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Old 04-17-2015, 08:46 AM #1
Sam_Skunkman
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RECENT interesting findings

https://link.springer.com/article/10....722-015-0254-2
I CAN'T POST THE FULL LINK, TRY ADDING THE SECOND ONE BELOW TO THE END OF THE FIRST BELOW. THAT WORKS.
https://link.springer.com/article/
10.1007%2Fs10722-015-0254-2

Size matters: evolution of large drug-secreting resin glands
in elite pharmaceutical strains of Cannabis sativa (marijuana)

Ernest Small . Steve G. U. Naraine
Genet Resour Crop Evol

----------------------------------------------
https://link.springer.com/article/10....722-015-0253-3
https://link.springer.com/article/
10.1007%2Fs10722-015-0253-3

OR
https://link.springer.com/article/
10.1007/s10722-015-0253-3#page-1

Expansion of female sex organs in response to prolonged
virginity in Cannabis sativa (marijuana)

Ernest Small . Steve G. U. Naraine
Genet Resour Crop Evol

Abstract
Female flowers of Cannabis sativa in wild-growing populations and in hemp plantations are almost always well supplied with pollen. The style-stigma portion of the pistils of such plants was found to average only about 3 mm in length and to invariably be two-branched. By contrast, “buds” (congested female inflorescences), the standard form of marijuana now produced in the illicit and medicinal marijuana sectors, are protected against pollen. This report documents that in the absence of pollen, the style-stigma parts of virgin pistils expand notably, average over 8 mm in length, and tend to develop more than two branches and to increase in girth. From an evolutionary viewpoint, this expansion of pollen-receptive tissue is an apparent adaptation for increasing the probability of fertilizing the females when males are extremely scarce. From a practical viewpoint, the expanded presence of stigma tissues may be both advantageous and disadvantageous. The high-THC secretory gland heads of Cannabis tend to fall away from marijuana buds, significantly decreasing pharmacological potency, but many gland heads become stuck to the receptive papillae of the stigmas, reducing the loss. Although stigmas constitute a small proportion of marijuana, their distinctive chemistry could have health effects.
-------------------------------------------------

https://www.sciencedirect.com/science...31942215001181
Sequence heterogeneity of cannabidiolic- and tetrahydrocannabinolic acid-synthase in Cannabis sativa L. and its relationship with chemical phenotype
Chiara Onofri a, Etienne P.M. de Meijer b, Giuseppe Mandolino a,
a Consiglio per la ricerca in agricoltura e l’analisi dell’economia agraria, Centro di Ricerca per le Colture Industriali, via di Corticella 133, 40128 Bologna, Italy
b GW Pharmaceuticals PLC, Ground Floor South Wing, Kingsgate House, Newbury Road, Andover SP10 4DU, United Kingdom

Received 2 October 2014, Revised 21 March 2015, Available online 9 April 2015

Abstract
Sequence variants of THCA- and CBDA-synthases were isolated from different Cannabis sativa L. strains expressing various wild-type and mutant chemical phenotypes (chemotypes). Expressed and complete sequences were obtained from mature inflorescences. Each strain was shown to have a different specificity and/or ability to convert the precursor CBGA into CBDA and/or THCA type products. The comparison of the expressed sequences led to the identification of different mutations, all of them due to SNPs. These SNPs were found to relate to the cannabinoid composition of the inflorescence at maturity and are therefore proposed to have a functional significance. The amount of variation was found to be higher within the CBDAS sequence family than in the THCAS family, suggesting a more recent evolution of THCA-forming enzymes from the CBDAS group. We therefore consider CBDAS as the ancestral type of these synthases.
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Does this mean THC synthase evolved or mutated from CBD synthase? Maybe man helped select the few that were THC synthase however they came to be, and spread them worldwide? I would not be surprised......
Man selects for THC and against CBD, in the case of Ganja, he selects for high resin contents regardless of Cannabinoid for Hashish, for hemp until recently there was little selections for Cannabinoids until after Cannabis became illegal less then 100 years ago, and very recently for CBD for mediacal varieties.
-SamS
------------------
https://onlinelibrary.wiley.com/doi/1...bph.12944/full


Are cannabidiol and Δ9-tetrahydrocannabivarin negative modulators of the endocannabinoid system? A systematic review

John M McPartland1, Marnie Duncan, Vincenzo Di Marzo and Roger G Pertwee
Article first published online: 8 JAN 2015

© 2014 The British Pharmacological Society
Issue
British Journal of Pharmacology
Volume 172, Issue 3, pages 737–753, February 2015

Abstract
Based upon evidence that the therapeutic properties of Cannabis preparations are not solely dependent upon the presence of Δ9-tetrahydrocannabinol (THC), pharmacological studies have been recently carried out with other plant cannabinoids (phytocannabinoids), particularly cannabidiol (CBD) and Δ9-tetrahydrocannabivarin (THCV). Results from some of these studies have fostered the view that CBD and THCV modulate the effects of THC via direct blockade of cannabinoid CB1 receptors, thus behaving like first-generation CB1 receptor inverse agonists, such as rimonabant. Here, we review in vitro and ex vivo mechanistic studies of CBD and THCV, and synthesize data from these studies in a meta-analysis. Synthesized data regarding mechanisms are then used to interpret results from recent pre-clinical animal studies and clinical trials. The evidence indicates that CBD and THCV are not rimonabant-like in their action and thus appear very unlikely to produce unwanted CNS effects. They exhibit markedly disparate pharmacological profiles particularly at CB1 receptors: CBD is a very low-affinity CB1 ligand that can nevertheless affect CB1 receptor activity in vivo in an indirect manner, while THCV is a high-affinity CB1 receptor ligand and potent antagonist in vitro and yet only occasionally produces effects in vivo resulting from CB1 receptor antagonism. THCV has also high affinity for CB2 receptors and signals as a partial agonist, differing from both CBD and rimonabant. These cannabinoids illustrate how in vitro mechanistic studies do not always predict in vivo pharmacology and underlie the necessity of testing compounds in vivo before drawing any conclusion on their functional activity at a given target.

