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RECENT interesting findings

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
One more new one:
I find interesting as CBD has been reported to not bind to the CB1 yet it does modulate THC, this is how.
-SamS

http://www.ncbi.nlm.nih.gov/pubmed/26218440

Br J Pharmacol. 2015 Jul 27.
Cannabidiol is a negative allosteric modulator of the type 1 cannabinoid receptor.

BACKGROUND AND PURPOSE:
Cannabidiol has been reported to act as an antagonist of cannabinoid agonists at type 1 cannabinoid receptors (CB1 ). We hypothesized that cannabidiol can inhibit cannabinoid agonist activity through negative allosteric modulation of CB1 .
 
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R043

New member
I first would like to thank the OP for this initiative which I found quite interesting and informative.

Secondly regarding the view that some shared, that the OP should have take the time to vulgarize the different study seems to me as lacking perspective.

I personally (and would think others are sharing my point of view) would rather have access to the material the OP offered than nothing at all and if he does not have the time to proceed with a full explanation of each article as he stated, it seems to me it would makes more sens to thank him instead of implying that it would have been his responsibility to do so.

Finally, a bit earlier today while browsing the interwebs I have stumble upon an interesting abstract I collected and after reading this whole thread, though it might be appropriate to share it.

My apologies if most of you are already aware of this study of if it seems unreliable as I do not feel competent enough to judge of this by myself.


Planta Med. 2010 Nov;76(16):1938-9. doi: 10.1055/s-0030-1249978. Epub 2010 Jun 8.

Genetic identification of female Cannabis sativa plants at early developmental stage.

Techen N1, Chandra S, Lata H, Elsohly MA, Khan IA.
Author information

Abstract

Sequence-characterized amplified region (SCAR) markers were used to identify female plants at an early developmental stage in four different varieties of Cannabis sativa. Using the cetyl trimethylammonium bromide (CTAB) method, DNA was isolated from two-week-old plants of three drug-type varieties (Terbag W1, Terbag K2, and Terbag MX) and one fiber-type variety (Terbag Fedora A7) of C. sativa grown under controlled environmental conditions through seeds. Attempts to use MADC2 (male-associated DNA from Cannabis sativa) primers as a marker to identify the sex of Cannabis sativa plants were successful. Amplification of genomic DNA using MADC2-F and MADC2-R primers produced two distinct fragments, one with a size of approximately 450 bp for female plants and one for male plants with a size of approximately 300 bp. After harvesting the tissues for DNA extraction, plants were subjected to a flowering photoperiod (i.e., 12-h light cycle), and the appearance of flowers was compared with the DNA analysis. The results of the molecular analysis were found to be concordant with the appearance of male or female flowers. The results of this study represent a quick and reliable technique for the identification of sex in Cannabis plants using SCAR markers at a very early developmental stage.

© Georg Thieme Verlag KG Stuttgart · New York.

PMID:20533168[PubMed - indexed for MEDLINE]
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
https://www.esciencecentral.org/jou...s-in-cannabis-2329-6836-1000181.php?aid=57624

https://dx.doi.org/10.4172/2329-6836.1000181

Open access full text!!

Natural Products Chemistry & Research
Cannabinoids and Terpenes as Chemotaxonomic Markers in Cannabis
Elzinga S, Fischedick J, Podkolinski R, and Raber J

Abstract
In this paper, we present principal component analysis (PCA) results from a dataset containing 494 cannabis
flower samples and 170 concentrate samples analyzed for 31 compounds. A continuum of chemical composition
amongst cannabis strains was found instead of distinct chemotypes. Our data shows that some strains are much
more reproducible in chemical composition than others. Strains labeled as indica were compared with those labeled
as sativa and no evidence was found that these two cultivars are distinctly different chemotypes. PCA of “OG” and
“Kush” type strains found that “OG” strains have relatively higher levels of α-terpineol, fenchol, limonene, camphene, terpinolene and linalool where “Kush” samples are characterized mainly by the compounds trans-ocimene, guaiol, β-eudesmol, myrcene and α-pinene. The composition of concentrates and flowers were compared as well. Although the absolute concentration of compounds in concentrates is much higher, the relative composition of compounds between flowers and concentrates is similar.
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https://www.docdroid.net/teh6qCm/101007s12229-015-9157-3.pdf.html
You can download the pdf here, just push the download button at the site

https://link.springer.com/article/10.1007/s12229-015-9157-3
DOI:10.1007/s12229-015-9157-3

The Botanical Review
August 2015
Date: 19 Aug 2015
Evolution and Classification of Cannabis sativa (Marijuana, Hemp) in Relation to Human Utilization
Ernest Small

Abstract
Cannabis sativa has been employed for thousands of years, primarily as a source of a stem fiber (both the plant and the fiber termed “hemp”) and a resinous intoxicant (the plant and its drug preparations commonly termed “marijuana”). Studies of relationships among various groups of domesticated forms of the species and wild-growing plants have led to conflicting evolutionary interpretations and different classifications, including splitting C. sativa into several alleged species. This review examines the evolving ways Cannabis has been used from ancient times to the present, and how human selection has altered the morphology, chemistry, distribution and ecology of domesticated forms by comparison with related wild plants. Special attention is given to classification, since this has been extremely contentious, and is a key to understanding, exploiting and controlling the plant. Differences that have been used to recognize cultivated groups within Cannabis are the results of disruptive selection for characteristics selected by humans. Wild-growing plants, insofar as has been determined, are either escapes from domesticated forms or the results of thousands of years of widespread genetic exchange with domesticated plants, making it impossible to determine if unaltered primeval or ancestral populations still exist. The conflicting approaches to classifying and naming plants with such interacting domesticated and wild forms are examined. It is recommended that Cannabis sativa be recognized as a single species, within which there is a narcotic subspecies with both domesticated and ruderal varieties, and similarly a non-narcotic subspecies with both domesticated and ruderal varieties. An alternative approach consistent with the international code of nomenclature for cultivated plants is proposed, recognizing six groups: two composed of essentially non-narcotic fiber and oilseed cultivars as well as an additional group composed of their hybrids; and two composed of narcotic strains as well as an additional group composed of their hybrids.
 
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MrBelvedere

Active member
ICMag Donor
Thx, this has a lot of good info on trying to understand whorling...

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4423988/?report=classic

A Dynamical Phyllotaxis Model to Determine Floral Organ Number

Abstract
How organisms determine particular organ numbers is a fundamental key to the development of precise body structures; however, the developmental mechanisms underlying organ-number determination are unclear. In many eudicot plants, the primordia of sepals and petals (the floral organs) first arise sequentially at the edge of a circular, undifferentiated region called the floral meristem, and later transition into a concentric arrangement called a whorl, which includes four or five organs. The properties controlling the transition to whorls comprising particular numbers of organs is little explored. We propose a development-based model of floral organ-number determination, improving upon earlier models of plant phyllotaxis that assumed two developmental processes: the sequential initiation of primordia in the least crowded space around the meristem and the constant growth of the tip of the stem. By introducing mutual repulsion among primordia into the growth process, we numerically and analytically show that the whorled arrangement emerges spontaneously from the sequential initiation of primordia. Moreover, by allowing the strength of the inhibition exerted by each primordium to decrease as the primordium ages, we show that pentamerous whorls, in which the angular and radial positions of the primordia are consistent with those observed in sepal and petal primordia in Silene coeli-rosa, Caryophyllaceae, become the dominant arrangement. The organ number within the outmost whorl, corresponding to the sepals, takes a value of four or five in a much wider parameter space than that in which it takes a value of six or seven. These results suggest that mutual repulsion among primordia during growth and a temporal decrease in the strength of the inhibition during initiation are required for the development of the tetramerous and pentamerous whorls common in eudicots.
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
One more:

https://www.plosone.org/article/fet....1371/journal.pone.0133292&representation=PDF

The Genetic Structure of Marijuana and Hemp

Jason Sawler, Jake M. Stout, Kyle M. Gardner, Darryl Hudson, John Vidmar, Laura Butler, Jonathan E. Page, Sean Myles PLOS x
Published: August 26, 2015
DOI: 10.1371/journal.pone.0133292

