Tag: Breeding

Cannabis plants having modified expression of thca synthase

Patent: WO2016189384A1 
Inventor Ekaterina Alexandra Boudko, Thomas Shipley IV, Douglas Johnson
 https://patents.google.com/patent/WO2016189384A1/en

The invention relates to novel genetically modified plants and methods or materials, such as polynucleotides, expression cassettes, or vectors for producing the same. Moreover, the invention relates to altering the content of cannabinoids in plants and to medical compositions derived from such plants. In particular embodiments, the present invention relates to cannabis plants having modified expression of tetrahydrocannabinolic acid (THCA) synthase and methods of modifying the amount of delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) in cannabis by modifying expression of THCA synthase.

Abstract

*Qualitative and Quantitative Aspects of the Inheritance of Chemical Phenotype in Cannabis

Giuseppe Mandolino, Manuela Bagatta, Andrea Carboni, Paolo Ranalli, Etienne de Meijer

March 2003 Journal of Industrial Hemp 8(2):51-72

DOI: 10.1300/J237v08n02_04

Four crosses were made between Cannabis plants with “pure” CBD and THC chemotypes. The F1 plants obtained were self-fertilised to produce F2s. Chemotypical distributions were analysed by gas-chromatography. A segregation analysis of the different F2 progenies obtained showed that chemotype, estimated as CBD/THC ratio, behaves as a qualitative character, and a model for a single locus B, with two co-dominant alleles, BD and BT is described. The CBD/THC ratios in the F1 offsprings were found to be significantly different in the heterozygous plants from the different pedigrees. The amount of CBD plus THC in the same pedigrees was also described. Heterosis was found to be a common feature, but not a general one, of cannabinoid accumulation in the F1s. Distribution of the values of cannabinoid content in classes was found to be normal. RAPD markers linked to the segregating chemotypes (“pure” CBD and “pure” THC) were identified by bulk segregant analysis, and the degree of linkage of these markers with the chemotype was described.

Abstract

Gene duplication and divergence affecting drug content in Cannabis sativa

George D. Weiblen, Jonathan P. Wenger, Kathleen J. Craft, Mahmoud A. ElSohly, Zlatko Mehmedic, Erin L. Treiber, M. David Marks

New Phytologist 17 July 2015

DOI: 10.1111/nph.13562

Cannabis sativa is an economically important source of durable fibers, nutritious seeds, and psychoactive drugs but few economic plants are so poorly understood genetically. Marijuana and hemp were crossed to evaluate competing models of cannabinoid inheritance and to explain the predominance of tetrahydrocannabinolic acid (THCA) in marijuana compared with cannabidiolic acid (CBDA) in hemp. Individuals in the resulting F2 population were assessed for differential expression of cannabinoid synthase genes and were used in linkage mapping. Genetic markers associated with divergent cannabinoid phenotypes were identified. Although phenotypic segregation and a major quantitative trait locus (QTL) for the THCA/CBDA ratio were consistent with a simple model of codominant alleles at a single locus, the diversity of THCA and CBDA synthase sequences observed in the mapping population, the position of enzyme coding loci on the map, and patterns of expression suggest multiple linked loci. Phylogenetic analysis further suggests a history of duplication and divergence affecting drug content. Marijuana is distinguished from hemp by a nonfunctional CBDA synthase that appears to have been positively selected to enhance psychoactivity. An unlinked QTL for cannabinoid quantity may also have played a role in the recent escalation of drug potency.

Abstract

Chemotaxonomy Of Cannabis I. Crossbreeding Between Cannabis Sativa And C. Ruderalis, With Analysis Of Cannabinoid Content

JOHN A. BEUTLER, ARA H. DER MARDEROSIAN
Economic Botany, 1978; 32(4) 387-394
DOI: 10.1007/BF02907934

A controlled cross between Cannabis sativa L. and C. ruderalis Janisch. gave progeny intermediate in both cannabinoid content and morphology. The progeny fell into two distinct populations, those whose tetrahydrocannabinol (THC) content was closer to the C. sativa parent (greater than 60% of total cannabinoids) and those whose THC content was closer to the C. ruderalis parent (less than 40% of total cannabinoids). The lower THC group was twice as frequent as the other group. Earliness of flowering, number of flowers, and height characteristics were intermediate between the parents.