-------------------------------

https://www.sciencedirect.com/science...3194221200475X
Try this , to correct it remove the empty space between the last w and the . in the http below
https://www .sciencedirect.com/science/article/pii/S003194221200475X


Phytochemistry
Volume 87, March 2013, Pages 51–59


Analysis of cannabinoids in laser-microdissected trichomes of medicinal Cannabis sativa using LCMS and cryogenic NMR
Nizar Happyanaa, b, Sara Agnoletc, Remco Muntendamd, Annie Van Dame, Bernd Schneiderc, Oliver Kaysera, ,
a Department of Technical Biochemistry, Technical University of Dortmund, Technische Biochemie, Emil-Figge-Strasse 66, D-44227 Dortmund, Germany
b Department of Chemistry, Bandung Institute of Technology, Jl. Ganesha 10, Bandung 40132, Indonesia
c Research Group Biosynthesis/NMR, Max Planck Institute for Chemical Ecology, Hans-Knöll-Straße 8, 07745 Jena, Germany
d Department of Pharmaceutical Biology, University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands
e Mass Spectrometry Core Facility, Center for Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV, Groningen, The Netherlands

Abstract
Trichomes, especially the capitate-stalked glandular hairs, are well known as the main sites of cannabinoid and essential oil production of Cannabis sativa. In this study the distribution and density of various types of Cannabis sativa L. trichomes, have been investigated by scanning electron microscopy (SEM). Furthermore, glandular trichomes were isolated over the flowering period (8 weeks) by laser microdissection (LMD) and the cannabinoid profile analyzed by LCMS. Cannabinoids were detected in extracts of 25–143 collected cells of capitate-sessile and capitate stalked trichomes and separately in the gland (head) and the stem of the latter. Δ9-Tetrahydrocannabinolic acid [THCA (1)], cannabidiolic acid [CBDA (2)], and cannabigerolic acid [CBGA (3)] were identified as most-abundant compounds in all analyzed samples while their decarboxylated derivatives, Δ9-tetrahydrocannabinol [THC (4)], cannabidiol [CBD (5)], and cannabigerol [CBG (6)], co-detected in all samples, were present at significantly lower levels. Cannabichromene [CBC (8)] along with cannabinol (CBN (9)) were identified as minor compounds only in the samples of intact capitate-stalked trichomes and their heads harvested from 8-week old plants. Cryogenic nuclear magnetic resonance spectroscopy (NMR) was used to confirm the occurrence of major cannabinoids, THCA (1) and CBDA (2), in capitate-stalked and capitate-sessile trichomes. Cryogenic NMR enabled the additional identification of cannabichromenic acid [CBCA (7)] in the dissected trichomes, which was not possible by LCMS as standard was not available. The hereby documented detection of metabolites in the stems of capitate-stalked trichomes indicates a complex biosynthesis and localization over the trichome cells forming the glandular secretion unit.


-----------------------------------------


https://www.sciencedirect.com/science...26669013000526
Industrial Crops and Products
Volume 46, April 2013, Pages 269–273

Ethephon application stimulats cannabinoids and plastidic terpenoids production in Cannabis sativa at flowering stage
Hakimeh Mansouri, , Fatemeh Salari, Zahra Asrar
Received 27 November 2012, Accepted 4 January 2013, Available online 1 March 2013


Abstract
We studied the effect of ethephon on levels of the major cannabinoids (tetrahydrocannabinol and cannabidiol) and chlorophyll, carotenoids and α-tocopherol in Cannabis sativa at productive stage. Results revealed that ethephon increased THC content of leaf in male and female plants and of male flowers. However, ethephon unable to enhancing THC content in female flowers. Treatment with etheohon increased CBD content in male and female leaf and female flowers. The treatment of male flowers with low ethephon concentration caused an increase, and those treated with high ethephon concentration resulted in a decrease in CBD content. The lowest level of ethephon (1 μM) enhanced chlorophyll a, b and total chlorophyll in male and female plants. Both sexes treated with ethephon showed an increase in carotenoids content, but 1 μM ethephon had the stronger effect in this regards. Male and female plants had a higher content of α-tocopherol when treated with ethephon. These results showed ethephon is a suitable treatment for increasing cannabinoids and α-tocopherol in productive stage of cannabis and there was not a relation between primary and secondary terpenoids.

Highlights
► Ethephon treatment has considerable effects on increasing of cannabinoids in male and female cannabis. ► There is no correlation between cannabinoids and other plastidial terpenoids. ► Using of ethephon treatment in flowering stage do not adverse effect on plant growth.



https://www.ncbi.nlm.nih.gov/pubmed/26000707

J Nat Prod. 2015 May 22. [Epub ahead of print]
Isolation and Pharmacological Evaluation of Minor Cannabinoids from High-Potency Cannabis sativa.
Radwan MM1, ElSohly MA1, El-Alfy AT1, Ahmed SA1, Slade D1, Husni AS1, Manly SP1, Wilson L1, Seale S1, Cutler SJ1, Ross SA1.


