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Development of AUTO strains through the overexpression of CONSTANS gene

Darpa

Member
Hi folk,

Since the introduction of original autoflowering strains in decades past, significant improvements have been made to these genetics. However autoflowering hybrid, as actually created by Seed Banks and breeder, will never compare with photoperiod strain in term of potency, terpene profile and yield, because of its Ruderalis predecessor. However, overexpression of CONSTANS genes on top quality photoperiodic strain could resolve this problem. This could be a revolution in early or auto flowering strain development. Maybe I should file a patent, so this idea is not stolen by big LP or corporations in this new era of legal cannabis business… (I think I’m safe, I don’t think they will read this post)


IDEA: Development of early flowering phenotype and\or photoperiod insensitive strain through the overexpression of CONSTANS genes.

The photoperiod pathway is a genetically conserved pathway that affects flowering in distantly related angiosperms. Plants sense changes in day length (= photoperiod) as a reliable seasonal cue to regulate important developmental transitions such as flowering. Integration of various external light information into the circadian clock-controlled mechanisms enables plants to precisely measure photoperiod changes in the surrounding environment. The core mechanism of photoperiodic measurement is regulation of CONSTANS (CO) activity, which takes place in phloem companion cells in leaves.

Recent advances in research indicate that the core of the day-length measurement mechanism lies in the circadian regulation of CONSTANS (CO) expression and the subsequent photoperiodic induction of the expression of FLOWERING LOCUS T (FT) gene, which might encode a major component of florigen.

In response to florigen signaling, the shoot apical and axillary meristem is converted into an inflorescence meristem. It stops producing leaves and instead produces bracts (degenerated leaf-like organs) with floral meristems at the axils of the bracts.

Transgenic cannabis plants carrying CO fused with the cauliflower mosaic virus 35S promoter (35S::CO) would flowered earlier than the control phenotype and will probably be completely insensitive to length of day. This would be the strategy to develop the autoflowering strains of quality never seen before.

This could be an awesome project for someone already performing TC.

Anyone interested? (I’m personally still waiting for my laminar flow cabinet)


Darpa
 
M

metsäkana

IMG_3602.jpg

61 north latitude outdoor autoflower :) yeah we are intrested about super duper auto, grape size resin:biggrin: :lurk:
 

blackone

Active member
Veteran
I don't know if I'm willing to agree with the postulate that autoflowers are not able to rival photoperiod plants in quality.
It will not take many more outcrossings on the ones we already have to remove every trace of the ruderalis genome, except for the autoflowering gene and the small percentage of genes that are linked to it.
Cannabis has 20 chromosomes. This means only 5% of the genes are even on the same chromosome, and only a minor percentage of those are actually linked.

Of course there could still be superior genes on the linked loci in photoperiod plants, but I've grown autoflowers with top quality buds in terms of both terpenes and potency.

Still your idea if it works has a lot of merit. I wonder if this would be just a breeding tool, or if perhaps this could also lead to a tool to trigger clones from photoperiod plants?

Because one thing autoflowers don't have unfortunately is the ability to select superior phenos for cloning. Even the best photoperiod strains I've grown have had pretty different phenos, where selecting the right one has been the difference between "good" and "stellar" product.
 
M

metsäkana

https://pbs.twimg.com/media/Cb1jBjNVAAAfEnn.jpg:large

here sum pic from memhisto genetics 360 analytical lab test if its legit the potency and terps seem to be there.. but yeah are the phenos popping similar from pack i have not tried memhistos gear its too expensive for my outdoor there is so much change to them die from cold and crap.. i wish i could i might be able to afford 1 pack for next year to try.. and compare to my own stuff



https://www.instagram.com/p/BgpNEi2g_XO/?taken-by=mephisto.genetics
 

TychoMonolyth

Boreal Curing
I think you're looking for something like this Thai dominant Northern Lights.

These sativa expressing Northern Lights Autoflower plants (regular) grew 10 to 12 feet tall and produced 1-2.5 lbs per plant.

My experiments on two patches, while not scientific, point to the importance of early morning sunrise. Morning sun earlier = turning on metabolism early and those plants grew huge. Plants that got shade in the early morning didn't thrive as much (but still 6 footers).

Plants with early morning shade also got hit with a blast of full sun at about 9am. It would be interesting to see if a gradual, more natural, increase in light, coming out of lights out, would make a difference than to simply shock the plants with a flip of a switch and blasting them with light for another 18 hours, followed by yet another flick of the switch into instant darkness. Damn.... that's gotta be hard on them.

This is a side branch
picture.php


Look at the wheel barrow for size comparison. This is about 3/4 of 1 plant.
picture.php


1.5 liter water bottle
picture.php


June 1st.
picture.php


Another thing was that as the first flowers became ready to harvest, new flower kept coming up top. So they just kept growing and growing. The top cola was super think, but I never found more than a couple wet grams of mold. But the mold was on the outside where a branch broke and was laying on a pile of sticks.

I let the males have their way with the girls and they produced 5 lbs of potent flower and a half bucket of seeds from 2 plants.
 

