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CANNABIS DNA PROJECT

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Sam_Skunkman

"RESIN BREEDER"
Moderator
Veteran
Cannabis Colleagues,

Greetings from the International Hemp Association in Amsterdam. We are writing to tell you about an exciting research program using modern DNA analysis to build a family tree for Cannabis, and to request your participation.

We are collaborating with a US-based group working on a definitive large-scale phylogenetic study of the evolution of Cannabis. Key researchers include Mowgli Holmes, the chief scientific officer at Phylos Bioscience, Rob DeSalle, a professor of evolutionary biology at Columbia University and curator at the American Museum of Natural History.

Presently, the team is building a high-resolution map of the Cannabis genome, based on a modern hybrid THC/CBD strain, using PacBio long-read Next-Gen Sequencing (NGS). The map will serve as a reference key for analysis of thousands of other accessions using an SNP (Single Nucleotide Polymorphism) NGS protocol called GBS (Genotyping By Sequencing) allowing high-resolution characterization of each accession.

So far, the group has sequenced over 400 drug-type Cannabis samples. DNA extraction equipment is installed in nine Cannabis testing labs in six US states, and we continue to gather modern hybrid drug varieties. In the next month the group will finish collecting and sequencing our first 1000 samples. In order to collect data from ancient seeds the group is modifying protocols used for sequencing fragmented Pleistocene DNA samples, developing protocols to work with single seeds, and adding a Whole Genome Amplification step to increase DNA yield.

In two or three months we hope to have a provisional relationship map worked out that we can put on the web and will let us ID modern strains and hopefully at least test theories concerning the evolution of Cannabis.

Within six months we will analyze additional modern cultivars plus traditional landraces and possibly herbarium sheets and archeological materials. From there on we will add to the live database which will continue to grow as more samples are received although the architecture should not change. Genome data will be studied via network theory to address the issues of hybridization and reticulation in the phylogeny.

Cannabis is an incredibly varied genus made up of a myriad of local landrace varieties and modern cultivars as well as their feral and wild relatives. This research will generate a huge amount of sequence data and unique SNPs spread over many thousands of samples, and we feel confident we can resolve the evolution of Cannabis under domestication.

To further our study and create as complete an evolutionary tree as possible we need seeds collected overseas (or reproduced domestically) and have not been interbred with modern drug hybrids. We are interested in ALL Cannabis whether grown for fiber, seed or drug production as well as feral and wild populations. We can now collect sufficient high-quality DNA for analysis from a single seed. And there is no requirement to grow the seeds, so we can also use dead seeds. Many conscious travelers, marijuana users and growers collected seeds that they never got around to sowing, and now years later they are dead. Dead seeds are useless to growers and breeders, but they still contain valuable genetic information that can provide us with deeper insights into Cannabis’s evolution. It is also legal for us to send dead non-viable seeds to our lab in the USA by post.

If you have any seeds you feel may be of interest (living or dead) and want to contribute to this fascinating research feel free to contact us, just PM SamS at IC. We will provide a Netherlands PO Box for you to send the seed samples to us and we will provide an optional questionnaire about each batch of seeds. Batches can range from just a single to 25+ seeds.

As research progresses we will share data with you about any accessions you provide. Upon completion the results will change how we all look at Cannabis, and we will better understand the heritage of modern Cannabis cultivars. Growers will be able to see how their varieties (as well as traditional landraces sent by contributors) fit into the big picture – to determine the landrace origins of modern hybrids (ex., Jamaican, Mexican, Colombian, etc.) and explore deeper evolutionary relationships. Fascinating, eh?
If you have any questions please feel free to contact us.

All the best,

Rob Clarke and Sam Skunkman



Many people have asked us questions about our projects. The questions below are the ones we hear again and again. We want our work to be good for everyone involved, and we intend to be transparent about how and why we’re doing it.


PHYLOS BIOSCIENCE DNA PROJECT FAQ

Why is Phylos sequencing the DNA of all these Cannabis strains?

