# Melatonin application improves turfgrass under stress (drought/heat)



## tneicna (May 6, 2019)

Melatonin is a well-known antioxidant, normally associated with human intake, but there is also a novel application for turfgrasses. [It] is a natural compound in organisms and has been observed in the seeds, roots, shoots, leaves, and fruits of all plant species and in various organs of animals (Hattori et al. 1995; Stehle et al. 2011; Zhang et al. 2013; Xu et al. 2016).

According to Du, Wang, and Huang (2009), the antioxidant activity of turfgrasses is altered under heat and drought stress, and a newer paper by Nur Alam et al. (2018) shows that a foliar application of Melatonin improves Tall Fescue growth, antioxidant enzyme activities, total chlorophyll content, and leaf water %. I was curious to see how this would behave on a Midnight Kentucky Bluegrass Monostand currently under heat stress. I purchased a bag of bulk Melatonin and mixed it with water at the rate of 3000mg per 1/4 gallon (I wanted to use Bacteriostatic Water but maybe next time)

I do not recommend trying this on a large patch of lawn. I sprayed my front area and left the other heat stressed area alone.

Conclusions are interesting.

Image #1 is the 'treated' area and image #2 is untreated.



Untreated, clear signs of heat stress:



Citations:

Du HM, Wang ZL, Huang BR (2009) Differential responses of warm-season and cool-season turfgrass species to heat stress associated with antioxidant enzyme activity. Journal of the American Society for Horticultural Science 134, 417-422.

Alam MN, Zhang L, Yang L, Islam MR, Liu Y, Luo H, Yang P, Wang Q, Chan Z (2018) Transcriptomic profiling of tall fescue in response to heat stress and improved thermotolerance by melatonin and 24-epibrassinolide. BMC Genomics 19, 224-237.

Alam Mohammad Nur, Wang Yanping, Chan Zhulong (2018) Physiological and biochemical analyses reveal drought tolerance in cool-season tall fescue (Festuca arundinacea) turf grass with the application of melatonin. Crop and Pasture Science 69, 1041-1049.

Arnao MB, Hernandez-Ruiz J (2007) Melatonin promotes adventitious- and lateral root regeneration in etiolated hypocotyls of Lupinus albus L. Journal of Pineal Research 42, 147-152.

Bi A, Fan J, Hu Z, Wang G, Amombo E, Fu J, Hu T (2016) Differential acclimation of enzymatic antioxidant metabolism and photosystem II photochemistry in tall fescue under drought and heat and the combined stresses. Frontiers of Plant Science 7, 453


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## g-man (Jun 15, 2017)

This is very interesting. Have you noticed any negative side effects?


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## SNOWBOB11 (Aug 31, 2017)

Pretty cool. It does look like the top image shows less heat stress. I too am interested if you've seen any negative effects.


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## social port (Jun 19, 2017)

^ same for me. I'm also interested in why you are not recommending this treatment in large areas. Is it simply because this is an experimental approach and you are playing it safe?


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## tneicna (May 6, 2019)

social port said:


> ^ same for me. I'm also interested in why you are not recommending this treatment in large areas. Is it simply because this is an experimental approach and you are playing it safe?


Yes, I just don't want people to go blindly do anything to their laws.

I'm [going to] experiment with different dosages to see if there's a much profound effect. My next test (in 2 weeks when the heat comes back) will be 10,000mg per 1/4 gallon of water on the same plots as #1

I was reading over the literature and there is strong evidence that it is safe for all turfgrasses, but I want to be sure.


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## tneicna (May 6, 2019)

g-man said:


> This is very interesting. Have you noticed any negative side effects?


Nothing (yet?) - No discoloration, nothing. There's also no smell with it either, unlike Primo and other PGRs.

I just checked it about an hour ago and it's much much darker - And temps were 85F with high humidity.


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## tneicna (May 6, 2019)

FYI. 50G of bulk powder is $30. That is cheaper than using PGRs to keep the turf from withering away.

If this doesn't have any side effects, I'll end up using that instead of PGR/TE (primo maxx)


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## 440mag (Jan 29, 2018)

tneicna said:


> FYI. 50G of bulk powder is $30. That is cheaper than using PGRs to keep the turf from withering away.


