Advanced seed germination techniques

[BigBozat]

@Diggaz -

Sorry about the lack of clarity... and, no, I wasn't making a mistake/confusing terms (rather, just wasn't being clear enough, I guess, in my subject matter)... I was talking about different things...

'Scarification' is the mechanical etching/scuffing up of the seed coat... I mentioned such in my first post in re: my quick & dirty list of seed germ techniques...

'Stratification' is an old gardening/horticultural term that relates to the cold treatment that Goat referenced...
e.g., see here

My recollection was that 'stratification' (i.e., cold treating the seeds for some period before attempting to germinate) was pertinent mainly (only?) to perennials, shrubs & trees... but cannabis is an annual, so was wondering about the applicability of cold treatment to cannabis generally?

 

[Goatstroker]

@Diggaz -

Sorry about the lack of clarity... and, no, I wasn't making a mistake/confusing terms (rather, just wasn't being clear enough, I guess, in my subject matter)... I was talking about different things...

'Scarification' is the mechanical etching/scuffing up of the seed coat... I mentioned such in my first post in re: my quick & dirty list of seed germ techniques...

'Stratification' is an old gardening/horticultural term that relates to the cold treatment that Goat referenced...
e.g., see here

My recollection was that 'stratification' (i.e., cold treating the seeds for some period before attempting to germinate) was pertinent mainly (only?) to perennials, shrubs & trees... but cannabis is an annual, so was wondering about the applicability of cold treatment to cannabis generally?

I figure it like this, see...

Cannabis, all wild-like, will produce seed in autumn. The seeds chill out over winter, either right there or in some bird doo or what have ya, then they spring like bunnies in, well, spring time.

So that's about that I suppose, I just give my seeds a longer winter before the spring time. I'm normally not a big supporter of winter, but this seemed logical to me and the seeds don't seem to mind.

I also double jar my seeds and vacuum seal the outer jar, but I don't know why. Just seemed like the ting to do and that's sorta how I operate so ahead and groove on that or whatever.

Peace

 

[BigBozat]

I figure it like this, see...

Cannabis, all wild-like, will produce seed in autumn. The seeds chill out over winter, either right there or in some bird doo or what have ya, then they spring like bunnies in, well, spring time.

So that's about that I suppose, I just give my seeds a longer winter before the spring time. I'm normally not a big supporter of winter, but this seemed logical to me and the seeds don't seem to mind.

I also double jar my seeds and vacuum seal the outer jar, but I don't know why. Just seemed like the ting to do and that's sorta how I operate so ahead and groove on that or whatever.

Peace

Your method is legit.
It's virtually THE definition of stratification in botany, AFAIK.

Just wondering if it's always applicable for all strains, or if it's only relevant to certain ones (cold tolerant, native to high altitudes and/or regions with seasonally cold weather)?

The vacuum seal I would guess is to... control moisture? protect against mold? Don't know for sure... anyone?

Wondering: how much does it depend on genetics?
Have you ever had a hoped-for grow disappoint cuz the seeds didn't like the fridge treatment? Was it a different strain than what you've otherwise run? How so?

Sorry if I ask too many Qs...
LMAO ... I'm a newb, after all...

 

[Goatstroker]

I vacuum seal to protect from fridge moisture, yes.
I figured seeds aren't probably held in a vacuum in nature, though I've admittedly yet to tour the inside of a bird...anyone who has, please chime in here.

So anyhoo I got myself a little jar and I take all my breeder pack seeds and put them in little plastic vial dealies, label them on occasion, then seal that little jar and put it in a quart sized jar and vacuum seal it. I use toilet paper to keep the jars from rattling 'cause I'm crafty and shit.

No problems thus far, but I've only been at this seed storage thing a year or so. Others here have done similar for longer so I'll be 6 on the side from here.

 

[Diggaz]

Good discussion here, thanks for the info. My bad BigB, you may be new here but it sounds as if you have a scientific background.

 

[BigBozat]

Good discussion here, thanks for the info. My bad BigB, you may be new here but it sounds as if you have a scientific background.

