Studies on Cannabis sativa and Cannabinoids

A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Physiology and Metabolism".

Deadline for manuscript submissions: closed (20 February 2023) | Viewed by 75913

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TU Dortmund University, 44227 Dortmund, Germany
Interests: plant secondary metabolites; designer organisms; metabolic engineering; medicinal plants
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Special Issue Information

Dear Colleagues, 

In recent years, the worldwide hype of cannabis and cannabis-related products is observable. This cannabis hype on the one hand entailed broad cannabis-related research since many research teams all over the world stepped into this hot topic. Today, many different varieties are already sequenced and transcriptomic data are available. Together with metabolic studies, the biosynthetic routes and the genetics get more and more understood. What is still missing are well designed studies to elucidate the medicinal impact of single cannabinoids. So far, only a limited number of serious studies are available. To ensure a long-term success of cannabis in medicine, much more information is needed. On the other hand, there are some reasons one could be worried, since many trends, provoked by the global money glut, are alarming, such as the efforts and ideas to have cannabinoid and especially THC-containing soft drinks or even alcoholic drinks. This will inevitably lead to a new discussion regarding the legalization of cannabis as recreational drug. The delayed effect of cannabinoids after oral consumption, especially in combination with alcohol, will lead to many overdoses, not to mention the enormous risks of consuming cannabinoids, particularly during early adolescence. It is therefore of enormous importance that scientists from all over the world find answers and exploit the potential of cannabis for human health and not let this unique opportunity pass by. 

In this context, the aim of this Special Issue is to bring together new trends, developments, and especially research papers, in order to get closer to the goal of using cannabis for the benefit of human health.

Dr. Felix Stehle
Guest Editor

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Keywords

  • Cannabis sativa
  • Cannabinoids
  • Minor cannabinoids
  • Entourage effect
  • Metabolic engineering
  • Metabolomics
  • Transcriptomics
  • Gene regulation
  • Medicinal cannabis

Published Papers (16 papers)

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15 pages, 1853 KiB  
Article
Moving Away from 12:12; the Effect of Different Photoperiods on Biomass Yield and Cannabinoids in Medicinal Cannabis
by Tyson James Peterswald, Jos Cornelis Mieog, Razlin Azman Halimi, Nelson Joel Magner, Amy Trebilco, Tobias Kretzschmar and Sarah Jane Purdy
Plants 2023, 12(5), 1061; https://doi.org/10.3390/plants12051061 - 27 Feb 2023
Cited by 4 | Viewed by 6039
Abstract
The standard practice to initiate flowering in medicinal cannabis involves reducing the photoperiod from a long-day period to an equal duration cycle of 12 h light (12L)/12 h dark (12D). This method reflects the short-day flowering dependence of many cannabis varieties but may [...] Read more.
The standard practice to initiate flowering in medicinal cannabis involves reducing the photoperiod from a long-day period to an equal duration cycle of 12 h light (12L)/12 h dark (12D). This method reflects the short-day flowering dependence of many cannabis varieties but may not be optimal for all. We sought to identify the effect of nine different flowering photoperiod treatments on the biomass yield and cannabinoid concentration of three medicinal cannabis varieties. The first, “Cannatonic”, was a high cannabidiol (CBD)-accumulating line, whereas the other two, “Northern Lights” and “Hindu Kush”, were high Δ9-tetrahydrocannabinol (THC) accumulators. The nine treatments tested, following 18 days under 18 h light/6 h dark following cloning and propagation included a standard 12L:12D period, a shortened period of 10L:14D, and a lengthened period of 14L:10D. The other six treatments started in one of the aforementioned and then 28 days later (mid-way through flowering) were switched to one of the other treatments, thus causing either an increase of 2 or 4 h, or a decrease of 2 or 4 h. Measured parameters included the timing of reproductive development; the dry weight flower yield; and the % dry weight of the main target cannabinoids, CBD and THC, from which the total g cannabinoid per plant was calculated. Flower biomass yields were highest for all lines when treatments started with 14L:10D; however, in the two THC lines, a static 14L:10D photoperiod caused a significant decline in THC concentration. Conversely, in Cannatonic, all treatments starting with 14L:10D led to a significant increase in the CBD concentration, which led to a 50–100% increase in total CBD yield. The results show that the assumption that a 12L:12D photoperiod is optimal for all lines is incorrect as, in some lines, yields can be greatly increased by a lengthened light period during flowering. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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23 pages, 2682 KiB  
Article
The Cannabis Plant as a Complex System: Interrelationships between Cannabinoid Compositions, Morphological, Physiological and Phenological Traits
by Erez Naim-Feil, Aaron C. Elkins, M. Michelle Malmberg, Doris Ram, Jonathan Tran, German C. Spangenberg, Simone J. Rochfort and Noel O. I. Cogan
Plants 2023, 12(3), 493; https://doi.org/10.3390/plants12030493 - 21 Jan 2023
Cited by 3 | Viewed by 4664
Abstract
Maintaining specific and reproducible cannabinoid compositions (type and quantity) is essential for the production of cannabis-based remedies that are therapeutically effective. The current study investigates factors that determine the plant’s cannabinoid profile and examines interrelationships between plant features (growth rate, phenology and biomass), [...] Read more.
