Plant tissue culture: The benefits & why producers are using it to supplement traditional propagation [Webinar]

As the cannabis market matures, cannabis producers are realizing that they can't rely on the traditional mother clone system forever. This method has a lot of pitfalls and brings with it a number of risks.

Once out of reach for many cannabis operators, tissue culture research in the cannabis industry has now advanced to a level where tissue culture services are available at an affordable price. Building out small-scale tissue culture labs within an existing cannabis facility is also possible for with the proper guidance.

In March, we ran a webinar all about tissue culturing in cannabis. In the webinar, we explored the issues with conventional propagation and cultivation, ways that tissue culture is helping growers reach their yield and quality goals and how to get started.

Re-watch the full webinar here, or read our write-up below.

What is Cannabis Tissue Culture?

Also referred to as TC, tissue culture is growing plants in an in-vitro setting. You take an explant (a piece of plant from a mother plant) and grow it in an artificial medium then produce a clone in an artificial lab environment. With the growing medium, you can control the nutrients and growth hormones and allow for directionality in how the plant grows.

Nodes are taken from the mother plant and go through a chemical process to eliminate contamination. Then they're tested to make sure they’re clean. Then they're placed in an agar media supplemented with macro and micronutrients, vitamins and hormones - everything required for plant growth.

The containers are sterile, and fine particulate HEPA filters the air in the growing chamber.

After three to four weeks, you take the grown plants, cut them back into nodes, and repeat the process. On average, you can 5x the number of clones of a single cultivar over a month. The next step is to make the unrooted cutting into clones ready to be shipped to producers.

In cannabis, tissue culture allows for a lot of different possibilities. The environment is very sterile, which enables growers to clean their cannabis plants, keep a clean culture of genetics, complete pheno hunting and much more.

Why cannabis cultivators are turning to tissue culture?

Lack of space for genetic storage and labour involved in taking care of mothers

Many cannabis producers store their genetics in grow rooms as mother plants. Mother plants can take up quite a bit of space, especially if your cannabis company is sitting on hundreds of different genetics. Producers might keep three to four mothers of each cultivar to retain the genetics. These plants need to be taken care of and replanted every three months.

Most companies will have mothers for genetics they aren't actively producing, and they don't know when that specific cultivar might become more popular and when they might take it into production. Until then, these plants are hanging out in the mother room.

Not to mention, storing your mothers in your facility is a huge gamble. You can lose years of work and entire cultivars due to a virus or pathogen outbreak through your facility. It happens to even the best growers.

Viral load and issues with plant health

Viruses and pathogens can be passed along throughout propagation, impacting the yield and quality of your crop. Once your cultivar is impacted, it's hard to do anything about it, and your best cultivar can stop performing as it once did.

Not only can TC allow you to store a clean culture, but it can also help you clean up your genetics to get them back to peak performance.

Skill, labour and time required for pheno hunting and unpredictability of results

Pheno hunting is incredibly time-consuming and labour-intensive. Typically producers sprout hundreds of seeds and need to grow them to flower. Then they would have to complete analytical tests to see the cannabinoid and terpene profiles of the cultivar before choosing which ones to keep.

On top of that, it can be very risky because seeds could come from a contaminated source, and you might know even know until they get a virus test.

Why should you care about tissue culture as a cannabis producer?

There are so many applications of tissue culture in the cannabis industry. The most common and valuable, free up space and reduce operational risk.

Micropropagation

In traditional propagation, mother plants get worn down over time, the growth of the clones changes, and growers lose consistency from batch to batch. The deterioration of quality can come from stress and many other factors.

Tissue culture clones are disease free, have more vigour and will be most like the original phenotype. Tissue culture is the best way to give cannabis producers a chance for batch consistency. Not to mention, TC clones have more tolerance against pathogens and produce better yields.

Cleaning your genetics

Sending your genetics to get cleaned is another application of TC. A plant cutting is cleaned through a chemical process and tested to ensure it's free from pathogens. Then it would go through the multiplication cycle to create new clones for your facility.

This is a great way to improve the performance of a cultivar that has been impacted by pathogens or viruses at your facility, and it wouldn't be possible without TC.

Pheno hunting

Pheno hunting with TC is much quicker and more efficient. Seeds are started in the tissue culture, ensuring the environment is sterile and the plants are clean. They can screen the seeds for viruses and pathogens before they use them.

A TC service provider can use genomic selection on the pheno hunts. Instead of flowering each seed or cutting, producers can select specific traits using genomic sequencing. This method can save a lot of time and money because you don't have to flower each cultivar to see if it has the traits you want.

From there, you can select five to 10 unique seedlings and then flower them, and you're sure those 10 are the ones you're looking for. You can pick the best three and move on.

This way, you can identify the novel cultivars and even do custom breeding, crossing different cultivars together and using genomic selection to determine the winners.

Because this type of pheno hunting method is so efficient, producers can identify new strains based on the market demand and quickly bring them into production.

With long-term genetic storage, you can preserve your pheno hunt and ensure you have a reliable way to store them safely.

Genetic storage

Space is one of the most critical aspects for a lot of cultivators. Most can't physically store hundreds or thousands of genetics in a mother room, but having a large selection can give producers a competitive advantage.

There are also only so many cultivars one operator can have in production at a single given point, which means most mother plants aren't being used and are taking up space and resources. There is no way to know when that particular phenotype will be desirable in the market, and you'll need that genetic again.

As mentioned earlier, if producers store all their genetics in one facility, there is a risk of disease and losing the entire cultivar. It's extremely risky.

TC service providers such as Klonetics offer cold storage that can keep genetics for multiple months or indefinitely in liquid nitrogen. Producers can reduce their mother stock in the facility while maintaining hundreds of genetics in a clean area, available for propagation whenever needed.

