasound-assisted extraction increased cannabinoid concentration from 13.2−39.2%. Considering reference ground samples, tetrahydrocannabinolic acid increased from 17.9 (g 100 g dry matter−1) to 28.5 and 20 with extraction efficiencies of 159.2% and 111.4% for ultrasound-assisted and microwave-assisted extraction, respectively. Principal component analyses indicate that the first two principal components accounted for 96.6% of the total variance (PC1 = 93.2% and PC2 = 3.4%) for ultrasound-assisted extraction and 92.4% of the total variance (PC1 = 85.4% and PC2 = 7%) for microwave-assisted extraction. Sample-to-solvent ratios significantly (p < 0.05) influenced the secondary metabolite profiles and yields for ultrasound-assisted extracts, but not microwave-assisted extracts.

Development of a Nuclear Magnetic Resonance Method and a Near Infrared Calibration Model for the Rapid Determination of Lipid Content in the Field Pea (Pisum sativum).

Pisum sativum is a leguminous crop suitable for cultivation worldwide. It is used as a forage or dried seed supplement in animal feed and, more recently, as a potential non-traditional oilseed. This study aimed to develop a low-cost, rapid, and non-destructive method for analyzing pea lipids with no chemical modifications that would prove superior to existing destructive solvent extraction methods. Different pea accession seed samples, prepared as either small portions (0.5 mm2) of endosperm or ground pea seed powder for comparison, were subjected to HR-MAS NMR analyses and whole seed samples underwent NIR analyses. The total lipid content ranged between 0.57-3.45% and 1.3-2.6% with NMR and NIR, respectively. Compared to traditional extraction with butanol, hexane-isopropanol, and petroleum ether, correlation coefficients were 0.77 (R2 = 0.60), 0.56 (R2 = 0.47), and 0.78 (R2 = 0.62), respectively. Correlation coefficients for NMR compared to traditional extraction increased to 0.97 (R2 = 0.99) with appropriate correction factors. PLS regression analyses confirmed the application of this technology for rapid lipid content determination, with trends fitting models often close to an R2 of 0.95. A better robust NIR quantification model can be developed by increasing the number of samples with more diversity.

A comprehensive study on the effect of light quality imparted by light-emitting diodes (LEDs) on the physiological and biochemical properties of the microalgal consortia of Chlorella variabilis and Scenedesmus obliquus cultivated in dairy wastewater.

The effect of light wavelengths on the physiological, biochemical and lutein content of the microalgal consortia Chlorella variabilis and Scenedesmus obliquus was evaluated using different light sources. Among different light treatments, cool-white fluorescent light produced the highest biomass of 673 mg L-1 with a specific growth rate of 0.75 day-1 followed by blue (500 mg L-1; 0.73 day-1). The chlorophyll content was enhanced under blue light (10.7 mg L-1) followed by cool fluorescent light (9.3 mg L-1), whereas the lutein productivity was enhanced under cool fluorescent light (7.22 mg g-1). Protein content of the microalgal consortia was enhanced under all light treatments with the highest protein accumulation under cool-white fluorescent light (~56% of dry mass) closely followed by amber light (52% of dry mass), whereas the carbohydrate content was higher under amber light (~35% of dry mass). The results revealed that the consortia could grow well on diluted dairy wastewater thereby reducing the cost of algal production when compared with the use of inorganic media and a two-phase culture process utilizing cool fluorescent and amber light could be employed for maximizing algal biomass and nutrient composition with enhanced lutein production. The study also emphasizes on the economic efficiency of LED lights in terms of biomass produced based on the modest electricity consumed and the importance of using amber light for cultivating microalgae for its nutrient content which has seldom been studied.

Proteome modifications on tomato under extreme high light induced-stress.

