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How to improve resiliency of in-vitro plants?

Ammonia burn
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Never heard of, never seen... What are the symptoms? Who and how has analyzed the connection between supposed damage & ammonia?
 
_Maq_ said:
Also, there's no connection with ammonium/ammonia, as all farmers (incl. submerged rice) worldwide know. They don't fear to fertilize with manure, urea and ammonia, but no matter what nitrogen fertilizer they use, they still spend enormous sums for fungicides.
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_Maq_ said:
Never heard of, never seen... What are the symptoms? Who and how has analyzed the connection between supposed damage & ammonia?
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I am not interested in this particular quarrel, but fertilizer burn from too much nitrogen and ammonia specifically is definitely a thing farmers are concerned about. I am not sure if or how all the mechanisms translate to submerged plants, but ammonium toxicity in plants is a well documented phenomenon and not an invention of this hobby.
 
ElleDee said:
ammonium toxicity in plants is a well documented phenomenon and not an invention of this hobby
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Well, I can contribute myself. This one is a good summary: https://kronzucker.org › wp-content › uploads › 2019 › 08 › amtox.pdf
Many substances may be toxic, incl. those we use for fertilization. I don't see why ammonium is exceptionally popular among them. I don't see why whenever there's a problem with plants the (almost) universal response is 'ammonium'. There are innumerable other ways how to get into trouble.
 
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Simon Cole said:
The point about ABA as a phytohormone is that it could potentially be added as a preservative. This would keep the in-vitro plant in a dormant state and avoid wilting until opened, but conversely could cause existing plants to shut their stomata and suffer stress if it is released locally into the water column in high concentrations, because in-vitro plants could be positively laced with ABA for all we know.
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Simon Cole said:
This does sound like a crazy conspiracy, so I apologise, because it is exactly that.
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From <this publication in 2011>:

"During the last 20–30 years, ABA has attracted the interest of many plant tissue culture specialists... ​
...ABA can act as anti-transpirant during the acclimatization of tissue culture-raised plantlets and reduces relative water loss of leaves during the ex vitro transfer of plantlets even when non-functional stomata are present. ​
...[ABA is] used as a growth retardant in plant tissue culture.​
...ABA has also received much attention for their involvement in abiotic tolerance and disease resistance."​
​
I did kind of expect something like this, because being lazy, I have often left them in their tubs for weeks in reasonable lighting, and noticed negligible new growth.
I don't really see how producers can avoid them bursting out of tubs when they are distributed and put on sale without something like ABA added.
Lots of the producers I that favoured actually sent them in packets directly to my door, probably without any ABA, and they tended to do really well.
I think it is fairly inevitable that they are grown with a selection of different PGRs, nutrients, biocides, and other additives.
 
Simon Cole said:
From <this publication in 2011>:

"During the last 20–30 years, ABA has attracted the interest of many plant tissue culture specialists... ​
...ABA can act as anti-transpirant during the acclimatization of tissue culture-raised plantlets and reduces relative water loss of leaves during the ex vitro transfer of plantlets even when non-functional stomata are present. ​
...[ABA is] used as a growth retardant in plant tissue culture.​
...ABA has also received much attention for their involvement in abiotic tolerance and disease resistance."​
​
I did kind of expect something like this, because being lazy, I have often left them in their tubs for weeks in reasonable lighting, and noticed negligible new growth.
I don't really see how producers can avoid them bursting out of tubs when they are distributed and put on sale without something like ABA added.
Lots of the producers I that favoured actually sent them in packets directly to my door, probably without any ABA, and they tended to do really well.
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Interesting article about ABA. Direct-to-consummer TC cups are a unique and kind of strange product, but I guess it would make sense for producers to ship out their cups with media that would support the longest shelf life, whatever that takes. I wish I had better insight, but it's out of my wheelhouse.

As for not bursting out of the cups, every plant has its own growth rate, but most things can sit for at least 6 weeks. There is a period where plants are the strongest though, and that's after they have recovered from being divided and before they get overcrowded and phenolics start to build up.