Agronomist

Active member
Transgenic cannabis plants carrying CO fused with the cauliflower mosaic virus 35S promoter (35S::CO) would flowered earlier than the control phenotype and will probably be completely insensitive to length of day. This would be the strategy to develop the autoflowering strains of quality never seen before.

Darpa

Transgenic Cannabis plants?!

Do you have a paper that shows a protocol that can produce such plants?
As far as i am concerned, No one had been able to produce such a product.
I will be more then happy to see a paper on transgenic Cannabis.
 

MJPassion

Observer
ICMag Donor
Veteran
[FONT=Arial, Helvetica, sans-serif]Maybe I should file a patent, so this idea is not stolen by big LP or corporations in this new era of legal cannabis business… (I think I’m safe, I don’t think they will read this post)[/FONT][FONT=Arial, Helvetica, sans-serif]
[/FONT]
Are you sure?
You know a lot of breeders occupy space here at IC Mag and that Sam Skunkman (a mod here) is associated with GW Pharma, right?


Also, when your project is completed please let the world know how you did what you did so those of us that oppose transgenic food and medicine actually have a choice as to whether we use it or not.


Put down the CRISPR and walk away slowly!
 

Darpa

Member
Hi friends!

Thanks for all the imputs. First of all, I know that transgenic plant is a controversial subject and I respect everybody opinion on that topic. However it’s just a question of time before big pharma or Licensed Producers start using genetic engineering techniques on this plant.

That been said, this thread is for scientific discussion only, and I do not encourage any person to experiment with gene transfers nor gene overexpression on this plant outside of research purpose. Releasing a GMO plant to the public would be illegal (at least under Canadian Federal Laws), and mostly completely immoral.

Thanks to Metsakana, I realized that there is high quality autoflowering strain out there. Mephisto Genetic strain lab results are outstanding (25.45% THC-TOTAL, 2.13% TERPENE-TOTAL -https://archive.analytical360.com/m/archived/493401). I am curious if these results are representative of the genetic or if theses line show some inconsistence in term of THC and terpene levels. It seem that some company like Mephisto are doing an awesome job in their autoflowering breeding project.

Mephisto: « In 2016, we will be releasing at least 4 brand new varieties for a new ‘artisanal line’ – which are new varieties developed from the current Mephisto lines outcrossed to elite photoperiods and then bred through 5 generations. We are soon to be trialling the f3 generations.»


Transgenic Cannabis plants?!

Do you have a paper that shows a protocol that can produce such plants?
As far as i am concerned, No one had been able to produce such a product.
I will be more then happy to see a paper on transgenic Cannabis.

Hi Agronomist,
There aren’t many scientific publication available regarding transgenic Cannabis plants. However, the protocol for producing such plant wouldn’t differ from existing transformation protocol.

There are already dozens of patent filed for such transformation on Cannabis.

Example:
Patent CN107630034A: Agrobacterium mediated mariguana transgenosis method

The present invention is to solve the technical problem, overcoming the disadvantages of the prior art, a way to provide faster access to cannabis Agrobacterium-mediated transgenic varieties improved method.

Or this on filed by the National Research Council Of Canada. Patent WO2015196275A1: Cannabichromenic acid synthase from cannabis sativa

Nucleic acid molecules from cannabis have been isolated and characterized and encode polypeptides having cannabichromenic acid synthase activity. Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds.

Here is another paper on transgenic Cannabis: Tissue culture and Agrobacterium-mediated transformation of hemp (Cannabis sativa L.)

M. Feeney, & Punja, Z. (2003). Tissue Culture and Agrobacterium-Mediated Transformation of Hemp (Cannabis sativa L.). In Vitro Cellular & Developmental Biology. Plant, 39(6), 578-585. Retrieved from https://www.jstor.org/stable/4293671

Here is an interesting article about the ongoing movement in cannabis genetic research.

Investors rush to patent genetically modified cannabis molecules https://www.cbc.ca/news/business/cannabis-genetic-biotech-patents-gmo-1.4854746

I was quite surprised to hear the position of big Cannadian LP:

GMO skeptics

«Other cannabis companies aren't so keen on the genetic engineering revolution. Bruce Linton, chief executive of Canopy Growth, one of Canada's largest cannabis firms, told CBC his company has meet with half a dozen biotech experts who pitched them on genetic modification, but Canopy has stayed away from the technology.»

Quite surprised, since Tweed filed a patent for protecting the creation of novel genetically modified cannabis plant having modified expression of THCA synthase.

Abstract
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.

https://patents.google.com/patent/WO2016189384A1/en

That been said, let’s not turn this thread in a GMO debate…

Cheer


Darpa
 

Only Ornamental

Spiritually inspired agnostic mad scientist
Veteran
Didn't read every post in detail... anyway, CO and FT are involved in flowering of long day or rather short night plants such as A. thaliana. What exactly induces flowering under non-inducing conditions in short day respectively long night plants remains elusive (at least it did till last year) except for rice and that's a monocot and therefore fairly different from cannabis. Hence, your plans for a GMO cannabis plant may not work. Sure, you could knock-in the responding genes but that doesn't mean they are expressed when and where they should. A safer way would be, given that the 'flowering genes/proteins' become known in cannabis, to increase these instead (not necessarily by overexpression ;) ).
 

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