Link? WHERE are you finding if?

I find this fascinating, fer sure!


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## tneicna (May 6, 2019)

440mag said:


> tneicna said:
> 
> 
> > FYI. 50G of bulk powder is $30. That is cheaper than using PGRs to keep the turf from withering away.
> ...


You can find it at any bulk supplier, or google 'bulk melatonin'

I don't recommend buying a lot, you can test it with a small amount (1G?) on a lawn.


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## Drewmey (Oct 3, 2018)

About how many sf did you spread the 3000mg on? Also would be awesome if you could use a Grey card or a white piece of paper in the photos next time. It would help with confirming that white balance is not at play in a color shift.

Looks promising!


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## Thick n Dense (May 7, 2019)

Not trying to discredit your work but it almost looks like the 2 pictures are taken either at different times of day(days) or in the opposite direction than the sun.

The top picture just looks soooooo much darker.

Can you like confirm or deny this ? It would say a lot if taken in the same time period and in the same direction.

Also, do both the areas get the same regimin?


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## tneicna (May 6, 2019)

Thick n Dense said:


> Not trying to discredit your work but it almost looks like the 2 pictures are taken either at different times of day(days) or in the opposite direction than the sun.
> 
> The top picture just looks soooooo much darker.
> 
> ...


They were both taken within 30 seconds of each. They receive the same treatments as the rest of my lawn (Fert, etc).

The first picture is facing north with the sun was directly overhead. 
The second picture is facing west with the sun above me. The sun was directly overhead

Both pictures were taken @ 1pm (which is when the sun, in my area, is directly up and not at angles to [try] and avoid any strange pictures)


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## tneicna (May 6, 2019)

Drewmey said:


> About how many sf did you spread the 3000mg on? Also would be awesome if you could use a Grey card or a white piece of paper in the photos next time. It would help with confirming that white balance is not at play in a color shift.
> 
> Looks promising!


Another thing is I will have to take a picture at the same times every day when calculating the angle of the sun + cloud cover as well. Yesterday was a perfect day.

I sprayed about 1000sqft total with about 1/4 gallon of mixed M, then added another 1/4 gallon of water because I was scared that 3000mg would have been /too/ strong and I wanted to dilute it slightly.


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## ryeguy (Sep 28, 2017)

tneicna said:


> FYI. 50G of bulk powder is $30. That is cheaper than using PGRs to keep the turf from withering away.
> 
> If this doesn't have any side effects, I'll end up using that instead of PGR/TE (primo maxx)


I haven't read the studies you linked, but your summaries didn't mention anything about growth regulation. What do these two have to do with each other?


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## tneicna (May 6, 2019)

ryeguy said:


> tneicna said:
> 
> 
> > FYI. 50G of bulk powder is $30. That is cheaper than using PGRs to keep the turf from withering away.
> ...


PGR's like Primo Maxx appear to improve heat stress in turfgrasses, specifically cool season ones by altering antioxidant status and others. (Ervin and Zhang, 2003). (Please see http://archive.lib.msu.edu/tic/golfd/article/2004may67.pdf)

Also see "Plant Growth Regulators Can Enhance the Recovery of Kentucky Bluegrass Sod from Heat Injury" Zhang et al. (2003)

I was trying something else instead of PGR to keep my lawn from being too stressed in the Transition Zone Heat, as the biological half-life of Primo Maxx is completely dependent on the air temperature. (Plus P.M. is expensive)


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## thegrassfactor (Apr 12, 2017)

tneicna said:


> Melatonin is a well-known antioxidant, normally associated with human intake, but there is also a novel application for turfgrasses. [It] is a natural compound in organisms and has been observed in the seeds, roots, shoots, leaves, and fruits of all plant species and in various organs of animals (Hattori et al. 1995; Stehle et al. 2011; Zhang et al. 2013; Xu et al. 2016).
> 
> According to Du, Wang, and Huang (2009), the antioxidant activity of turfgrasses is altered under heat and drought stress, and a newer paper by Nur Alam et al. (2018) shows that a foliar application of Melatonin improves Tall Fescue growth, antioxidant enzyme activities, total chlorophyll content, and leaf water %. I was curious to see how this would behave on a Midnight Kentucky Bluegrass Monostand currently under heat stress. I purchased a bag of bulk Melatonin and mixed it with water at the rate of 3000mg per 1/4 gallon (I wanted to use Bacteriostatic Water but maybe next time)
> 
> ...