NP... and, no I don't have any scientific background... I'm just a nerd who reads a lot, loves to learn/figure sh!t out. Happened to have taken some Intro Horticulture waaaaaaaaaaaaay back when I was in college, so I had a vague memory of the stratification/cold treatment...

I still want to know whether cold treatment is really effective for cannabis (i.e., does it actually improve germination rate? are the germinated seedlings healthier? does it improve root growth? etc.)?

My recollection is that cold treatment/stratification is meant for & effective for [only?] perennials, trees & shrubs, particularly native cultivars of cold weather environs.

Cannabis, however, is a short-day (long-night, actually) flowering annual, not a perennial, tree or shrub. Moreover, many cannabis cultivars - think equatorial sativas, among others - are native to warm weather environments, where the logic of cold treatment to break seed dormancy would seem to fail...

Goat's description sounds a bit more like a storage strategy than a method for improving seed germination. Sure, makes total sense to me to use cold in storage to preserve viability... we've all stashed breeder's packs in the fridge to store them... but does it do anything in terms of breaking seed dormancy or otherwise enhancing (not just preserving) seed germination?

IDK... I'm interested/curious, and would be delighted to be convinced with valid evidence [pref. more than unsubstantiated subjective anecdote]... but, based on what [little] I do know, I am a bit skeptical.

Has anyone around here ever run any side-by-side test(s) to see if - all other variables being equal - cold treatment of seeds improves germation success rates or germination speed or the quality of subsequent root/shoot/leaf/etc. development, yadda yadda yadda [insert your success metric for germination enhancement here]?

How much, if any, improvement was there in whatever success metric was targeted?

Has anyone experimented to see if it is strain dependent (e.g., some strains prefer cold treatment, some a warm treat ment, yet others an alternatine treatment of warm/cold or cold/warm)?

I haven't... mostly cuz I can't afford to risk expensive beans.

 

[BigBozat]

@FlaCaEnt - More directly in re: your stated goal(s) of "taproot vigor, and thickness , as well as lush healthy first set of false leaves and first set of true leaves", I found this:

Promotive Effects of DCPTA on Seedling Development and Growth...

Abstract:
"Compared with controls, seed treatment using 30 μM DCPTA significantly (P = 0.05) enhanced the rates of root
and hypocotyl elongation and seedling dry weight. Enhanced hypocotyl development by DCPTA showed a significant
linear correlation (r = 0.83) with the increased taproot yield of mature plants grown from DCPTA-treated seeds.
The harvestable taproot yield and harvest index of plants grown from seeds treated with 30 μM DCPTA were increased
109% and 38%, respectively, as compared with controls."


Haven't tried this myself (among other things: where do I get DCPTA?)- actually, hadn't even seen/heard/thought of this before FlaCaEnt raised the question.

Didn't see any refs to DCPTA elsewhere on the forums, either... which is surprising given the deep & broad knowledge that seems prevalent here. Maybe shaggyballs or others with experimental proclivities over in the PGR threads would take on?

FWIW, I am really stunned that, e.g., spurr hasn't covered DCPTA over in his PGR thread.

 

[BigBozat]

Just what you were looking for<<<

Nice!

Sorry I didn't click this link earlier... I kinda leaned organic/soil when I started doing this, stumbled across Jeff Loewenfells (Teaming W/ Microbes/Nutrients), and this looks right up that alley!

Thanks for the link!

 

[BigBozat]

Algae

@MJtI - Algae... hmmm.. Ok... but, a week in water? Ain't ya risking drowning the poor bastards?

 

[MJtheIndicator]

I love you Koi

I love you Koi

@MJtI - Algae... hmmm.. Ok... but, a week in water? Ain't ya risking drowning the poor bastards?

I can dig it, but youth craves oxygen and algae have photosynthetic machinery ultimately derived from cyanobacteria that produce oxygen as a by-product of photosynthesis. Nature's bubble stone, its what kelp does. I mostly like the idea of bulbous seeds in cahoots with fat Koi, fancy bright white and orange nurses. Such a majestic auspicious creature the nishikigoi

 

[BigBozat]

"More even germination" meaning consistency in numbers. This product is geared toward farmers who plant acres of seeds.
I assume this is what you were confused about?

No doubt soaking in water is more technical than plugging a seed directly in the dirt.

Here DL these and listen while driving, I like to listen while Im in the garden.
Maybe you have already Ive posted this multiple times but this is the stuff Im referring to.


2013 Soil & Nutrition Conference: Putting Principles into Practice
John Kempf: Part 1/ Part 2/Part 3/Part 4http://www.bionutrient.org/audio/2013_soil_nutrition_conference/d-01-31-2013-JohnKempf.mp3

Dan Kittredge

http://www.bionutrient.org/audio/2013_soil_nutrition_conference/g-02-01-2013-DerekandDan.mp3

http://www.bionutrient.org/audio/2013_soil_nutrition_conference/h-02-01-2013-DanDerekandJohn.mp3

http://www.bionutrient.org/audio/2013_soil_nutrition_conference/i-02-01-2013-DanDerekandJohn.mp3

Hey thanks for posting these... I blazed by them when you first posted on this thread, but later after I was trundling around in the AEA thread I recognized and came back & checked 'em out...

I love the IC Forums!

 

[BigBozat]

Here's my smorgasbord of [advanced?] seed germination techniques:

1 - Seed scarification (scuffing)

2 - 12-hour pre-soak in a solution of:
- pH neutral water (chlorine & chloramine free) (1 Gal.)
- Gibberelic Acid (75 ppm = roughly 1/8th tsp/gal)
- Wright's Liquid Smoke - Hickory (contains karrakinolides) (7.5 tsp/gal)
- L-Glycine Amino Acid (synergistic interaction w/ karrakinolides) (1/4 gram)
- L-Aspartic Acid (1/4 gram)
- L-Glutamic Acid (1/4 gram)
- L-Phenylaninine (1/4 gram)
- L-Lysine Amino Acid (1/4 gram)
- OptiFlora PreBiotic Complex (contains fructooligosaccharides, Vitamin E) (1 tbsp)
- Dirt M.D. Activated Liquid Humic/Fulvic Acid (1/4 tsp)
- Botanicare / Fulvex Trace Mineral Extract (1/4 tsp)
- Botanicare / Liquid Karma (1/4 tsp)
- Microbe Life / Nourish-L Liquid Organic Rare Earth Humus Conditioner & Cypress Lignin (1/4 tsp)
- Flora USA/HB-101 (1/2 tsp/gal.)

3 - Fresh banana peels in the enclosed germinating environment (produces ethylene)

Spme links in re: Karrikinolides:
http://www.ncbi.nlm.nih.gov/pubmed/21280622
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0095109

2nd Revised edition:

Substitute Coconut water in lieu of distilled water in #2
Add to #2: Neptune's Harvest Liquid Seaweed (up to 8 tsp/gal)
After the 12-hour pre-soak, coat seeds in Aloe Vera gel before planting in germination media
And, as per my earlier revision: Substitute Etehphon for the banana peels in #3

 

[messn'n'gommin']

You might find these interesting.

Light and Seed Germination in Diverse Taxa
http://www.kew.org/science-conserva...y/projects/light-and-seed-germination-diverse

Smoke Induced Germination
http://www.plantphysiol.org/cgi/content/full/128/4/1167

 

[BigBozat]

You might find these interesting.

Light and Seed Germination in Diverse Taxa
http://www.kew.org/science-conserva...y/projects/light-and-seed-germination-diverse

Smoke Induced Germination
http://www.plantphysiol.org/cgi/content/full/128/4/1167

Nice! Thanks for the links...

Read the abstract on the 1st, but I'll have to study the 1st one a bit more (question whether light-induced germination is relevant for cannbis seeds) before commenting...


The second corresponds well with other research I've seen in re: karrakinolides in smoke & their effect on seed germination (thus, why Wright's liquid smoke is in my seed germ pre-soak formula)... will have to study further... the literature cites in it are a bit dated relative to some more recent research by Flematti which points to karrakinolides as the point of action for breaking seed dormancy, not NO2 or any of the other speculations the authors cite in the abstract...

 

[BigBozat]

Nice! Thanks for the links...

Read the abstract on the 1st, but I'll have to study the 1st one a bit more (question whether light-induced germination is relevant for cannbis seeds) before commenting...


The second corresponds well with other research I've seen in re: karrakinolides in smoke & their effect on seed germination (thus, why Wright's liquid smoke is in my seed germ pre-soak formula)... will have to study further... the literature cites in it are a bit dated relative to some more recent research by Flematti which points to karrakinolides as the point of action for breaking seed dormancy, not NO2 or any of the other speculations the authors cite in the abstract...

Here are some links re: karrakinolides & their effect on seed germination:
http://www.sciencemag.org/content/305/5686/977.abstract
This one is pretty much the definitive reference on karrakinolides by the guy who disovered them (Flematti)... registration is free for full access... all sorts of links to other papers that cite this one, including:

http://hortsci.ashspublications.org/content/47/10/1453.full.pdf+html
This one suggests karrakinolides also help protect from pythium blight... paywall, but I'll see if I can find the full text elsewhere...

http://pubs.acs.org/doi/abs/10.1021/jf1041728



The paper is of interest becuz it highlights the synergistic (I hate that word, but don't have a better one handy) effects between karrakinolides & l-glycine (ergo, why l-glycine is in my formula)

http://www.actahort.org/members/showpdf?booknrarnr=813_20
This one includes Flematti - the guy generally credited w/ discovering karrakinolides - as one of the authors... sorry it's behind a paywall (it's also on PubMed), but maybe there's a free source out there somewhere... the abstract references some application rates (which may lead me to revise my formula)

http://onlinelibrary.wiley.com/doi/10.1002/jpln.201200488/abstract
Another paywal (sigh)... this one's not so much about seed germ, per se, but references effects of foliar spraying on growth (interesting)

http://link.springer.com/article/10.1007/s10725-012-9685-3#
The link to Springer allows you to view [at least parts of] the full .PDF...

http://www.caws.org.au/awc/2010/awc201012991.pdf
Finally, no paywall... unfortunately, this one's less relevant, as it's primarily looking at using karakinolides' seed germ effects to control weeds (rather than to enhance seed germ on crop seeds)


That's enough for now...

 

[BigBozat]

I also suspect that karrakinolides explain much of the 'mystery' of the fertility of Terra Preta soils/Bio Char (albeit not all, e.g., soil structure, drainage/water holding, carbon source, etc.)

 

[MJtheIndicator]

I'm don't know this from experience…

I'm don't know this from experience…

2nd Revised edition:

Substitute Coconut water in lieu of distilled water in #2
Add to #2: Neptune's Harvest Liquid Seaweed (up to 8 tsp/gal)
After the 12-hour pre-soak, coat seeds in Aloe Vera gel before planting in germination media
And, as per my earlier revision: Substitute Etehphon for the banana peels in #3

I usually notice Thiamine (vitamin B1) as part of the germination process, is there a reason why there doesn't seem to be any outright B1 in your germination technique?

 

[BigBozat]

I usually notice Thiamine (vitamin B1) as part of the germination process, is there a reason why there doesn't seem to be any outright B1 in your germination technique?

LOL, yeah... prolly just cuz I I already got too much going on w/ all the other sh!t I got in what is an evolving/experimental formua that I haven't done much of any research on B1's effects on seed germ...

Seriously, what little I have trundled across seems (at best) ambiguous to me... some sources seem to indicate sheer snake oil, others suggest there may be some benefit in re: germination but indicate there are substantial unknowns... The research that was done in the 1930s that led to the vitamin B1 'myth' [if such it is] was done in a lab, and it's results have not been generally/consistently replicated in the field... the benefits of Superthrive, eg, really come from NAA, not the B1; and similar 'magic formulas' that tout B1 in 'em really are conflating B1 w/ other things in their formula (often seaweed extracts)...