A few reasons. First we have some scientific questions we just want answered. We want to know how Cannabis has evolved, what its history was like, and how it has co-evolved with humans. And we want to know what domestication does to the shape of evolution.

We also want to understand today’s crazy mix of hybrid strains. We want to know where they came from, why they’re so different, and what makes each one unique.

But the reason that will probably affect most people is that we want to change the Cannabis industry into a modern, legitimate marketplace, where people actually know what they’re getting. We think this is the only way.


Will Phylos make and sell genetically modified Cannabis?

No. Absolutely not.


Will someone else make and sell genetically modified Cannabis?

Maybe. But it won’t be us. And it won’t make much sense to do it, because it is so easy to create amazing strains of Cannabis using traditional breeding techniques.


Will Phylos patent the strains that are submitted? Doesn’t sequencing the DNA of something let you control it or patent it?

We won’t patent the strains that are submitted. We don’t want to, and we couldn’t. Here are a few important facts about patents and plants:

* Sequencing the DNA of something does NOT let you patent it. In fact, after the court cases <http://www.supremecourt.gov/opinions/12pdf/12-398_1b7d.pdf>Assoc. for Molecular Pathology v. Myriad Genetics and <http://www.supremecourt.gov/opinions/11pdf/10-1150.pdf>Mayo Collaborative Services v. Prometheus Laboratories, Inc., it is no longer possible to patent naturally occurring DNA sequences at all.

* Cannabis strains cannot be patented, at least not now. To apply for a plant patent you have to declare that you successfully grew your new Cannabis variety somewhere on U.S. soil. The patent office will not grant claims that rest on violations of federal law.

* Intellectual property protection over a plant variety can also be obtained through the USDA Plant Variety Protection Office. But only for plants on their official list. Cannabis is not on that list, and won’t be until it is federally legal.

* Only things that are NEW can be patented. Once something is public and known about, it’s too late. Once something has been sold commercially, it’s too late. Even if Cannabis strains could be patented, all the existing strains are now in the public domain, and will stay there forever. Which is a good thing.


But doesn’t Monsanto want to patent all the Cannabis strains in the world and then make it illegal for me to grow any of them?

Probably. But existing strains can’t be patented.



What will Phylos do with the information it collects from sequencing the DNA of different Cannabis varieties?

We will use it to construct a map of all the different strains. We will publish this and make the data freely available to the research community. The individual data from each sample will also be freely available to the person who submitted it.
 
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harry74

Active member
Veteran
I think was my idea but I haven´t got the logistics so is great to hear that someone is doing this.

The portuguese were for long time in India around Goa.
http://en.wikipedia.org/wiki/Portuguese_India

I don´t know if Indian genetics could somehow arrive to Brazil but the link is there.

¿Could we take some of that brazilian genetics and trough DNA testing trace their origins?

No idea really, but would be interesting.

A worldwide Cannabis DNA library?
https://www.icmag.com/ic/showthread.php?t=290988
 
S

stix~

:tiphat: max respect Samskunk .. amazingly interesting proyect .. best o luck!!! wish ya needed a helper jeje :dance013:

i doubt i can contribute any helpful hybrids....however , i'm looking forward to sendin all the landraces in my seedstash.... New Guinea, thai79 , mexican oaxacan , Costa.rican Sativa , Colorado p.bud , & a couple more :ying:

good vibes!!!
 

Mustafunk

Brand new oldschool
Veteran
Fantastic project, I'm sure many members of the community would love to take part and support this. The results of such an intensive research will definitely help us to gain a better understanding of this plant and its evolution, providing answers, that's what it is all about. I'm looking forward to see this happening.

Best of luck with the project.
Vibes.
 
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Betterhaff

Well-known member
Veteran
Presently, the team is building a high-resolution map of the Cannabis genome, based on a modern hybrid THC/CBD strain, using PacBio long-read Next-Gen Sequencing (NGS). The map will serve as a reference key for analysis of thousands of other accessions using an SNP (Single Nucleotide Polymorphism) NGS protocol called GBS (Genotyping By Sequencing) allowing high-resolution characterization of each accession.

