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:D Genetic Preservation :D - Breeding

acespicoli

Well-known member
Any pictures of that red Thai The Real Seed Company at harvest?. Love that structure
Have yet to do that project, if your interested in doing the seed run your welcome to it.
It is special, wish I had time and favorable conditions .

https://therealseedcompany.com/2023/05/thai-stick-landraces-good-ganja/
The photo above is a lower branch of a Lao / ‘Thai Stick’ ganja landrace collected by Angus c. 2012 and pushed well into senescence indoors by a customer.

Strong aromas such as musk, tobacco, dark chocolate, and overripe fruit are typical of this type of Central Lao cannabis.
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@Roms
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@Roms
It is possible this ‘Red Thai’ line is related to a prized ’70s Lao/Isaan strain that was grown in this region and known to smokers as ‘Sa Daeng’ or ‘Red Ganja’.



The Red Thai originally came from The Real Seed Company.
Bodhi, famous strain hunter and breeder, collected the seeds directly at source during his 2014 yunnan china and northern laos seed safari. A joint preservation project with the Mitseedco then allowed the team at Khalifa Genetics to preserve this rare strain while improving its sexual stability indoors.

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Is this the one your interested in ?
If your resources are booked as well I will keep it in mind
:huggg:
 
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acespicoli

Well-known member
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This new Acapulco Gold release is sold with full written permission from Bodhi Seeds (@plantmoreseeds) who these Acapulco Gold seeds come from! It is my personal goal to bring back pure landrace seeds to the community as much as possible so they can be enjoyed and preserved, rather than be lost to time.
 
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acespicoli

Well-known member
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$2 seeds must be catching on @tom_hill ;)






Having a female BD cut what would be the best bx to bring the line back to regular
the NL Hash Plant or Ortega ?

Maybe a random affie ? :thinking:
The PM resistance of BD is mandatory in the line this must remain
Im thinking its mostly from the nl side
I have some huge single column Ortega bud stick regular seed some Deep Chunk affie maybe Balkhi
The ortega is on hand and these NL would keep it true to type? In a bx8 to the cut ?
Ideas??? reg BD seeds have been elusive...I want to keep as many male genetics in the lines as possible

Why is a reversed female not genetically identical to a true male?
What DNA/genes is missing in the chemical reversal ?

The Y chromosome is larger than the X chromosome, and the female plant’s haploid genome is estimated to be 818 Mb in size, while the male plant’s genome is estimated to be 843 Mb10,11.

Male pollen travels great distances by wind while the female remains in place to be out-crossed
With various male pollen from perhaps many separate donors from diverse wild populations
All the while helping to reduce inbreeding depression and genetic bottle necks ?

The prevalence of the XY system found in 44 out of 48 species may reflect the predominance of the evolutionary pathway from gynodioecy towards dioecy. All dioecious species have the potential to evolve sex chromosomes, and reversions back from dioecy to various forms of monoecy, gynodioecy, or androdioecy have also occurred. Such reversals may occur especially during the early stages of sex chromosome evolution before the lethality of the YY (or WW) genotype is established.


A final future area of investigation may be into the use of true male pollen in testing the sterility of the triploid C. sativa plants. Previous research has indicated that the pollen from reversed female C. sativa plants is frequently less viable than the pollen from true genetic males [38].
 
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GMT

The Tri Guy
Veteran
Since all you really need is the Y chromosome, and since that's recognisable 🤔, while normally a fruitless exercise, this could be the perfect time for the old cubing method. Pick the Ortega, and start backcrossing the best boys back to your true cut for a few generations. The autosome should work itself out while keeping the boy/girl thing.
Just a thought mate😉.
 

Hammerhead

Disabled Farmer
ICMag Donor
Veteran
The Red Thai originally came from The Real Seed Company.
Bodhi, famous strain hunter and breeder, collected the seeds directly at source during his 2014 yunnan china and northern laos seed safari. A joint preservation project with the Mitseedco then allowed the team at Khalifa Genetics to preserve this rare strain while improving its sexual stability indoors.
View attachment 18869795
Is this the one your interested in ?
If your resources are booked as well I will keep it in mind
:huggg:

Is this the same as Red Thai F4 from The Real Seed Company pic?. For a 30+ week strain, it didn't fill in like I thought it would. OOps, I might be looking at the wrong pic?.
 

acespicoli

Well-known member
I have missed you here @GMT
Since all you really need is the Y chromosome, and since that's recognisable 🤔, while normally a fruitless exercise, this could be the perfect time for the old cubing method. Pick the Ortega, and start backcrossing the best boys back to your true cut for a few generations. The autosome should work itself out while keeping the boy/girl thing.
Just a thought mate😉.
Glad you saw this @GMT always a pleasure reading your insights :huggg:
 
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Reactions: GMT

acespicoli

Well-known member
Is this the same as Red Thai F4 from The Real Seed Company pic?. For a 30+ week strain, it didn't fill in like I thought it would. OOps, I might be looking at the wrong pic?.
The one I have is a repro from the RSC its called Red Thai to distinguish it from the others
It is a bit confusing several of the sources work the same line although they do appear dissimilar
Im sure its worth a go what it lacks in yield it more than makes up for it in effect it what I have been told

The kinda high that you dont get from modern hybrids,
There were two separate mango thai releases, :thinking:
these should be exactly like the top two pictures leathery tobacco
He said 26 weeks another grower ran them in 18 so to veg or not to veg ?
Longer flowering would produce the desired effect with a veg of 8 weeks maybe
Id veg min 4 weeks on my first run looking for alternating phyllotaxy to signal flip
12/12 or darker from seed can be a trigger for hermaphrodite stress
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Strain Details

13 seeds per pack

The Red Thai originally came from The Real Seed Company. The plants won’t begin flowering under two months of age. The flowering time is 25+ weeks. so a minimum of 33 weeks is needed to properly grow this one. There are two variations in taste and high i found.