This is actually extremely fascinating. Diving further into this, the role it plays on a wide variety of plants really increases its appeal in my opinion. Clearly this is another product that works by hormonally stimulating the plant into a state of regeneration and defense.



The influence on IAA is particularly interesting because of what promotes the development of IAA in the plant: tryptophan.

First, the role of IAA:



> IAA enters the plant cell nucleus and binds to a protein complex composed of a ubiquitin-activating enzyme (E1), a ubiquitin-conjugating enzyme (E2), and a ubiquitin ligase (E3), resulting in ubiquitination of Aux/IAA proteins with increased speed.Aux/IAA proteins bind to auxin response factor (ARF) proteins, forming a heterodimer, suppressing ARF activity. In 1997 it was described how ARFs bind to auxin-response gene elements in promoters of auxin regulated genes, generally activating transcription of that gene when an Aux/IAA protein is not bound.[10]
> 
> IAA inhibits the photorespiratory-dependent cell death in photorespiratory catalase mutants. This suggests a role for auxin signalling in stress tolerance.




So also through a series of pathways we can also reach melatonin production from tryptophan (as well as IAA), which would make sense why melatonin has an influence on IAA levels in the plant.

Now in terms of growth REGULATION effects, I'm not seeing much of that as stated in this piece:



In C3 plants (cool season grasses), you'll see better cold tolerance (greener, longer into winter), decreased cell membrane damage (frost/freeze damage), and increase antioxidant enzymes (purge cell mutations associated with stress). You actually have an INCREASE in growth, tillering, and turf quality. Increased chlorophyll retention (darker color), photosynthesis efficiency (increased carbon sequestration/decreased light requirement [anecdotal evidence also shows Trinexapac also improves turf performance in decreased light], decreased senescence (aging) - _decreased ABA, ABA biosynthesis genes, ABA signalling whatever factors_- which is interesting because as ABA concentrations decrease, so does senescence.



And the deeper you get, the more interesting it gets - my goodness.

Here on PRG:



> More recently, the protective role of melatonin in heat-induced senescence of the perennial ryegrass (Lolium perenne L.) has been demonstrated (J. Zhang et al., 2017). * Melatonin increases heat-induced growth inhibition, delays leaf senescence and downregulates senescence-associated genes (LpSAG12 and Lph36)*. It also increases tiller numbers, cell membrane stability, chlorophyll retention and photosynthesis rate (Table 2). Melatonin-treated plants increase CK levels during heat stress. Thus, isopentenyladenine and trans-zeatin riboside levels, which decrease in heat stress, recovered in melatonin-treated plants, showing a positive correlation between endogenous levels of melatonin and CKs. In contrast, under non-stress conditions, the application of melatonin did not alter CKs levels. As a novelty, the authors demonstrated that two CK biosynthesis genes (LpIPT2 and LpOG1) were upregulated by melatonin in stress conditions. Also, two types of transcription factor (A-ARRs and B-ARRs) involved in the CK signalling pathways were down- and upregulated, respectively. Taken together, the above demonstrates that the alleviation of heat stress by melatonin is the result of an interesting cross-talk between melatonin and CKs (J. Zhang et al., 2017)


So when faced with cultural stress - heat in this instance - melatonin _increases_ growth regulation (as part of plant preservation). Though, there is no data over whether or not it will have a direct effect on giberrelic acid. So from how I'm interpreting this, when the plant is in ideal growing conditions, you will in effect have growth surges, but conversely, while under stress, melatonin will self regulate the growth as an expression of survival - through the ABA pathways.



> In heat-induced senescence in perennial ryegrass leaves, exogenous melatonin decreased the ABA content, delaying the senescence process (J. Zhang et al., 2017). The decrease in ABA was correlated with the downregulation of two ABA biosynthesis genes (LpZEP and LpNCED1), which were upregulated by heat stress, although melatonin suppressed this effect. Melatonin also suppressed the heat-induced upregulation of two ABA signalling transcription factor genes (LpABI3 and LpABI5) and of two senescence-associated genes (Table 2). Thus, the response of ABA in heat-induced senescence was delayed by melatonin through the decrease of ABA biosynthesis and the downregulation of signalling pathway factors, while inducing the biosynthesis of CKs that act as senescence retardant agents (J. Zhang et al., 2017).


