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By Bob Axmear This is an overall description of tissue culturing hostas. It is meant to give you a rough idea of how to go about it. You might need to do a little more research on your own. I am going to add to it as things happen, how to remove the explants, etc when they come up. Also Carol Stiff is very helpful and conducts seminars and sells tc products in small quantities. Also a good place for much of the supplies is PhytoTechnology Laboratories, you can buy prepared media there if you want to go that route. A good beginner book is Plants from Test Tubes. Some TC lists are here. Some CDs, I found Carol's informative for beginners, Dr. Ronald A. de Fossard is for more advanced people.
At this time I am going to show you 3 plants I treated with Bap 10. This is the hormone that causes dormant buds to break open and start growing, these are what you use to collect your explants(to remove living tissue especially to a medium for tissue culture). I treated these three plants on 5-26-2007. I used the hormone at a 3% solution. This means 30ml per liter or 1 oz in a quart of water or fractions of these amounts for making small amounts of solution. Also add a drop or two of surfactant, dish soap works fine. I just use a spray bottle and apply to the leaves and especially to the crown of the plant, down between the leaves. Within about 10 days you should see all kinds of new shoots coming up at the base of the plants.
Within about 10 days you should see all kinds of new shoots just starting to poke out of the soil.
Many buds to remove for their mersistem
Bud removed from the plant with a little of the base tissue. I use a hooked scalpel to do this but whatever works for you.
You need to clean the bud of the outside leaf tissue down to the remaining meristem and a little of the base, it peels like an onion. Make sure it is as clean and white as you can get it by trimming any discoloration off of the base tissue. That is easy to do, just shave the sides of the base with your scalpel.
In the coming days I will add more photos and a general media recipe or protocol for hostas, this will be for initiation and multiplication of the explants. You can also find recipes in the search engines for this. I will add a basic list of tools and equipment you will need as well soon.
1000 ml beaker or graduated cylinder
Scalpel and blades, buy assortment of blade styles, hooked ones seem to work best for removing explant. Make sure blade fits handle size as well
6" or 8" Forceps
Scale
PH Paper or Meter
Take 1 to 4
The protocol isn't too complicated and if someone wants to continue on I can sure explain it easily enough. Here is a link to one so you can see it. Also here is a link to some videos online. Another
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