Despite its cultivation as a source of food, fibre and medicine, and its global status as the most used illicit drug, the genus Cannabis has an inconclusive taxonomic organization and evolutionary history. Drug types of Cannabis (marijuana), which contain high amounts of the psychoactive cannabinoid Δ9-tetrahydrocannabinol (THC), are used for medical purposes and as a recreational drug. Hemp types are grown for the production of seed and fibre, and contain low amounts of THC. Two species or gene pools (C. sativa and C. indica) are widely used in describing the pedigree or appearance of cultivated Cannabis plants. Using 14,031 single-nucleotide polymorphisms (SNPs) genotyped in 81 marijuana and 43 hemp samples, we show that marijuana and hemp are significantly differentiated at a genome-wide level, demonstrating that the distinction between these populations is not limited to genes underlying THC production. We find a moderate correlation between the genetic structure of marijuana strains and their reported C. sativa and C. indica ancestry and show that marijuana strain names often do not reflect a meaningful genetic identity. We also provide evidence that hemp is genetically more similar to C. indica type marijuana than to C. sativa strains.
 
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oldchuck

Active member
Veteran
Man, this taxonomy tangle is fascinating but it's also nuts:

"We also provide evidence that hemp is genetically more similar to C. indica type marijuana than to C. sativa strains."

Linnaeus originally defined Cannabis Sativa as hemp. Now these guys are saying hemp is genetically more like Cannabis Indica which Lamarck thought came from India but now is said to be actually Cannabis Afganica.

Clearly, Cannabis is a global genetic stewpot but the fact remains that Cannabis from everywhere is willing and able to breed freely with Cannabis from everywhere else. That makes it a single species in my humble opinion.
 

Only Ornamental

Spiritually inspired agnostic mad scientist
Veteran
...
Clearly, Cannabis is a global genetic stewpot but the fact remains that Cannabis from everywhere is willing and able to breed freely with Cannabis from everywhere else. That makes it a single species in my humble opinion.
Hi oldchuck
Look at cacti and orchids, many breed freely with each other (obviously cactus with cactus and orchid with orchid). Still, they are only loosely related and comprise very different genera.
 

Donn

Member
Sure, and orchid taxonomy too - not something I know a lot about, but happened this week to be reading about Brazilian Laelia species, which were then reclassified as Sophronitis, but that only lasted a few years and then they got shuffled over to Cattleya. Lots of very popular orchids are Laeliocattleya hybrids - but maybe with parents that are now both Cattleya? so in principle that needs to be straightened out too. And likewise the Brassolaeliocattleya hybrids are usually a parent that was removed from Brassavola some time back, and now they're really Ryncholaeliocattleya.

It's wonderful that they're able to make these giant strides with their taxonomy, but it doesn't really serve the trade very well, I'm guessing. The same could be said for the sativa/indica/afghanica thing. Sometimes I think there ought to be some stiff fee attached to changing a botanical name that's in wide use.
 

MrBelvedere

Active member
ICMag Donor
Plant taxonomy is just us humans making a lame attempt at trying to assign words to differentiate plants, nothing more, nothing less.
 

oldchuck

Active member
Veteran
I'm reading that genetic study,[FONT=Arial, Helvetica, sans-serif] The Genetic Structure of Marijuana and Hemp, cited above. My understanding of it is limited but the following strikes me as a good reason to consider Cannabis a single species:[/FONT]

The average FST between hemp and marijuana is 0.156 (S1 Fig), which is similar to the degree of genetic differentiation in humans between Europeans and East Asians [9]. Thus, while cannabis breeding has resulted in a clear genetic differentiation according to use, hemp and marijuana still largely share a common pool of genetic variation.


The genetic difference between Europeans and East Asians does not stike me as sufficient to declare them separate species.
 

Only Ornamental

Spiritually inspired agnostic mad scientist
Veteran
I understand it in such a way that the nowadays cannabis and hemp varieties are a topsy-turvy of crosses and hybrids between many different 'species'. The seemingly closer relationship of hemp and 'indica' (aka BLD) might be the reason for BLH or broad leaflet hemp commonly associated with southern ecotypes (which BTW are common ancestors of many modern hemp varieties).
One remark: The CAN57 they mention is a Syrian collection designated as Cannabis sp. and NOT as hemp (C. sativa)! Morphologically, it's very similar to an 'indica' hash plant with a stout branched stature, absent stretch, broad leaflets, and the typical 'hash' smell. As a 'wild' collection it might be anything from hemp over multi-purpose variety to pure drug cultivar.
 