In the absence of any better/clearer info, and in the presence of many other things that seem more promising and/or better understood, and given that I have limited resources to investigate / analyze / experiment, I simply haven't seen investigating B1, per se, as worth my time [yet?]... cost/benefit ratio and all that, n'est-ce pas?

 

[MJtheIndicator]

I'm don't know this from experience…

I'm don't know this from experience…

LOL, yeah... prolly just cuz I I already got too much going on w/ all the other sh!t I got in what is an evolving/experimental formua that I haven't done much of any research on B1's effects on seed germ...

Seriously, what little I have trundled across seems (at best) ambiguous to me... some sources seem to indicate sheer snake oil, others suggest there may be some benefit in re: germination but indicate there are substantial unknowns... The research that was done in the 1930s that led to the vitamin B1 'myth' [if such it is] was done in a lab, and it's results have not been generally/consistently replicated in the field... the benefits of Superthrive, eg, really come from NAA, not the B1; and similar 'magic formulas' that tout B1 in 'em really are conflating B1 w/ other things in their formula (often seaweed extracts)...

In the absence of any better/clearer info, and in the presence of many other things that seem more promising and/or better understood, and given that I have limited resources to investigate / analyze / experiment, I simply haven't seen investigating B1, per se, as worth my time [yet?]... cost/benefit ratio and all that, n'est-ce pas?

I agree the data is often archaic and conflated within synthetic formulas touting "natural" foundation, but I still wonder why its connection to germination is redundant. I'll try and dig up some concrete agro data on B1 and see if we can't for once bury it or progress.

 

[MJtheIndicator]

I'm don't know this from experience…

I'm don't know this from experience…

Myth and rumors: Researchers knew that thiamine was normally found in roots (agreed), so they put thiamine in the culture medium and found that root growth did occur (unfortunately this control environment forgoes B1 in the seedling stage, they were too obsessed and focused on root growth). Vitamin B1 is manufactured in plant leaves and sent to the roots (this is nonsense because we know it occurs in seed), but if roots are cut off and placed in a petri plate, vitamin B1 stimulates growth of the roots when it saturates the culture medium (this last statement is also unstable because it ignores chloroplast).

Planting trees in a soil environment, however, is vastly different from a laboratory culture (why? we plant trees in labs frequently). Most important, gardeners aren't in the habit of cutting off the root system when planting, (wrong… transplanting is the very core of this phenomena). Several studies using intact mums, apple trees, orange trees, pine, tomato, beans, pepper, corn, pear, watermelon and squash have failed to demonstrate that vitamin B1 treatments provide any type of growth response. (Cox failed to back this sweeping statement up with anything, hence he’s playing the “I told ya so” game.)

Some "root stimulator" products contain a rooting hormone and fertilizer along with vitamin B1. These materials may increase rooting and growth, not the vitamin B1. (And again he points to the synthetic, but does not back up as to why he feels B1 has nothing to do with it, i.e., armchair science)

The bottom line: While root stimulator products are not necessary for transplant success (notice this contradiction if one considers his earlier statement surrounding “gardener habits”), if you do use one, make sure it contains a rooting hormone and fertilizer rather than just vitamin B1. (Oh, so it is beneficial then?) The vitamin B1 is for marketing purposes rather than actual effect. (Wait, but you just said…)

So says Robert Cox, Horticulture Agent, Colorado State University Cooperative Extension? He seems a bit on the arm-chair routine, panning for street-cred or just plain suffering from “I told ya so” syndrome. Besides, I don’t agree with Cox regarding Sulfur.

Thiamine??? All organisms require it as a cofactor in its form as thiamin pyrophosphate (TPP) for the activity of key enzymes of central metabolism.