BACKGROUND: Abiotic stress reduces photosynthetic yield and plant growth, negatively impacting global crop production and is a major constraint faced by agriculture. However, the knowledge on the impact on plants under extremely high irradiance is limited. We present the first in-depth proteomics analysis of plants treated with a method developed by our research group to generate a light gradient using an extremely intense light. METHODS: The method consists of utilizing light emitting diodes (LED) to create a single spot at 24,000 µmol m- 2 s- 1 irradiance, generating three light stress levels. A light map and temperature profile were obtained during the light experiment. The proteins expressed in the treated tomato (Solanum lycopersicum, Heinz H1706) leaves were harvested 10 days after the treatment, allowing for the detection of proteins involved in a long-term recovery. A multiplex labeled proteomics method (iTRAQ) was analyzed by LC-MS/MS. RESULTS: A total of 3994 proteins were identified at 1% false discovery rate and matched additional quality filters. Hierarchical clustering analysis resulted in four types of patterns related to the protein expression, with one being directly linked to the increased LED irradiation. A total of 37 proteins were found unique to the least damaged leaf zone, while the medium damaged zone had 372 proteins, and the severely damaged presented unique 1003 proteins. Oxygen evolving complex and PSII complex proteins (PsbH, PsbS, PsbR and Psb28) were found to be abundant in the most damaged leaf zone. This leaf zone presented a protein involved in the salicylic acid response, while it was not abundant in the other leaf zones. The mRNA level of PsbR was significantly lower (1-fold) compared the control in the most damaged zone of the leaf, while Psb28 and PsbH were lower (1-fold) in the less damaged leaf zones. PsbS mRNA abundance in all leaf zones tested presented no statistically significant change from the control. CONCLUSIONS: We present the first characterization of the proteome changes caused by an extreme level of high-light intensity (24,000 µmol m- 2 s- 1). The proteomics results show the presence of specific defense responses to each level of light intensity, with a possible involvement of proteins PsbH, Psb28, PsbR, and PsbS.

Photobiology eye safety for horticultural LED lighting: Transmittance performance of eyewear protection using high-irradiant monochromatic LEDs.

Light emitting diodes have slowly gained market share as horticultural lighting systems in greenhouses due to their rapid improvement in color performances and light outputs. These advancements have increased the availability of the full spectrum of visible wavelengths and the corresponding irradiance outputs available to plants. However, light emitting diodes owners have limited information on the proper options for personal eyewear protection as the irradiance levels have increased. The objective of this study was to measure the light transmittance performance of 12 eyewear protection including welding goggles, safety goggles, polarized glasses, and sunglasses across the human visible spectrum (380-740 nm) up to an irradiance level of 1500 W·m-2 from high-irradiant light emitting diodes assemblies. Based on the spectral measurements, certain transmitted spectra exhibited spectrum shifts or an alteration in the bimodal distribution which were different than the light emitting diodes spectra, due to the uneven transmittance efficiencies of the glasses. As for the measured transmittance percentages in two experiments, each type of eyewear protection showed distinct transmittance performances, and the performance of the tested eyewear protection was not impacted by irradiance but was dependent on the wavelength. The mean light transmittance was 1.77% for the welding glasses, 13.12% for the polarized glasses, 15.27% for the safety goggles, and 27.65% for the sunglasses. According to these measured results and the spectral weighting exposure limits from the International Electrotechnical Commission 62471 and EU directive 2006/25, consumers and workers using horticultural lighting can select welding goggles or polarized glasses, to limit the possible ocular impact of the high irradiance of monochromatic light in electrical lighting environment. Sunglasses and safety goggles would not be advised as protection, especially if infrared radiation was used.

Comparative Shotgun Proteomic Analysis of Wastewater-Cultured Microalgae: Nitrogen Sensing and Carbon Fixation for Growth and Nutrient Removal in Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii was batch-cultured for 12 days under continuous illumination to investigate nitrogen uptake and metabolic responses to wastewater processing. Our approach compared two conditions: (1) artificial wastewater containing nitrate and ammonia and (2) nutrient-sufficient control containing nitrate as sole form of nitrogen. Treatments did not differ in final biomass; however, comparison of group proteomes revealed significant differences. Label-free shotgun proteomic analysis identified 2358 proteins, of which 92 were significantly differentially abundant. Wastewater cells showed higher relative abundances of photosynthetic antenna proteins, enzymes related to carbon fixation, and biosynthesis of amino acids and secondary metabolites. Control cells showed higher abundances of enzymes and proteins related to nitrogen metabolism and assimilation, synthesis and utilization of starch, amino acid recycling, evidence of oxidative stress, and little lipid biosynthesis. This study of the eukaryotic microalgal proteome response to nitrogen source, availability, and switching highlights tightly controlled pathways essential to the maintenance of culture health and productivity in concert with light absorption and carbon assimilation. Enriched pathways in artificial wastewater, notably, photosynthetic carbon fixation and biosynthesis of plant hormones, and those in nitrate only control, most notably, nitrogen, amino acid, and starch metabolism, represent potential targets for genetic improvement requiring targeted elucidation.

