The Protocol To Tissue Culture Peperomia spp.
25 Oct 2022

The Protocol To Tissue Culture Peperomia spp.

Anjali Singh, MS

As a content and community manager, I leverage my expertise in plant biotechnology, passion for tissue culture, and writing skills to create compelling articles, simplifying intricate scientific concepts, and address your inquiries. As a dedicated science communicator, I strive to spark curiosity and foster a love for science in my audience.

Anjali Singh, MS
Table of Contents


Peperomia is one of the largest genera of basal angiosperms. It consists of 1500-1700 species and belongs to the family of Piperaceae and is native to southern Florida, Mexico, and the Caribbean in the United States. However, they are also found in Africa, Oceania, and Southern Asia.

Peperomia species have different adaptations. They are lithophytes (grow on rock or rock crevices), epiphytes (grow on other plants), and many others are xerophytes (drought-tolerant) and geophytes (having underground tubers).

The leaves of succulent Peperomias are thick, fleshy, and waxy with shapes ranging from oval, lance-shaped, or heart-shaped. Further, some varieties have green or striped leaves, marbled or bordered with pale green, red, or gray, and they have red petioles.

The plant is well known for its beautiful small foliage plants best suited to greenhouses and terrariums. Many of the plants are succulents and are widely used as indoor plants for decoration purposes. Some examples of Peperomia species include P. argyreia, P. caperata, P. nitida, P. umbilicata, and P. obtusifolia.

In this article, we talk about the propagation technique of Peperomia and how tissue culture provides an edge over conventional approaches for commercial-scale propagation of the plant.

Tissue Culture of Peperomia spp.

Because of their variant appearances, Peperomia species are quite popular among gardeners. The plants are also easy to care for as they are low maintenance, slow-growing, and can be planted all year long.

Vegetatively Peperomia can be grown either using seeds or stem cuttings. However, there are many shortcomings of the techniques if grown for commercial purposes, such as pathogen attack, slow growth, and the need for many mother plants. Often propagating plants using vegetative techniques can lead to their extinction.

The solution to these challenges is offered by the advanced tissue culture technique. It only requires a few tissues to regenerate the whole plant. Further, it provides a way to protect your plants against pathogens as they are grown in complete aseptic conditions. Also, in a very small space, hundreds and thousands of plants can be regenerated using the technique.

Procedure to Tissue Culture Peperomia

The given procedure is taken from the study of El-Naggar, Hany & Osman, and Amira. (2014). Micro Propagation and Organogenesis of Peperomia obtusifolia. Asian Journal of Crop Science. 6. 58-66. 10.3923/ajcs.2014.58.66.

  • Collect single node stem segments from a healthy mother plant of Peperomia obtusifolia.
  • Wash the explant with home detergent and rinse them using distilled water three times.
  • Dip shoot cuttings in 70% ethyl alcohol for 30 seconds.
  • Wash the explant with distilled water.
  • Dip the explant in 20% Clorox bleach for 10 minutes.
  • Wash the explant three times using distilled water.
  • Prepare a shooting media using MS salt and vitamins supplemented with 30 grams/liter sucrose and 8 grams/liter agar.
  • Adjust the pH of the medium to 5.8.
  • Autoclave the medium at 121 degrees Celsius and 15 lbs for 20 minutes.
  • Pour media into culture tubes.
  • Culture single stem node segment in the media containing tubes.


  • After a few weeks, some growth will be obtained. Now, the plant is ready to transfer to shoot proliferation media.
  • Prepare shoot proliferation media using basal media as prepared above supplemented with kinetin at 5 mg/L combined with NAA at 1 mg/L.
  • Incubate cultures under cool white fluorescent lights at an intensity of 66-52 micromol per meter square per second for 16 hours and 8 hours in the dark.
  • After 3 months plants will grow up to 2-3 cm with a couple of leaves. Then, transfer the plant to a rooting medium.
  • Prepare a rooting medium using MS medium, vitamins, 30 grams/liter sucrose, 5 grams/liter agar, and 0.5 grams/liter NAA.
  • Adjust the pH of the medium to 5.8.
  • Autoclave the medium at 121 degrees Celsius and 15 lbs for 20 minutes.
  • Pour media into culture tubes.
  • Transfer the regenerated shoots to the rooting media.
  • Incubate cultures under cool white fluorescent lights at an intensity of 66-52 micromol per meter square per second for 16 hours and 8 hours in the dark.
  • After in vitro plants have proper roots, take them off using tissue paper and wash their roots gently to remove any stick medium.
  • Transplant the plants in a 6 cm pot containing a mixture of peat and perlite in a 2:1 ratio.


Plant Cell Technology is helping tissue culturists worldwide by providing unique and world-class products and services that smoothen their process. The PCT Store has MS media, agar, gellan gum, Plant Preservative Mixture (PPM), culture vessels, Biocoupler(TM), and masks in its store to facilitate your processes.

And, that’s not it! Plant Cell Technology also offers consultation services to culturists of all sizes that help to get instant solutions to your tissue culture problems.

You can either book a one-on-one consultation call or a physical visit to your lab. We help you at every step of the tissue culture process, ranging from establishing a tissue culture lab to preventing contamination problems or any specific challenges in your process.

So, visit today and learn more about our products and services and how they help you excel in your tissue culture processes.

Happy Culturing!

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