Sativied

Well-known member
Veteran
But I want intersex tests, that is what I could use and a lot of other people also.
-SamS

Testlabamsterdam (.nl) claims to offer, translated from their Services page, gender tests to "determine gender and perform a genetic check (M/F/Hermaphrodite)". 100e for 10 samples. Don't know how and if reliable but they've been offering that service for a while already.

----------

Full pdf: www. docdroid.net/teh6qCm/101007s12229-015-9157-3.pdf.html

For this one:
http://link.springer.com/article/10....229-015-9157-3


The Botanical Review
August 2015
Date: 19 Aug 2015
Evolution and Classification of Cannabis sativa (Marijuana, Hemp) in Relation to Human Utilization
Ernest Small

Abstract
Cannabis sativa has been employed for thousands of years, primarily as a source of a stem fiber (both the plant and the fiber termed “hemp”) and a resinous intoxicant (the plant and its drug preparations commonly termed “marijuana”). Studies of relationships among various groups of domesticated forms of the species and wild-growing plants have led to conflicting evolutionary interpretations and different classifications, including splitting C. sativa into several alleged species. This review examines the evolving ways Cannabis has been used from ancient times to the present, and how human selection has altered the morphology, chemistry, distribution and ecology of domesticated forms by comparison with related wild plants. Special attention is given to classification, since this has been extremely contentious, and is a key to understanding, exploiting and controlling the plant. Differences that have been used to recognize cultivated groups within Cannabis are the results of disruptive selection for characteristics selected by humans. Wild-growing plants, insofar as has been determined, are either escapes from domesticated forms or the results of thousands of years of widespread genetic exchange with domesticated plants, making it impossible to determine if unaltered primeval or ancestral populations still exist. The conflicting approaches to classifying and naming plants with such interacting domesticated and wild forms are examined. It is recommended that Cannabis sativa be recognized as a single species, within which there is a narcotic subspecies with both domesticated and ruderal varieties, and similarly a non-narcotic subspecies with both domesticated and ruderal varieties. An alternative approach consistent with the international code of nomenclature for cultivated plants is proposed, recognizing six groups: two composed of essentially non-narcotic fiber and oilseed cultivars as well as an additional group composed of their hybrids; and two composed of narcotic strains as well as an additional group composed of their hybrids.
 

Buckowens

Member
Sam,
I really like the idea of the intersex tests. But there is one thing that is bugging me about how accurate these tests may be.

I have grown thousands of plants, hundreds of clone only varieties and one thing is always true about the plants, they never act the same. Some varieties that are known for their predisposition for intersex traits, and some that are know for how solid they preform (fewer intersex tendencies), yet every now and again I will get minor male expressions to full blown early male flowering. Or in some cases nothing at all.
How will these tests take into account abiotic or environmental stresses that may cause these expressions?
Not all hermaphrodite plants are hermaphrodite all the time, so what are the perimeters for these tests? If I am screening seeds from a chemdog line or sour diesel line for example, what are the odds that the tests will indicate that all plants are intersex or none of the plants re intersex?
Thanks,
Paco
 

Stradel

Member
Oouu so those wild hemp in my country from 2 to 5 meters long thin leafs and clear optimistic no burnout high when i make milk and coockies from them are indicas muhahahaha this is a sadness laugh because i always think this are sativa caracteristics im so stupid omg all thos years i didnt know the difrence betwen ind and sats i am so confused now.:))))
 

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
Here are a few more, new and old. A lot are Molecular Biology, not really my forte. If anyone has links to the full papers post them and I will add the links.
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Purification and characterization of cannabidiolic-acid synthase from Cannabis sativa L. Biochemical analysis of a novel enzyme that catalyzes the oxidocyclization of cannabigerolic acid to cannabidiolic acid

Futoshi Taura, S Morimoto, Yukihiro Shoyama
Faculty of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812, Japan.
Journal of Biological Chemistry . 08/1996; 271(29):17411-6. DOI:*10.1074/jbc.271.29.17411