Smoke report on the red thai
The high starts in the head then goes down the body, bringing a blissful slightly relaxed feeling. Euphoria builds up and gets you going with its balanced energy. smoke more and drift away into your head of thoughts, a fun happy high. its effects lasting 3-4 hrs. The taste is a really nice complex lemon/citrus/frankincense with strong spice background.

There are rarer fatter leaved plants which show some red color in the stems. It formed tighter buds then the other one. The taste is at first citrusy then goes into a incense like tobacco taste that fades into an earthy chocolate. The initial high is an energy boost, your mood elevates with positive vibrations and thoughts. Then you slowly relax into bliss and may fall asleep, lasting about 3 hrs.

Original info from RSC 
Genetics: Pure Southeast Asian Ganja Plant 
Latitude: 15°N 
Regional Maturation: late December to early February 
Height: 4m or more 
Aroma: rich and ‘leathery’ like good tobacco, but nicer! 
Characteristics: long red pistils on dark calyxes, big yields, great ‘Thai’ flavour

This is a big ganja strain grown in Northeast Thailand (‘Isaan’) and Lao. Buds are typically a deep reddish-brown to mauve-brown and have unusual long red pistils (‘hairs’). Smoke is particularly smooth and rich. The precise aroma is impossible to describe: something like leathery pipe tobacco, with hints of chocolate, coffee and musk. These are large plants, capable of going over 4 metres outdoors. A superior plant to the ‘Mekong’, though unfortunately only limited stock is available at present. The high is very euphoric and long-lasting. It is possible this ‘Red Thai’ line is related to a prized ’70s Lao/Isaan strain that was grown in this region and known to smokers as ‘Sa Daeng’ or ‘Red Ganja’.
 
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acespicoli

Well-known member
Monoecy (/məˈniːsi/; adj. monoecious /məˈniːʃəs/)[1] is a sexual system in seed plants where separate male and female cones or flowers are present on the same plant.[2] It is a monomorphic sexual system comparable with gynomonoecy, andromonoecy and trimonoecy.[3]
Monoecy often co-occurs with anemophily.[2] It can prevent self-pollination in an individual flower but cannot prevent self-pollination between male and female flowers on the same plant.[4]: 32 
Monoecy in angiosperms has been of interest for evolutionary biologists since Charles Darwin.[5]

A dioecious plant, Cannabis sativa has two sex chromosomes (X and Y). The genome sizes of the diploid female and male plants were determined to be 1636 and 1683 Mbp, respectively, by flow cytometry. By the karyotype analysis, the X and Y chromosomes were found to be submetacentric and subtelocentric, respectively. The Y chromosome had the largest long arm with a satellite in the terminal of its short arm. Conspicuous condensation was specifically observed in the long arm and satellite of the Y chromosome during the prometaphase to metaphase stages. These results indicate that the Y chromosome, especially in its long arm, specifically differentiates in Cannabis sativa and might contribute to the sex determination

Dioecy, in which individual plants have either male (staminate) or female (pistillate) flowers, ensures that outbreeding will always occur.

At some point, a DNA change happened in one of the pair, creating the SRY gene. This is the gene on the Y chromosome that gets the ball rolling in turning an embryo into a male. Shortly after this happened, the X and the Y chromosomes stopped recombining with each other to keep the new SRY gene from changing.

Polymorphism, as related to genomics, refers to the presence of two or more variant forms of a specific DNA sequence that can occur among different individuals or populations. The most common type of polymorphism involves variation at a single nucleotide (also called a single-nucleotide polymorphism, or SNP).



Dutch Flowers' Metal Haze Description​

Logo Dutch Flowers

Experienced smokers know nothing comes close to the potency of a true Haze. Growers also know this, but long flowering periods, unruly phenotypes, and the infamous 1-in-5-crapshoot chance of finding a special female have held many back. Breeding an indoor-friendly Haze strain that consistently delivers superior potency has been the very focus of DF breeding efforts since its inception. To that end, the team has collected and grown enough clones and crosses to setup a small Haze museum. Synthesizing our best Haze genetics into one line posed a considerable challenge.

In early trials, proven clones passed down their potency genes on to the next generations using an unacceptable, unpredictable domino pattern. To develop a superior seed line, males were tested the hard way, growing out their offspring to recognize the best possible pollen source. Winning donors pollinated our choicest females, and their offspring was then heavily selected at the second generation. Recombinant individuals displaying the desired potency, phenotype, flowering and yield traits were identified and inbred into separate lines. Three years after the project begun, the resulting lines were finally blended, giving rise to our Metal Haze, a hybrid that received high praise from skeptic industry testers.

Named after the metal-lit room that keeps our Haze clone collection alive, Metal Haze displays the brain-warping quality that fueled the legend. It is a modern, trippy cerebral sativa, with classic psychoactive soar and scant body cues. Connoisseurs report a deadly, creeper with no ceiling, combination of effects. The phenotype is fully suitable for indoor growing, with a branching habit resulting in an abundance of dense, oily, attractive buds. Aromas range from animal sour spice to sweet piney mint, often accompanied by rotten meat undertones. Overindulging the Metal Haze is not recommended for the sativa sensitive or the inexperienced. It is a deceptively smooth smoke that starts out uplifting and super-active -ideal for musicians and creative types- and climbs ever higher in a succession of intense waves, taking perception to uncharted lands. We trust the virtues bred into Metal Haze to earn its place in discriminating gardens, and satisfy demanding sativa aficionados

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In biology, polymorphism is the occurrence of two or more clearly different morphs or forms, also referred to as alternative phenotypes, in the population of a species. To be classified as such, morphs must occupy the same habitat at the same time


3.3. Discovery of Highly Predictive Markers​

The comparison of sequences allowed us to identify highly reliable markers of plant type. We sequenced both the THCAS and the CBDAS genes and selected the most discriminating SNP loci (highest predictive values) from among those determined to be statistically significant, including 47 SNPs in THCAS and 40 in CBDAS. We found that some SNPs were heterozygous in drug-type samples. We decided to focus on homozygous SNPs because they are most confident and reliable for our work and future diagnostic analyses. Considering previously described four diagnostic SNPs in THCA synthase gene [15], we found that three of them (pos953, pos1035 and pos1079) were heterozygous in some drug-type samples.