Environmental Impact of Outdoor Cannabis Production.

Environmental impacts of cannabis production are of increasing concern because it is a newly legal and growing industry. Although a handful of studies have quantified the impacts of indoor production, very little is known about the impact of outdoor cannabis agriculture. Outdoor production typically uses little direct energy but can require significant fertilizer and other inputs due to dissipative losses via runoff and mineralization. Conversely, fertilizer high in nitrogen can be counterproductive, as it produces flowers with decreased cannabinoid content. This study has two aims: (1) To identify reduced-fertilizer regimes that provide optimal cannabis flower yields with reduced inputs and (2) to quantify how this shifts greenhouse gas emissions, resource depletion (fossil and metal), terrestrial acidification, and the eutrophication potential of outdoor cannabis production. Primary data from a fertilizer response trial are incorporated into a life-cycle assessment model. Results show that outdoor cannabis agriculture can be 50 times less carbon-emitting than indoor production. Dissemination of this knowledge is of utmost importance for producers, consumers, and government officials in nations that have either legalized or will legalize cannabis production.

The use of LEDs for the stomatal response, light compensation points, and storage of spinach and kale.

The spectral composition of some light-emitting diodes (LEDs) reportedly results in higher crop yield, prevents wilting, and reduces thermal damage to plants. The use of LEDs for postharvest storage and shelf-life extension has been limited, but the potential of this technology will allow for greater applications in horticulture and the food industry. In this experiment, 'Winterbor' kale (Brassica oleracea) and 'Melody' spinach (Spinacia oleracea) plants were measured for the light compensation point and stomatal response under 14 different wavelengths of light ranging from 405 to 661 nm. Data collected from these measurements were used to select two different wavelengths of LEDs and determine the proper irradiance levels for an LED irradiance storage test on spinach and kale. Treatments comprising blue, red, and amber lights were effective at increasing the stomatal opening, while the green light resulted in reduced stomatal opening. For spinach, the light response curve showed that light compensation points at 500 nm and 560 nm were 65.3 and 64.7 µmol m-2 s-1, respectively. For kale, the light compensation points at 500 nm and 560 nm were 50.8 and 44.1 µmol m-2 s-1, respectively. For the storage test experiment at room temperature, kale and spinach were stored under four different treatments: dark treatment (control), standard white fluorescent light, 500 nm, and 560 nm LED wavelengths. For spinach, the moisture content was 70.1% at 560 nm and 53.7% for dark, moisture losses of 41.5% under the 560-nm treatment and 52.0% for the dark treatment. The fresh basis moisture content was 74.6% at 560 nm and 59.3% in the dark. Moisture loss under the 560 nm treatment was 39.6% while the dark treatment had a 54.0% moisture loss. A visual assessment scale was monitored, 560 nm resulted in the top visual quality for kale compared to the other treatments with the lowest visual quality under the dark treatment at day 4. For spinach, the visual quality for 560 nm treatment was statistically the standard white fluorescent light and 500 nm, with poor-quality product occurring by day 4 and the lowest-quality product occurring at day 5. The LED treatments improved the shelf life of spinach and kale, likely as a result of stomatal aperture closure, photosynthetic rate near the light compensation point and stability of the atmospheric moisture content. This study provides valuable information on the extension of the shelf life of leafy greens during storage. Reducing fresh produce waste in grocery stores will increase revenue, thereby benefiting the Canadian economy while providing social and environmental benefits that entail increased food security and reduced food waste.

Light emitting diode effect of red, blue, and amber light on photosynthesis and plant growth parameters.

The visible light spectrum (400-700 nm) powers plant photosynthesis and innumerable other biological processes. Photosynthesis curves plotted by pioneering photobiologists show that amber light (590-620 nm) induces the highest photosynthetic rates in this spectrum. Yet, both red and blue light are viewed superior in their influence over plant growth. Here we report two approaches for quantifying how light wavelength photosynthesis and plant growth using light emitting diodes (LEDs). Resolved quantum yield spectra of tomato and lettuce plants resemble those acquired earlier, showing high quantum utilization efficiencies in the 420-430 nm and 590-620 nm regions. Tomato plants grown under blue (445 nm), amber (595 nm), red (635 nm), and combined red-blue-amber light for 14 days show that amber light yields higher fresh and dry mass, by at least 20%. Principle component analysis shows that amber light has a more pronounced and direct effect on fresh mass, whereas red light has a major effect on dry mass. These data clarify amber light's primary role in photosynthesis and suggest that bandwidth determines plant growth and productivity under sole amber lighting. Findings set precedence for future work aimed at maximizing plant productivity, with widespread implications for controlled environment agriculture.

