Tissue Culture Kill Tank

[Art]

I need some help. My name is Art and I kill tissue culture plants.

As an homage to @Vin's famous Rotala Kill Tank thread, I've also titled this thread using the "kill tank" designation as it fits with what has been happening to me lately. I do need help to figure it out and, hopefully, others will also benefit.

I have a new tank (~30 gallons) with the dimensions similar to a 60p. It was started on May 31, 2023 using the dark start method where I inoculated the tank with Dr. Tim's One and Only and dosed PO4 for about 2 weeks. Then I planted heavily.

I have been able to convert the emersed plants I started with and the submersed ones are doing just fine. However, each and every one of the tissue culture plants I have planted have died within a week or two. I don't understand why.

Possible reasons - I think​

1. New tank syndrome - the tank is not over the year old point so maybe it is still cycling in some what that is impacting the sensitive tissue culture plants. Yes, I'm using Aqua Soil but I'm testing for ammonia and I'm getting zero reading. I know @Dennis Wong has mentioned this as a cause.
2. Poor quality tissue culture plants - I have purchased from several retailers and the plants are coming from different growers. However, they plants are still at stage 1 of development. This means that they have yet to develop a strong enough root system. When I get them, I almost want to incubate them somewhere until they move into stage 2 and have a better chance at life in an aquarium.
3. Wrong aquarium conditions - something is off with my key parameters such as CO2, light or fertilization. While my set up may not be ideal for young tissue culture plants, the other plants are doing just fine. Maybe the lighting is too intense for them and I should shade them and slowly move them to more light?
4. Poor aquarist technique - sometimes it's not the tool but the person using the tool. That's me. Maybe my technique just sucks and I need to be coached up.

That's what I can think of.

Do you have any techniques with tissue culture plants that gets you a higher chance of success?

 

[Dennis Wong]

I think the tank needs more time to mature. Ammonia cycling is just the first step, but I wouldn't put delicate TC, Bucep or Chai in the tank even after a dark start. When the tank is running stable with no algae, and stuff is growing well - that's a matured tank. Other common causes are poor TC stock and possibly low CO2. Fert / parameters very far down the list unless they are off a long way, but if so, then it will show in other plants

 

[HardeeParty]

A good 50% of my tanks were originally tissue cultures. I’ve always floated them in a shallow breeder box at the top of the tank in a low-flow area wedged between a few pieces of lava rock (to lock the orientation). I also trim the darkened roots and any leafs in poor shape. It keeps them closest to the light, but still acclimating to the water. Depending on the plant a minimal amount will melt, but quickly shows new roots (less than a week). I’ll dose a little FE to the box and once I see rounded new growth I’ll plant. Whole process takes about 10 days.

 

[FrankZ]

This is great. So far my best method is “poke and pray”. Just poke ‘em in and pray they grow.

AR of all types and mermaid weed have been 100% successful every time I’ve tried them. Full grown form is a different story for both of these for me.

Hottonia turned to mush in 24 hours but looked great in the gel.

Pink flamingo crypt was eaten by a large severum before I dried off my arm off from planting, but that doesn’t count LOL.

The rest I have tried were very undramatic in their demise.

 

[BigWave]

A good 50% of my tanks were originally tissue cultures. I’ve always floated them in a shallow breeder box at the top of the tank in a low-flow area wedged between a few pieces of lava rock (to lock the orientation). I also trim the darkened roots and any leafs in poor shape. It keeps them closest to the light, but still acclimating to the water. Depending on the plant a minimal amount will melt, but quickly shows new roots (less than a week). I’ll dose a little FE to the box and once I see rounded new growth I’ll plant. Whole process takes about 10 days.

This in every essence of it. @Jhardee85 is 100% correct. Never lost one doing this technique.
Edit: I will aslo note depending upon what manufacturer the TC is comming from plays a huge role. Tropica plants seem to do pretty well along with Didennerle and AquaFlora. If the TC came from Asia such as Kiri or some of the others good luck. Ive seen quite often TC's comming from state side sources to be not quite up to par.