Abstract

Cannabinoid research in the 2010s

Mauro Maccarrone and Steve PH Alexander
British Journal of Pharmacology 165(8) 2409-2410
DOI: 10.1111/j.1476-5381.2012.01930.x

Cannabis sativa is possibly the plant with the longest history of cultivation by man (Russo, 2007). It has long been exploited for its fibre; as a biomass converter, it has exceptional utility. For most people, however, there is the association of cannabis with ‘recreational drugs’, which has lead to the profusion of names associated with the plant and extracts thereof (marijuana,hashish, bhang, weed, grass, etc.). The ‘modern’ scientific era of cannabis research was prompted by the discovery of the major psychoactive ingredient in cannabis extracts (Gaoni and Mechoulam, 1964). This was, of course, D9-tetrahydrocannabinol or THC. Raphael Mechoulam has numerous publications, filled with seminal observations, including the identification of the two ‘best’ candidates for endogenous cannabinoid molecules: anandamide (Devane et al., 1992) and 2-arachidonoylglycerol (Mechoulam et al., 1995). He has become something of an icon in the cannabis field, with this issue of BJP containing a series of original articles prompted by a symposium held in Jerusalem in November 2010 to celebrate his 80th birthday. The first issue, entitled ‘Cannabinoids in Biology and Medicine’, containing primarily reviews, was published in August 2011 (https://onlinelibrary.wiley.com/doi/…sue-7/issuetoc). Current research incannabinoid-related areas is vibrant, with the added focus of TRPV1 ion channels, PPAR nuclear receptors and the ‘orphan’ G-protein coupled receptors, GPR18, GPR55 and GPR119, as molecular targets of cannabinoids and cannabinoid-like molecules. Furthermore, the identification of endogenous agonists at cannabinoid receptors which lead to the demonstration of multiple routes for synthesis and transformation of these endocannabinoids has added to the molecular targets available for potential exploitation.

Abstract

Variations of 9-THC content in single plants of hemp varieties


Klemens Mechtler, Josef Bailer, Karl de Hueber
Industrial Crops and Products. 19(1), 19-24
DOI: 10.1016/S0926-6690(03)00077-3

Within a given plant population, the concentration of any constituent is expected to vary within a certain bandwidth. To test the distribution of 9-tetrahydrocannabinol (THC) in hemp populations, a number of single plants were taken from populations of five well-known hemp accessions (Fasamo, Beniko, Bialobrzeskie, Félina 34, Kompolti) and a Hungarian provenance. The quantitative analysis of single plants delivered a set of 30–61 THC and Cannabidiol (CBD) values for each of the six hemp accessions under consideration. The distribution of THC within a number of hemp plants often shows no Gaussian distribution, the different varieties have quite characteristic distributions of THC. Most single-plant values are close together, the variation, however, differing from variety to variety. In addition, single plants are found with THC values far outside this bandwidth.

Abstract

*The inheritance of chemical phenotype in Cannabis sativa L. (V): regulation of the propyl-/pentyl cannabinoid ratio, completion of a genetic model

