How to take tissue culture?

[badatusernames]

In Bjorn's recent videos featuring Dave Bogan and Phil Julian, they both mentioned that starting willowleaf ficus from tissue culture resulted in larger bases. I hadn't heard of tissue cultures before, and was curious if anyone here had experience propagating in this manner and how they go about it?

 

[Crazy Diamond]

Interesting for sure, but probably not very practical in a small aficionado scale, you will need laboratory material I think and that wont be cheap, also learn the techniques and stuff

 

[rockm]

A lot of work for little return for the backyard hobbyist:

TISSUE CULTURE OF WOODY PLANTS - Earth-Kind® Landscaping Earth-Kind® Landscaping

Texas A&M University - Academic analyses and information on horticultural crops ranging from fruits and nuts to ornamentals, viticulture and wine.

aggie-horticulture.tamu.edu

 

[badatusernames]

Good to know! Ok, might be a tad much haha

 

[penumbra]

In Bjorn's recent videos featuring Dave Bogan and Phil Julian, they both mentioned that starting willowleaf ficus from tissue culture resulted in larger bases. I hadn't heard of tissue cultures before, and was curious if anyone here had experience propagating in this manner and how they go about it?

Tissue culture is somewhat analogous to test tube propagation. It is not something the average person would do.
I am certain there is an easier way to grow a larger base on your ficus.

 

[Wires_Guy_wires]

Tissue culture is somewhat analogous to test tube propagation. It is not something the average person would do.
I am certain there is an easier way to grow a larger base on your ficus.

Yes, it's exactly that.
Weird thing to say you grew a nebari that way, but a good radial nebari is what I've found to happen almost always in shallow containers in tissue culture.
Back in the days we changed deep and long testtubes for hockey puck shaped containers (about twice as tall/high as a hockey puck) and it produced even rooting all around.


It's in essence the same to place a tile underneath a rootbase of a cutting; the roots have no place to go but outwards.

 

[boguz]

In Bjorn's recent videos featuring Dave Bogan and Phil Julian, they both mentioned that starting willowleaf ficus from tissue culture resulted in larger bases. I hadn't heard of tissue cultures before, and was curious if anyone here had experience propagating in this manner and how they go about it?

i am also looking for some stuffs about tissue culture for bonsai. its an old post but is it possible to share that video?

 

[badatusernames]

i am also looking for some stuffs about tissue culture for bonsai. its an old post but is it possible to share that video?

Nah…. It’s somewhere in his paid archives, I don’t have access any more but you aren’t able to download them anyways (plus he needs to get paid for his content!)

 

[Glaucus]

You need sterile conditions and the right mixture of plant hormones. It is not very practical. It is ideal when you need to create tens of thousands of plants and you have time to generate hundreds to fine-tune the protocol.
You want a laminar flow hood and autoclave to do it properly.
There have been some cases where the same genetic stock behaves different in terms of epigenetics from traditional cuttings.
Really doubt there is much benefit to anyone here doing it for bonsai.
Getting plant material to become sterile is not that easy. Bacteria and fungi also live inside plants. You need to nuke non-sterile tissues first, killing most of them, so you get cell clumps that are completely sterile and growing.
Then when you have sterile cell lines, you can propagate them over and over and try different hormone mixtures to get the correct root, leaf, or shoot growth.

 

[EHA]

You need sterile conditions and the right mixture of plant hormones. It is not very practical. It is ideal when you need to create tens of thousands of plants and you have time to generate hundreds to fine-tune the protocol.
You want a laminar flow hood and autoclave to do it properly.
There have been some cases where the same genetic stock behaves different in terms of epigenetics from traditional cuttings.
Really doubt there is much benefit to anyone here doing it for bonsai.
Getting plant material to become sterile is not that easy. Bacteria and fungi also live inside plants. You need to nuke non-sterile tissues first, killing most of them, so you get cell clumps that are completely sterile and growing.
Then when you have sterile cell lines, you can propagate them over and over and try different hormone mixtures to get the correct root, leaf, or shoot growth.

100% Echo what Glaucus say. I work as a scientist in Biotechnology and we have a tissue culture division. You really need a laboratory set up and as stated endophytes (internal bacteria in the plant) can be difficult at times. The quantity of plant hormones to use for micropropagation also needs to be researched and optimized. Even different cultivars from the same species sometimes react differently. We produce more than 800 000 plant clones at a time and for that kind of numbers the only way to go is plant tissue culture. I do not think it is practical for bonsai and definitely not attainable at home unless you have the know how and deep pockets to buy all the equipment needed.

 

[EHA]

100% Echo what Glaucus say. I work as a scientist in Biotechnology and we have a tissue culture division. You really need a laboratory set up and as stated endophytes (internal bacteria in the plant) can be difficult at times. The quantity of plant hormones to use for micropropagation also needs to be researched and optimized. Even different cultivars from the same species sometimes react differently. We produce more than 800 000 plant clones at a time and for that kind of numbers the only way to go is plant tissue culture. I do not think it is practical for bonsai and definitely not attainable at home unless you have the know how and deep pockets to buy all the equipment needed.

double posted. Apologies.

 

[Port Farmer]

Nah…. It’s somewhere in his paid archives, I don’t have access any more but you aren’t able to download them anyways (plus he needs to get paid for his content!)