 

[Art]

I think the tank needs more time to mature. Ammonia cycling is just the first step, but I wouldn't put delicate TC, Bucep or Chai in the tank even after a dark start. When the tank is running stable with no algae, and stuff is growing well - that's a matured tank. Other common causes are poor TC stock and possibly low CO2. Fert / parameters very far down the list unless they are off a long way, but if so, then it will show in other plants

Thanks @Dennis Wong. Low CO2 can be ruled out. I'm at about a 1.6 pH drop, stable.

Why do you think maturity matters with tissue culture plants but not others? I really they are very young plants but what's the difference with a young offshoot cutting?

For example, I can plant a new shoot that I broke off of a larger plant. This shoot will grow well albeit also being young and not having an established root system.

Why are tissue cultured plants different?

 

[Art]

A good 50% of my tanks were originally tissue cultures. I’ve always floated them in a shallow breeder box at the top of the tank in a low-flow area wedged between a few pieces of lava rock (to lock the orientation). I also trim the darkened roots and any leafs in poor shape. It keeps them closest to the light, but still acclimating to the water. Depending on the plant a minimal amount will melt, but quickly shows new roots (less than a week). I’ll dose a little FE to the box and once I see rounded new growth I’ll plant. Whole process takes about 10 days.

So you are growing them out in a separate container. How do you acclimatize them to the water?

 

[Art]

So far my best method is “poke and pray”. Just poke ‘em in and pray they grow.

This has been my method and it isn't working for me.

I just keep thinking that there has to be a better way. More and more plant growers are selling the tissue culture plants because of the ease of shipping and storage. However, how many of these plants are melting in people's aquariums?

Are tissue culture plants really the solution we thought they would be? I don't think so at this point.

 

[HardeeParty]

So you are growing them out in a separate container. How do you acclimatize them to the water?

It’s a fry/breeding isolation box that’s in the same tank, so they acclimate in the same water, just closer to the light source.

 

[spawnerjp]

I need some help. My name is Art and I kill tissue culture plants.

As an homage to @Vin's famous Rotala Kill Tank thread, I've also titled this thread using the "kill tank" designation as it fits with what has been happening to me lately. I do need help to figure it out and, hopefully, others will also benefit.

I have a new tank (~30 gallons) with the dimensions similar to a 60p. It was started on May 31, 2023 using the dark start method where I inoculated the tank with Dr. Tim's One and Only and dosed PO4 for about 2 weeks. Then I planted heavily.

I have been able to convert the emersed plants I started with and the submersed ones are doing just fine. However, each and every one of the tissue culture plants I have planted have died within a week or two. I don't understand why.

Possible reasons - I think​

1. New tank syndrome - the tank is not over the year old point so maybe it is still cycling in some what that is impacting the sensitive tissue culture plants. Yes, I'm using Aqua Soil but I'm testing for ammonia and I'm getting zero reading. I know @Dennis Wong has mentioned this as a cause.
2. Poor quality tissue culture plants - I have purchased from several retailers and the plants are coming from different growers. However, they plants are still at stage 1 of development. This means that they have yet to develop a strong enough root system. When I get them, I almost want to incubate them somewhere until they move into stage 2 and have a better chance at life in an aquarium.
3. Wrong aquarium conditions - something is off with my key parameters such as CO2, light or fertilization. While my set up may not be ideal for young tissue culture plants, the other plants are doing just fine. Maybe the lighting is too intense for them and I should shade them and slowly move them to more light?
4. Poor aquarist technique - sometimes it's not the tool but the person using the tool. That's me. Maybe my technique just sucks and I need to be coached up.

That's what I can think of.

Do you have any techniques with tissue culture plants that gets you a higher chance of success?

I normally let them float for a week or two

 

[Art]

I normally let them float for a week or two

In the cup with the agar or do you rinse the agar out and let them float without the cup?

 

[BigWave]

In the cup with the agar or do you rinse the agar out and let them float without the cup?

Remove the agar. All it does is cause algae outbreaks and polute your water. Yes as said above just float them. Even if they were contained withing a large makeshift feeding ring type deal.
From my understanding each type of plant requires a different technique when the agar is being created. Some reqire more of this nutrient less of this hormone etc. I dont know much about actual plant biology but have a hunch most of the plantlets are so young and havnt developed the right parts to be able to convert to submersed right off the bat. Gues its aken to the sink or swim saying. You either drown or start swimming or in this case grow real quick.