That is freakin' SLICK.

And even more interesting is that there is no real poor performance data out about it, but we only have 5 years of data:



> However, it must be taken into account that only molecular studies between melatonin and plant hormones have been carried out since 2014 (Weeda et al., 2014). Many more experiments are necessary, for example using plant hormone mutants. Unfortunately, only one ABA-deficient mutant in barley has been used in treatments with melatonin (X. Li et al., 2106). Regarding melatonin transgenic lines, there are many transgenic melatonin-deficient or melatonin-overaccumulating lines (Arnao and Hernández-Ruiz, 2015b). Unfortunately, there are no studies with them and plant hormones yet. One of the most promising fields of study related to melatonin in plants is that of endophytic relationships. The direct relationship between melatonin levels and the decrease in ROS due to activation of the redox defences (enzymes and metabolites) in Vitis sp. suggests that high levels of endogenous melatonin induced by endophytic microbes, such as Bacillus amyloliquefaciens, B. thuringiensis, B. cereus, Agrobacterium tumefaciens and Pseudomonas fluorescens, among others (or their own levels of melatonin), work in unison to cope with oxidative stress, acting as an antistress agent, and also as a root-promoting agent.


As with all data, it's over what plant species and cultivars can you mimic the results as they may not all share the same receptors:



> Identification of a possible melatonin receptor is a priority task to clarify the mechanisms that activate the responses in plants. Thus, although its double action (as an antioxidant agent and as a modulator of gene expression) seems evident, it still poses questions concerning its transport between cells (free or protein assisted, taking into account that it is an amphipatic molecule) and between tissues and organs (through the xylem and phloem), its possible conjugation with carbohydrates or amino acids, the signal transduction elements involved and the possible sharing with plant hormone mechanisms, among others. Although in this work there are many indications of connections between plant hormones and melatonin, there is still a long way to go before any firm conclusions can be reached. However, future studies on melatonin and individual plant hormones will undoubtedly throw light on the degree of interaction that exists at cellular and tissular level, and possible synergies or antagonisms in the mediated responses.


Who's ordering melatonin for their lawn?

_I am_

I guess next up will be the comparison between melatonin vs kelp vs bacteria strains stated in study vs combined.

https://academic.oup.com/aob/article/121/2/195/4560814

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855980/


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## tneicna (May 6, 2019)

thegrassfactor said:


> ...


Bulk Melatonin is cheap. I can obtain 1 kilogram for the same price as a full jug of Primo.

Dosage is now the question. And other questions:

- The purity of Melatonin. Need HPLC Report/Mass Spec analysis. Impurities/fillers might be toxic to the lawn.
- The half life of Melatonin varies depending on factors like the above. 
- Upregulation of SOD/GSH/etc in the turf, does this happen for prolonged periods (>1 day?) 
- Dosage. I went crazy, but is it not enough, or too much?

- Need to list benefits (all of them that have been documented in the literature). We know that SOD2 is upregulated with M application, as well as various other genes.

IMO, this is comparable to PGR (Primo Maxx) but 'safer' as it's more natural.


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## adgattoni (Oct 3, 2017)

tneicna said:


> Drewmey said:
> 
> 
> > About how many sf did you spread the 3000mg on? Also would be awesome if you could use a Grey card or a white piece of paper in the photos next time. It would help with confirming that white balance is not at play in a color shift.
> ...


So ~3g per 1ksqft? I'm seeing 50g for $37 shipped. At 3g per 1ksft that's about $2.22 per 1ksqft per app. Somewhat pricey. Curious what the efficacy would be at 1g per 1ksqft and how long it lasts.


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## thegrassfactor (Apr 12, 2017)

tneicna said:


> Bulk Melatonin is cheap. I can obtain 1 kilogram for the same price as a full jug of Primo.
> 
> Dosage is now the question. And other questions:


I will start with the high end of rates on the chart used in PRG. Interestingly, PRG requires more TNEX (but TNEX is decreasing GA, rather than decreasing ABA)for regulation than Bermuda, so I want to see what rates closer to a "toxic effect" would look like. Would it mimic the rate differences if Bermuda? If M increases photosynthesis effeciency, would it indeed require a lower rate since Bermuda is a C4?