Separate Effects of Sodium on Germination in Saline-Sodic and Alkaline Forms at Different Concentrations.

Salinity negatively impacts crop productivity, yet neutral and alkali salt stresses are not often differentiated. To investigate these abiotic stresses separately, saline and alkaline solutions with identical concentrations of sodium (12 mM, 24 mM and 49 mM) were used to compare the seed germination, viability and biomass of four crop species. Commercial buffers containing NaOH were diluted to generate alkaline solutions. The sodic solutions tested contained the neutral salt NaCl. Romaine lettuce, tomato, beet, and radish were seeded and grown hydroponically for 14 days. A rapid germination was observed for alkaline solutions when compared to saline-sodic solutions. The highest plant viability recorded (90.0%) was for the alkaline solution, containing 12 mM Na+, and for the control treatment. Plant viability, with a value of 49 mM Na+ in saline-sodic and alkaline solutions, was the lowest (50.0% and 40.8% respectively), and tomato plants did not germinate. EC values were higher for the saline-sodic solutions than the alkaline solutions, yielding greater fresh mass per plant for all species, with the exception of beets grown in alkaline solution, with a value of 24 mM Na+. The fresh mass of romaine lettuce grown in the 24 mM Na+ saline-sodic solution was significantly greater than romaine lettuce grown in the alkaline solution with the same sodium concentration.

Effect of Amber (595 nm) Light Supplemented with Narrow Blue (430 nm) Light on Tomato Biomass.

Full-spectrum light-emitting diodes (LEDs) mainly comprising 460-nm + 595-nm light are becoming a mainstay in the horticulture industry, and recent studies indicate that plant productivity under white LEDs is higher than combined blue and red LED lighting. Different light properties (wavelength and bandwidth) in full-spectrum light, particularly for the blue and amber light regions, have only partly been explored. This research aimed to characterize the effects of amber + blue light wavelengths and bandwidths on tomato (Solanum lycopersicum cv. Beefsteak) growth, morphology, and production efficiency. Tomato seedlings were subjected to four different light treatments for 60 days: narrow amber light (595 nm), narrow blue + narrow amber light (430 nm + 595 nm) with a 1:10 ratio, white LED (455 nm + 595 nm), and a high-pressure sodium (HPS) lamp (control). The highest mean fresh mass yield occurred with the narrow blue + narrow amber light (479 g), followed by white LED at 20% less, HPS at 34% less, and narrow amber at 40% less. Dry mass and plant height were similar among light treatments. Supplementing narrow amber light with 430-nm blue light led to a 20% increase in chlorophyll content. Findings indicate that narrow amber light is more efficient in biomass accumulation than broad amber light and that precise selection of different blue and amber wavelengths can greatly impact the growth and development of tomato seedlings. This energy-efficient narrow-wavelength combination shows improvement over white LED lighting for maximizing tomato growth.

Eff-3DPSeg: 3D Organ-Level Plant Shoot Segmentation Using Annotation-Efficient Deep Learning.

Reliable and automated 3-dimensional (3D) plant shoot segmentation is a core prerequisite for the extraction of plant phenotypic traits at the organ level. Combining deep learning and point clouds can provide effective ways to address the challenge. However, fully supervised deep learning methods require datasets to be point-wise annotated, which is extremely expensive and time-consuming. In our work, we proposed a novel weakly supervised framework, Eff-3DPSeg, for 3D plant shoot segmentation. First, high-resolution point clouds of soybean were reconstructed using a low-cost photogrammetry system, and the Meshlab-based Plant Annotator was developed for plant point cloud annotation. Second, a weakly supervised deep learning method was proposed for plant organ segmentation. The method contained (a) pretraining a self-supervised network using Viewpoint Bottleneck loss to learn meaningful intrinsic structure representation from the raw point clouds and (b) fine-tuning the pretrained model with about only 0.5% points being annotated to implement plant organ segmentation. After, 3 phenotypic traits (stem diameter, leaf width, and leaf length) were extracted. To test the generality of the proposed method, the public dataset Pheno4D was included in this study. Experimental results showed that the weakly supervised network obtained similar segmentation performance compared with the fully supervised setting. Our method achieved 95.1%, 96.6%, 95.8%, and 92.2% in the precision, recall, F1 score, and mIoU for stem-leaf segmentation for the soybean dataset and 53%, 62.8%, and 70.3% in the AP, AP@25, and AP@50 for leaf instance segmentation for the Pheno4D dataset. This study provides an effective way for characterizing 3D plant architecture, which will become useful for plant breeders to enhance selection processes. The trained networks are available at https://github.com/jieyi-one/EFF-3DPSEG.

Relationship between Total Antioxidant Capacity, Cannabinoids and Terpenoids in Hops and Cannabis.

Efficient determination of antioxidant activity in medicinal plants may provide added value to extracts. The effects of postharvest pre-freezing and drying [microwave-assisted hot air (MAHD) and freeze drying] on hops and cannabis were evaluated to determine the relationship between antioxidant activity and secondary metabolites. The 2,2-diphenyl-1-picrylhydrazine (DPPH) reduction and ferric reducing ability of power (FRAP) assays were assessed for suitability in estimating the antioxidant activity of extracted hops and cannabis inflorescences and correlation with cannabinoid and terpene content. Antioxidant activity in extracts obtained from fresh, undried samples amounted to 3.6 Trolox equivalent antioxidant activity (TEAC) (M) dry matter-1 and 2.32 FRAP (M) dry matter-1 for hops, in addition to 2.29 TEAC (M) dry matter-1 and 0.25 FRAP (M) dry matter-1 for cannabis. Pre-freezing significantly increased antioxidant values by 13% (DPPH) and 29.9% (FRAP) for hops, and by 7.7% (DPPH) and 19.4% (FRAP) for cannabis. ANOVA analyses showed a significant (p < 0.05) increase in total THC (24.2) and THCA (27.2) concentrations (g 100 g dry matter-1) in pre-frozen, undried samples compared to fresh, undried samples. Freeze-drying and MAHD significantly (p < 0.05) reduced antioxidant activity in hops by 79% and 80.2% [DPPH], respectively and 70.1% and 70.4% [FRAP], respectively, when compared to antioxidant activity in extracts obtained from pre-frozen, undried hops. DPPH assay showed that both freeze-drying and MAHD significantly (p < 0.05) reduced the antioxidant activity of cannabis by 60.5% compared to the pre-frozen samples although, there was no significant (p < 0.05) reduction in the antioxidant activity using the FRAP method. Greater THC content was measured in MAHD-samples when compared to fresh, undried (64.7%) and pre-frozen, undried (57%), likely because of decarboxylation. Both drying systems showed a significant loss in total terpene concentration, yet freeze-drying has a higher metabolite retention compared to MAHD. These results may prove useful for future experiments investigating antioxidant activity and added value to cannabis and hops.

Temporal analysis of xylose fermentation by Scheffersomyces stipitis using shotgun proteomics.

Proteomics and fermentation technology have begun to integrate to investigate fermentation organisms in bioprocess development. This is the first shotgun proteomics study employed to monitor the proteomes of Scheffersomyces stipitis during xylose fermentation under oxygen limitation. We identified 958 nonredundant proteins and observed highly similar proteomes from exponential to early stationary phases. In analyzing the temporal proteome, we identified unique expression patterns in biological processes and metabolic pathways, including alternative respiration salicylhydroxamic acid (SHAM) pathway, activation of glyoxylate cycle, expression of galactose enzymes, and secondary zinc-containing alcohol dehydrogenase and O-glycosyl hydrolases. We identified the expression of a putative, high-affinity xylose sugar transporter Xut1p, but low-affinity xylose transporters were absent. Throughout cell growth, housekeeping processes included oxidative phosphorylation, glycolysis, nonoxidative branch of the pentose phosphate pathway, gluconeogenesis, biosynthesis of amino acids and aminoacyl total RNA (tRNA), protein synthesis and proteolysis, fatty acid metabolism, and cell division. This study emphasized qualitative analysis and demonstrated that shotgun proteomics is capable of monitoring S. stipitis fermentation and identifying physiological states, such as nutrient deficiency.