Abstract
Seven new naturally occurring hydroxylated cannabinoids (1-7), along with the known cannabiripsol (8), have been isolated from the aerial parts of high-potency Cannabis sativa. The structures of the new compounds were determined by 1D and 2D NMR spectroscopic analysis, GC-MS, and HRESIMS as 8α-hydroxy-Δ9-tetrahydrocannabinol (1), 8β-hydroxy-Δ9-tetrahydrocannabinol (2), 10α-hydroxy-Δ8-tetrahydrocannabinol (3), 10β-hydroxy-Δ8-tetrahydrocannabinol (4), 10α-hydroxy-Δ9,11-hexahydrocannabinol (5), 9β,10β-epoxyhexahydrocannabinol (6), and 11-acetoxy-Δ9-tetrahydrocannabinolic acid A (7). The binding affinity of isolated compounds 1-8, Δ9-tetrahydrocannabinol, and Δ8-tetrahydrocannabinol toward CB1 and CB2 receptors as well as their behavioral effects in a mouse tetrad assay were studied. The results indicated that compound 3, with the highest affinity to the CB1 receptors, exerted the most potent cannabimimetic-like actions in the tetrad assay, while compound 4 showed partial cannabimimetic actions. Compound 2, on the other hand, displayed a dose-dependent hypolocomotive effect only.

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Old 04-17-2015, 04:59 PM #2
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Sam…those science direct links are not working, says the page does not exist…maybe entered wrong or password protected??
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Old 04-17-2015, 08:54 PM #3
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Quote:
Originally Posted by Betterhaff View Post
Sam…those science direct links are not working, says the page does not exist…maybe entered wrong or password protected??
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Old 04-17-2015, 09:05 PM #4
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Thanks Sam...interesting stuff.
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Old 04-17-2015, 09:17 PM #5
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Good lookin' out on the cannabivarin article and current studies involving attenuation.
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Old 05-15-2015, 06:04 AM #6
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The biochemistry reading on cannabinoid synthesis frames CBDA as mutated THCA genes...basically, THCA genes that are not functional. This isn't a clear contradiction, but may be.

There are no actual wild populations of cannabis now...only renaturalized ones.

What about the cannabinoid knock out gene which blocks all cannabinoid biosynthesis? Why does it exist and why is it uncommon?

Abstract
Sequence variants of THCA- and CBDA-synthases were isolated from different Cannabis sativa L. strains expressing various wild-type and mutant chemical phenotypes (chemotypes). Expressed and complete sequences were obtained from mature inflorescences. Each strain was shown to have a different specificity and/or ability to convert the precursor CBGA into CBDA and/or THCA type products. The comparison of the expressed sequences led to the identification of different mutations, all of them due to SNPs. These SNPs were found to relate to the cannabinoid composition of the inflorescence at maturity and are therefore proposed to have a functional significance. The amount of variation was found to be higher within the CBDAS sequence family than in the THCAS family, suggesting a more recent evolution of THCA-forming enzymes from the CBDAS group. We therefore consider CBDAS as the ancestral type of these synthases.
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Old 05-15-2015, 08:18 AM #7
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wat. Cool Article, thanks.
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Old 05-29-2015, 02:46 PM #8
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New added article.. At top post last article in list.
-SamS
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Old 06-19-2015, 03:46 PM #9
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THE FIRST TWO ARE NOT CANNABIS SPECIFIC, but interesting


https://journals.plos.org/plosone/art...l.pone.0128808

Large-Scale Evolutionary Analysis of Genes and Supergene Clusters from Terpenoid Modular Pathways Provides Insights into Metabolic Diversification in Flowering Plants
Johannes A. Hofberger, Aldana M. Ramirez, Erik van den Bergh, Xinguang Zhu, Harro J. Bouwmeester, Robert C. Schuurink, M. Eric Schranz

Abstract
An important component of plant evolution is the plethora of pathways producing more than 200,000 biochemically diverse specialized metabolites with pharmacological, nutritional and ecological significance. To unravel dynamics underlying metabolic diversification, it is critical to determine lineage-specific gene family expansion in a phylogenomics framework. However, robust functional annotation is often only available for core enzymes catalyzing committed reaction steps within few model systems. In a genome informatics approach, we extracted information from early-draft gene-space assemblies and non-redundant transcriptomes to identify protein families involved in isoprenoid biosynthesis. Isoprenoids comprise terpenoids with various roles in plant-environment interaction, such as pollinator attraction or pathogen defense. Combining lines of evidence provided by synteny, sequence homology and Hidden-Markov-Modelling, we screened 17 genomes including 12 major crops and found evidence for 1,904 proteins associated with terpenoid biosynthesis. Our terpenoid genes set contains evidence for 840 core terpene-synthases and 338 triterpene-specific synthases. We further identified 190 prenyltransferases, 39 isopentenyl-diphosphate isomerases as well as 278 and 219 proteins involved in mevalonate and methylerithrol pathways, respectively. Assessing the impact of gene and genome duplication to lineage-specific terpenoid pathway expansion, we illustrated key events underlying terpenoid metabolic diversification within 250 million years of flowering plant radiation. By quantifying Angiosperm-wide versatility and phylogenetic relationships of pleiotropic gene families in terpenoid modular pathways, our analysis offers significant insight into evolutionary dynamics underlying diversification of plant secondary metabolism. Furthermore, our data provide a blueprint for future efforts to identify and more rapidly clone terpenoid biosynthetic genes from any plant species.


https://journals.plos.org/plosone/art...l.pone.0129598

Transcriptomic Analysis Reveals Possible Influences of ABA on Secondary Metabolism of Pigments, Flavonoids and Antioxidants in Tomato Fruit during Ripening

* Wangshu Mou,
* Dongdong Li,
* Zisheng Luo,
* Linchun Mao,
* Tiejin Ying
Transcriptomic Analysis Reveals Possible Influences of ABA on Secondary Metabolism of Pigments, Flavonoids and Antioxidants in Tomato Fruit during Ripening

Abstract
Abscisic acid (ABA) has been proven to be involved in the regulation of climacteric fruit ripening, but a comprehensive investigation of its influence on ripening related processes is still lacking. By applying the next generation sequencing technology, we conducted a comparative analysis of the effects of exogenous ABA and NDGA (Nordihydroguaiaretic acid, an inhibitor of ABA biosynthesis) on tomato fruit ripening. The high throughput sequencing results showed that out of the 25728 genes expressed across all three samples, 10388 were identified as significantly differently expressed genes. Exogenous ABA was found to enhance the transcription of genes involved in pigments metabolism, including carotenoids biosynthesis and chlorophyll degradation, whereas NDGA treatment inhibited these processes. The results also revealed the crucial role of ABA in flavonoids synthesis and regulation of antioxidant system. Intriguingly, we also found that an inhibition of endogenous ABA significantly enhanced the transcriptional abundance of genes involved in photosynthesis. Our results highlighted the significance of ABA in regulating tomato ripening, which provided insight into the regulatory mechanism of fruit maturation and senescence process.



https://journals.plos.org/plosone/art...l.pone.0110638

Diversity Analysis in Cannabis sativa Based on Large-Scale Development of Expressed Sequence Tag-Derived Simple Sequence Repeat Markers
Chunsheng Gao , Pengfei Xin , Chaohua Cheng, Qing Tang, Ping Chen, Changbiao Wang, Gonggu Zang, Lining Zhao


Abstract
Cannabis sativa L. is an important economic plant for the production of food, fiber, oils, and intoxicants. However, lack of sufficient simple sequence repeat (SSR) markers has limited the development of cannabis genetic research. Here, large-scale development of expressed sequence tag simple sequence repeat (EST-SSR) markers was performed to obtain more informative genetic markers, and to assess genetic diversity in cannabis (Cannabis sativa L.). Based on the cannabis transcriptome, 4,577 SSRs were identified from 3,624 ESTs. From there, a total of 3,442 complementary primer pairs were designed as SSR markers. Among these markers, trinucleotide repeat motifs (50.99%) were the most abundant, followed by hexanucleotide (25.13%), dinucleotide (16.34%), tetranucloetide (3.8%), and pentanucleotide (3.74%) repeat motifs, respectively. The AAG/CTT trinucleotide repeat (17.96%) was the most abundant motif detected in the SSRs. One hundred and seventeen EST-SSR markers were randomly selected to evaluate primer quality in 24 cannabis varieties. Among these 117 markers, 108 (92.31%) were successfully amplified and 87 (74.36%) were polymorphic. Forty-five polymorphic primer pairs were selected to evaluate genetic diversity and relatedness among the 115 cannabis genotypes. The results showed that 115 varieties could be divided into 4 groups primarily based on geography: Northern China, Europe, Central China, and Southern China. Moreover, the coefficient of similarity when comparing cannabis from Northern China with the European group cannabis was higher than that when comparing with cannabis from the other two groups, owing to a similar climate. This study outlines the first large-scale development of SSR markers for cannabis. These data may serve as a foundation for the development of genetic linkage, quantitative trait loci mapping, and marker-assisted breeding of cannabis.

https://journals.plos.org/plosone/art...l.pone.0085118

Molecular Cytogenetic Characterization of the Dioecious Cannabis sativa with an XY Chromosome Sex Determination System
Mikhail G. Divashuk, Oleg S. Alexandrov, Olga V. Razumova, Ilya V. Kirov, Gennady I. Karlov

Abstract
Hemp (Cannabis sativa L.) was karyotyped using by DAPI/C-banding staining to provide chromosome measurements, and by fluorescence in situ hybridization with probes for 45 rDNA (pTa71), 5S rDNA (pCT4.2), a subtelomeric repeat (CS-1) and the Arabidopsis telomere probes. The karyotype has 18 autosomes plus a sex chromosome pair (XX in female and XY in male plants). The autosomes are difficult to distinguish morphologically, but three pairs could be distinguished using the probes. The Y chromosome is larger than the autosomes, and carries a fully heterochromatic DAPI positive arm and CS-1 repeats only on the less intensely DAPI-stained, euchromatic arm. The X is the largest chromosome of all, and carries CS-1 subtelomeric repeats on both arms. The meiotic configuration of the sex bivalent locates a pseudoautosomal region of the Y chromosome at the end of the euchromatic CS-1-carrying arm. Our molecular cytogenetic study of the C. sativa sex chromosomes is a starting point for helping to make C. sativa a promising model to study sex chromosome evolution.



https://journals.plos.org/plosone/art...l.pone.0099641

Understanding Cultivar-Specificity and Soil Determinants of the Cannabis Microbiome
Max E. Winston , Jarrad Hampton-Marcell, Iratxe Zarraonaindia, Sarah M. Owens, Corrie S. Moreau, Jack A. Gilbert, Josh Hartsel, Suzanne J. Kennedy, S. M. Gibbons