Scores based on homozygous SNPs in the THCAS gene had an AUC of 100% for any of the following 25 loci: “pos136”, “pos137”, “pos154”, “pos221”, “pos269”, “pos287”, “pos300”, “pos355”, “pos383”, “pos385”, “pos409”, “pos412”, “pos418”, “pos424”, “pos494”, “pos505”, “pos612”, “pos678”, “pos699”, “pos744”, “pos749”, “pos763”, “pos862”, “pos864”, and “pos869”. Scores based on homozygous SNPs in the CBDAS gene were also evaluated and were found to have an AUC of 100% for any of the following eight loci: “pos407”, “pos545”, “pos583”, “pos588”, “pos613”, “pos637”, “pos688”, and “pos704”. Of these SNP-based scores, only one locus among the 33 selected loci (Figure 3A, B) was sufficient for discriminating between the two subgroups.
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Furthermore, we selected other statistically significant markers, particularly the deletion/insertion polymorphism identified in CBDAS, using the sequence from a fiber-type variety (Carmen KJ469374) as the reference sequence. Specifically, we detected a deletion of four bases from positions 153–156 and an insertion of three bases at position 755 (AAC) in the drug-type varieties.

A score based on the deletion/insertion polymorphism of CBDAS was calculated by assigning 1.1 points to the deletion chosen at position 154 (drug-type varieties) and −1 point to the insertion at position 755 + 3 (drug-type varieties). The possible values of the score were thus −1, 0, 0.1, and 1.1. The AUC was 99.87% (95% CI: 99.65–100.00%) in this case, and the threshold of 0 (score > 0, indicating classification as a drug-type variety) showed 100% sensitivity (95% CI: 100.00–100.00%) and 95.56% specificity (95% CI: 88.37–100.00%). Therefore, the CBDAS deletion/insertion was also able to discriminate between the two cannabis subgroups (we also found a deletion/insertion in the THCAS gene that was discarded because the AUC score was 75%). The use of Benjamini and Hochberg correction [29] guaranteed that the expected proportion of falsely detected mutations was below 5%. The empirical results confirmed that all selected SNPs were highly discriminating between marijuana and hemp. Sensitivity and specificity were above 95% for several thresholds. Sensitivity analysis revealed that these outstanding results were not dependent on the scoring system used. The AUCs for these genetic markers reached 100% even when only the CBDAS deletion polymorphism was tested together with one of the 33 SNPs listed above. Any score based on mutations that perfectly separated the two subgroups of cannabis achieved an AUC of 100% and possibly 100% sensitivity and specificity. Our suggested scores also took into account the mutation prevalence in our empirical data to maximize positive and negative predicted values. The mutations most reliably separating the two subgroups were in positions 269, 494, 749, and 763 on THCAS; in positions 637 and 583 (A/T polymorphism) on CBDAS; and a CBDAS deletion of four bases in positions 153–156 (CGTA) in drug-type genotypes (a very early stop codon resulting in a truncated protein).


3.4. Predicted Protein Sequence of THCAS and CBDAS​

To test the functional meaning of the significant mutations, two artificial genes were constructed: A THCAS gene (containing the 25 selected SNPs of Figure 3A) and a CBDAS gene (containing eight SNPs plus the insertion/deletion; Figure 3B). Both genes were translated using standard genetic code from the MEGA6 platform. Polymorphisms in the primary structure of the protein were then investigated (Figure 4), and the effect of amino acid substitutions assessed according to the expected impact on secondary and tertiary structures and the mutation effect on the biological function of the protein
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With this aim, the chemical nature of the amino acids’ side chains was taken into account: changes from hydrophobic to charged structures (and vice versa) were considered of high impact, while changes from hydrophobic to polar, or from rigid/ring to fluid structures, were considered of moderate impact. The mutated THCAS gene gave a protein with 18 amino acid changes caused by 19 non-synonymous mutations. As expected, most of the SNPs were functional with a high non-synonymous/synonymous ratio (3.2), and 10 out of the 18 amino acid changes involved moderate to severe alteration of side chain characteristics and were thus likely to impact secondary and tertiary structures. In particular, we found that mutations in nucleotide positions 269, 494, 749, and 763 cause amino acid changes (respectively, positions 90, 165, 250, and 255) that are likely to trigger severe alterations in the protein chain. To predict whether these amino acid variations also affected protein function, we performed an in silico functional analysis with PROVEAN. This analysis revealed that the mutation at the THCA protein position 165 had a high probability to be deleterious (Figure 4A, in red). Based on this, it is reasonable to assume that the catalytic activity of THCAS is low or null in fiber-type genotypes. On the other hand, the translation of CBDAS indicated that the 4 bp deletion caused severe amino acid changes (in detail, the deletion of Leu51 and Val52) that were predicted to be deleterious for the protein function. Furthermore, a very early stop codon in position 583 of drug-type genotypes may cause a truncated protein of 195 amino acids instead of 544. The PROVEAN analysis showed that the two mutations at positions 136 (Arg/His) and 182 (Gly/Ala) were also deleterious (Figure 4B, in red). In all these cases, the corresponding CBDAS truncated and/or mutated protein would make the enzyme completely inactive.
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4. Discussion​