ABSTRACT We identified a unique enzyme that catalyzes the oxidocyclization of cannabigerolic acid to cannabidiolic acid (CBDA) in Cannabis sativa L. (CBDA strain). The enzyme, named CBDA synthase, was purified to apparent homogeneity by a four-step procedure: ammonium sulfate precipitation followed by chromatography on DEAE-cellulose, phenyl-Sepharose CL-4B, and hydroxylapatite. The active enzyme consists of a single polypeptide with a molecular mass of 74 kDa and a pI of 6.1. The NH2-terminal amino acid sequence of CBDA synthase is similar to that of Delta1-tetrahydrocannabinolic-acid synthase. CBDA synthase does not require coenzymes, molecular oxygen, hydrogen peroxide, and metal ion cofactors for the oxidocyclization reaction. These results indicate that CBDA synthase is neither an oxygenase nor a peroxidase and that the enzymatic cyclization does not proceed via oxygenated intermediates. CBDA synthase catalyzes the formation of CBDA from cannabinerolic acid as well as cannabigerolic acid, although the kcat for the former (0.03 s-1) is lower than that for the latter (0.19 s-1). Therefore, we conclude that CBDA is predominantly biosynthesized from cannabigerolic acid rather than cannabinerolic acid.

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

Tetrahydrocannabinolic acid synthase, the enzyme controlling marijuana psychoactivity is secreted into the storage cavity of the glandular trichomes.

Supaart Sirikantaramas, Futoshi Taura, Yumi Tanaka, Yu Ishikawa, Satoshi Morimoto, Yukihiro Shoyama
Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582 Japan.
Plant and Cell Physiology . 10/2005; 46(9):1578-82. DOI:*10.1093/pcp/pci166

ABSTRACT Tetrahydrocannabinolic acid (THCA) synthase is the enzyme responsible for the production of tetrahydrocannabinol (THC), the psychoactive component of marijuana (Cannabis sativa L.). We suggest herein that THCA is biosynthesized in the storage cavity of the glandular trichomes based on the following observations. (i) The exclusive expression of THCA synthase was confirmed in the secretory cells of glandular trichomes by reverse transcription-PCR (RT-PCR) analysis. (ii) THCA synthase activity was detected in the storage cavity content. (iii) Transgenic tobacco expressing THCA synthase fused to green fluorescent protein showed fluorescence in the trichome head corresponding to the storage cavity. These results also showed that secretory cells of the glandular trichomes secrete not only metabolites but also biosynthetic enzyme.


----------------------------------------------------------------------------
http://www.jbc.org/content/282/28/20739.full
Identification and Characterization of Cannabinoids That Induce Cell Death through Mitochondrial Permeability Transition in Cannabis Leaf Cells
doi: 10.1074/jbc.M700133200
July 13, 2007 The Journal of Biological Chemistry, 282, 20739-20751.
Satoshi Morimoto 1 , Yumi Tanaka, Kaori Sasaki, Hiroyuki Tanaka, Tomohide Fukamizu, Yoshinari Shoyama, Yukihiro Shoyama and Futoshi Taura

Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan

Cannabinoids are secondary metabolites stored in capitate-sessile glands on leaves of Cannabis sativa. We discovered that cell death is induced in the leaf tissues exposed to cannabinoid resin secreted from the glands, and identified cannabichromenic acid (CBCA) and Δ1-tetrahydrocannabinolic acid (THCA) as unique cell death mediators from the resin. These cannabinoids effectively induced cell death in the leaf cells or suspension-cultured cells of C. sativa, whereas pretreatment with the mitochondrial permeability transition (MPT) inhibitor cyclosporin A suppressed this cell death response. Examinations using isolated mitochondria demonstrated that CBCA and THCA mediate opening of MPT pores without requiring Ca2+ and other cytosolic factors, resulting in high amplitude mitochondrial swelling, release of mitochondrial proteins (cytochrome c and nuclease), and irreversible loss of mitochondrial membrane potential. Therefore, CBCA and THCA are considered to cause serious damage to mitochondria through MPT. The mitochondrial damage was also confirmed by a marked decrease of ATP level in cannabinoid-treated suspension cells. These features are in good accord with those of necrotic cell death, whereas DNA degradation was also observed in cannabinoid-mediated cell death. However, the DNA degradation was catalyzed by nuclease(s) released from mitochondria during MPT, indicating that this reaction was not induced via a caspase-dependent apoptotic pathway. Furthermore, the inhibition of the DNA degradation only slightly blocked the cell death induced by cannabinoids. Based on these results, we conclude that CBCA and THCA have the ability to induce necrotic cell death via mitochondrial dysfunction in the leaf cells of C. sativa.

