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Experience With Tissue Cultures?

J

JesterJoker

sorry if this post is mis-categorized I would think it goes under grow tech probably?
 
I'm looking at starting to do tissue cultures, from original strains of peppers.(so no need to keep a large original plant taking up space or worrying about cross pollination and keeping seed)
 
I found a few suppliers pretty much places like http://www.carolina.com/ and  http://www.planttc.com/ other places selling kits/media, but not specifically stated for peppers?
I can find a pressure cooker fairly easy, or a used autoclave so I should have the tools needed to make media.  
 
But I wanted to know what the starter material for making media, specifically for peppers, for agar and the solution and hormones for rooting/vegetative growth.
If anyone can help that would be great. I also once I get started, I want to setup a tissue culture club to share pepper varieties because we all know seed does not always grow true.
 
tissue cultures would be the best for true genetics.
 
 
Hopefully someone has some background info/material I can read up on, anything I find I will publish online and make it free resource.
 
I would think that big-time planters that buy seed would profit from this sort of information and be able to use it to their advantage, for growing true product instead of relying on eBay seed sellers and mislabeled or hybridized seeds.  Or they can also grow hybrids 100% the same all the time with it, without worrying about genetic differences.
 
 
Yes, many planters would benefit from this but it isn't the most consistent method out there.What I've read in most publications is that most plants that come from cultures tend to mutate.
 
I have 20 years experience with tissue culture. Mostly mammalian and insect cells, but I`ve also done plant cultures, as well as germinating orchid seeds in tissue culture.
 
Here is a very brief guide to what you can do.  http://csmbio.csm.jmu.edu/biology/renfromh/pop/pctc/cloning.htm
 
The techniques are relatively straight forward, but learning sterile technique is the first hurdle you have to jump. You can do it in an area with low air flow on a table, but you will see contamination with fungus/bacteria. A small portable culture hood would be far better, but they are not cheap. You will also need an incubator to control temps and humidity precisely. 
 
thanks, that is just the material I have been looking for.
I just now need to find the materials from that information for me to use for the cultures
 
JesterJoker said:
sorry if this post is mis-categorized I would think it goes under grow tech probably?
 
I'm looking at starting to do tissue cultures, from original strains of peppers.(so no need to keep a large original plant taking up space or worrying about cross pollination and keeping seed)
 
I found a few suppliers pretty much places like http://www.carolina.com/ and  http://www.planttc.com/ other places selling kits/media, but not specifically stated for peppers?
I can find a pressure cooker fairly easy, or a used autoclave so I should have the tools needed to make media.  
 
But I wanted to know what the starter material for making media, specifically for peppers, for agar and the solution and hormones for rooting/vegetative growth.
If anyone can help that would be great. I also once I get started, I want to setup a tissue culture club to share pepper varieties because we all know seed does not always grow true.
 
tissue cultures would be the best for true genetics.
 
 
Hopefully someone has some background info/material I can read up on, anything I find I will publish online and make it free resource.
 
I would think that big-time planters that buy seed would profit from this sort of information and be able to use it to their advantage, for growing true product instead of relying on eBay seed sellers and mislabeled or hybridized seeds.  Or they can also grow hybrids 100% the same all the time with it, without worrying about genetic differences.
 
Click to expand...
https://www.youtube.com/watch?v=hgOqTyiI_30 looks like it has been done there are some pepper plant pics in the beginning of this video
 
@ nigel, im curious, beyond heavy duty breeding projects, what uses do you find for this?
 
can you store a culture at low temps for long times? like years at a time? can you store plant cells at cryogenic temperatures for longer periods of time?
 
queequeg152 said:
@ nigel, im curious, beyond heavy duty breeding projects, what uses do you find for this?
 
can you store a culture at low temps for long times? like years at a time? can you store plant cells at cryogenic temperatures for longer periods of time?
Click to expand...
I think the use is primarily a scientific tool for labs who study plant metabolism, signaling and structure. It`s obviously very useful if a lab want to make transgenic plant cells or plants, too. Give the difficulty of adding or subtracting genes from plant cells/plants (it`s relatively straightforward, but a bit hit and miss), maintaining clonal populations of cell material is also essential in a laboratory setting. It allows the researchers to manipulate the plant cells for basic research or applied research. 
 
The other thing this has been used for is producing thousands of plants of highly endangered species from all sorts of different genera. Or even plants where only one or no individual is known to be left in the wild. While what`s left are clones, it does allow their study with the possibility of introducing a small amount of genetic diversity. Encephalartos woodii is a cycad from southern Africa where only one plant is known. It`s a male, as Cycads are ancient, having both male and female plants. Studying clones has lead to a way to convert some clones to females (almost but not quite, actually) and hence start a breeding project. Not ideal, genetically, but it`s a start.
 