> - The purity of Melatonin. Need HPLC Report/Mass Spec analysis. Impurities/fillers might be toxic to the lawn.
> - The half life of Melatonin varies depending on factors like the above.
> - Upregulation of SOD/GSH/etc in the turf, does this happen for prolonged periods (>1 day?)
> - Dosage. I went crazy, but is it not enough, or too much?


My assumption will be "if it's safe for consumption, it's safe for turf" - I'm willing to take that blanket risk.

My assumption will also be that duration observed effects are varied. Regulation occurring during heatstress will be greater than growth stimulation during peak growth. It will obviously grow out of the plant much faster during peak growth than while under regulation.



> - Need to list benefits (all of them that have been documented in the literature). We know that SOD2 is upregulated with M application, as well as various other genes.
> 
> IMO, this is comparable to PGR (Primo Maxx) but 'safer' as it's more natural.


I will disagree with comparison to Primo. It may have regulation effects, but through completely different means and pathways. Also, the regulation provided by M, according to the article I read, would only occurring during periods of stress. For cool season grasses,this would offer summer regulation but during the extreme vegetative period of spring, it would likely contribute to the growth - at least that's how I read the study, and I may have that completely wrong! And for Bermuda, at what temperature would "heat stress" (as opposed to drought)lead to ABA expression that M could then regulate?

I'm going to assume that this will have greater on turf quality rather than regulation. But boy oh boy, wouldn't it be awesome if it did offer an alternative to TNEX.


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## tneicna (May 6, 2019)

thegrassfactor said:


> I will disagree with comparison to Primo. It may have regulation effects, but through completely different means and pathways. Also, the regulation provided by M, according to the article I read, would only occurring during periods of stress. For cool season grasses,this would offer summer regulation but during the extreme vegetative period of spring, it would likely contribute to the growth - at least that's how I read the study, and I may have that completely wrong! And for Bermuda, at what temperature would "heat stress" (as opposed to drought)lead to ABA expression that M could then regulate?
> 
> I'm going to assume that this will have greater on turf quality rather than regulation. But boy oh boy, wouldn't it be awesome if it did offer an alternative to TNEX.


Let me clarify. I used trinexapac-ethyl (Primo) to /prevent/ heat stress on my lawn in the >90F degree times during summer. Primo significantly alters the antioxidant status by altering SOD2, and others in turfgrasses. If I can use a cheaper product that does the same, and then some, yes.

"trinexapac-ethyl application increased SOD, APX, POX, and CAT activities, especially under drought stress conditions. Correlation analysis showed that there were significantly positive correlation between turf quality and RWC and negative correlation between turf quality and proline content, EL, MDA and SSC in both species. The use of TE seems to have enhanced resistance to drought stress in both species by improving proline, SSC and antioxidant activities." via Etemadi et al. (2015)

Interestingly, SOD activity with TE appears to increase just as much as M application.

Sure, TE/PM suppresses growth during the active growing season, but during the baking summer months, I was more interested in lawn survival.


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## tneicna (May 6, 2019)

Melatonin seems to affect Bermudagrass in the same way as [any?] cool season grasses. I'm reading over a paper about it now.


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## tneicna (May 6, 2019)

adgattoni said:


> tneicna said:
> 
> 
> > Drewmey said:
> ...


I was shotgunning the rate. It might have been /too/ much (not harmful, though?). I need for the temperature to get back into the 90s in the next week so I can retest it on another part of the lawn with different application rates to see which is most effective.

Perhaps a rate of 10uM is the best one, and >=20uM is just a waste of money with no further improvement.


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## tneicna (May 6, 2019)

Ok, so some other notes

- The Melatonin product I was using had an use-by-date of 9/14.. it may have lost much of it's purity(?) 
- Most of the papers I have read over indicate that 10uM to 20uM is the best bet. It's similar to an application rate of PGRs, but with dilution and bioavailability, I don't know. 20uM might be the best bet.


