How To...

What Is Tissue Culturing?

This article appears in Volume 7 – Issue 3 of SKUNK Magazine.

 

TISSUE CULTURING consists of preserving plant genetics by taking smaller cuttings and growing them in a sugar and nutrient-rich media.   Tissue culture doesn’t change the genetic material or the organism—rather, it denotes multiplying the genetic material. Unlike traditional seed or cloning you do not have to wait approx 6 months to take a new cutting, you can take a new cutting within 2-4 weeks and start the process over again. Research has shown that if you start your plant from a tissue culture and then multiply them in your lab, as opposed to multiplying them from clippings, your plants can hold the same genetic material for 20 years.

Other Than Yield, Why Do We Want to Control Plants Through Culturing?

We all know that medical marijuana has eased the effects of many illness and chronic pain for many years. Staph have been linked to diseases stemming from molds and fungi in cloned plants. One of the best parts about tissue culturing is you can now get rid of contaminated cannabis plants. Tissue culturing will allow you to breed out viruses, molds, bacteria and yeasts that may be hibernating in your plant giving you a TRUE CLEAN medical product.  We are now capable of producing therapeutic medicine that will give optimal benefits with consistency for the consumer with minimal side effects.   If we generate a cannibinoid with little to no disease and minimal side effects, we will be providing contaminant-free products, which in turn will give a higher quality of medicine. This specifically will help people with a high sensitivity to molds, fungi, and yeasts.  Most importantly, all plant and nutrient research has come from tissue culture research.

MOTHERS

  • Always have a shortage
  • One strain/mother
  • Not always contaminant free
  • Not cost effective

CUTTINGS

  • Can have a poor survival rate
  • Use more light, water, room
  • Vulnerable to diseases
  • Can only cut so many at a time

SEEDS

  • Lack uniformity
  • Use more light, water, room
  • Take longer to mature
  • Not always true to type

TISSUE CULTURING

  • Never a shortage of mothers
  • Less light, water, room
  • Uniform plants
  • Contaminant, virus free plant
  • Preserve plant genetics
  • Less maintenance
  • Can produce plants from stem leaf or callus
  • Brighter, fuller
  • Up to 25% more resistant to disease

The hardest part about tissue culturing is contamination risks and consistency.  Remember one particle of dust can carry 100s of molds, bacteria or fungus spores.  As longs as you know how to keep yourself, your work area and your storage area super clean, you can tissue culture.

TISSUE CULTURING

The first step to tissue culturing is clean, clean, clean.  Clean the area you will be working in by spraying and wiping down all counter surfaces. (If using a BBF Box, thoroughly wipe down inside with 25% alcohol.)

Spray and wipe down the following with your sterile napkins and place in the clean area:

  • Plate
  • Tools
  • Tool Jar (approx 2 to 4 in. tall and narrow)
  • Burner
  • Lighter
  • Sterile water jars
  • Your jars to be used for propagation

Arrange your work area to your liking.

If using a plastic tub for your clean area, make sure to cover area while clipping your cuttings.  If you’re using a BBF Box be sure to shut the door before taking your cuttings.

Next Step Is to Make Your Media

You will need the following:

  • Sugar
  • BBF Hormones
  • Ph Tester kit
  • 1 liter measuring cup
  • BBF MS Salts
  • BBF Agar
  • Mixer
  • 1,000L distilled water

We recommend using BillBerry Farms Tissue Culture Set.  This set is made specifically for your cannabis plant.

Using a blender, mix the all of the ingredients together. PH the media and cook in the microwave following the BBF instruction manual.  When finished pour an equal amount into your choice of test tubes, tubs, or jars.  Place your containers, napkins, tools, water to sterilize into a pressure cooker and cook at 20psi for NO more than 20 min.  This process will remove any contaminants and sterilize your media.  Once cooled, place into your clean work area or into your clean storage area for future use. 

Taking Your Cuttings

(these instructions are for using the BBF set, other media may require smaller cuttings)

You want to choose the biggest, fullest and healthiest, pest-free plant.

  1. Pick a branch, starting at the top of the branch move down 2 nodes and cut. Be sure to leave yourself some stem to work with.
  2. Trim all sun leaves, extra leaf stems and any extra leafs that you do not need.  Place into a jar filled with water.  Node should be no smaller that 5/8” in and no longer that 1 ½” Plantlet should have no more than 2 nodes. (See photo below)
  3. Repeat until you have a significant amount of plantlets.
  4. Wash plants with water, bleach, and soap solutions.  Rinse with alcohol to remove any left over bleach or soap residue, then a final rinse with sterile water.  Place into your work area. 
  5. Before entering your work area wash hands and arms with warm soapy water. Here you will proceed to placing your nodes into the media filled vessels.
  6. Spray hands and work area with 70% alcohol, wipe with sterilized paper towels.
  7. Spray plate with 70% alcohol and wipe with sterilized paper towels
  8. Run tools over the flame (to burn off the alcohol, a burner is not required.) and place on the plate (for 15 sec) to cool down before touching your plant.  Note:  If you grab a plant with hot tools you take a chance of burning the plant.
  9. Once cooled. open the plantlet jar with one hand and with the tweezers in the other hand remove one plant, close lid, place plant  on plate.
  10. Using the knife cut 1/16th to 1/8th of the stem and any tips or leaves that could have been burned in the cleaning progress.
  11. With one hand grab a media filled vessel and open using one hand, with the tweezers in the other hand pick up the plant and place in the medium gel, stem down. (Depending on jar size place 1 to 3 plants per jar.)  Close lid completely.   Place in a storage area/clean room 
  12. The storage area/clean room needs to be a sealed area and always kept clean.  The only thing that should be in this area is your vessels to reduce the contaminations hazards.  Use florescent lighting, no more than 2500 lux per shelf for 16 hours a day.
  13. Maintain temperature between 75-85 degrees.  Average 78 degrees depending on health of plant.
  14. Maintain humidity between 45-65%

BillBerry Farms cannabis tissue culture roots

To move from stage I to stages II or III follow the directions above but omitting the washing instructions.

When your plantlet is ready to move to out of stage 3 remove from jar carefully by breaking up the gel (making sure not to break any roots.  Under soft running tap water rinse off any gel that is remaining on the plantlet’s roots. 

Place into the growing media of your choice.

The principles of tissue culturing are all around us in nature and the way we can learn is by our own research and that of other growers and their experiences. We should learn about the normal requirements of soil composition and nutrients, light, and temperature for a specific plant species. We should study its form, growth, habits, and how it reproduces. These are only some of the clues that we can utilize in learning how to tissue culture. With this information we can turn to specific formulas and principles unique to tissue culturing to help us produce the best quality medicine.

 

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