Another use is to germinate Orchid seeds. They will only germinate in cell culture, as they have no "on-board" nutrient systems. In the wild they co-exists with mycorrhizal fungi that provide for the seeds needs. 
 
Cultured material - stems, callouses etc, can be stored at around 4°C for up to a year, I think. Yes, you can store cell cultures in liquid nitrogen, although I do not know how long they remain viable. 
 
I am a very, very long way from being an expert with plant cell culture. 
 
interesting stuff for sure... regarding what nigel said with respect to propigating rare species... i just recently read a story where... a local gov, or w/e needed to remove a rare albino tree, of which only a handful are known to exist.
 
apparently they took 1000's of clones of this tree in anticipation of removing the thing. never knew albino plants existed tbh. im sure its markedly different than albinism in mamals etc tho...
 
btw can a tissue culture be used to save a plant that's infected with a virus? sounds like a long shot, but ever heard of such a thing? i have read(i think?) that several species of orchid have virus's that produce fascinating colors/patterns. maby im thinking of tulips? 
 
ive always loved cultures, fungi especially. i bought all these books and a monster hepa filter weighing like 70 lbs, just never got to doing anything with any of it.
lol i think i just wanted to play junior science cadet with the petree dishes and loops etc.
seems like a skill worth cultivating.
 
I would be up to trading rare plant types with people when I start.
 
I might even start a tissue culture group if I get into it big, im definitely going to share anything I learn with others free.
 
JesterJoker said:
I would be up to trading rare plant types with people when I start.
 
I might even start a tissue culture group if I get into it big, im definitely going to share anything I learn with others free.
Click to expand...
 
I considered doing this awhile back. A biology professor friend of mine gave me good instructions for a recipe (don't have the copy with me atm). We looked into what part of the plant to use. It looked quite difficult for capsicum. I found a scientific article that used cotyledons. Let me know what parts work for you. I didn't give it a shot.
 
Edit: It could be easy for all I know. I have no experience and the biology professor wasn't experienced in capsicum tissue cultures. If I did the experiment, I would have tried the cotyledons since they work and various other parts of the plant.
 
 
I found a pdf that would be helpful for us.
 
Here is the link:
https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=13&cad=rja&uact=8&ved=0CGkQFjAM&url=http%3A%2F%2Fir.canterbury.ac.nz%2Fbitstream%2F10092%2F6747%2F1%2Fbodhipadma_thesis.pdf&ei=Ji5GU4XbDPHUsAT34IAw&usg=AFQjCNG0E3kMoADy165f45A3ypvg-runlg&sig2=4NPwElnGJcmhMU7aYLc__g&bvm=bv.64507335,d.cWc
 
Why not just propagate from cuttings?  You get true to type (genetically identical) plants at a fraction of the cost and time.
 
There is a misconception that tissue culture will produce millions of plants in a fraction of the time that conventional techniques can.  The truth is that the actual vegetative output of a plant remains the same in TC i.e. if it is a slow grower it will be slow in TC.
 
Also to calculate the exact recipe needed for proliferation of the callous is a very time-consuming process.  There is no "one recipe" that fits all.  Sure, there are three basic recipes (Knudsen K etc) for the growing medium (gel) but that is not all that needed.  Growth stimulants are also needed.  I helped a post-grad student with her work on Scadoxus (I supplied propagation material) - she took forever to eventually compile the exact concentrations needed for each stimulant.  These recipes are strictly guarded!  And they are often not generic for a Genus - individual species might require a tweaked version.
 
Time is also an issue.  I once wanted some very rare Agapanthus TC'd.  The dude asked how many immature inflorescences (the meristem that is required) I had for him to work with - I replied "Ten".  He laughed and said he wouldn't consider it with less than 1000!  It just takes too long and is too risky to maintain and grow the callous that is needed to produce a large numer of plants - this is an expensive process.  I canned the project - if I had a 1000 I wouldn't need TC!!!!
 
Then there's hardening off!  Jeepers creepers that is another nightmare.  And most often plants out of TC flower immediately.  In some plants this is a serious reduction of vigour and losses are great.
 
Having said all of that I would like to see how this pans out.  It would be a very interesting study.  I do have to wonder why it hasn't been done before though.
 
It's been done with capsicum or do you mean members on the forum? Someone around here might have done it with capsicum. I was considering doing it for a fun personal science project (I'm a strange person). It's infinitely easier to clone cuttings. I clone mine by just sticking the suckers in soil and keeping their leaves wet. The main stem of my F1 hybrid rotted, so I had to clone it using my stick it in the soil and spray bottle method, lol. It didn't lose any leaves. If I used tissue culture instead, it would have taken much longer to get the same growth. I could have gotten a lot more plants though. I can see it being useful if you need to grow a lot of F1s.
 
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