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## thegrassfactor (Apr 12, 2017)

@tneicna big thank you for bringing this up. For whatever reason, this has really motivated me to do something other than live in my own anxiety.

So this is what I decided to do:

I picked out the driest spot in my yard, the high point, that has been subjected to everything from bobcats to cars to kids to general neglect. It is not under irrigation, and we are currently 2 weeks without rain and averaging 90 during the heat of the day over that time period. It's hot and dry.

It has been 6 weeks since any fertilizer has been applied to the lawn.



I broke the lawn up using bricks as dividers into several sections and made various applications using the following:
4gallon backpack sprayer regulated with a CF valve to 29psi. I'm using TeeJet AI11004 nozzles and applying approximately 1Gallon/M of diluted material. I'm mixing for 1000sqft, but only treating inside the designated area. Each rate change, the backpack tank and spray line is washed, purged, and reprimed with the new rate.

The turf is common bermuda, reel mowed at .75, unregulated, unirrigated, pH 5.8, and drought stressed.

Section 1, I applied 1Gram/M of melatonin.



Section 2, 2Grams/M of melatonin



Section 3, 3Grams/M of melatonin



At those three rates, it really covers the gammit. To really throw some heat at it, I went ahead and added soluble kelp extract (nova scotia, organic approach) at 1oz in combination with the 3Grams/M melatonin

1oz Kelp+3Grams Melatonin/M



And because i thought it would be cool to see the limit, perhaps to the point of causing damage:

2oz Kelp + 6Grams Melatonin/M



And of course we have a control, RIP to the tidbit of the shed that made it in the photo:



Alrighty, folks. Let's see what happens. The whole point of this is to try and see _something_ and from there dial in rates, turf types, etc. (I did apply the stupid highest rate to drought stressed fescue).


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## 440mag (Jan 29, 2018)

thegrassfactor said:


> @tneicna big thank you for bringing this up. For whatever reason, this has really motivated me to do something other than live in my own anxiety. ...
> Alrighty, folks. Let's see what happens. The whole point of this is to try and see _something_ and from there dial in rates, turf types, etc. (I did apply the stupid highest rate to drought stressed fescue).


 :thumbup: YOUDA MAN! :thumbup: YOUDA MAN! :thumbup: YOUDA MAN! :

And, +1 @tneicna Thank You for kicking this off - plant micro-biology Rocks!


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## social port (Jun 19, 2017)

440mag said:


> YOUDA MAN! YOUDA MAN! YOUDA MAN! :
> 
> And, +1 @tneicna Thank You for kicking this off - plant micro-biology Rocks!


Here, here!!
I've been checking out melatonin prices as well.


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## 440mag (Jan 29, 2018)

social port said:


> Here, here!!
> I've been checking out melatonin prices as well.


@social port (heck, I guess this goes for everyone  ) If you don't mind, would you please post up any better links you find? My Google foo is usually pretty good but, I lack severely when it comes to finding supplements like these (melatonin, humic, iron, etc.)


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## social port (Jun 19, 2017)

440mag said:


> social port said:
> 
> 
> > Here, here!!
> ...


Hey 440mag,
I was thinking about this
https://www.amazon.com/BulkSupplements-Melatonin-Powder-100-grams/dp/B00JV46FD2
Simple. From Amazon. A little under 38 bucks for 100 grams.
That's to get things started. There might be a better long-term solution.


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## ctrav (Aug 18, 2018)

Very interesting topic. I look forward following but I did doze off for a sec...


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## LawnSolo (Jul 17, 2018)

@thegrassfactor

*Carbon-M* incoming  That would be sweet! if it works


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## adgattoni (Oct 3, 2017)

@thegrassfactor @tneicna

Have either of you guys seen any research related to Tryptophan? In one of the earlier referenced images from these studies, there was mention of how tryptophan ultimately produces melatonin in the plant. Tryptophan seems to be much cheaper than melatonin - is there any mention of using this instead?


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## tneicna (May 6, 2019)

thegrassfactor said:


> @tneicna big thank you for bringing this up. For whatever reason, this has really motivated me to do something other than live in my own anxiety.


I think the problem is dosage and/or half-life. There's conflicting information in the literature regarding the half-life of Melatonin.