Abstract
Understanding microbial partnerships with the medicinally and economically important crop Cannabis has the potential to affect agricultural practice by improving plant fitness and production yield. Furthermore, Cannabis presents an interesting model to explore plant-microbiome interactions as it produces numerous secondary metabolic compounds. Here we present the first description of the endorhiza-, rhizosphere-, and bulk soil-associated microbiome of five distinct Cannabis cultivars. Bacterial communities of the endorhiza showed significant cultivar-specificity. When controlling cultivar and soil type the microbial community structure was significantly different between plant cultivars, soil types, and between the endorhiza, rhizosphere and soil. The influence of soil type, plant cultivar and sample type differentiation on the microbial community structure provides support for a previously published two-tier selection model, whereby community composition across sample types is determined mainly by soil type, while community structure within endorhiza samples is determined mainly by host cultivar.



https://journals.plos.org/plosone/art...l.pone.0120016

Impacts of Surface Water Diversions for Marijuana Cultivation on Aquatic Habitat in Four Northwestern California Watersheds
Scott Bauer , Jennifer Olson , Adam Cockrill, Michael van Hattem, Linda Miller, Margaret Tauzer, Gordon Leppig

Abstract
Marijuana (Cannabis sativa L.) cultivation has proliferated in northwestern California since at least the mid-1990s. The environmental impacts associated with marijuana cultivation appear substantial, yet have been difficult to quantify, in part because cultivation is clandestine and often occurs on private property. To evaluate the impacts of water diversions at a watershed scale, we interpreted high-resolution aerial imagery to estimate the number of marijuana plants being cultivated in four watersheds in northwestern California, USA. Low-altitude aircraft flights and search warrants executed with law enforcement at cultivation sites in the region helped to validate assumptions used in aerial imagery interpretation. We estimated the water demand of marijuana irrigation and the potential effects water diversions could have on stream flow in the study watersheds. Our results indicate that water demand for marijuana cultivation has the potential to divert substantial portions of streamflow in the study watersheds, with an estimated flow reduction of up to 23% of the annual seven-day low flow in the least impacted of the study watersheds. Estimates from the other study watersheds indicate that water demand for marijuana cultivation exceeds streamflow during the low-flow period. In the most impacted study watersheds, diminished streamflow is likely to have lethal or sub-lethal effects on state-and federally-listed salmon and steelhead trout and to cause further decline of sensitive amphibian species.

-----------

And one for OO,

https://www.researchgate.net/publicat...nabis_sativa_L.)

Sex chromosomes and quantitative sex expression in monoecious hemp (Cannabis sativa L.)


Anne-Michelle Faux

Alice Berhin

Nicolas Dauguet

P. Bertin
Euphytica 03/2014; 196(2).
ABSTRACT Hemp (Cannabis sativa) has a highly variable sexual phenotype. In dioecious hemp, the sex is controlled by heteromorphic sex chromosomes according to an X-to-autosomes equilibrium. However, in monoecious hemp, the sex determinism remains widely unknown and has never been related to a quantitative approach of sex expression. The present paper aims to contribute to the comprehension of the sex determinism in monoecious hemp by assessing the genotypic variability of its sex expression and establishing its sex chromosomes. Five monoecious and one dioecious cultivars were grown in controlled conditions under several photoperiods. The monoecy degree of 194 monoecious plants was recorded at each node by a figure ranging from 0 (male flowers only) to 6 (female flowers only). The genome size of 55 plants was determined by flow cytometry. The DNA of 115 monoecious plants was screened with the male-associated marker MADC2. The monoecy degree varied significantly among monoecious cultivars from 3.36 ± 2.28 in ‘Uso 31’ to 5.70 ± 0.81 in the most feminised ‘Epsilon 68’. The variation of monoecy degree among cultivars remained consistent across trials despite a significant “cultivar × trial” interaction and partly agreed with their earliness. The genome size of monoecious plants (1.791 ± 0.017 pg) was not different from that of females (1.789 ± 0.019 pg) but significantly lower than that of males (1.835 ± 0.019 pg). MADC2 was absent from all monoecious plants. These results strongly support that cultivars of monoecious hemp have the XX constitution and that their sex expression has a genetic basis.

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https://www.ncbi.nlm.nih.gov/pubmed/26048837

Talanta
Volume 140, 1 August 2015, Pages 150–165

1H NMR and HPLC/DAD for Cannabis sativa L. chemotype distinction, extract profiling and specification
Wieland Peschel, , Matteo Poli

Highlights

1H NMR (DMSO-d6) key signals allow distinction of THC, CBD and CBG type cannabis.

We detect neutral/acidic cannabinoids and phenolics co-compounds via HPLC/DAD.

Non-heated extract profiles suggest consideration of cannabinoid acids.

Cell viability reduction correlated with the total cannabinoid content but not THC.