Simon Cole said:
I think it is fairly inevitable that they are grown with a selection of different PGRs, nutrients, biocides, and other additives.
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Oh yeah, those things make the magic possible.
 
_Maq_ said:
Never heard of, never seen... What are the symptoms? Who and how has analyzed the connection between supposed damage & ammonia?
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I've just read through this thread and some of your posts could be interpreted as a little confrontational. It's not really in the spirit of the forum. Please take a look at the forum rules and guidelines for guidance.

In "The Ecology of the Planted Aquarium, 2nd Ed", Diana Walstad (2003), she states that ammonia toxicity can reduce growth or kill plants, but species vary in tolerance. Sensitive species would be harmed by about 1 mg/1 NH4. For instance, Stratiotes aloides showed decay and destruction of plant tissue when exposed to only 0.9 mg/l NH4

Is the amount of ammonia leached from fresh AS greater than this?
If so which species, other than the above, would suffer harm?
And are in vitro cultures of these or other species even more vulnerable?
 
I've just read through this thread and some of your posts could be interpreted as a little confrontational.
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My fault, I apologize. I should have posted the question more narrowly, referring to my recent problem. If I described the situation and development, I think ammonia/ium would not appear in the discussion as a possible cause.
Generally, I don't deny that ammonium in excess (and esp. as a sole source of N) can harm the plants. Symptoms are described as chlorosis and reduced growth. However
1) I don't know of any evidence (or at least reasonable indications) that melting of in-vitro plants is regularly or at least occasionally caused by elevated ammonium,
2) I've never heard of 'ammonia burn' in connection with plants.
 
In "The Ecology of the Planted Aquarium, 2nd Ed", Diana Walstad (2003), she states that ammonia toxicity can reduce growth or kill plants, but species vary in tolerance. Sensitive species would be harmed by about 1 mg/1 NH4. For instance, Stratiotes aloides showed decay and destruction of plant tissue when exposed to only 0.9 mg/l NH4
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And again in a post by Diana herself…
www.plantedtank.net

High ammonia from Ada Aqua soil

www.plantedtank.net www.plantedtank.net

In her book Diane references her source material at the end of every chapter. Here are just two to be going on with...

1. Bennett AC. Toxic effects of aqueous ammonia etc, on root growth In: Carson EW (ed) The plant Root and its Environment. Univ. Press of Virginia. pp. 670-683.
2. Santamaria L, etal. 1994. The influence of ammonia on the growth and photosynthesis of Ruppia Drepanensis Tineo from Donana National Park (SW Spain). Hydrobiologia. pp. 275-276: 219-231.
 
Hi all,
_Maq_ said:
1) I don't know of any evidence (or at least reasonable indications) that melting of in-vitro plants is regularly or at least occasionally caused by elevated ammonium,
2) I've never heard of 'ammonia burn' in connection with plants.
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<"Google scholar"> is your friend.

There is quite a lot of scientific work on <"TAN tolerance"> in plants used for the phytoremediation of animal slurries etc. Basically what people want to find are the plants that can grow at the highest ammonia loadings.

Xueping Tian, et al. (2021) "Ammonium detoxification mechanism of ammonium-tolerant duckweed (Landoltia punctata) revealed by carbon and nitrogen metabolism under ammonium stress" Environmental Pollution, 277,

cheers Darrel
 
_Maq_ said:
Recently, I've got a cup of in-vitro Rotala indica (aka Ammannia Bonsai). They looked like healthy. I've divided that batch between one of my aquaria and a paludarium. In both of them, the Rotalas started to melt, and from them the melting spread further, to unrelated plants which were healthy until then.
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I've tried potassium sorbate 1 mg/L. It didn't help.
Acriflavin worked better. But many plants were lost or heavily damaged before that.
 
This <study> suggested that 30 seconds of exposure to 0.5 ppm chlorine can kill 100% of Phythium aphanidermatum zoospores (Oxidation reduction potential (ORP) 748-790 mV and pH 6.3) - lower pH levels in that study appeared to be more effective. Presumably this would be true for most Oomycetes (water moulds), and anecdotally this method is already employed successfully by many hydroponic growers. Tap water is a rather handy source of chlorine. One <publication> from the 1980s suggested that Spiked water milfoil Myriophyllum spicatum growth was unaffected by intermittent residual chlorine concentrations below 1 ppm (for three 2 hour periods daily, up to 4 days). Obviously, you would need to check for more recent research in order to validate these findings.

If you think about how we typically do a wet start - we plant the in-vitro plants, but will be changing the water every couple of days for about a week, before gradually dropping off to once per week in order to avoid high ammonium levels, only adding livestock when we are certain the ammonia and nitrite levels are safe. It does strike me that most die-back seems to occur during this initial period. Perhaps that is because water mould zoospore levels are reduced by each water change, and at that point in time they are not so much of a problem for in-vitro plants. If during that initial period, we simply avoided water conditioners and applied a suitable chlorine concentration, perhaps the instances die-back would reduce.
 
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Hi, I've just read through this thread and was goig to start a new conversation but thought it might be better to add my question to this one....

I've bought some tissue culture plants and am going to add them to a brand new set up (low tech, no CO2, hard water area, Tropica substrate capped with sand).

I've been reading that in vitro plants can be particularly sensitive to ammonia from the soil in new set ups... Is there anything recommended to minimise this risk, such as not adding them to the tank immediately, or leaving them free flowing for a while, would this make any difference?
 
Simon Cole said:
This <study> suggested that 30 seconds of exposure to 0.5 ppm chlorine can kill 100% of Phythium aphanidermatum zoospores (Oxidation reduction potential (ORP) 748-790 mV and pH 6.3) - lower pH levels in that study appeared to be more effective. Presumably this would be true for most Oomycetes (water moulds), and anecdotally this method is already employed successfully by many hydroponic growers. Tap water contains around 4 ppm chlorine, so it is a rather handy source of chlorine. One <publication> from the 1980s suggested that Spiked water milfoil Myriophyllum spicatum growth was unaffected by intermittent residual chlorine concentrations below 1 ppm (for three 2 hour periods daily, up to 4 days). Obviously, you would need to check for more recent research in order to validate these findings.

If you think about how we typically do a wet start - we plant the in-vitro plants, but will be changing the water every couple of days for about a week, before gradually dropping off to once per week in order to avoid high ammonium levels, only adding livestock when we are certain the ammonia and nitrite levels are safe. It does strike me that most die-back seems to occur during this initial period. Perhaps that is because water mould zoospore levels are reduced by each water change, and at that point in time they are not so much of a problem for in-vitro plants. If during that initial period, we simply avoided water conditioners and applied a suitable chlorine concentration, perhaps the instances die-back would reduce.
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A very interesting take on chlorine. I doubt if any hobbyist would decide to omit the water conditioner when seeing their precious plants struggle. Maybe we need to experiment more!

I know that sulfur is used by organic growers as a “natural” fungicide. Either sprayed or burned in a kind of lantern. Would that be an option for aquatic plants?

See e.g.:
academic.oup.com

Elemental sulphur as an induced antifungal substance in plant defence

Abstract. Man's oldest fungicide has probably long functioned in this role in plants, as a natural component of induced antifungal defence. Elemental sulph
academic.oup.com academic.oup.com
 
@Laoshan I did look into using sulphur to treat oomycetes, but I did not find very much evidence that it would be effective. My mind was blow when I found out that chlorine would be effective at such low levels. The concentration of most tap water could be enough to stop an infection in its tracks! As you say, it's a very simple fix.
 
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Hi all,
Hufsa said:
How much chlorine do you guys have in your tap water?
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In the UK we have <"less than 0.5 ppm">. 0.5 ppm is about the level of chlorine at which you get an obvious taste and smell.

I'd predict that the <"level will go up in UK tap water">, purely because we aren't investing anything like the money we should in infrastructure and our present government:
  • Is a kleptocracy which is successfully asset-stripping the country for personal gain or
  • Making difficult decisions, at a time of economic strife, before the Brexit bonus kicks in*.
* Please delete as you feel is appropriate.

cheers Darrel
 
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