Research The nexian phalaris breeding programme

[Ivan Illich]

Thanks! I appreciate the quick answer.
This is a quite a solid recurrent selection setup, Some things remain unclear to me however. A few follow-ups on what I think are some key things to point out/find out here:

How many plants enter each breeding cycle, and how many contribute pollen and seed, are these sampled?
Since the plant is self-incompatible and tetraploid you can't inbreed to fix traits, so a population size is really your main defense against losing traits. Removing the bottom 50% low producers each round is a good intensity, but population has to be kept high enough. Please note here!!!: If you keep 100 plants but only a handful end up as pollen donors (because a few produce copious pollen and dominate, or they produce ripe pollen first), your Ne is set by that handful, not by 100.

Have you tracked whether group means for metabolite are actually moving across cycles, or is the gain assumed? What measures have been taken to minimize the effect of environment on alkaloid content? This is to make sure that the effects you are seeing actually have a genetic foundation instead of an environmental one.

The selected plants are then grouped according to their chemotype profiles using the following classification:
Type 1: DMT-dominant
Type 2: DMT and 5-MeO-DMT
Type 3: 5-MeO-DMT and 5-MeO-NMT

Plants are subsequently cross-pollinated within their respective groups. Where, exceptional individuals may be used for introgression between groups to combine desirable traits.

Since the metabolite profiles likely represent the final branches in the metabolic pathway, it is probably not very useful to separate your plants in these chemotype groups yet and just aim for higher total alkaloid content instead. As this represents the total flux capacity of the metabolic pathways that can produce these compounds. If the effective breeding population is sufficiently large, you will keep all these chemo types at the end of that ride, then you select from there. Even keeping gramine in might actually be good, that is if it is yet another tip of the fork of the N,N-DMT or 5-Meo-DMT biosynthesis pathway. However it is super promising to see that it can be selected out relatively quickly from a population! I'll see if I can dive into some literature to find out what the bio synthetic pathways look like for these compounds.

It makes sense that the lineages are unknown from your wild samples. However, have population level lineages been kept, as soon as you started breeding and crossing?
moreover:
- The amount of generations for example,
- which plants were taken for the next generation,
- how many plants were kept? (Although getting the actual pollinators would be amazing).
Basically anything that will help me see important aspects of the breeding process

Really looking forward to hearing what you guys think! It's been a while for me when I had to use good old breeding genetics, so if I made some errors, or you just have general disagreements, let me know!

Thanks a lot for initiating this wonderful project.

Much love

 

[TransistorBass]

I'm simply asking out of interest.

I don't understand why you don't like acid base? Tnt tek is just another form of alcohol Tek.

 

[Grasshoppers]

The Evolution of a Underground Project
This project began years ago, sparked by decades-old rumors in the psychedelic community and literature on dead sheep on Phalaris pastures. It started with gardeners, hobbyists, and plant enthusiasts collecting seeds, exchanging ideas, and conducting positive bioassays that induced life-changing psychedelic journeys.

Eventually, we moved toward Thin-Layer Chromatography (TLC). By testing what we grew, we began exerting selective breeding pressure, leading to even more intriguing discoveries.
There is no written genealogical tree—no standardization, no formal documentation. Only anecdotes. It isn’t standardized today, and perhaps it never will be. The number of plants entering each breeding cycle is strictly limited by the testing capacity of myself and my fellow grasshoppers.

This work has evolved far beyond average gardening. Today, chemical testing makes up the majority of the breeding effort. While the exact number is unknown, a few hundred plants enter a new cycle each year.


The Current State & Breeding Dynamics
Progress is undeniable. Potency means are way up, and low-yielding plants have become rare in the breeding line. In fact, some of our selections are approaching the potency of Psychotria viridis. However, this plant is notoriously difficult to work with.

Here is what we are learning and navigating right now:

• Environmental Variables: Plants must be compared locally—grown in the same conditions and sampled during the same season—to limit the massive impact of environmental factors.
• Testing Rigor: Repeated testing is highly recommended before selecting a single plant. We hope to reduce the recommended number of tests per plant once we better understand the alkaloid fluctuations.
• Genetic Challenges: Self-incompatibility is not a reliable trait in these hybrids (Putievsky, Oram, Malafant 1980). Furthermore, extreme profiles (Type 1 and Type 3) could easily get lost in the massive Type 2 middle ground of the tetraploid Hardy-Weinberg equilibrium. Because these extremes are vital for future exploration, separating them into distinct breeding groups is essential, even if temporary.
• Keep moving forward: We currently recommend a heavy culling rate of about 80%.

Where We Are Headed
Right now, our focus is to deeply understand these chemical profiles: how they respond to climate cues, how to streamline the testing/growing process, and how to map the exact psychedelic effects of the different types and subtypes. Of course, we are also continuously acquiring wild germplasm to expand the gene pool.
In the future, we will certainly see a more structured, planned approach—though it can never be completely steered, given the independent nature of the grasshoppers involved.


Join the Search
Every gardener is welcome to join us. We need more hands plowing the soil to raise promising seedlings, and more eyes finding wild germplasm to enrich our gene pool.
There are still so many unanswered questions, and so many things left to try.
I am curious to hear your thoughts and deeply appreciate your contributions to this space. Your posts will be read.
Welcome to the journey if you choose to join us.

The project is just beginning.