Maintaining specific and reproducible cannabinoid compositions (type and quantity) is essential for the production of cannabis-based remedies that are therapeutically effective. The current study investigates factors that determine the plant’s cannabinoid profile and examines interrelationships between plant features (growth rate, phenology and biomass), inflorescence morphology (size, shape and distribution) and cannabinoid content. An examination of differences in cannabinoid profile within genotypes revealed that across the cultivation facility, cannabinoids’ qualitative traits (ratios between cannabinoid quantities) remain fairly stable, while quantitative traits (the absolute amount of Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabichromene (CBC), cannabigerol (CBG), Δ9-tetrahydrocannabivarin (THCV) and cannabidivarin (CBDV)) can significantly vary. The calculated broad-sense heritability values imply that cannabinoid composition will have a strong response to selection in comparison to the morphological and phenological traits of the plant and its inflorescences. Moreover, it is proposed that selection in favour of a vigorous growth rate, high-stature plants and wide inflorescences is expected to increase overall cannabinoid production. Finally, a range of physiological and phenological features was utilised for generating a successful model for the prediction of cannabinoid production. The holistic approach presented in the current study provides a better understanding of the interaction between the key features of the cannabis plant and facilitates the production of advanced plant-based medicinal substances. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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10 pages, 1549 KiB  
Article
Cannabigerol Activates Cytoskeletal Remodeling via Wnt/PCP in NSC-34: An In Vitro Transcriptional Study
by Ivan Anchesi, Federica Betto, Luigi Chiricosta, Agnese Gugliandolo, Federica Pollastro, Stefano Salamone and Emanuela Mazzon
Plants 2023, 12(1), 193; https://doi.org/10.3390/plants12010193 - 3 Jan 2023
Viewed by 2082
Abstract
Cannabigerol (CBG) is a non-psychoactive phytocannabinoid present in the Cannabis sativa L. plant. In our study, CBG at the concentration of 10 µM was used to treat NSC-34 motor neuron-like cells. The aim of the study was to evaluate the effects of CBG [...] Read more.
Cannabigerol (CBG) is a non-psychoactive phytocannabinoid present in the Cannabis sativa L. plant. In our study, CBG at the concentration of 10 µM was used to treat NSC-34 motor neuron-like cells. The aim of the study was to evaluate the effects of CBG on NSC-34 cells, using next-generation sequencing (NGS) technology. Analysis showed the activation of the WNT/planar cell polarity (PCP) pathway and Ephrin-Eph signaling. The results revealed that CBG increases the expression of genes associated with the onset process of cytoskeletal remodeling and axon guidance. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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15 pages, 3316 KiB  
Article
Single-Molecule Real-Time Sequencing of Full-Length Transcriptome and Identification of Genes Related to Male Development in Cannabis sativa
by Hui Jiang, Ying Li, Mingbao Luan, Siqi Huang, Lining Zhao, Guang Yang and Gen Pan
Plants 2022, 11(24), 3559; https://doi.org/10.3390/plants11243559 - 16 Dec 2022
Viewed by 1599
Abstract
Female Cannabis sativa plants have important therapeutic properties. The sex ratio of the dioecious cannabis is approximately 1:1. Cultivating homozygous female plants by inducing female plants to produce male flowers is of great practical significance. However, the mechanism underlying cannabis male development remains [...] Read more.
