Tissue culture

Several years ago there was an orchid breeding greenhouse locally that we used to visit. While there I would watch them tissue cloning orchids under hoods. They were using microscopes and placing small bits of plant material in petri dishes and placing the petri dishes into incubators. I thought: Good Lord, you must have to have at least a Ph D to tackle that.

Over the years the process has evolved so that it is possible to do tissue culture at home, so I thought I would give it a try. As I needed a sink and clean environment, I elected to wait until my wife was going to be out for a while and use the kitchen for my laboratory.

I used an opaque storage container for a hood. The storage container had to be sterilized with 90% alcohol, along with everything else used in the process, A small section of green growing branch tip was removed from a Mission and Pendolino olive tree. It was necessary to wash the plant material in detergent, alcohol, beach with detergent and finally sterile water. Rinsing was also required several times between each wash.

The sterile water rinse was performed under the hood, along with all trimming and placing the plant material into the vials containing the nutrient gel. The vials have been placed in a sterilized container under a small T5 grow light, where in a few weeks new plants should begin to develop. That is if I have the process correct, but if not I will try again, as I found it fascinating. It should be possible to produce hundreds, or thousands, of plants from a tiny piece of the plant.

Here is a link with more information, and youtube also has several videos on Plant Tissue Culture for anyone interested.

http://hort201.tamu.edu/YouthAdventureProgram/TisueCulture/TissueCulture.html

 

That looks very intriguing - look forward to seeing results in due course,
Jenny

 

Cor

 

double cor

 

Very interesting, wouldn't mind trying that, don't think my wife would like me turning our kitchen into a laboratory.

 

Thank you for the link really interesting

 

You might find the book "Plants from Test Tubes" helpful
www.amazon.com/Plants-Test-Tubes-Introduction-Micropropogation/dp/1604692065

 

Couple of other links which may be of interest were posted in this thread:

http://gardenerscorner.co.uk/forum/threads/ultimate-conservatory-garden.55051/#post-744952

 

Here is my first attempt. It is not impressive, however, after only three weeks, a small piece of Gardenia Veitchii is now developing into four new plants. It is a fascinating process to say the least.

 

I'm impressed Hydro. 4 lovely Gardenias...
Jenny

 

I thought an update would be of interest; the Gardenia Veitchii clone is still multiplying. This process is so interesting that I have set up a permanent space for micro-propagation in the basement and gave my wife back her kitchen.

 

Priceless! Well done, and wishing you luck with your new hobby

 

I have found that tissue culture techniques are excellent for starting hard to germinate seeds. The top photo, taken a few weeks ago, is of a Bougainvillea seed in vitro. The seeds planted in soil have not done anything, while the one started in protocol is now potted and in the greenhouse, as seen in the second photo.

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This is fascinating @hydrogardener I'd love a go at this can I ask what media you used? was it a commercial media or a homemade agar? I have found a simple mix on this site for the latter https://www.omnisterra.com/botany/cp/slides/tc/tc.htm

• 1/8 cup table sugar
• 1 cup water
• 1/2 cup stock solution (Miracid diluted 1/4 tsp in 1 Gallon water)
• 1/2 inositol tablet (125mg)
• 1/4 vitamin tablet with thiamin
• 2 Tablespoons agar flakes

Which doesn't look to far beyond my means I have also read that once you are beyond the intial multiplication you can add coconut milk to the mix to encourage more growth and then also malt extract once you have moved onto rooting.

also how did you go about sterilising your jars? I have found most instructions require the use of a pressure cooker however some mention just using a microwave but then requiring special lids for the baby jars?

all the best,

 

Ice,
The media I used was one I mixed with different growth hormones and lab grade agar. Different plants require different hormones, there is not one universal protocol for all plants. The formula you found will most likely work for easy to propagate plants like African Violet and such. Also, I used a large pressure cooker to sterilize the media. It would a good idea for you to visit: http://www.kitchenculturekit.com
The owner's name is Carol and she has a Ph. D. in this field. Ask her to email you her workshop handout and she will provide it at no charge. It will give you a step by step guide in getting started and it has a recipe similar to the one you have above. She has slide shows on her site that will answer your questions regarding microwaving, and yes you can use one. You will also find information on caps and vessels on her site. Carol is passionate about TC and will answer any question that you send her. I know, I ask questions frequently and receive prompt answers. Good Luck.