 

[Art]

Yes, I've been using the sink or swim technique, unsuccessfully. I think next I'm going to try Tropica or ADA tissue culture rather than the Asian ones I've been using. Let's see if the source of the tissue culture plants really does make a difference.

 

[spawnerjp]

In the cup with the agar or do you rinse the agar out and let them float without the cup?

out in jar without the gel

 

[Count Krunk]

Did you try a new TC source yet?

 

[Art]

Did you try a new TC source yet?

Not yet. Giving the tank a chance to mature some more before trying again with a different supplier.

By the way, THANK YOU for setting up your achievements! You're the first member that I see that has done that!

 

[qwedfg]

I’m surprised to hear people having trouble with tissue culture plants. I am a noob but never really had issues with tissue culture plants but I never had hard plants I guess. I just got a bunch in and usually just trim the roots and stick it in the aqua soil. I got some rotala pearl that was so bad it got refunded but still sent in my order and it’s slowly recovering.

 

[Grobro]

I feel like I am all over your forum now so I apologize if I am commenting way too much. I am really enjoying these topics.

I am nowhere near the plant expert like Dennis and others. I can offer my take on the topic as someone who has spent a lot of time with tissue cultures. I use practically 80-90% cultures for my layouts because I am more confident I can mitigate algae using them versus plants that were grown in farm tanks. In fact I had one issue where I bought some plants from a facebook group that came with Gibba and that Gibba ended up in both my tank and my son's tank probably through pinsettes or net. Super frustrating. Anyhow..

This tissue culture issue is super dependent upon

1. Cycle - where you are in the startup cycle
2. Soil - aquasoil? if so how much soil you are working with / how much of an ammonia spike is present
3. Water changes - frequency and method, disruptive and heavy or smaller with higher frequency
4. Quality of tissue cultures - when you look at the cultures themselves, are they recently placed or they are or have they grown a bit
5. How densely planted you went
6. Type / species of plant culture

If you use amazonia, especially if you use a high soil to water ratio, your tank is going to be an ammonia inferno for plants during the first 30 days at least, even if you do a decent amount of water changes.

Solutions:
1. Plant after your tank is cycled (this means you drain the entire thing to plant, don't disrupt the water too much, save some of the water with bacteria in it to refill it. This will ensure you don't nuke your tc's. Or if you want to plant straight away, do frequent water changes, I am talking every day for the first two weeks. ADA used to have a recommendation sheet on water changes with dosing instructions. I'll see if I can find it.
2. If you use a ton of amazonia or something nutrient rich, versus something inert, that is going to fry your tissues unless you water change a lot. If you use something inert you will probably be fine to plant straight away.
3. Every day for the first 2 weeks at least 20% but consistent with the parameters. Slowly add the water back and try not to disrupt the soil and plants. Also you can't do frequent water changes, then just stop. You have to slowly taper down in frequency.
4. Look for tc's that appear a little grown in but not browning. Sometimes tissue cultures look so young they have little chance unless the tank is at the 6-9 month stage.
5. Plant them densely. A great example is Filipe Oliveira.
6. Some cultures just shouldn't be planted yet - like Dennis said, super sensitive plants will burn. You need to wait until the water is balanced more.

I have wasted a lot of money frying tissue cultures and have found that it depends largely on the soil and the cycle. Also the type of plant. I could plant a field of glosso or monte carlo and have no problems. Same with most crypts and some stems, even if the crypts melt they come back strong. Stems seem to always survive when planted densely.

I think the best option is to plant after 30 days but I am way too impatient for that, so I go for it and sometimes I lose a culture or too but overall end up successful if I am rigorous with my husbandry.

 

[Mr.Shenanagins]

I’ve got some tissue cultures myself. The tank has been running since the beginning of the year so definitely matured. I just purchased Java fern petite and hydrocotyle trip mini. The mini I glue to small stones to keep them down. So far, it appears the hydrocotyle is starting to grow! Haven’t had luck in the past with tissue cultures either so excited to see it take off.