Female Cannabis sativa plants have important therapeutic properties. The sex ratio of the dioecious cannabis is approximately 1:1. Cultivating homozygous female plants by inducing female plants to produce male flowers is of great practical significance. However, the mechanism underlying cannabis male development remains unclear. In this study, single-molecule real-time (SMRT) sequencing was performed using a mixed sample of female and induced male flowers from the ZYZM1 cannabis variety. A total of 15,241 consensus reads were identified, and 13,657 transcripts were annotated across seven public databases. A total of 48 lncRNAs with an average length of 986.54 bp were identified. In total, 8202 transcripts were annotated as transcription factors, the most common of which were bHLH transcription factors. Moreover, tissue-specific expression pattern analysis showed that 13 MADS transcription factors were highly expressed in male flowers. Furthermore, 232 reads of novel genes were predicted and enriched in lipid metabolism, and qRT-PCR results showed that CER1 may be involved in the development of cannabis male flowers. In addition, 1170 AS events were detected, and two AS events were further validated. Taken together, these results may improve our understanding of the complexity of full-length cannabis transcripts and provide a basis for understanding the molecular mechanism of cannabis male development. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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13 pages, 2237 KiB  
Article
Profiling Cannabinoid Contents and Expression Levels of Corresponding Biosynthetic Genes in Commercial Cannabis (Cannabis sativa L.) Cultivars
by Ae Lim Kim, Young Jae Yun, Hyong Woo Choi, Chang-Hee Hong, Hyun Joo Shim, Jeong Hwan Lee and Young-Cheon Kim
Plants 2022, 11(22), 3088; https://doi.org/10.3390/plants11223088 - 14 Nov 2022
Cited by 5 | Viewed by 3100
Abstract
Cannabis (Cannabis sativa L.) is widely cultivated and studied for its psychoactive and medicinal properties. As the major cannabinoids are present in acidic forms in Cannabis plants, non-enzymatic processes, such as decarboxylation, are crucial for their conversion to neutral active cannabinoid forms. [...] Read more.
Cannabis (Cannabis sativa L.) is widely cultivated and studied for its psychoactive and medicinal properties. As the major cannabinoids are present in acidic forms in Cannabis plants, non-enzymatic processes, such as decarboxylation, are crucial for their conversion to neutral active cannabinoid forms. Herein, we detected the levels of cannabidivarin (CBDV), cannabidiol (CBD), cannabichromene (CBC), and Δ9-tetrahydrocannabinol (Δ9-THC) in the leaves and vegetative shoots of five commercial Cannabis cultivars using a combination of relatively simple extraction, decarboxylation, and high-performance liquid chromatography analyses. The CBDV, CBC, and Δ9-THC levels were 6.3–114.9, 34.4–187.2, and 57.6–407.4 μg/g, respectively, and the CBD levels were the highest, ranging between 1.2–8.9 μg/g in leaf and vegetative shoot tissues of Cannabis cultivars. Additionally, correlations were observed between cannabinoid accumulation and transcription levels of genes encoding key enzymes for cannabinoid biosynthesis, including CsCBGAS, CsCBDAS, CsCBCAS, and CsTHCAS. These data suggest that the high accumulation of cannabinoids, such as CBC, Δ9-THC, and CBD, might be derived from the transcriptional regulation of CsCBGAS and CsCBDAS in Cannabis plants. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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21 pages, 4288 KiB  
Article
Genome-Wide Identification and Expression Analysis of Wall-Associated Kinase (WAK) Gene Family in Cannabis sativa L.
by Hülya Sipahi, Terik Djabeng Whyte, Gang Ma and Gerald Berkowitz
Plants 2022, 11(20), 2703; https://doi.org/10.3390/plants11202703 - 13 Oct 2022
Cited by 4 | Viewed by 2066
Abstract
Wall-associated kinases (WAKs) are receptors that bind pectin or small pectic fragments in the cell wall and play roles in cell elongation and pathogen response. In the Cannabis sativa (Cs) genome, 53 CsWAK/CsWAKL (WAK-like) protein family members were identified and characterized; their amino [...] Read more.