The aims of research on Cannabis varieties [1,34,35,36] have been mainly twofold: (i) to better understand the biochemical mechanisms regulating the actual chemical profile of plants (thus supporting both the toxic effects and the therapeutic applications) and (ii) to develop effective tools suitable for forensic investigations in order to counteract the illegal market and provide economic protection for the industrial cultivation of hemp. Despite efforts to establish genetic relationships, as well as to highlight genetic differences among plant varieties (with different chemical phenotypes and different psychoactive effects), these goals have remained challenging for the scientific community to date, particularly concerning the two most investigated genes in cannabis, THCAS and CBDAS. [13,19,23,24,37]).
Staginnus [16] developed a strategy to detect the BT/BT and BT/BD genotypes, independently from the developmental stage of the plant and the tissues examined, confirming the two chemotype classifications. In addition to the within-variety diversity observed by performing multidimensional scaling of Gower distances, we found a simple and reliable way for discriminating between fiber-type and drug-type samples.
Results emerging from this study showed significant genetic difference among two subgroups of Cannabis in two genes regulating accumulation of THC in this plant species, allowing discriminating between drug and fiber varieties. A first point of novelty in this work relates to the study of both THCAS and CBDAS genes. In fact, considering that their derived proteins compete for the same substrate, the involvement of both genes can provide a stronger and more robust discrimination between drug and fiber varieties. Nonetheless, another point of novelty related to the identification of 18 amino acid substitutions in alignment of the sequences of high-THC and low/absent-THC accessions. This information is essential to gain insights into the functionality of the enzyme. With this regard, four amino acid substitutions appeared to induce a decrease in THCAS activity in the fiber-type cannabis plants, and one of them was deleterious. Furthermore, the earlier stop codon at position 195 and the 4 bp deletion in the CBDAS sequence producing a frame-shift both cause a truncated protein and a non-functional enzyme in high-THC accessions. Assuming that the protein encoded by THCAS could still be active in fiber-type genotypes, the (shared) intermediate substrate cannabigerolic acid would be preferentially metabolized by the high-affinity CBDAS-encoded enzyme. Drug-type genotypes possess an opposite trend, having the CBDAS protein completely inactive and THCAS functional. In this condition, the substrate cannabigerolic acid could be transformed by THCAS only despite its relatively low affinity for the enzyme. Both these results were confirmed and validated at the metabolic level by the chemical analysis of cannabinoids (Table 1).
Finally, highly reliable markers were identified, including the CBDAS deletion polymorphism and the 33 identified SNPs (the eight loci in the CBDAS gene and 25 loci in the THCAS gene). We refer to the associated score as (d). The score achieved an AUC of 100%, sensitivity of 100% (95% CI: 100.00–100.00%), and specificity of 100% (95% CI: 83.33–100.00%) at the zero threshold. A boxplot of CBD and THC percentages is shown in Figure 5 (p < 0.001). These markers were not previously described by other authors such as Kojoma [10] and Rotherham-Harbison [15]. These markers are able to distinguish between varieties prior to the stage of plant maturity (when the synthesis and storage of cannabinoids begin). This will facilitate the early distinction of cannabis plants, as well as the selection of cannabis seeds according to their applications in the primary sector (i.e., the cultivation of hemp for textiles, cosmetics or the production of renewable energy) or pharmaceuticals (i.e., the production of cannabinoids for therapeutic use), while at the same time providing an effective tool for controlling the illicit drug market.

Figure 5
Box plot showing: CBD % (A) and THC % (B) for the drug-type and fiber-type plants based on the SNPs and deletions identified; score (C).
The future direction for this study will be to develop a rapid, highly reliable diagnostic test that maintains the lowest possible cost to expedite forensic investigations to suppress the illegal market while also providing economic protection for the industrial cultivation of hemp.

The absence of a Y chromosome does not appear to be sufficient to ensure a total lack of production of male flowers (Faux et al., 2014; Menzel, 1964; Razumova et al., 2016). Some Cannabis plants are monoecious, producing both male and female flowers on the same plant (Menzel, 1964). While monoecious plants are commonly referred to as hermaphrodites, the botanical definition of hermaphrodite requires staminate and carpellate parts on the same flower (Lebel-Hardenack & Grant, 1997), a phenomenon rarely seen in C. sativa.

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Selection on chemical traits plays a strong role in crop domestication, which is often characterized by recessive alleles at few loci1 (e.g., Cannabis sativa,2–5 Capsicum annuum,6 Coffea,7 Nicotiana tabacum,8 and Papaver somniferum9,10). Yet, understanding the genetic basis of chemical differences between crop and wild relatives is in its infancy. Cannabis sativa L. is a fascinating example of adaptive chemical divergence, producing more than 100 terpenophenolic secondary metabolites called cannabinoids.11–14 Although populations vary in cannabinoid composition, the most common cannabinoids include cannabidiol (CBD15,16), Δ-9-tetrahydrocannabinol (THC17), cannabigerol (CBG18), and cannabichromene (CBC19). Breeding efforts have largely focused on modifying the production of THC and CBD.20–23

Molecular data suggest that the stunning chemical diversification in cannabis cultivars is the result of recent breeding efforts.5,24,25 Cultivars of C. sativa are typically characterized based on both the ratio of THC:CBD and the overall abundance of these cannabinoids. Generally, psychoactive cultivars have high THC:CBD ratios and produce high quantities of cannabinoids. Fiber or oilseed cultivars (hemp) are characterized as low THC:CBD, and generally produce lower quantities of cannabinoids. Here, we ask: what genetic models describe cannabinoid expression within plants, and how many genes underlie cannabinoid differences between C. sativa cultivars?