Shotgun proteomic monitoring of Clostridium acetobutylicum during stationary phase of butanol fermentation using xylose and comparison with the exponential phase.

Economically viable production of solvents through acetone-butanol-ethanol (ABE) fermentation requires a detailed understanding of Clostridium acetobutylicum. This study focuses on the proteomic profiling of C. acetobutylicum ATCC 824 from the stationary phase of ABE fermentation using xylose and compares with the exponential growth by shotgun proteomics approach. Comparative proteomic analysis revealed 22.9% of the C. acetobutylicum genome and 18.6% was found to be common in both exponential and stationary phases. The proteomic profile of C. acetobutylicum changed during the ABE fermentation such that 17 proteins were significantly differentially expressed between the two phases. Specifically, the expression of five proteins namely, CAC2873, CAP0164, CAP0165, CAC3298, and CAC1742 involved in the solvent production pathway were found to be significantly lower in the stationary phase compared to the exponential growth. Similarly, the expression of fucose isomerase (CAC2610), xylulose kinase (CAC2612), and a putative uncharacterized protein (CAC2611) involved in the xylose utilization pathway were also significantly lower in the stationary phase. These findings provide an insight into the metabolic behavior of C. acetobutylicum between different phases of ABE fermentation using xylose.

Highly Efficient and Reproducible Genetic Transformation in Pea for Targeted Trait Improvement.

A reproducible tissue culture protocol is required to establish an efficient genetic transformation system in highly recalcitrant pea genotypes. High-quality callus with superior regeneration ability was induced and regenerated on optimized media enriched with copper sulfate and cytokinins, 6-benzylaminopurine and indole-3-acetic acid. This successful regeneration effort led to the development of a highly efficient transformation system for five pea genotypes using immature and mature seeds. The new transformation protocol included the addition of elevated glucose and sucrose concentrations for cocultivation and inoculation media to improve callus induction and regeneration, thus resulting in consistent transformation frequencies. Using the Agrobacterium strain AGL1, a transformation frequency of up to 47% was obtained for the pea genotype Greenfeast, using either of two different selection marker genes, PAT or NPT, sourced from two different vectors. Sixty-two transgenic pea events were able to survive kanamycin and phosphinothricin selection. A total of 30 transgenic events for Greenfeast, 15 for CN 43016, 9 for snap pea, and 5 for CN 31237 are reported herein. Two additional transgenic events were recovered from particle gun bombardment experiments. Quantitative RT-PCR analysis confirmed the transgenic status of pea plants, indicating elevated expression of relevant genes cloned into the transformation constructs.

Light Quality Impacts Vertical Growth Rate, Phytochemical Yield and Cannabinoid Production Efficiency in Cannabis sativa.

Light is one of the most crucial parameters for enclosed cannabis (Cannabis sativa) production, as it highly influences growth, secondary metabolite production, and operational costs. The objective of this study was to investigate and evaluate the impact of six light spectra on C. sativa ('Babbas Erkle Cookies' accession) growth traits and secondary metabolite (cannabinoid and terpene) profiles. The light spectra evaluated included blue (430 nm), red (630 nm), rose (430 + 630 nm, ratio 1:10), purple (430 + 630 nm, ratio 2:1), and amber (595 nm) LED treatments, in addition to a high-pressure sodium (HPS, amber-rich light) treatment as a control. All the LED light treatments had lower fresh mean inflorescence mass than the control (HPS, 133.59 g plant-1), and monochromatic blue light yielded the least fresh inflorescence mass (76.39 g plant-1). Measurement of Δ9-tetrahydrocannabinol (THC) concentration (%) and total yield (g plant-1) showed how inflorescence mass and THC concentration need to be analyzed conjointly. Blue treatment resulted in the highest THC concentration (10.17% m/m), yet the lowest THC concentration per plant (1.44 g plant-1). The highest THC concentration per plant was achieved with HPS (2.54 g plant-1). As with THC, blue light increased cannabigerol (CBG) and terpene concentration. Conversely, blue light had a lesser impact on cannabidiol (CBD) biosynthesis in this C. sativa chemotype. As the combined effects of the light spectrum on both growth traits and secondary metabolites have important ramifications for the industry, the inappropriate spectral design could cause a reduction in cannabinoid production (20-40%). These findings show promise in helping producers choose spectral designs that meet specific C. sativa production goals.