Wall-associated kinases (WAKs) are receptors that bind pectin or small pectic fragments in the cell wall and play roles in cell elongation and pathogen response. In the Cannabis sativa (Cs) genome, 53 CsWAK/CsWAKL (WAK-like) protein family members were identified and characterized; their amino acid lengths and molecular weights varied from 582 to 983, and from 65.6 to 108.8 kDa, respectively. They were classified into four main groups by a phylogenetic tree. Out of the 53 identified CsWAK/CsWAKL genes, 23 CsWAK/CsWAKL genes were unevenly distributed among six chromosomes. Two pairs of genes on chromosomes 4 and 7 have undergone duplication. The number of introns and exons among CsWAK/CsWAKL genes ranged from 1 to 6 and from 2 to 7, respectively. The promoter regions of 23 CsWAKs/CsWAKLs possessed diverse cis-regulatory elements that are involved in light, development, environmental stress, and hormone responsiveness. The expression profiles indicated that our candidate genes (CsWAK1, CsWAK4, CsWAK7, CsWAKL1, and CsWAKL7) are expressed in leaf tissue. These genes exhibit different expression patterns than their homologs in other plant species. These initial findings are useful resources for further research work on the potential roles of CsWAK/CsWAKL in cellular signalling during development, environmental stress conditions, and hormone treatments. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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19 pages, 3454 KiB  
Article
Impacts of Different Light Spectra on CBD, CBDA and Terpene Concentrations in Relation to the Flower Positions of Different Cannabis Sativa L. Strains
by Philipp Reichel, Sebastian Munz, Jens Hartung, Stiina Kotiranta and Simone Graeff-Hönninger
Plants 2022, 11(20), 2695; https://doi.org/10.3390/plants11202695 - 13 Oct 2022
Cited by 10 | Viewed by 5064
Abstract
Cannabis is one of the oldest cultivated plants, but plant breeding and cultivation are restricted by country-specific regulations. The plant has gained interest due to its medically important secondary metabolites, cannabinoids and terpenes. Besides biotic and abiotic stress factors, secondary metabolism can be [...] Read more.
Cannabis is one of the oldest cultivated plants, but plant breeding and cultivation are restricted by country-specific regulations. The plant has gained interest due to its medically important secondary metabolites, cannabinoids and terpenes. Besides biotic and abiotic stress factors, secondary metabolism can be manipulated by changing light quality and intensity. In this study, three morphologically different cannabis strains were grown in a greenhouse experiment under three different light spectra with three real light repetitions. The chosen light sources were as follows: a CHD Agro 400 ceramic metal-halide lamp with a sun-like broad spectrum and an R:FR ratio of 2.8, and two LED lamps, a Solray (SOL) and an AP67, with R:FR ratios of 13.49 and 4, respectively. The results of the study indicated that the considered light spectra significantly influenced CBDA and terpene concentrations in the plants. In addition to the different light spectra, the distributions of secondary metabolites were influenced by flower positions. The distributions varied between strains and indicated interactions between morphology and the chosen light spectra. Thus, the results demonstrate that secondary metabolism can be artificially manipulated by the choice of light spectrum, illuminant and intensity. Furthermore, the data imply that, besides the cannabis strain selected, flower position can have an impact on the medicinal potencies and concentrations of secondary metabolites. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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22 pages, 12158 KiB  
Article
Proanthocyanidins Alleviate Cadmium Stress in Industrial Hemp (Cannabis sativa L.)
by Ming Yin, Langlang Pan, Junfei Liu, Xiaojuan Yang, Huijuan Tang, Yuxin Zhou, Siqi Huang and Gen Pan
Plants 2022, 11(18), 2364; https://doi.org/10.3390/plants11182364 - 10 Sep 2022
Cited by 4 | Viewed by 1936
Abstract
Industrial hemp (Cannabis sativa L.), an annual herbaceous cash crop, is widely used for the remediation of heavy metal-contaminated soils due to its short growth cycle, high tolerance, high biomass, and lack of susceptibility to transfer heavy metals into the human food [...] Read more.