In 2003, de Meijer et al.2 proposed a genetic model to explain THC and CBD production in C. sativa populations. Cannabinoid yield per crop area was described as a complex trait that was the product of total above-ground biomass, the fraction of biomass composed of inflorescence, total cannabinoid content, and cannabinoid purity (equation 1 in Ref.2). To start acquiring data to test this model, de Meijer et al.2 elucidated the inheritance of THC:CBD ratios. Since THC and CBD production is influenced by the presence of enzymes that catalyze deacidification, the enzymes themselves seem to be controlled by simple Mendelian additivity at a synthase locus.2 Individuals possessing two BT alleles result in THC synthase (and THC) production. Individuals possessing two BD alleles result in CBD synthase (and CBD) production. Individuals with one BT and BD allele produce both THC and CBD. This hypothesis was supported by sequencing of THC synthase and CBD synthase genes, which exhibit 89% genetic similarity.26,27 Furthermore, de Meijer et al.28 proposed that a homozygous genotype possessing two loss-of-function alleles, B0, at the same synthase locus controlled the accumulation of the precursor CBG in adult plants. Finally, the expression of CBC was found to be environmentally sensitive to light and its expression did not fit an additive or dominance genetic model.4 Thus, de Meijer et al.4,28 described the inheritance of THC:CBD ratios and CBC production, and proposed a mathematical model describing cannabinoid abundance as a potentially more complex quantitative trait. Although de Meijer et al.4,28 examined the inheritance patterns of cannabinoids, the only genetic model tested was a simple additive model, whereas more complex genetic effects remained unexamined. Moreover, the de Meijer et al. model remained largely unexamined until the recent research by Weiblen et al.5 which identified two genes controlling the ratio of THC:CBD and a third gene controlling the abundance of cannabinoids, still relying on a relatively simple model of cannabinoid inheritance.

Although cannabinoid inheritance has only been tested against relatively simple additive models,2,4,5,28,29 both published data and anecdotal evidence suggest that more complex genetic models are necessary. Growers repeatedly describe differences among clones (suggesting environmental effects) and mothers (suggesting maternal effects). Furthermore, genetically uniform F1 offspring of parents homozygous for THC and CBD synthase alleles exhibited significant amounts of phenotypic variation (e.g., THC:CBD range: 0.91–1.48, figure 2 in Ref.2), suggesting a strong influence of environmental variation on cannabinoid expression profiles. Furthermore, THC and CBD expression patterns are reliant on multiple linked loci on chromosome 6 and gene duplication to produce tetrahydrocannabinolic acid (THCA) and cannabidiolic acid (CBDA) synthase sequences.5 In addition, selection on hemp cultivars for low THC concentrations, to meet government regulations, has failed to completely remove the expression of THC in most hemp cultivars.30 All this evidence suggests more complex inheritance patterns of several cannabinoids than could be described by simple additive or dominance genetic models.

[IMG alt="An external file that holds a picture, illustration, etc.
Object name is can.2018.0015_figure2.jpg"]https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7173683/bin/can.2018.0015_figure2.jpg[/IMG]
FIG. 2.
Results from replicate analyses of the genetic architecture of CBD concentration in Cannabis sativa. (A, C) Traditional line cross analysis plot of CBD concentration with respect to proportion of the genome inherited from THC-rich parent (P1). If CBD concentration was inherited in a Mendelian manner, the points should fall on the dotted line and the F1 and F2 data points should fall directly on top of each other. (B, D) Model-weighted parameter estimates for the genetic architecture underlying CBD concentration in C. sativa. Points are sized based on vi scores and indicate the magnitude of the genetic effects. Whiskers indicate the unconditional SEs. Only parameter estimate >0, where vi > 0.01 and wi > 0.001, was included. The genetic effects axis describes the weighted parameter estimates calculated by SAGA in R.31 Data presented in (A) and (B) are from de Meijer et al.,2 whereas data presented in (C) and (D) are from Staginnus et al.29
To predict the pace and direction of cannabinoid evolution in response to artificial selection, the genetic architecture that underlies variation in cannabinoid production must be understood. Two original studies2,29 compared chemotype data to either a single- or two-locus model with multiple alleles using chi-square tests to assess model fit, but did not consider alternative models. We use an information-theoretic (I-T) approach developed by Blackmon and Demuth31 to estimate the composite genetic effects (CGEs) contributing to variation in cannabinoid production among divergent psychoactive and fiber/oilseed cultivars, and their F1 and F2 offspring. In addition, to estimate the number of genes that drive the expression of THC:CBD chemotype and variation in the absolute production of CBD, THC, and CBC, we used the Castle–Wright estimator. Given that the studied lineages were a product of inbreeding and selection,32 we expected to confirm the previous findings that the difference in cannabinoid production among the cultivars examined was governed by few loci with mostly additive effects. However, the use of an I-T approach31 revealed several cannabinoids to be the product of multigene complexes and illuminated additional genetic effects contributing to cultivar differences that were undetected using previous approaches.
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The Composite Genetic Effects, Their Abbreviation, and a Brief Definition That Were Compared Using an Information Theory Approach with the Analysis of Line Cross Data
Name​
Abbreviation​
Definition​
Autosomal additive​
Aa​
When the combined effects of alleles at different loci are equal to the sum of their individual effects. Autosomal genes are located on one of the numbered, or non-sex, chromosomes.A3,A4​
Autosomal dominance​
Ad​
In dominant genetic models, a single allele copy of the mutation is enough to cause expression of the trait.A5​
Cytotype additive​
Ca​
Genetic variation in cytoplasmic genomes (i.e., the combined mitochondrial and plastid genomes) can influence trait expression.A3,A6​
Maternal effect additive​
Mea​
Maternal effects arise when the genetic and environmental characteristics of a mother influence the phenotype of her offspring, beyond the direct inheritance of alleles. In this case, transgenerational expression of induced genes with additive genetic effects.A3,A7​
Maternal effect dominance​
Med​
In this case, transgenerational expression of induced genes with dominance genetic effects.A3​
Autosomal additive by additive epistasis​
AaAa​
The single locus additive value for a given locus (e.g., A) changes depending on the genotype at a second locus (e.g., B) and vice versa. When an alternate allele at locus A is present, it will result in smaller phenotype values when the B-locus is homozygous for the primary genotype and will result in large phenotype values when the B-locus is homozygous for the alternate genotype.A8​
Autosomal dominance by dominance epistasis​
AdAd​
The single locus dominance genotype value for a given locus (e.g., A) changes depending on the genotype at a second locus (e.g., B) and vice versa. The single locus dominance genotypic value is underdominant when the alternate locus genotype is homozygous and overdominant when the alternate locus genotype is heterozygous.A8​