We suggest markers for drug distinction and pharmaceutical specification.
Abstract
The medicinal use of different chemovars and extracts of Cannabis sativa L. requires standardization beyond ∆9-tetrahydrocannabinol (THC) with complementing methods. We investigated the suitability of 1H NMR key signals for distinction of four chemotypes measured in deuterated dimethylsulfoxide together with two new validated HPLC/DAD methods used for identification and extract profiling based on the main pattern of cannabinoids and other phenolics alongside the assayed content of THC, cannabidiol (CBD), cannabigerol (CBG) their acidic counterparts (THCA, CBDA, CBGA), cannabinol (CBN) and cannflavin A and B. Effects on cell viability (MTT assay, HeLa) were tested. The dominant cannabinoid pairs allowed chemotype recognition via assignment of selective proton signals and via HPLC even in cannabinoid-low extracts from the THC, CBD and CBG type. Substantial concentrations of cannabinoid acids in non-heated extracts suggest their consideration for total values in chemotype distinction and specifications of herbal drugs and extracts. Cannflavin A/B are extracted and detected together with cannabinoids but always subordinated, while other phenolics can be accumulated via fractionation and detected in a wide fingerprint but may equally serve as qualitative marker only. Cell viability reduction in HeLa was more determined by the total cannabinoid content than by the specific cannabinoid profile. Therefore the analysis and labeling of total cannabinoids together with the content of THC and 2–4 lead cannabinoids are considered essential. The suitability of analytical methods and the range of compound groups summarized in group and ratio markers are discussed regarding plant classification and pharmaceutical specification.



https://www.ncbi.nlm.nih.gov/pubmed/23890639

Forensic Science International
Volume 231, Issues 1–3, 10 September 2013, Pages 208–212


Analysis of THCA synthase gene expression in cannabis: A preliminary study by real-time quantitative PCR
Fidelia Cascinia, Stella Passerottib, Ilaria Boschia

Abstract
In this paper we describe analyses performed by Real-Time Reverse-Transcriptase Polymerase Chain Reaction (real-time RT-PCR) on RNA of 12 samples, carried out for forensic purposes to investigate a correlation between tetrahydrocannabinol (THC) concentration in Cannabis and the tetrahydrocannabinol acid synthase (THCAS) gene expression.

Samples were obtained from an experimental cultivation of declared potency Cannabis variety seeds and from seizures. The Rubisco gene and the 26S ribosomal RNA gene were used as internal control genes for their constant expression and stability.

As results we found minor gene expression in samples from leaves of young plants.

Further, grouping results for cannabis samples with similar characteristics, we have found an increased relative expression in samples with the highest percentage of THC coming from seized sample and adult plants.



https://neuron.mefst.hr/docs/CMJ/issu...3/12808725.pdf

Encyclopedia of Forensic Sciences (Second Edition)
2013, Pages 382–386

Biology/DNA/Botany

Cannabis DNA Typing Methods
H. Miller Coyle

Abstract
This article focuses on Cannabis, primarily, the development of biomarkers to classify and uniquely identify marijuana. DNA methods such as short tandem repeat, amplified fragment length polymorphism, and single-nucleotide polymorphism are discussed with an emphasis on forensic data-basing applications and the benefits and limitations of each method. It also provides an overview of marijuana regarding the history of cultivation in the United States, associative evidence at crime scenes and current law enforcement concerns in the US, and microscopic and chemical methods for classification, geo-sourcing and high-throughput screening.


https://www.researchgate.net/profile/...i/publications

Studies in Natural Products Chemistry
Volume 45, 2015, Pages 17–57

Chapter 2 – Cannabis and Bioactive Cannabinoids
Federica Messina, Ornelio Rosati, Massimo Curini, M. Carla Marcotulli

Abstract
The therapeutic use of Cannabis dates back to ancient times and this plant has been used for centuries as remedy for a large number of diseases. Today it is well known that biological activity of Cannabis is related to the endocannabinoid system (ECS), a complex signaling network that comprises classical cannabinoid receptors (CB1 and CB2), arachidonic acid-derived ligands, and enzymes degrading the endocannabinoids anandamide and 2-arachidonoyl glycerol, namely fatty acid amide hydrolase and monoacylglycerol lipase. The modulation of the ECS activity turned out to be a therapeutic promise in a wide range of diseases. A problem to the development of Cannabis and cannabinoid medications is the psychoactive property of natural or synthetic agonists, mediated by CB1 receptor. This review deals with the literature analysis of the important biological activities of Cannabis and the efforts aimed to the discovery of natural and nonnatural selective cannabinoids.




https://www.ncbi.nlm.nih.gov/pubmed/24528581

Forensic Science International: Genetics
Volume 9, March 2014, Pages 61–65


Characterization of 15 STR cannabis loci: Nomenclature proposal and SNPSTR haplotypes
Laura Valverdea, Christian Lischkaa, Stefanie Scheipera, Johanna Nedelea, Rachel Challisa, Marian M. de Pancorbob, Heidi Pfeiffera, Stephan Köhnemanna,

Abstract
The standardization of methods for individualizing Cannabis sativa plants could offer new possibilities in the investigation of its illegal trade. Here we present the first nomenclature proposal for 15 cannabis STRs, which allows an initial standardization for performing comparisons between laboratories and generating genotype databases. Several alleles of the 15 STR loci have been sequenced. This has revealed that not all the STR loci are equally suitable for the individualization purposes. Moreover, several nucleotide variations have been detected both inside the repeat structure and/or in the flanking region. All the different SNPSTR haplotypes are presented and compared with the previous sequence raw data of the 15 STR loci. The SNPSTR data could considerably increase the informative value of the STRs, which could be very useful in complex cases.



https://www.sciencedirect.com/science/book/9780124186798

The Effects of Drug Abuse on the Human Nervous System
2014, Pages 387–422


Chapter Thirteen – Effects of Cannabis and Cannabinoids in the Human Nervous System
Harold Kalant