Industrial hemp (Cannabis sativa L.), an annual herbaceous cash crop, is widely used for the remediation of heavy metal-contaminated soils due to its short growth cycle, high tolerance, high biomass, and lack of susceptibility to transfer heavy metals into the human food chain. In this study, a significant increase in proanthocyanidins was found in Yunnan hemp no. 1 after cadmium stress. Proanthocyanidins are presumed to be a key secondary metabolite for cadmium stress mitigation. Therefore, to investigate the effect of proanthocyanidins on industrial hemp under cadmium stress, four experimental treatments were set up: normal environment, cadmium stress, proanthocyanidin treatment, and cadmium stress after pretreatment with proanthocyanidins. The phenotypes from the different treatments were compared. The experimental results showed that pretreatment with proanthocyanidins significantly alleviated cadmium toxicity in industrial hemp. The transcriptome and metabolome of industrial hemp were evaluated in the different treatments. Proanthocyanidin treatment and cadmium stress in industrial hemp mainly affected gene expression in metabolic pathways associated with glutathione metabolism, phenylpropanoids, and photosynthesis, which in turn altered the metabolite content in metabolic pathways of phenylalanine, vitamin metabolism, and carotenoid synthesis. The combined transcriptomic and metabolomic analysis revealed that proanthocyanidins mitigated cadmium toxicity by enhancing photosynthesis, secondary metabolite synthesis, and antioxidant synthesis. In addition, exogenous proanthocyanidins and cadmium ions acted simultaneously on EDS1 to induce the production of large amounts of salicylic acid in the plant. Finally, overexpression of CsANR and CsLAR, key genes for proanthocyanidins synthesis in industrial hemp, was established in Arabidopsis plants. The corresponding plants were subjected to cadmium stress, and the results showed that CsLAR transgenic plants were more tolerant to cadmium than the CsANR transgenic and wild-type Arabidopsis plants. The results showed that salicylic acid and jasmonic acid were increased in Arabidopsis overexpressing CsLAR compared to AT wild-type Arabidopsis, and levels of secondary metabolites were significantly higher in Arabidopsis overexpressing CsLAR than in AT wild-type Arabidopsis. These results revealed how proanthocyanidins alleviated cadmium stress and laid the foundation for breeding industrial hemp varieties with higher levels of proanthocyanidins and greater tolerance. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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10 pages, 2273 KiB  
Article
Overexpression of CsMIXTA, a Transcription Factor from Cannabis sativa, Increases Glandular Trichome Density in Tobacco Leaves
by Samuel R. Haiden, Peter V. Apicella, Yi Ma and Gerald A. Berkowitz
Plants 2022, 11(11), 1519; https://doi.org/10.3390/plants11111519 - 6 Jun 2022
Cited by 5 | Viewed by 4609
Abstract
Cannabinoids are synthesized in glandular stalked trichomes on the female flowers of Cannabis sativa (cannabis). The regulation of glandular trichome development has not been characterized in cannabis. We recently identified an R2R3-MYB transcription factor, CsMIXTA, which could be involved in trichome morphogenesis [...] Read more.
Cannabinoids are synthesized in glandular stalked trichomes on the female flowers of Cannabis sativa (cannabis). The regulation of glandular trichome development has not been characterized in cannabis. We recently identified an R2R3-MYB transcription factor, CsMIXTA, which could be involved in trichome morphogenesis in cannabis. Some homologous genes of CsMIXTA are known to function in glandular trichome initiation in other plant species. CsMIXTA is highly expressed in flower tissue compared to vegetative tissues. Interestingly, CsMIXTA is also highly expressed in trichomes isolated from female flower tissue. In addition, CsMIXTA is upregulated during the peak stages of female flower maturation in correlation with some cannabinoid biosynthetic genes. Transient expression in Nicotiana benthamiana showed that CsMIXTA is localized in the nucleus. Furthermore, yeast transcriptional activation assay demonstrated that CsMIXTA has transactivation activity. Overexpression of CsMIXTA in Nicotiana tabacum resulted in higher trichome density, larger trichome size, and more branching on stalked glandular trichomes. The results indicate that CsMIXTA not only promotes glandular trichome initiation in epidermal cells, but also regulates trichome development in tobacco leaves. In this report, we characterized the novel function of the first cannabis transcription factor that may be critical for glandular trichome morphogenesis. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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15 pages, 1568 KiB  
Article
Genome-Wide Identification and Expression Analysis of Homeodomain Leucine Zipper Subfamily IV (HD-ZIP IV) Gene Family in Cannabis sativa L.
by Gang Ma, Alice Kira Zelman, Peter V. Apicella and Gerald Berkowitz
Plants 2022, 11(10), 1307; https://doi.org/10.3390/plants11101307 - 13 May 2022
Cited by 5 | Viewed by 2624
Abstract
The plant-specific homeodomain zipper family (HD-ZIP) of transcription factors plays central roles in regulating plant development and environmental resistance. HD-ZIP transcription factors IV (HDZ IV) have been involved primarily in the regulation of epidermal structure development, such as stomata and trichomes. In our [...] Read more.