E. P. M. de Meijer . K. M. Hammond
May 2016 Euphytica 210(2)
DOI 10.1007/s10681-016-1721-3

In order to complete a genetic model for the inheritance of chemotype in Cannabis, this paper explores the regulation of the propyl-/pentyl cannabinoid ratio. Plants almost pure in compounds with a C5 side chain are by far the most common, and such a chemotype can be considered a wild-type condition. Mutant progenitors with higher levels of the rarer cannabinoid THC-C3 (tetrahydrocannabivarin) were identified. Their propyl cannabinoid proportion in the total cannabinoid fraction (PC3) ranged from 14 to 69 %, which, through selective inbreeding, could be increased to highly specific lineage maxima. Inbred plants with maximised PC3 derived from the different progenitors, were then crossed with a pure C5 wild type and the PC3 distribution patterns of the F2s examined. Distinct patterns, compatible with oligogenic and polygenic segregation appeared. It was hypothesised that the PC3 regulating loci of the six source progenitors would be at least partially different, complementary, and additive in their phenotypical effect. So, high PC3 offspring from the different lineages were mutually crossed. Inbred lines derived from multi-cross hybrid combinations reached unprecedented PC3 levels of up to 96 % which supports the hypothesis. For the regulation of C3/C5 ratios, a model of a multiple locus A 1–A 2–…A n is proposed, with the pentyl- and propyl cannabinoid pathway being enhanced by alleles A pe1−n and A pr1−n, respectively.

Abstract

*The inheritance of chemical phenotype in Cannabis sativa L. (IV): cannabinoid-free plants

E. P. M. de Meijer, E K. M. Hammond, A Sutton
Euphytica (2009) 168:95–112
DOI 10.1007/s10681-009-9894-7

A genetic factor that blocks the cannabinoid biosynthesis in Cannabis sativa has been investigated. Crosses between cannabinoid-free material and high content, pharmaceutical clones were performed. F1s were uniform and had cannabinoid contents much lower than the mean parental value. Inbred F2 progenies segregated into discrete groups: a cannabinoid-free chemotype, a chemotype with relatively low cannabinoid content and one with relatively high content, in a monogenic 1:2:1 ratio. In our model the cannabinoid knockout factor is indicated as a recessive allele o, situated at locus O, which segregates independently from previously presented chemotype loci. The genotype o/o underlies the cannabinoid-free chemotype, O/o is expressed as an intermediate, low content chemotype, and O/O is the genotype of the high content chemotype. The data suggests that locus O governs a reaction in the pathway towards the phenolic cannabinoid precursors. The composition of terpenoids and various other compound classes of cannabinoid-free segregants remains unaffected. Backcrossing produced cannabinoid-free homologues of pharmaceutical production clones with potential applications in pharmacological research. A new variant of the previously presented allele ‘B 0’, that almost completely obstructs the conversion of CBG into CBD, was also selected from the source population of the cannabinoid knockout factor.

Abstract

*The inheritance of chemical phenotype in Cannabis sativa L. (III): variation in cannabichromene proportion

E. P. M. de Meijer · K. M. Hammond · M. Micheler
Euphytica (2009) 165:293–311
DOI 10.1007/s10681-008-9787-1

The mechanism that controls the proportion of cannabichromene (CBC), a potential pharmaceutical, in the cannabinoid fraction of Cannabis sativa L. is explored. As with tetrahydrocannabinol (THC) and cannabidiol (CBD), CBC is an enzymatic conversion product of the precursor cannabigerol (CBG). CBC is reported to dominate the cannabinoid fraction of juveniles and to decline with maturation. This ontogeny was confirmed in inbred lines with different mature chemotypes. A consistent CBC presence was found in early leaves from a diverse clone collection, suggesting that CBC synthase is encoded by a fixed locus. Morphological variants possessing a ‘prolonged juvenile chemotype’ (PJC), a substantial proportion of CBC persisting up to maturity, are presented. PJC is associated with a reduced presence of floral bracts, bracteoles, and capitate-stalked trichomes. Genetic factors causing these features were independent of the allelic chemotype locus B that was previously postulated and regulates THC and CBD synthesis and CBG accumulation. In contrast to previously described Cannabis chemotypes, the cannabinoid composition of PJCs showed plasticity in that reduced light levels increased the CBC proportion. The ability of PJC plants to enable the production of pharmaceutical raw material with high CBC purity is demonstrated.

Abstract