Abstract
The endocannabinoid (EC) system, consisting of ECs, their synthesizing and degrading enzymes, specific transmembrane EC transporters and receptors, is located in both excitatory and inhibitory synapses of all the classical neurotransmitter types throughout the central and peripheral nervous systems, where it acts as a retrograde signaling mechanism to inhibit further release of transmitter. This form of synaptic plasticity is a major component of both rapid short-term and sustained long-term adaptive responses that underlie such processes as homeostasis, learning, memory, and extinction. The functional effects on any given pathway can be either inhibitory or excitatory, depending on whether excitatory (e.g., glutamatergic) or inhibitory (e.g., GABAergic) modulation normally predominates in that pathway. However, the dose-effect curves of EC activity are in many instances biphasic, because sustained strong activity leads to EC receptor desensitization and down-regulation, resulting in progressive loss or even reversal of the effect. Therefore the effects of cannabis and exogenous cannabinoids, of both plant and synthetic origin, are in many cases different from, or even opposite to, those of the EC system.

The functional effects of the EC system and of exogenous cannabinoids are compared with respect to neuronal growth and maturation, neuroprotection against toxic and traumatic damage, sensory pathways, nausea and vomiting, appetite and food intake, the sleep/wake cycle, affective responses and mood states, motor control, seizure activity and cognitive functions. Effects in laboratory animals are compared to those in humans, including both actual and potential therapeutic effects and adverse effects. The therapeutic effects in most instances correspond to the low-dose actions of the EC system, whereas the adverse effects generally correspond to the high-dose range. The exogenous cannabinoids are less selective in their actions than the EC system because they act on a much wider range of EC receptors throughout the nervous system. It is concluded that for most potential therapeutic applications the future will lie with the development of highly selective site-specific agents that act on individual components of the EC system, rather than on the whole system.



https://www.sciencedirect.com/science...79073815001449

Forensic Science International
Volume 251, June 2015, Pages 95–106


Evaluation of elemental profiling methods, including laser-induced breakdown spectroscopy (LIBS), for the differentiation of Cannabis plant material grown in different nutrient solutions
Moteaa M. El-Deftara, , , James Robertsona, , Simon Fosterb, , Chris Lennardc,

Highlights

Application of LIBS, ICP-MS, LA-ICP-MS and μXRF for the analysis of Cannabis plant.

Discrimination of Cannabis is evaluated based on their elemental composition data.

It is possible to correctly associate Cannabis with a particular nutrient.

ICP-MS, LA-ICP-MS and LIBS are suitable methods for the analysis of Cannabis plant.
Abstract
Laser-induced breakdown spectroscopy (LIBS) is an emerging atomic emission based solid sampling technique that has many potential forensic applications. In this study, the analytical performance of LIBS, as well as that of inductively coupled plasma mass spectrometry (ICP-MS), laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and X-ray microfluorescence (μXRF), was evaluated for the ability to conduct elemental analyses on Cannabis plant material, with a specific investigation of the possible links between hydroponic nutrients and elemental profiles from associated plant material. No such study has been previously published in the literature. Good correlation among the four techniques was observed when the concentrations or peak areas of the elements of interest were monitored. For Cannabis samples collected at the same growth time, the elemental profiles could be related to the use of particular commercial nutrients. In addition, the study demonstrated that ICP-MS, LA-ICP-MS and LIBS are suitable techniques for the comparison of Cannabis samples from different sources, with high discriminating powers being achieved. On the other hand, μXRF method was not suitable for the discrimination of Cannabis samples originating from different growth nutrients.




https://www.sciencedirect.com/science...21967314016173

Journal of Chromatography A
Volume 1370, 28 November 2014, Pages 200–215


Multidimensional analysis of cannabis volatile constituents: Identification of 5,5-dimethyl-1-vinylbicyclo[2.1.1]hexane as a volatile marker of hashish, the resin of Cannabis sativa L.
Marie Marchinia, Céline Charvozb, Laurence Dujourdyb, Nicolas Baldovinia, Jean-Jacques Filippia, ,

Highlights

Analysis of cannabis herb and hashish volatile constituents by HS-SPME-GC × GC–MS.

Identification of a new volatile marker of hashish.

Photolytic rearrangement of β-myrcene into hashishene.

Formation of photo-oxidation products during hashish manufacture.
Abstract
The volatile constituents of drug samples derived from Cannabis sativa L. were investigated by means of headspace solid phase microextraction (HS-SPME) and gas chromatography techniques (GC–MS, GC × GC–MS). Samples of cannabis herb and hashish showed clear differences in their volatile chemical profiles, mostly resulting from photo-oxidation processes occurring during the transformation of fresh cannabis herb into hashish. Most unexpectedly, we could demonstrate hashish samples as containing remarkable amounts of a rare and unusual monoterpene – 5,5-dimethyl-1-vinylbicyclo[2.1.1]hexane – among the volatile compounds detected in their headspaces. We gave evidence for the formation of this compound from the light induced rearrangement of β-myrcene during the manufacture of hashish. In view of its high abundance among volatile constituents of cannabis resin and its scarce occurrence in other natural volatile extracts, we propose to rename this specific monoterpene hashishene.



https://www.sciencedirect.com/science...90536X14000525

Egyptian Journal of Forensic Sciences
Available online 11 November 2014

In Press, Corrected Proof — Note to users


Open Access
GCMS analysis of Cannabis sativa L. from four different areas of Pakistan
Muhammad Tayyab, , Durre Shahwar