Implementing tissue culture in your cannabis operation

Deciding whether to build out your own tissue culture lab or to find a service provider to help you, depends on a few factors.

Consider building a lab if you're in a market where tissue culture services are unavailable. These services already exist in Canada and are a great way to get started.

To assess if you should start a lab or go with a service provider, you want to look at your team, your budget, and internal cannabis knowledge and decide what makes the most sense.

How to start a tissue culture lab

It's possible to build out a tissue culture lab on a small scale in a shipping container, for example. There is an initial capital investment for equipment. You'll need equipment such as:

• Laminar flow hood
• Autoclave
• PH EC meter
• Analytical balance

Apart from the capital investment, you need someone knowledgeable about tissue culture to run your lab. Finding someone can be challenging as there are not too many people with cannabis tissue culture experience at this time.

Partnering with consultants, like Atiyyah Ferouz at AgCann Consultancy, can help you set up your lab, provide you with media recipes, and support you for as long as it takes to initiate your cultivars.

For cannabis, some media recipes will work for most cultivars, but there will always be around 20% of cultivars that need tweaking to their recipes. Tissue culture media has thousands of variables, lots of different macros and micros, vitamins, amino acids, charcoal etc. Troubleshooting media is the most challenging part, and you need someone with experience. The tissue culture specialist can identify how to tweak tissue culture media based on the plant's appearance.

For most facilities, a TC lab will be part of production using nodal or meristem culture rather than an R&D lab. Even if you have a tissue culture lab, look at using a provider for long-term genetic banking, cryogenics, protoplast culture, or another tissue culture method with which you might have little personal experience.

If you want to learn more about building a tissue culture lab, we’d really recommend getting in touch with Atiyyah Ferouz at AgCann Consultancy.

Working with a tissue culture service provider

Working with a service provider is the most cost-effective and safe option to delve into tissue culture.

Service providers such as Kloentics offer various services depending on the producer's needs.

Ordering cannabis tissue culture clones

While operators can purchase well-known genetics straight from Klonetics, many producers opt to bring in their existing cultivars to Klonetics. Kloentics guarantees that those genetics are exclusively the producers and will only be used for the tissue culture service.

Whenever producers need clones, they can order them from Klonetics, and they are kept safe and sound in their biobanking storage.

Clean TC clones can be ready within a couple of months. However, if plants are virus-infected, it takes more time to clean up the genetics, around 4-6 months to receive the clones.

Tissue culture is not quite as fast as taking cuttings and having clones in two weeks. It takes some time to clean genetics and build up a stock if you want 2000 clones, for example. It takes a bit of extra planning.

Customers would need to pre-order ahead of time - further in advance than traditional cloning to allow the TC nodes to grow into clones. You need to be able to plan two to three crop cycles ahead and have solid production models.

A solid production model is vital to be organized enough to know when to order TC clones and prevent downtime in the facility.

Cannabis tissue culture clone hardening-off period

The hardening-off period is one of the more challenging aspects of the TC process and is one of the most critical points of the TC timeline.

The TC clones are grown in an artificial environment, and when introduced into a greenhouse (or outdoors), they have to adjust to new environmental conditions. If this step is done incorrectly, you risk losing all your clones.

Producers need to have a hardening-off protocol in place to help the TC clones adjust. Service providers will go on-site if the customer is local and explain the hardening-off protocols required.

For customers further away, a good provider should give you instructions on how to help the plants acclimatize to their new conditions. If hardening-off procedures are not followed, then there is a risk of killing off their clones.

Conclusion

The research around cannabis tissue culture is there, and service providers have optimized these methods to offer TC clones at a similar price to conventional clones.

Tissue culture is the final frontier for Hop Latent Viroid (HLVd) and many issues in cannabis cultivation. TC will grow in popularity worldwide because it's one of the only methods to deal with many problems that are coming up in growing cannabis on a commercial scale.

The Panelists

Atiyyah Ferouz, CEO, AgCann Consultancy

Atiyyah is a plant scientist with an extensive background in regulated industries. After completing her MSc in plant molecular biology, she began working at a GMP-certified food facility, gaining experience in QA, regulatory affairs, and product development.

Atiyyah then transitioned into the cannabis industry and has since held various roles that have honed her skills in cultivation, tissue culture, post-harvest processing, sales, and business development. She founded AgCann Consultancy in 2020.

Dr. Surender Khatodia, Chief Tissue Culture Scientist, Klonetics

Surender leads the scientific & technical operations of the commercial-scale cannabis micropropagation at Klonetics for cannabis tissue culture gen zero clones production.

He brings over 15 years of extensive experience in plant tissue culture and genomics work on various crops. During the last four years, he has made progressive developments in optimizing the methods of pathogen eradication for clean clone production, genetic testing and germ-plasm conservation of the popular cannabis cultivars.

Tom Smale, Founder & CEO, Growbox Consulting

Tom is the founder and CEO of Growbox Consulting. He has worked with multiple brands and facilities across Canada to bring high-quality craft cannabis to market while providing facility operations and compliance support.

Growbox Consulting was built from the grow room out and Tom and his team have been able to offer guidance and leadership from the growers perspective while understanding the many regulations and challenges that the industry faces on a day-to-day basis.

Austin Bell, Implementation Specialist, Elevated Signals

Austin works on the Customer Success team at Elevated Signals, helping cannabis operators implement software to optimize their operations and improve their products.

He has been working with the cannabis plant for the last 15 years in both the legacy and licensed markets. His experience is based around leading teams and operations in the production space, with an emphasis on extraction and cultivation. Austin brings a passion for the plant and its potential for healing to all of his cannabis work.