For quantitative traits such as cannabinoid concentrations, their expression may be determined by genetic factors, environmental factors, and parental environment. Thus, when assessing the inheritance of chemotypes, a diversity of genetic models must be considered.A3

In early trials, proven clones passed down their potency genes on to the next generations using an unacceptable, unpredictable domino pattern. To develop a superior seed line, males were tested the hard way, growing out their offspring to recognize the best possible pollen source. Winning donors pollinated our choicest females, and their offspring was then heavily selected at the second generation. Recombinant individuals displaying the desired potency, phenotype, flowering and yield traits were identified and inbred into separate lines. Three years after the project begun, the resulting lines were finally blended, giving rise to our Metal Haze, a hybrid that received high praise from skeptic industry testers.
 
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acespicoli

Well-known member
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acespicoli

Well-known member

Summary​



  • We recently described, in Cannabis sativa, the oldest sex chromosome system documented so far in plants (12–28 Myr old). Based on the estimated age, we predicted that it should be shared by its sister genus Humulus, which is known also to possess XY chromosomes.
  • Here, we used transcriptome sequencing of an F1 family of H. lupulus to identify and study the sex chromosomes in this species using the probabilistic method SEX-DETector.
  • We identified 265 sex-linked genes in H. lupulus, which preferentially mapped to the C. sativa X chromosome. Using phylogenies of sex-linked genes, we showed that a region of the sex chromosomes had already stopped recombining in an ancestor of both species. Furthermore, as in C. sativa, Y-linked gene expression reduction is correlated to the position on the X chromosome, and highly Y degenerated genes showed dosage compensation.
  • We report, for the first time in Angiosperms, a sex chromosome system that is shared by two different genera. Thus, recombination suppression started at least 21–25 Myr ago, and then (either gradually or step-wise) spread to a large part of the sex chromosomes (c. 70%), leading to a degenerated Y chromosome.
@Sam_Skunkman




Cannabis Hops Hybrid Welcome to Jurassic Pot™​

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Welcome to JURASSIC POT. The first blog in this series details the background of a potentially new cannabis-hop hybrid, the story about the breeder, why we think these plants could be the real deal, and the testing we plan to do in the future.​

The speciation of cannabis and its origins are challenging and highly debated topics. However, it is scientifically understood that cannabis and humulus (hops) are closely related as they diverged from their ancestral species millions of years ago. The family of Cannabaceae contains 170 species and eleven distinct genera. Cannabis and humulus are the only plants related to each other of all the Cannabaceae genera; they aren’t cousins – they’re siblings! For those that enjoy exploring the evolution, divergence, and research of cannabis and hops, I recommend Cannabis Evolution and Ethnobotany by Mark and Merlin. This extensive text goes the most in-depth on this niche subject matter.​

Cannabaceae genera

cannabaecaea

Why cannabis and hops?

For over a decade, I’ve had an affinity for beer and weed, especially when combining the two since they work and play together nicely. Early in my career, I was still unsure which area to focus on, but I eventually decided to develop Interpening instead of a THC-infused beer company in 2013. Still, I federally trademarked “Cannabrew” after consulting cannabis and beer industry experts on the product and name. I could go on forever about flavor profiles, plant characteristics, brew designs, alcohol-free THC beers; you name it.

Before diverging into the plant types, consider this. Have you ever experienced an IPA beer and weed that both tasted and smelled dank? The reason is because of the similarities between cannabis and hops, including the aromatic profiles they muster up. The plant bodies and root systems are different, but their leaf, flower, and chemical makeup are incredibly similar.
If cannabis and beer are so damn closely related, why hasn’t there been a hybrid type yet? The OG’s from back in the day tell me, “everyone has tried crossing cannabis and hops in the backyard since the 1970s, and it just doesn’t work”. And with today’s insane technology in any field, why don’t we smash these beauties together in a lab? What harm could that do – easy peasy – right?

Except this is apparently where the Hop Latent Viroid originated.

An unsuccessful hybridization between cannabis and hops is where the disease jumped over into cannabis and is wreaking havoc across the industry today.

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Welcome to Jurassic Pot™

Where did the little hybrid dino Jurassic Pot come from? It all started with a guy named Kaly from Germany who wanted to hide weed from a Karen next door by disguising it as a different-looking plant. Kaly tried breeding cannabis and hops together unsuccessfully until he came across an uncommon hops species with enough genetic material to cross over in the late 1990s. It was not Kaly’s goal to create a new species of cannabis. Instead, he aimed to create a unique smokable hop plant with THC and CBD.
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I interviewed Kaly’s business partner and son, André, regarding how and why Kaly crossed hops with cannabis since Kaly doesn’t speak English. André sent me Kalys original notes with leaf presses of the early crosses, pictures of some of the first plants, and the seeds of five types. As someone who has grown and heavily researched cannabis and hops, the story, notes, and plants seem pretty telling, especially since there isn’t anything like them aside from the similar appearance of Ducksfoot.
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Cannabis-Hops Skepticism?