The plant-specific homeodomain zipper family (HD-ZIP) of transcription factors plays central roles in regulating plant development and environmental resistance. HD-ZIP transcription factors IV (HDZ IV) have been involved primarily in the regulation of epidermal structure development, such as stomata and trichomes. In our study, we identified nine HDZ IV-encoding genes in Cannabis sativa L. by conducting a computational analysis of cannabis genome resources. Our analysis suggests that these genes putatively encode proteins that have all the conserved domains of HDZ IV transcription factors. The phylogenetic analysis of HDZ IV gene family members of cannabis, rice (Oryza sativa), and Arabidopsis further implies that they might have followed distinct evolutionary paths after divergence from a common ancestor. All the identified cannabis HDZ IV gene promoter sequences have multiple regulation motifs, such as light- and hormone-responsive elements. Furthermore, experimental evidence shows that different HDZ IV genes have different expression patterns in root, stem, leaf, and flower tissues. Four genes were primarily expressed in flowers, and the expression of CsHDG5 (XP_030501222.1) was also correlated with flower maturity. Fifty-nine genes were predicted as targets of HDZ IV transcription factors. Some of these genes play central roles in pathogen response, flower development, and brassinosteroid signaling. A subcellular localization assay indicated that one gene of this family is localized in the Arabidopsis protoplast nucleus. Taken together, our work lays fundamental groundwork to illuminate the function of cannabis HDZ IV genes and their possible future uses in increasing cannabis trichome morphogenesis and secondary metabolite production. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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9 pages, 3561 KiB  
Article
Effects of Harpin and Flg22 on Growth Enhancement and Pathogen Defense in Cannabis sativa Seedlings
by Lauren B. Sands, Taylor Cheek, Joseph Reynolds, Yi Ma and Gerald A. Berkowitz
Plants 2022, 11(9), 1178; https://doi.org/10.3390/plants11091178 - 27 Apr 2022
Cited by 7 | Viewed by 4089
Abstract
Pathogen-associated molecular patterns, PAMPs, are a diverse group of molecules associated with pathogenic microbes and are known to activate immune response and in some cases enhance growth in plants. Two PAMPs, harpin and flg22, have shown these affects in various plant species. PAMPs [...] Read more.
Pathogen-associated molecular patterns, PAMPs, are a diverse group of molecules associated with pathogenic microbes and are known to activate immune response and in some cases enhance growth in plants. Two PAMPs, harpin and flg22, have shown these affects in various plant species. PAMPs are known to activate basal immunity, the ethylene signaling pathway, alter gene expression and change plant composition. Pretreatment with harpin enhanced hemp seedling resistance to Pythium aphanidermatum, while flg22 failed to induce the defense mechanism towards P. aphanidermatum. In the absence of the pathogen, both harpin and flg22 enhanced seedling growth when compared to the water control. Ethylene is a hormone involved in both plant defense signaling and growth. Both harpin and flg22 pretreatment induced certain ethylene responsive genes but not all the genes examined, indicating that harpin and flg22 act differently in ethylene and potentially defense signaling. In addition, both harpin and flg22 induced CsFRK1 and CsPR1, two marker genes for plant innate immunity. Both PAMPs can enhance growth but likely induce different defense signaling pathways. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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16 pages, 1448 KiB  
Article
Impact of Harvest Time and Pruning Technique on Total CBD Concentration and Yield of Medicinal Cannabis
by Danilo Crispim Massuela, Jens Hartung, Sebastian Munz, Federico Erpenbach and Simone Graeff-Hönninger
Plants 2022, 11(1), 140; https://doi.org/10.3390/plants11010140 - 5 Jan 2022
Cited by 19 | Viewed by 15522
Abstract
The definition of optimum harvest and pruning interventions are important factors varying inflorescence yield and cannabinoid composition. This study investigated the impact of (i) harvest time (HT) and (ii) pruning techniques (PT) on plant biomass accumulation, CBD and CBDA-concentrations and total CBD yield [...] Read more.