I believe it is in the 3 part tropical series.

Not sure how legit, but this you tube channel is all about tissue culture propagation at home. https://www.youtube.com/@plantsinjars

 

[nuttiest]

You need sterile conditions and the right mixture of plant hormones. It is not very practical. It is ideal when you need to create tens of thousands of plants and you have time to generate hundreds to fine-tune the protocol.
You want a laminar flow hood and autoclave to do it properly.
There have been some cases where the same genetic stock behaves different in terms of epigenetics from traditional cuttings.
Really doubt there is much benefit to anyone here doing it for bonsai.
Getting plant material to become sterile is not that easy. Bacteria and fungi also live inside plants. You need to nuke non-sterile tissues first, killing most of them, so you get cell clumps that are completely sterile and growing.
Then when you have sterile cell lines, you can propagate them over and over and try different hormone mixtures to get the correct root, leaf, or shoot growth.

Are the new cell lines then more prone to disease? Is the transition to pots high success rate?

 

[Glaucus]

Are the new cell lines then more prone to disease? Is the transition to pots high success rate?

Not sure. I think you mean that plant tissues and then the mature plants derived from them would completely lack endophytes, and thus have less health.
But I assume there would still be endophytes that cannot grow in cell cultures and that do not take over the growth medium and contaminate the plant tissue culture.
It seems some do however. I am not sure that if a plant tissue grows nicely and seemingly 'sterile' in a tissue culture, there really are no bacteria colonizing inside the plant any longer.

In general, the issue with tissue culture isn't really going from a tissue culture back to a health plant that can grow in soil or substrate. Though of course that takes some transitioning. Often also because there is a different protocol for cell division and another one for creating a plant with roots that can grow in soil/substrate. And it will depend a lot on the species of plant.
But this isn't the impractical hurdle.

I believe in house plants there are a lot of people that believe that tissue culture plants are inferior. But this seems mostly based on myths. There is this reasoning that if you have a plant cell line in basically stem cell clumps in growth media for many generations, there are some epigenetic and eventually also genetic changes that make it a different type of plant that a genetic line of plants that always grow outdoors and are propagated by seeds. But evolution will be very slow. And one can go back to the original (epi)genetics of a plant that grows outdoors and start a new cell line. Not sure if for the commercial plant tissue culture propagators observe this and terminate lines after months or years. Theoretically they would be immortal, but there's some technical research papers on how these cell lines do change more quickly than maybe expected.

 

[nuttiest]

I am not sure that if a plant tissue grows nicely and seemingly 'sterile' in a tissue culture, there really are no bacteria colonizing inside the plant any longer.

That means there is room for changing gene expression still even in clones as they can lose more endophytes over generations?
I feel like we see that in cuttings of cuttings, and for strength some need to go back to seed. But oddly you can take a new growth cutting of a diseased plant and make a disease free new plant sometimes.
Anyway, it just fascinates me how tissue done now, when it used to just be part of a nursery job - cut and stick every week and a whole field of pots of new plants to replace sales. A lower percentage in the numbers game, but no need for a biologist!
How many sterile hoods to purchase or workers for 800,000 plants, lol.

 

[EHA]

That means there is room for changing gene expression still even in clones as they can lose more endophytes over generations?
I feel like we see that in cuttings of cuttings, and for strength some need to go back to seed. But oddly you can take a new growth cutting of a diseased plant and make a disease free new plant sometimes.
Anyway, it just fascinates me how tissue done now, when it used to just be part of a nursery job - cut and stick every week and a whole field of pots of new plants to replace sales. A lower percentage in the numbers game, but no need for a biologist!
How many sterile hoods to purchase or workers for 800,000 plants, lol.

That's the beauty of tissue culture. We grow the little plants in vesels in semi solid media. Each vesel can easily hold 1000 plus plants in clumps. So 4 double laminar flows and 12 staff and its easy to handle. Hardening the plants outside in a glasshouse after the tissue culture process is the biggest logistical problem. Luckily we outsource that to commercial nurseries. Planting 800 000 little plants in speedling trays are not an easy and quick task. We do this type of operation for the agricultural industry where farmers need thousands of clones of the same cultivar to plant. I guess for the horticultural and home garden industry your numbers and aims would be totally different.

 

[Glaucus]

Rooting cuttings is labour intensive for both the traditional method or the tissue culture. Because the labour cost are the major factor lab costs can actually be soaked up, because tissue culture must be more expensive.
Why and when it is commercially viable to do tissue culture, I am not sure. But I saw a quotation for 1 euro per plug for a company that produces plants though tissue culture.
It may actually be that one company that has people producing cuttings full time regardless of the reason, that then sell them to nurseries with automated nurseries that try to keep the number of labourers as low as possible, is the most economically viable.
A nursery with an automated greenhouse likely won't have enough labourers under contract to do short bursts of intense cutting takings during those periods of the season where cuttings are produced. Also, in Europe this allows countries with lower wages to produce the cuttings, like Portugal and Poland, for nurseries in Germany or the Netherlands.

 

[nuttiest]

I guess that's how we get the perfectly similar strawberries. And Netherlands outsourcing bulb slices would be interesting.