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Cannabinoids Production by Hairy Root Cultures of Cannabis sativa L.
Sayed Farag, Oliver Kayser*
American Journal of Plant Sciences, 2015, 6, 1874-1884
Technische Biochemie, Fachbereich Bio- und Chemieingenieurwesen, Technische Universität Dortmund, Dortmund, Germany
Abstract:
Tetrahydrocannabinol (THC) derivatives are used clinically as analgesic, anti-inflammatory, appetite stimulant, anti-emetic and anti-tumor cannabinoids. THC and its related compounds are at present obtained by extraction from intact Cannabis plants or chemical synthesis, but plant cell cultures may be an alternative source of production. In the present study, hairy root cultures of C. sativa (Cannabaceae) were induced by incubation of aseptically grown callus culture with solid B5 medium supplemented with 4 mg/l naphthaleneacetic acid in darkness at 25 ̊C. Hairy root growth profiles in shake flask, increased periodically during 35 days of growth cycle. The cannabinoid contents produced in minor levels and remained below 2.0 μg/g dry weight. The contents of can-
nabinoid were analyzed by liquid chromatography and confirmed by mass spectrometry.

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

Production of D9-tetrahydrocannabinolic acid from cannabigerolic acid by whole cells of Pichia (Komagataella) pastoris expressing D9-tetrahydrocannabinolic acid synthase from Cannabis sativa L.
Bastian Zirpel . Felix Stehle . Oliver Kayser
Biotechnol Lett (2015) 37:1869–1875
DOI 10.1007/s10529-015-1853-x
Abstract
Objective The D9-tetrahydrocannabinolic acid synthase (THCAS) from Cannabis sativa was expressed
intracellularly in different organisms to investigate the potential of a biotechnological production of D9-
tetrahydrocannabinolic acid (THCA) using whole cells. Results Functional expression of THCAS was ob-
tained in Saccharomyces cerevisiae and Pichia (Ko- magataella) pastoris using a signal peptide from the
vacuolar protease, proteinase A. No functional expression was achieved in Escherichia coli. The highest
volumetric activities obtained were 98 pkat ml-1 (intracellular) and 44 pkat ml-1 (extracellular) after
192 h of cultivation at 15 C using P. pastoris cells. Low solubility of CBGA prevents the THCAS appli-
cation in aqueous cell-free systems, thus whole cells were used for a bioconversion of cannabigerolic acid
(CBGA) to THCA. Finally, 1 mM (0.36 g THCA l-1) THCA could be produced by 10.5 gCDW l-1 before
enzyme activity was lost.
Conclusion
Whole cells of P. pastoris offer the capability of synthesizing pharmaceutical THCA production

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

Molecular analysis of genetic fidelity in Cannabis sativa L. plants grown from synthetic (encapsulated) seeds following in vitro storage
Biotechnology Letters
December 2011, Volume 33, Issue 12, pp 2503-2508
Hemant Lata, Suman Chandra , Natascha Techen, Ikhlas A. Khan, Mahmoud A. ElSohly

Abstract
The increasing utilization of synthetic (encapsulated) seeds for germplasm conservation and propagation necessitates the assessment of genetic stability of conserved propagules following their plantlet conversion. We have assessed the genetic stability of synthetic seeds of Cannabis sativa L. during in vitro multiplication and storage for 6 months at different growth conditions using inter simple sequence repeat (ISSR) DNA fingerprinting. Molecular analysis of randomly selected plants from each batch was conducted using 14 ISSR markers. Of the 14 primers tested, nine produced 40 distinct and reproducible bands. All the ISSR profiles from in vitro stored plants were monomorphic and comparable to the mother plant which confirms the genetic stability among the clones. GC analysis of six major cannabinoids [Δ9-tetrahydrocannabinol, tetrahydrocannabivarin, cannabidiol, cannabichromene, cannabigerol and cannabinol] showed homogeneity in the re-grown clones and the mother plant with insignificant differences in cannabinoids content, thereby confirming the stability of plants derived from synthetic seeds following 6 months storage.

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

Recent Advances in Cannabis sativa Research: Biosynthetic Studies and Its Potential in Biotechnology.