The definition of optimum harvest and pruning interventions are important factors varying inflorescence yield and cannabinoid composition. This study investigated the impact of (i) harvest time (HT) and (ii) pruning techniques (PT) on plant biomass accumulation, CBD and CBDA-concentrations and total CBD yield of a chemotype III medical cannabis genotype under indoor cultivation. The experiment consisted of four HTs between 5 and 11 weeks of flowering and three PTs-apical cut (T); removal of side shoots (L) and control (C), not pruned plants. Results showed that inflorescence dry weight increased continuously, while the total CBD concentration did not differ significantly over time. For the studied genotype, optimum harvest time defined by highest total CBD yield was found at 9 weeks of flowering. Total CBD-concentration of inflorescences in different fractions of the plant’s height was significantly higher in the top (9.9%) in comparison with mid (8.2%) and low (7.7%) fractions. The T plants produced significantly higher dry weight of inflorescences and leaves than L and C. Total CBD yield of inflorescences for PTs were significantly different among pruned groups, but do not differ from the control group. However, a trend for higher yields was observed (T > C > L). Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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11 pages, 1936 KiB  
Article
Potential of Impedance Flow Cytometry to Assess the Viability and Quantity of Cannabis sativa L. Pollen
by Hamza Rafiq, Jens Hartung, Lisa Burgel, Georg Röll and Simone Graeff-Hönninger
Plants 2021, 10(12), 2739; https://doi.org/10.3390/plants10122739 - 13 Dec 2021
Cited by 1 | Viewed by 2874
Abstract
Over the last decade, efforts to breed new Cannabis sativa L. cultivars with high Cannabidiol (CBD) and other non-psychoactive cannabinoids with low tetrahydrocannabinol (THC) levels have increased. In this context, the identification of the viability and quantity of pollen, which represents the fitness [...] Read more.
Over the last decade, efforts to breed new Cannabis sativa L. cultivars with high Cannabidiol (CBD) and other non-psychoactive cannabinoids with low tetrahydrocannabinol (THC) levels have increased. In this context, the identification of the viability and quantity of pollen, which represents the fitness of male gametophytes, to accomplish successful pollination is of high importance. The present study aims to evaluate the potential of impedance flow cytometry (IFC) for the assessment of pollen viability (PV) and total number of pollen cells (TPC) in two phytocannabinoid-rich cannabis genotypes, KANADA (KAN) and A4 treated with two different chemical solutions, silver thiosulfate solution (STS) and gibberellic acid (GA3). Pollen was collected over a period of 8 to 24 days after flowering (DAF) in a greenhouse experiment. Impedance flow cytometry (IFC) technology was used with Cannabis sativa to assess the viability and quantity of pollen. The results showed that the number of flowers per plant was highest at 24 DAF for both genotypes, A4 (317.78) and KAN (189.74). TPC induced by STS was significantly higher compared to GA3 over the collection period of 8 to 24 DAF with the highest mean TPC of 1.54 × 105 at 14 DAF. STS showed significantly higher viability of pollen compared to GA3 in genotype KAN, with the highest PV of 78.18% 11 DAF. Genotype A4 also showed significantly higher PV with STS at 8 (45.66%), 14 (77.88%), 18 (79.37%), and 24 (51.92%) DAF compared to GA3. Furthermore, counting the numbers of flowers did not provide insights into the quality and quantity of pollen; the results showed that PV was highest at 18 DAF with A4; however, the number of flowers per plant was 150.33 at 18 DAF and was thus not the maximum of produced flowers within the experiment. IFC technology successfully estimated the TPC and differentiated between viable and non-viable cells over a period of 8 to 24 DAF in tested genotypes of Cannabis sativa. IFC seems to be an efficient and reliable method to estimate PV, opening new chances for plant breeding and plant production processes in cannabis. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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24 pages, 5527 KiB  
Article
Impact of Three Different Light Spectra on the Yield, Morphology and Growth Trajectory of Three Different Cannabis sativa L. Strains
by Philipp Reichel, Sebastian Munz, Jens Hartung, Achim Präger, Stiina Kotiranta, Lisa Burgel, Torsten Schober and Simone Graeff-Hönninger
Plants 2021, 10(9), 1866; https://doi.org/10.3390/plants10091866 - 9 Sep 2021
Cited by 12 | Viewed by 5280
Abstract
Cannabis is one of the oldest cultivated plants, but plant breeding and cultivation are restricted by country specific regulations. Plant growth, morphology and metabolism can be manipulated by changing light quality and intensity. Three morphologically different strains were grown under three different light [...] Read more.