If you’re skeptical about these cannabis-hop hybrids, you’re not alone. After posting our first Instagram post about these plants, many people commented and privately messaged us, saying they don’t believe a cannabis hop hybrid is authentic, that Kaly is just another industry scam, and asked for the evidence. We want to take on the research about these plants because we aren’t 100% sure they are cannabis hop crosses either. To be sure, we will research their chemical and genetic profiles to understand better what these plants are and if they may have any valuable research to offer. Please share insights on “Kaly-types” or evidence for the cross in our comments below!
cannabis hops

cannabis hops

cannabis hops


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Basic / Breeders Info​


Logo KalySeeds
Legitimo (aka jap.hop) is a mostly indica variety from KalySeeds and can be cultivated outdoors. KalySeeds' Legitimo is a THC dominant variety and is/was never available as feminized seeds. The Jap.hop is a hybrid from 1998, between Purple Star and japanese Hop. He has long grown faint-vigorous and stunted, in 2008 it was back to normal in growth. This hybrid is the ancestor of all PAC strains.

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or those that enjoy exploring the evolution, divergence, and research of cannabis and hops, I recommend Cannabis Evolution and Ethnobotany by Mark and Merlin. This extensive text goes the most in-depth on this niche subject matter.​


Phylogenetic analyses of modern and ancient Cannabis samples revealed that H. japonicus is more closely related to both ancient and modern samples of Cannabis, than H. lupulus [21, 22]. Successful grafting between C. sativa, H. japonicus, and H. lupulus further underscores the close relationship between these species [23]. The xanthohumol and bitter acid pathways in hop and the cannabinoid pathway in Cannabis contain enzymes that perform analogous reactions and accept similar precursor structures [24], which is important for interpreting the evolution of genes involved in these pathways. For all three pathways, the first reaction involves type III polyketide synthases and malonyl-CoA, and the second reaction involves aromatic prenyltransferases and isoprenoid structures.

Humulus japonicus (syn. H. scandens, Antidesma scandens) is known by the common name of "Japanese hop",[4][5] or "wild hop".[6]

Originally native to East Asian countries such as China, Japan, Korea, and extending its habitat to Vietnam, it was imported to North America in the late 19th century as an ornamental. Since its arrival in North America, it has spread widely. It can be found throughout the Northeastern U.S. and eastern Canada, and considered an invasive species in North America. It also features on the list of invasive alien species of Union Concern since 2019. This means it can no longer be imported in the European Union.[2] Additionally, it has become illegal to plant it, breed it, transport it, or bring it into the wild in all Member States.[3]
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Organic Japanese Hop (Humulus Japonicus)

The japanese hop with the scientific name Humulus japonicus belongs to the family of hemp (Cannabaceae). Its origin leads to China, Japan and Korea aswell as the tropical Asia with Vietnam.
During the late 1500s it was imported to the USA as neophyte and was used there as a tonic or ornamental wine. Different from the Cannabis plant, this kind of hemp does not contain any cannabinoids. It can grow up to a height of four meters in size with the habitus of a small tree or an understory shrub.
As a climbing plant it needs some walls or ropes to grow at. The stem is rough-textured and covered with downward pointing short and sharp prickles. Thus it should be paid attention to it when the plant is planted or transplanted. Do not do this without using gloves!
The leaves are also rough-textured and grow paired. They look like hands with a lobed shape and a size of ten to fifteen centimeters. The little yellow to greenish flowers can be seen from July to August. Male and female flowers can be found on separate plants. The seeds have a light-brown color marked with some darker specks and grow and mature in September. As herbal plant it is used in China and Japan. There a brew of all parts of this pant is drunken for the strengthening of bladder and genitals.
This plant prefers shady places to grow, they should be kept dry. Exceptions are pot-planted plants. They need humid and moisture soils.

Found growing wild near streams ;)
Is there a genetic possibility of cross ?

Genome size in Humulus lupulus L. and H. japonicus Siebold and Zucc. (Cannabaceae)​


Aleksander Grabowska-Joachimiak, Elwira Śliwińska, Magdalena Piguła, Urszula Skomra, Andrzej J. Joachimiak



Abstract​



We analysed chromosome lengths, karyotype structure, and nuclear DNA content (flow cytometry) in diploid (2n=20) and triploid (2n=30) European H. lupulus var. lupulus, American H. lupulus var. neomexicanus (2n=20) and Japanese ornamental hop, H. japonicus (F/2n=16; M/2n=17). Diploid female representatives of H. lupulus var. lupulus and H. l. var. neomexicanus differed in total length of the basal chromosome set (23.16 µm and 25.99 µm, respectively) and nuclear 2C DNA amount (5.598 pg and 6.064 pg) but showed similar karyotype structure. No deviation from the additivity, both in chromosome length and 2C DNA amount was evidenced in triploid monoecious H. lupulus (2n=30, XXY). H. japonicus showed different karyotype structure, smaller basal chromosome set (F/18.04 µm, M/20.66 µm) and lower nuclear DNA amount (F/3.208 pg and M/3.522 pg). There are first evaluations of nuclear genome size in diploid, not commercial representative of European H. lupulus var. lupulus and American H. lupulus var. neomexicanus and first attempt to determine the absolute male and female genome size in two Humulus species.


The therapeutic efficacy of medicinal cannabis is mainly dependent on cannabinoids, which are endemic metabolites unique to C. sativa8, among which THC and cannabidiol (CBD) are the main chemical cannabinoid compounds.

Although cannabis has considerable economic and medical value, information about its genome is limited. While a genomic draft was published recently, in 20119, the splicing of this draft was neither of good quality nor complete, thus hindering its usefulness.

Cannabis is mostly dioecious, with a diploid genome (2n = 20) containing nine pairs of autosomes and one pair of sex chromosomes (female plants (XX) and male plants (XY)). The Y chromosome is larger than the X chromosome, and the female plant’s haploid genome is estimated to be 818 Mb in size, while the male plant’s genome is estimated to be 843 Mb10,11. However, the published genome is for cloned C. sativa and was assembled using SOAPdenovo software to obtain a genome of approximately 786 Mb9. It shows a contig N50 = 2.8 kb and scaffold N50 = 16.2 kb, and genome annotations are missing. Additionally, the genome assembly quality is poor since it contains incomplete assembly of gene regions and repeat sequences.