There's also liquid melatonin on Amazon, but I have not researched it too much. Could it be as simple as placing the powder on the lawn in a non-solution form?

Either way, it might be possible that you will have to do 24 hour/48/72 hour treatments due to the above. Again, this is all experimental.

There are other things I am investigating, such as Pharmaceutical grade probiotics as a turf stimulant (WIP), but my expertise is in Geophysics and not turf


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## tneicna (May 6, 2019)

Also; I think Liquid Melatonin might be a better application, just apply it with water in a tank sprayer. It's quite possible the Molecules for Melatonin are too large for a proper foliar application, so they will need to be diluted with something like Bacteriostatic Water if you want to walk on the highly experimental side.


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## Jconnelly6b (Mar 4, 2018)

Matt did you use well or city water? I did a quick google search and didn't come up with much, but I would not be surprised in chlorinated city water had an effect on a melatonin cocktail.


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## tneicna (May 6, 2019)

adgattoni said:


> @thegrassfactor @tneicna
> 
> Have either of you guys seen any research related to Tryptophan? In one of the earlier referenced images from these studies, there was mention of how tryptophan ultimately produces melatonin in the plant. Tryptophan seems to be much cheaper than melatonin - is there any mention of using this instead?


I'll look over the literature and get back to you about it, give me a few days.


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## thegrassfactor (Apr 12, 2017)

No discernable difference in turf quality between the plots that can be attributed to anything other than the rain. I'll be making my next application shortly. @Jconnelly6b I'd mix with distilled water, but i'm afraid even many irrigation systems may have chlorinated water moving through it which could could add a variable at scale that would make it not applicable.



@adgattoni there is limited data on tryptophan, but this presentation from VA Tech does show some data surrounding it:

http://mogic.org/wp-content/uploads/page/8/mogic-biostim-greens-ervin.pptx


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## tneicna (May 6, 2019)

Are you using raw powder? Or are you tank mixing it with water?

I think I am going to try a different approach, liquid MT from amazon. BSW + Pharma grade MT is going to be too expensive.

The molecular weight of raw MT is probably too large for proper foliage application directly, even mixed with water.


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## thegrassfactor (Apr 12, 2017)

tneicna said:


> Are you using raw powder? Or are you tank mixing it with water?
> 
> I think I am going to try a different approach, liquid MT from amazon. BSW + Pharma grade MT is going to be too expensive.
> 
> The molecular weight of raw MT is probably too large for proper foliage application directly, even mixed with water.


I'm using raw powder mixed with water.

I sift some over a small area to see if any noticeable effects.


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## tneicna (May 6, 2019)

thegrassfactor said:


> tneicna said:
> 
> 
> > Are you using raw powder? Or are you tank mixing it with water?
> ...


I went back and looked over the papers, they are dissolving the MT into Ethanol+Dilution before application. So it will likely need to be in a similar form.

From Zhang et al (2017):

"The melatonin was dissolved in ethanol, temporarily stored in 20C and diluted in dH2O prior to application."


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## thegrassfactor (Apr 12, 2017)

Would the purpose of ethanol be to increase solubility? Forgive me, I'm not a chenist


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## tneicna (May 6, 2019)

thegrassfactor said:


> Would the purpose of ethanol be to increase solubility? Forgive me, I'm not a chenist


The molecular size of raw Melatonin is likely too large to penetrate the leaf structure, so it needs to be dissolved into a liquid form.

There's liquid MT on Amazon which I purchased and going to tank mix it to a rate of 20uM in a controlled test.

Here are some more interesting data from the aforementioned paper. For PRG (P. Ryegrass):


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## Jconnelly6b (Mar 4, 2018)

I gather the mixing with ethanol before mixing with dH2O ( presumably deionized water) would be the same concept as us mixing FeSO4 with hot water in a pre-mix before adding to a large sprayer tank.


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## gm560 (Feb 22, 2018)

Did anyone do this last year? Saw some buzz around it but curious if anyone integrated it into a program and the results. This mini heatwave has me thinking about heat stress on the lawn....


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## 440mag (Jan 29, 2018)

social port said:


> 440mag said:
> 
> 
> > social port said:
> ...


@social port Thank YOU!!!!!! :thumbup:


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