Cannabis is one of the oldest cultivated plants, but plant breeding and cultivation are restricted by country specific regulations. Plant growth, morphology and metabolism can be manipulated by changing light quality and intensity. Three morphologically different strains were grown under three different light spectra with three real light repetitions. Light dispersion was included into the statistical evaluation. The light spectra considered had an influence on the morphology of the plant, especially the height. Here, the shade avoidance induced by the lower R:FR ratio under the ceramic metal halide lamp (CHD) was of particular interest. The sugar leaves seemed to be of elementary importance in the last growth phase for yield composition. Furthermore, the last four weeks of flowering were crucial to influence the yield composition of Cannabis sativa L. through light spectra. The dry flower yield was significantly higher under both LED treatments compared to the conventional CHD light source. Our results indicate that the plant morphology can be artificially manipulated by the choice of light treatment to create shorter plants with more lateral branches which seem to be beneficial for yield development. Furthermore, the choice of cultivar has to be taken into account when interpreting results of light studies, as Cannabis sativa L. subspecies and thus bred strains highly differ in their phenotypic characteristics. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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15 pages, 2112 KiB  
Perspective
New Insight into Ornamental Applications of Cannabis: Perspectives and Challenges
by Mohsen Hesami, Marco Pepe, Austin Baiton, Seyed Alireza Salami and Andrew Maxwell Phineas Jones
Plants 2022, 11(18), 2383; https://doi.org/10.3390/plants11182383 - 13 Sep 2022
Cited by 30 | Viewed by 7703
Abstract
The characteristic growth habit, abundant green foliage, and aromatic inflorescences of cannabis provide the plant with an ideal profile as an ornamental plant. However, due to legal barriers, the horticulture industry has yet to consider the ornamental relevance of cannabis. To evaluate its [...] Read more.
The characteristic growth habit, abundant green foliage, and aromatic inflorescences of cannabis provide the plant with an ideal profile as an ornamental plant. However, due to legal barriers, the horticulture industry has yet to consider the ornamental relevance of cannabis. To evaluate its suitability for introduction as a new ornamental species, multifaceted commercial criteria were analyzed. Results indicate that ornamental cannabis would be of high value as a potted-plant or in landscaping. However, the readiness timescale for ornamental cannabis completely depends on its legal status. Then, the potential of cannabis chemotype Ⅴ, which is nearly devoid of phytocannabinoids and psychoactive properties, as the foundation for breeding ornamental traits through mutagenesis, somaclonal variation, and genome editing approaches has been highlighted. Ultimately, legalization and breeding for ornamental utility offers boundless opportunities related to economics and executive business branding. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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12 pages, 472 KiB  
Systematic Review
A Systematic Review on Cannabinoids for Neuropathic Pain Administered by Routes Other than Oral or Inhalation
by Jose-Manuel Quintero, German Pulido, Luis-Fernando Giraldo, Marta-Ximena Leon, Luis-Eduardo Diaz and Rosa-Helena Bustos
Plants 2022, 11(10), 1357; https://doi.org/10.3390/plants11101357 - 20 May 2022
Cited by 5 | Viewed by 3601
Abstract
The use of cannabis and cannabinoid products for the treatment of neuropathic pain is a growing area of research. This type of pain has a high prevalence, limited response to available therapies and high social and economic costs. Systemic cannabinoid-based therapies have shown [...] Read more.
The use of cannabis and cannabinoid products for the treatment of neuropathic pain is a growing area of research. This type of pain has a high prevalence, limited response to available therapies and high social and economic costs. Systemic cannabinoid-based therapies have shown some unwanted side effects. Alternative routes of administration in the treatment of neuropathic pain may provide better acceptance for the treatment of multiple pathologies associated with neuropathic pain. To examine the efficacy, tolerability, and safety of cannabinoids (individualized formulations, phytocannabinoids, and synthetics) administered by routes other than oral or inhalation compared to placebo and/or conventional medications in the management of neuropathic pain. This systematic review of the literature reveals a lack of clinical research investigating cannabis by routes other than oral and inhalation as a potential treatment for neuropathic pain and highlights the need for further investigation with well-designed clinical trials. There is a significant lack of evidence indicating that cannabinoids administered by routes other than oral or inhaled may be an effective alternative, with better tolerance and safety in the treatment of neuropathic pain. Higher quality, long-term, randomized controlled trials are needed to examine whether cannabinoids administered by routes other than inhalation and oral routes may have a role in the treatment of neuropathic pain. Full article
(This article belongs to the Special Issue Studies on Cannabis sativa and Cannabinoids)
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