The cannabis genome has been sequenced9, but the sequenced plant came from a cultivated variety. Generally, cultivated varieties lose substantial genetic diversity through successive bottlenecks due to domestication and selection for traits to increase yield under intensive human cultivation12. Therefore, wild-type varieties are an important source of genetic diversity for molecular breeding. In this report, we performed genomic sequencing, assembly, annotation, and evolutionary analysis in wild-type varieties of C. sativa. The genetic data obtained in this study will be a valuable resource for future studies assessing the pharmacology, chemical constituents, cultivation, and genetic improvement of the traits of these plants and could be used as a reference in future population genetic studies of C. sativa.
 
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acespicoli

Well-known member
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In Buddhism, kammaṭṭhāna is a Pali word (Sanskrit: karmasthana) which literally means place of work. Its original meaning was someone's occupation (farming, trading, cattle-tending, etc.) but this meaning has developed into several distinct but related usages all having to do with Buddhist meditation.

Tao or Dao came from Chinese, where it signifies the way, path, route, road





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@dubi 🙏 :huggg: :thank you:


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acespicoli

Well-known member
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Pua Mana Pakalolo Hawaiian Duckfoot Pic Info​


Introducing this year's Male and Female breeding pair now formally wed and isolated: Her Royal Highness along with Her Royal Subject, the King, and a worthy King he has proven to be: They were both chosen for their genetic hardiness, similarities, and genetic stability, so the traits shared between them are expressed consistently for posterity. I will address them frequently to see if I can assist them in anyway. This is our only breeding pair. That is the theory, now for the practice... 😉 This is a picture of Hawaiian Pakalolo Heaven. 🙂 for more info, check out www.puamanaohana.us This is a licensed grow for medicinal purposes and my best at preserving these rare Hawaiian Landrace Genetics. *As far as the appetites of Kings go... I may end up breeding another female as well... oh my goodness... we will see which one, if any, I will allow for further breeding... but, he may have to sacrifice a limb to do it.
*Update: This morning, by the light of the morning sun, I determined that the Queen is in fact a Pink Hawaiian Duckfoot. Her flowers are now beginning to turn a Pinkish hue in the morning sun with white tips. Truly, she is blushing after her first night with the King. 😃 Yayyy! 😃 I have a P.H.D. hahaha! 😃 There's no such thing as a hole in your hat, for example. hahaha! 😃 I've decided to name this year's Queen, Kakahiakala. Kala is this year's King. *Only the one breeding pair will ever carry these names for now and in the future. They are I.B.L. (brother and sister). They are the first of their name but, God willing, will not be the last of their names. We refer to all their siblings or progeny as P.H.D. if they display their P.H.D. traits. The P.H.D. are the "mutated" or I should say "adapted" descendants of the E.D.B. from www.puamanaohana.us



~ 10 x REGULAR 'Ano 'Ano (seeds) ~

HAWAIIAN DUCKFOOT

Hawaiian Duckfoot has been cultivated here in Hawai'i for as long as medicated memory can record.

This old school landrace Hawaiian indica strain is still a Big Island favorite within our underground grower community.

Hawaiian Duckfoot is given to growers just starting out, so they can learn to master a faster flowering Hawaiian indica, prior to tackling mo challenging Hawaiian Sativa strains like: Maui Wowie or Kona Gold.

Hawaiian Duckfoot has prominent "Duck foot" leaves and enjoys both indoor and outdoor cultivation here in Hawai'i.

Hawaiian Duckfoot is a go to strain for pain, according to our Kūpuna (Wise Elders) who have kept this potent Hawaiian Indica strain in circulation for Hanauna (Generations).

As similar Duckfoot characteristics can be found in a few Australian strains, we believe Hawaiian Duckfoot has been here in Hawai'i nei for over 200 years -- brought by sea and passed around the globe by people who know good Pakalōlō -- on a quack pot quest to spread Aloha worldwide!

Phylos Bioscience Genome sequencing show us that Hawaiian Duckfoot is one of the most ancient strains in their entire database.

Pua Mana 'Ohana is very thankful this strain still exists to this day.

Mahalo (Thanks) to our Kūpuna (Wise Elders) we are overjoyed to be able to provide this powerful Hawaiian medicine to you and your 'Ohana (Family)!!

Mahalo Ke Akua = Thank God

Yield: Nui = Big, Large
Flavor: Tropical, Hashy, Earthy
Flower time: 9 weeks
Photoperiod: Hawaiian 11/13 or Traditional 12/12
Indoor/Outdoor: Can/Can

#HawaiianDuckfoot

~ Pua Mana 'Ohana & Pua Mana 1st Hawaiian Pakalōlō Seed Bank release our rare endangered Hawaiian Genetics for souvenir & preservation purposes ~

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NLzer0
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Duckfoot
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acespicoli

Well-known member
On site collections gather as many seeds as possible
Sharing field collections >200 seeds when possible as needed for reproductions
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Part of: McPartland JM, Small E (2020) A classification of endangered high-THC cannabis (Cannabis sativa subsp. indica) domesticates and their wild relatives.
PhytoKeys 144: 81-112. https://doi.org/10.3897/phytokeys.144.46700



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Every nation is responsible for the current state of our world.

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acespicoli

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The term Central Asia refers to a region in Asia between the Caspian Sea and Western China. Central Asia includes Afghanistan, Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan, and Uzbekistan, which are all predominantly Muslim countries and, with the exception of Afghanistan, were all former Soviet republics.
An estimated 74 million people live in Central Asia (in 2020).

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acespicoli

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