I’m curious as to what strain is the base line and why? I’m sure it will come out as the research develops but just trying to rap my head around what would be chosen. Very interesting project and long overdue.
 

high life 45

Seen your Member?
Veteran
I was waiting for this to happen! The genome of hops has been mapped out, so it was only a matter of time for cannabis to follow. Be interesting to see the genetic foundation of OGs
 
Cannabis Colleagues,

If you have any seeds you feel may be of interest (living or dead) and want to contribute to this fascinating research feel free to contact us, just PM SamS at IC. We will provide a Netherlands PO Box for you to send the seed samples to us and we will provide an optional questionnaire about each batch of seeds. Batches can range from just a single to 25+ seeds.

Howdy Sam,

I'm excited to see your findings. Mapping the family tree and evolution of cannabis will be a revealing project.

I know you say dead seeds are fine, but this seems like a prime opportunity to pool old genetics. Do you have any special tech to revive the oldest of seeds so you can germinate and self the lines?

I wish you could've had a chance to revive some of those 2,700 yr. old Gushi seeds. Jurrassic park style.

Thanks!
-pH
 

bigherb

Well-known member
Veteran
Your efforts and dedication are appreciated . This is very exciting thanks for sharing . Can't wait too see the progress and findings

Thanks for allowing the community to help

Also what Ec said

Best wishes brother's

1luvbigherb
 

oldchuck

Active member
Veteran
Ahh... Thank you Sam. That is a very interesting company.

I'm wondering how an ordinary woods hippie might be able to use their services.
 

Chimera

Genetic Resource Management
Veteran
A chinese group recently published similar work on this topic using data from 115 hemp varietals:

http://www.plosone.org/article/info:doi/10.1371/journal.pone.0110638

Diversity Analysis in Cannabis sativa Based on Large-Scale Development of Expressed Sequence Tag-Derived Simple Sequence Repeat Markers

Chunsheng Gao, Pengfei Xin, Chaohua Cheng, Qing Tang, Ping Chen, Changbiao Wang, Gonggu Zang, Lining Zhao

Abstract

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

Fig 2 of the mentionned paper shows a dendrogram based on cluster analysis of SSR markers that shows the various clades of the studied hemp cultivars.

The linked study is probably the best work in this field to date.

The problem with the previous study, and the project the Phylos guys are undertaking is that the genetic information is not tied to phenotypic nor chemotypic data sets, making it almost useless information for breeding at this point. The chinese group's work provides one hell of a basis to work with for those that have phenotypic data. The phylos guys are also talking about using herbaria samples, which is fraught with myriad problems, and in all honesty is quite doubtful that the herbaria work is likely (again imo).

This type of in silico work using dead seeds, without growing plants is rather academic; interesting to know the heritage story, but not particularly useful for those currently working with the plant. If you don't have phenotypic/chemotypic data, you don't have much. The linked paper is more valuable to those that are working in the same field imho, and in conunction with some other ongoing work, it has great potential and lays one heck of a foundation. FInally, the future looks promising.

Still waiting to hear from you Sam about the thing with the thing and that guy at the place...

Bada bing,
-Chimera
 
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oldchuck

Active member
Veteran
This is valuable work and will get done by somebody, sometime. I can't see that it is any threat and may be very useful to us small time growers but, so far, I can't quite see how. The Chinese study Chimera cites seems pretty useless. The Phylos people say they want their work published in a way useful to a grower but don't elaborate.
 
Interesting endeavor. Id love to follow along. Will results be posted anywhere in particular?

I'm most interested in finding out which of the purple expressions were first to show up in the gene pool.
 
B

Bob Green

400 drug types and growing. I wonder how many of these are related. I also am wondering what acient seeds he will be testing to match up the WLD/NLD cannabis he has collected.

I wonder how long till we learn how to revive dead seeds by extracting DNA. There has to be amazing plants in old dead seeds.
 
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