“In humans, deficiency is widespread particularly in populations where polished rice is a major component of the diet. Considerable progress has been made on the elucidation of the biosynthesis pathway within the last few years enabling concrete strategies for biofortification purposes to be devised, with a particular focus here on genetic engineering.”
-Examining strategies to facilitate vitamin B1 biofortification of plants by genetic engineering, Lucille Pourcel, Michael Moulin, and Teresa B. Fitzpatrick 2013

I asses that a fair amount of historical research is attached to the motivation of increasing B1 in consumed plant life (bio-fix/damage control). Specifically, thiamin biosynthesis occurs in the chloroplast, with this in mind we can ask how vital is it during germination (yet another point scored on the side of speculation). Then again predominant vitamers of B1 are TMP and TPP, both phosphates are generally agreed as advantageous to early growth. This would show a motive pointed at the biofortification of plants and not humans exclusively. For thiamine’s biosynthesis significant progress has been made over the last few years (comprehensive reviews, see Goyer, 2010; Rapala-Kozik, 2011). TMP is not directly phosphorylated to TPP, it is assumed to be subsequently dephosphorylated to thiamine by broad specificity phosphatases. Indeed, a broad substrate acid phosphatase has been isolated from maize.

So this got me to thinking of micro-nutrients and the norm as is concerning them. The average focus is on chelates, but never if ever B1. Yet… I ran across this from 2000 - Effect of thiamine on the cadmium-chelating capacity of thiol compounds. Tandon SK1, Prasad S. The thiamine coadministration enhanced the efficacy of MFA and MTA in reducing hepatic and renal burden of cadmium and that of DMSA and DEDMS in mobilizing hepatic cadmium. If thiamine can do this adversely to cadmium, I can’t agree that its not a potential shield for new plant growth. And another study from 1996 - Thiamine (vitamin B1) seed treatment enhances germination and seedling growth of bean (Phaseolus vulgaris L.) exposed to soaking injury:

Seed treatment with thiamine mononitrate significantly increased germination rate of bean (Phaseoh vulgaris L.) seedlings germinated at high soil moisture levels and low night-temperature (5°C). Beneficial effects of thiamine applications on germination were also observed in soil-free germination tests when the seeds were submerged in water for 1-4 d at 18OC. Optimum response was achieved at thiamine concentrations of 3 mM. applied during the first 24 h of imbibition. As an early response to the thiamine treatment. Vital staining with triphenyltetrazolium chloride revealed a lower proportion of damaged tissue of the embryonic axes. In later stages of seedling development, thiamine seed treatment resulted in a lower proportion of abnormal seedlings with malformations of the shoot apex and of the primary leaves. The beneficial effect of thiamine was more pronounced when the temperature during submergence was kept at 2SoC compared to 5°C. There was no response to thiamine applications when the germinating seeds were exposed to low temperature treatments only. The thiamine effect could be mimicked by raising the osmotic potential of the incubation medium. indicating a relationship to soaking injuries. The results suggest that normal seedling development may be impaired by thiamine deficiency in the embryonic tissue induced by soaking injury, whereas low temperatures are not involved as a stress-factor.

So back to that bit about phosphates, concerning maturation thiamine is stored in unphosphorylated form even in mature seeds, yet it depends on the thiamine binding proteins or TBPS. IN 1975 and thru ’87 they found that seedlings first utilize thiamine that is stored in seeds and through analyses this total thiamine seed content does not change or decrease during seed germination. This is occurring 2-4 days after inhibition as seedlings commence thiamine biosynthesis. TPK (two-pore-domain potassium channel) activity progressively increases during seed germination and growth. As stated before TDP is crucial i.e, fundamental to mitochondrial function. TMP needs green tissues as transporter, roots. Now I start to see how potassium and B1 are synergistic.

Maybe of importance for those venturing into thiamine amendment is the concern for its antagonists: caffeic acid, chlorogenic acid, tannic acid, quercetin and rutin. With all this rambling over with, I feel B1 lacks a contemporary focus because science always tended to pigeon hole its growth factors as relates yeast which is essentially the redundant inconclusivity surrounding studies of B1 and plant germination.