 

[aizoaceous]

Since the plant is self-incompatible and tetraploid you can't inbreed to fix traits,

To emphasize @Grasshoppers's note, the genus Phalaris is reported to contain both consistently self-compatible species (e.g. P. brachystachys) and consistently self-incompatible species (e.g. P. arundinacea). From previous discussion P. aquatica appears to be inconsistent, with examples of both.

I assume you're not emasculating for your crosses, so it's possible that some of them are actually selfed? Have you tried deliberately selfing any of your plants, both as potentially interesting descendants and as a test of whether emasculation would have an effect? Manual emasculation seems impossibly tedious, but hot water was reported to work on P. canariensis.

This is really interesting work. @Grasshoppers potentially has the most diverse collection of Phalaris anywhere in the world, if commercial breeding programs (for forage grass or alpiste) discard most of that diversity when they first select for low alkaloids.

 

[Grasshoppers]

Thank you.

We did not perform emasculation. Self-fertilization is likely occurring. If no additional germplasm is introduced, self-incompatibility should be restored over successive generations. However, this topic would certainly merit a more thorough investigation.

I was not aware of the hot water method for emasculation, so thank you for bringing it to my attention.

Emasculation will also be essential for future attempts to hybridize P. aquatica and P. arundinacea, which I believe is one of the most important directions for future research.

 

[neurobloom]

Yes hot water emasculation is well covered up in agronomic literature.

Arudinacea was hybridized with aquatica using hot water emasculation and the offspring was crossed back with aquatica again to obtain water logging tolerant Aquatica cultivar according to CSIRO but the cultivar was dismissed for causing high rates of sheep staggers.

I think the reverse can be done to migrate aquatica genetics into arudinacea. I think this is better approach than maintaining the original hybrid which would almost certainly have issues one way or another like very low seed count per panicle or completely absent seeds or poor seed viability.

It could also exhibit other genetic flaws like poorly formed panicles or mutated weird looking leaves.

 

[Ivan Illich]

Nice! I'll look into the water method, as well as that self-fertilization paper!
I am also making an overview of the bio synthetic pathways for the metabolites that are found in Phalaris spp.
I am still affiliated with my university, so if you guys need something more specific like papers or some database, let me know.

Genetic Challenges: Self-incompatibility is not a reliable trait in these hybrids (Putievsky, Oram, Malafant 1980). Furthermore, extreme profiles (Type 1 and Type 3) could easily get lost in the massive Type 2 middle ground of the tetraploid Hardy-Weinberg equilibrium. Because these extremes are vital for future exploration, separating them into distinct breeding groups is essential, even if temporary.

For you current breeding line, do you have numbers of the ratio's between type 1, 2 and 3? Or all they all kind of equally occurring?
As for the seeds you have for that line, they are at the population level, or from a single individual?

Also @Grasshoppers, I appreciate the thorough message, however I notice a lot of LLM-style embellishments in the text. The content is most important, but I think a human touch would be nice . I suppose being a bit more direct with communication can't go wrong with projects like these!

 

[Ivan Illich]

There is quite a good genome available for P. arundinacea! Could be really useful.

Phalaris arundinacea genome assembly lpPhaArun1.hap1.1

- Phalaris arundinacea

www.ncbi.nlm.nih.gov

 

[Transform]

appreciate the thorough message, however I notice a lot of LLM-style embellishments in the text. The content is most important, but I think a human touch would be nice . I suppose being a bit more direct with communication can't go wrong with projects like these!

AFAIK, @Grasshoppers is a collective, none of whom have English as their native language. (In fact, you might be surprised at the number of ESOL Nexians who are active here…) Ergo, if there's any user who I'd give generous slack with use of LLM "AI" as an aid to composition, it's these guys

 

[Ivan Illich]

I understand, whatever works best of course! I am not against use of it.
The text just felt a bit off to me, perhaps I shouldn't have commented on it. I hope I didn't rub anyone in the wrong way. Sorry!

I am not surprised on the amount of ESOL's here, I am one myself too.

 

[Transform]

I understand, whatever works best of course! I am not against use of it.
The text just felt a bit off to me, perhaps I shouldn't have commented on it. I hope I didn't rub anyone in the wrong way. Sorry!

I am not surprised on the amount of ESOL's here, I am one myself too.

It's fine, quite understandable, in fact!

Also, it's very exciting to have you here with your background in plant bioscience - and I can barely wait to see what you'll manage to bring to the project! Do you anticipate you'll be applying TLC testing to your specimens as well?

 

[Ivan Illich]

It's fine, quite understandable, in fact!

Also, it's very exciting to have you here with your background in plant bioscience - and I can barely wait to see what you'll manage to bring to the project! Do you anticipate you'll be applying TLC testing to your specimens as well?

I have done a few TLC's in the past for a course where we are allowed to extract a compound of choice from a plant of choice (most fun course ever, we achieved 99% pure mitragynine from kratom). However I have never done it for tryptamines and at home, so perhaps I can learn the proper technique here from the experts!
My main worry for this project however, is that I do not have a garden. I live in a student dormitory. However I am a nearly graduated student (A few more months to go!!!) so my living situation is likely going to change rather soon.

It's rather a shame because I love gardening, my parents allowed me to grow a cannabis plant in their garden and I grew this monstrosity of a plant!

 

[Ivan Illich]

on a sidenote, am I allowed to share books in here? (Copyrighted material)
I have a nice book pdf called "Principles of Plant Genetics and Breeding"

 

[Ivan Illich]

I guess your love emoji means yes, so here it is: