Tissue culture

Discussion in 'Propagation This Month' started by hydrogardener, Dec 27, 2013.

  1. hydrogardener

    hydrogardener Total Gardener

    Joined:
    May 10, 2013
    Messages:
    1,005
    Gender:
    Male
    Occupation:
    Retired
    Location:
    Brunswick, New York, USA
    Ratings:
    +4,731
    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

    [​IMG]

    [​IMG]
     
    • Like Like x 7
    • Informative Informative x 2
    • Jenny namaste

      Jenny namaste Total Gardener

      Joined:
      Mar 11, 2012
      Messages:
      18,324
      Gender:
      Female
      Occupation:
      retired- blissfully retired......
      Location:
      Battle, East Sussex
      Ratings:
      +31,022
      That looks very intriguing - look forward to seeing results in due course,
      Jenny
       
      • Agree Agree x 3
      • Phil A

        Phil A Guest

        Ratings:
        +0
        Cor :):dbgrtmb:
         
        • Agree Agree x 1
        • Spruce

          Spruce Glad to be back .....

          Joined:
          Apr 10, 2009
          Messages:
          8,731
          Gender:
          Male
          Ratings:
          +12,318
        • Ian Taylor

          Ian Taylor Total Gardener

          Joined:
          Nov 1, 2013
          Messages:
          2,228
          Gender:
          Male
          Occupation:
          Maintenance Manager, Oddfellows on the Park.
          Location:
          Cheadle Hulme
          Ratings:
          +2,751
          Very interesting, wouldn't mind trying that, don't think my wife would like me turning our kitchen into a laboratory.
           
        • sal73

          sal73 Total Gardener

          Joined:
          Sep 4, 2011
          Messages:
          2,833
          Gender:
          Male
          Location:
          Bedford
          Ratings:
          +2,998
          Thank you for the link really interesting
           
        • Kristen

          Kristen Under gardener

          Joined:
          Jul 22, 2006
          Messages:
          17,534
          Gender:
          Male
          Location:
          Suffolk, UK
          Ratings:
          +12,667
          • Like Like x 1
          • Kristen

            Kristen Under gardener

            Joined:
            Jul 22, 2006
            Messages:
            17,534
            Gender:
            Male
            Location:
            Suffolk, UK
            Ratings:
            +12,667
            • Like Like x 1
            • hydrogardener

              hydrogardener Total Gardener

              Joined:
              May 10, 2013
              Messages:
              1,005
              Gender:
              Male
              Occupation:
              Retired
              Location:
              Brunswick, New York, USA
              Ratings:
              +4,731
              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.

              [​IMG]
               
              • Like Like x 5
              • Informative Informative x 1
              • Jenny namaste

                Jenny namaste Total Gardener

                Joined:
                Mar 11, 2012
                Messages:
                18,324
                Gender:
                Female
                Occupation:
                retired- blissfully retired......
                Location:
                Battle, East Sussex
                Ratings:
                +31,022
                I'm impressed Hydro. 4 lovely Gardenias...:wub2:
                Jenny
                 
              • hydrogardener

                hydrogardener Total Gardener

                Joined:
                May 10, 2013
                Messages:
                1,005
                Gender:
                Male
                Occupation:
                Retired
                Location:
                Brunswick, New York, USA
                Ratings:
                +4,731
                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.

                [​IMG]
                 
              • Kristen

                Kristen Under gardener

                Joined:
                Jul 22, 2006
                Messages:
                17,534
                Gender:
                Male
                Location:
                Suffolk, UK
                Ratings:
                +12,667
                Priceless! Well done, and wishing you luck with your new hobby :)
                 
              • hydrogardener

                hydrogardener Total Gardener

                Joined:
                May 10, 2013
                Messages:
                1,005
                Gender:
                Male
                Occupation:
                Retired
                Location:
                Brunswick, New York, USA
                Ratings:
                +4,731
                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.

                [​IMG][/URL]

                [​IMG]
                 
                • Like Like x 1
                • Informative Informative x 1
                • IceColdRum

                  IceColdRum Cacti & Herb Mad

                  Joined:
                  Aug 9, 2014
                  Messages:
                  506
                  Gender:
                  Male
                  Occupation:
                  Customer Service Advisor
                  Location:
                  Totland Bay, Isle of Wight
                  Ratings:
                  +837
                  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,
                   
                • hydrogardener

                  hydrogardener Total Gardener

                  Joined:
                  May 10, 2013
                  Messages:
                  1,005
                  Gender:
                  Male
                  Occupation:
                  Retired
                  Location:
                  Brunswick, New York, USA
                  Ratings:
                  +4,731
                  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.
                   
                  • Informative Informative x 1
                  Loading...

                  Share This Page

                  1. This site uses cookies to help personalise content, tailor your experience and to keep you logged in if you register.
                    By continuing to use this site, you are consenting to our use of cookies.
                    Dismiss Notice