Abstract
Cannabis is most frequently used drug of abuse not only in Pakistan but also in the whole world. Its use is mounting drastically every year. GCMS allows analysis of Cannabis sativa which shows divergence of the constituents of this plant. Prevalence of this plant can be identified through knowledge of its constituents. In this way we can obstruct the production if we know the region in which it is produced. GCMS is a useful technique for the comparison of constituents of this drug of abuse which will assist the investigator concerning the origin of plant. Comparison also aids in the understanding and acquaintance of similarities of different samples of cannabinoids.




https://link.springer.com/article/10....738-011-0874-x

Acta Physiologiae Plantarum
March 2012, Volume 34, Issue 2, pp 743-750
Date: 02 Dec 2011
In vitro germplasm conservation of high Δ9 -tetrahydrocannabinol yielding elite clones of Cannabis sativa L. under slow growth conditions
Hemant Lata, Suman Chandra, Zlatko Mehmedic, Ikhlas A. Khan, Mahmoud A. ElSohly

Abstract
Germplasm conservation of a high Δ9-tetrahydrocannabinol yielding variety of Cannabis sativa L. was attempted using synthetic seed technology and media supplemented with osmotic agents. Explants of nodal segments containing single axillary bud were excised from in vitro proliferated shoot cultures and encapsulated in high-density sodium alginate (230 mM) hardened by 50 mM CaCl2. The ‘encapsulated’ (synthetic seeds) and ‘non-encapsulated’ nodal segments were stored at 5, 15 and 25°C for 8, 16 and 24 weeks and monitored for the re-growth and survival frequency under the tissue culture conditions (16-h photoperiod, 25°C) on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ 0.5 μM). ‘Encapsulated’ nodal segments could be stored at low temperature 15°C up to 24 weeks with maximum re-growth ability and survival frequency of 60%. Similar to ‘encapsulated’ cultures, the highest re-growth in ‘non-encapsulated’ cultures was observed in the explants kept at 15°C without osmotic agents. Furthermore, the effect of osmotic agents mannitol and sorbitol (2 and 4% w/v, added individually and in combination to the media at culture room conditions i.e. 25°C) on non-encapsulated shoot cultures was also evaluated. A considerable decrease in re-growth and survival was observed in the cultures treated with osmotic agents. Among the cultures treated with different concentrations of osmotic agents, the highest rate of re-growth and survival was observed at the lowest concentration of 2% sorbitol and 2% mannitol individually added to the media. Well-developed plantlets regenerated from ‘encapsulated’ nodal segments were successfully acclimatized inside the growing room with 90% survival frequency. Gas chromatography-flame ionization detection (GC-FID) was used to compare the chemical profile and the concentration of the different cannabinoids (cannabidiol, cannabichromene, cannabigerol, cannabinol, Δ9-tetrahydrocannabinol and tetrahydrocannabivarin) of the plants grown from ‘encapsulated’ nodal segments to that of the donor plant. The data showed similar cannabinoid profile and insignificant differences in the cannabinoids content between the two types of plants. This study is of high significance since the encapsulation technology would allow the prolonged storage (thus reducing the cost of labor) of high-yielding C. sativa germplasm selected for the isolation of THC, a high-value bulk active pharmaceutic.




https://link.springer.com/article/10....705-011-1168-8

Arch Virol DOI 10.1007/s00705-011-1168-8
Complete sequence of a cryptic virus from hemp (Cannabis sativa)
Angelika Ziegler • Jaroslav Matousˇek •
Gerhard Steger • Jorg Schubert
Received: 9 August 2011 / Accepted: 29 October 2011
Springer-Verlag 2011
Abstract
Hemp (Cannabis sativa) was found to be a
useful propagation host for hop latent virus, a carlavirus.
However, when virus preparations were analysed by elec-
tron microscopy, along with the expected filamentous
particles, spherical particles with a diameter of around
34 nm were found. RNA from virus preparations was
purified, and cDNA was prepared and cloned. Sequence
information was used to search databases, and the greatest
similarity was found with Primula malacoides virus 1, a
putative new member of the genus Partitivirus. The full
sequences of RNA 1 and RNA 2 of this new hemp cryptic
virus were obtained.

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Old 06-19-2015, 07:20 PM #10
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https://journals.plos.org/plosone/art...l.pone.0099641

Understanding Cultivar-Specificity and Soil Determinants of the Cannabis Microbiome
Max E. Winston , Jarrad Hampton-Marcell, Iratxe Zarraonaindia, Sarah M. Owens, Corrie S. Moreau, Jack A. Gilbert, Josh Hartsel, Suzanne J. Kennedy, S. M. Gibbons

Abstract
Understanding microbial partnerships with the medicinally and economically important crop Cannabis has the potential to affect agricultural practice by improving plant fitness and production yield. Furthermore, Cannabis presents an interesting model to explore plant-microbiome interactions as it produces numerous secondary metabolic compounds. Here we present the first description of the endorhiza-, rhizosphere-, and bulk soil-associated microbiome of five distinct Cannabis cultivars. Bacterial communities of the endorhiza showed significant cultivar-specificity. When controlling cultivar and soil type the microbial community structure was significantly different between plant cultivars, soil types, and between the endorhiza, rhizosphere and soil. The influence of soil type, plant cultivar and sample type differentiation on the microbial community structure provides support for a previously published two-tier selection model, whereby community composition across sample types is determined mainly by soil type, while community structure within endorhiza samples is determined mainly by host cultivar.
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