Supaart Sirikantaramas, Futoshi Taura, Satoshi Morimoto, Yukihiro Shoyama
Graduate School of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan.
Current pharmaceutical biotechnology. 09/2007; 8(4):237-43. DOI:*10.2174/138920107781387456
ABSTRACT:
Cannabinoids, consisting of alkylresorcinol and monoterpene groups, are the unique secondary metabolites that are found only in Cannabis sativa. Tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabichromene (CBC) are well known cannabinoids and their pharmacological properties have been extensively studied. Recently, biosynthetic pathways of these cannabinoids have been successfully established. Several biosynthetic enzymes including geranylpyrophosphate:eek:livetolate geranyltransferase, tetrahydrocannabinolic acid (THCA) synthase, cannabidiolic acid (CBDA) synthase and cannabichromenic acid (CBCA) synthase have been purified from young rapidly expanding leaves of C. sativa. In addition, molecular cloning, characterization and localization of THCA synthase have been recently reported. THCA and cannabigerolic acid (CBGA), its substrate, were shown to be apoptosis-inducing agents that might play a role in plant defense. Transgenic tobacco hairy roots expressing THCA synthase can produce THCA upon feeding of CBGA. These results open the way for biotechnological production of cannabinoids in the future.

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

Production of Δ1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris

Futoshi Taura, Emi Dono, Supaart Sirikantaramas, Kohji Yoshimura, Yukihiro Shoyama,
Satoshi Morimoto
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
Biochemical and Biophysical Research Communications. 10/2007; 361(3):675-80. DOI:*10.1016/j.bbrc.2007.07.079

ABSTRACT Delta(1)-Tetrahydrocannabinolic acid (THCA) synthase is the enzyme that catalyzes the oxidative cyclization of cannabigerolic acid into THCA, the acidic precursor of Delta(1)-tetrahydrocannabinol. We developed a novel expression system for THCA synthase using a methylotrophic yeast Pichia pastoris as a host. Under optimized conditions, the transgenic P. pastoris secreted approximately 1.32nkat/l of THCA synthase activity, and the culture medium, from which the cells were removed, effectively synthesized THCA from cannabigerolic acid with a approximately 98% conversion rate. The secreted THCA synthase was readily purified to homogeneity. Interestingly, endoglycosidase treatment afforded a deglycosylated THCA synthase with more catalytic activity than that of the glycosylated form. The non-glycosylated THCA synthase should be suitable for structure-function studies because it displayed much more activity than the previously reported native enzyme from Cannabis sativa as well as the recombinant enzyme from insect cell cultures.
----------------------------------------
 
Last edited:

Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
I hope for several intersex tests, one for intersex genes and one for stress related intersex expression, I assume they are not the same.
If intersex plants have intersex genes like male and female sex chromsome genes, they can be found, If stress is required to express intersex then the genes that allow intersex expression with stress can be found, I hope.
I am looking, as are others...
-SamS


Sam,
I really like the idea of the intersex tests. But there is one thing that is bugging me about how accurate these tests may be.

I have grown thousands of plants, hundreds of clone only varieties and one thing is always true about the plants, they never act the same. Some varieties that are known for their predisposition for intersex traits, and some that are know for how solid they preform (fewer intersex tendencies), yet every now and again I will get minor male expressions to full blown early male flowering. Or in some cases nothing at all.
How will these tests take into account abiotic or environmental stresses that may cause these expressions?
Not all hermaphrodite plants are hermaphrodite all the time, so what are the perimeters for these tests? If I am screening seeds from a chemdog line or sour diesel line for example, what are the odds that the tests will indicate that all plants are intersex or none of the plants re intersex?
Thanks,
Paco
 

Buckowens

Member
I hope for several intersex tests, one for intersex genes and one for stress related intersex expression, I assume they are not the same.
If intersex plants have intersex genes like male and female sex chromsome genes, they can be found, If stress is required to express intersex then the genes that allow intersex expression with stress can be found, I hope.
I am looking, as are others...
-SamS
Thanks Sam.
 

Tom Hill

Active member
Veteran
"The mean volume of the resin heads of the narcotic strains was more than four times larger than that of the industrial hemp strains. """""This is the first documented report of a consistent morphological separator of elite narcotic strains and non-narcotic plants."""" (Ernest Small , Steve G. U. Naraine 4/2015) "

^^^^^ A rather ambitious statement lol..

but good stuff Sam thank you..
 

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