Can This Genetic Breakthrough End Powdery Mildew

The change could potentially eliminate a major concern for growers, while ushering in a new era of cannabis farming that looks more like traditional agriculture.

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This story originally appeared on Marijuana Venture

An Israeli company hopes to have stable seeds for powdery mildew-resistant cannabis plants available to growers by the end of this year, potentially eliminating a major concern for growers and ushering in a new era of cannabis farming that looks more like traditional agriculture.

Using the 2020 Nobel Prize-winning CRISPR-Cas9 gene-editing technology, CanBreed scientists have successfully produced seeds that when grown, should be resistant to the powdery mildew fungus, a project more than three years in the making.

The technology has been used in other agricultural crops, but this is the first time the science has been applied to cannabis.

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“There is a well-known gene that is in most of agricultural crops; we identified that this gene also exists in cannabis as well and it has been proven that editing that gene in tomato or cucumber or whatever – a plethora of other crops – makes those plants resistant by default,” explains Ido Margalit, CanBreed’s CEO. “Cannabis has some miraculous properties, but it is just a plant.”

CanBreed was founded in 2017 after Margalit and his business partner, Dr. Tal Sherman, were let go from Syngenta when that company closed operations in Israel. With years of experience already using the CRISPR technology, the pair formed CanBreed with an eye on applying the technology to conventional crops. But difficulties raising money for tomatoes and soy turned their attention to the burgeoning cannabis industry.

“That was the first time we started to look at the cannabis business,” he says. “And us being agronomists, being professionals and coming from the business world, we very quickly identified two major gaps that exist in the cannabis business.”

First, they noticed that because cannabis is often grown from clones, there is no uniformity of the final product like in other industries, resulting in a lack of standardization. Second, the genetics of modern cannabis plants were adapted for growing in basements and other illicit locations, often with artificial lighting and were therefore not suited for mass production in greenhouses. That resulted in growers having to apply technology that increased growing costs.

“We immediately knew the solution to that problem was you had to produce cannabis from stable seeds, just like the rest of the agricultural sector, and you have to enhance it with agronomical traits that will allow the plant to grow very efficiently and at a low cost,” he says. “Now, if we have stable seeds enhanced with agronomical traits, growing cannabis should not cost you more than growing tomatoes.”

And Margalit says this is just the beginning. CanBreed is already at work developing autoflowering forms of cannabis and recently purchased a hemp farm in San Diego to begin work creating similar breakthroughs for hemp.

Marijuana Venture spoke with Margalit from Israel via Zoom about the company’s breakthrough, the technology they used and when the company’s seeds will be available to growers.

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Marijuana Venture (MV): Let’s start with the seeds. The idea here is that stable seeds like this are used across the board in other agricultural industries, right?

Ido Margalit (IM): Growing an agricultural crop is, by default, done from seeds. The only agricultural crops that are not grown from seeds are those crops which physiologically can’t be grown from seeds, like bananas or potatoes, (for which) you have to do vegetative reproduction. If a plant knows how to reproduce itself through seeds, then by default you grow the agricultural output from seeds.

Now, about four years ago, the professionals said, “You cannot do stable seeds from cannabis, therefore the only way you can get true-to-type offspring of cannabis is from cloning vegetative or tissue culture.” We said that’s not the case, there are similar plants in the industry from a genetic composition of cannabis, and they do sexual reproduction for agricultural output from stable seeds. So everybody said that cannot be done and then four years after, voila, we finish the development.

MV: The breakthrough itself deals with powdery mildew. What did you guys do?

IM: One of the problems that the cannabis breeding industry faces is the fact that there is not enough genetic diversity that you can do breeding, conventionally, of agronomical traits. Let’s say I’m a tomato breeding company and I find a tomato gene bank in Peru that has some sort of mutant that has a resistance to a virus or disease that I have found in Israel. I would order that mutant, have it sent to Israel and integrate it in my breeding program, and then in three to six years of trial and error, I manage to solve that. Now, in cannabis, since it was an illegal crop, there is zero science invested in it, meaning there are no gene banks in the world, there are no mutants in the world and even if there were some special and unique mutants, they could not travel across borders because of the fact that it’s illegal. So what that means is if you’re doing conventional breeding using cannabis, even in regions and territories where now investing in science and breeding is legal, you do not have this genetic diversity.

It’s going to take decades for the science community to catch up to where the genetic diversity is in regards to other commercial crops. So whoever says they’re going to do breeding conventionally of unique traits, that’s not going to happen. The only solution we have at hand is CRISPR.

CRISPR is a protein found about nine years ago in bacteria. It knows how to run on any DNA, and you can tell it like a GPS-guided missile exactly where in the DNA to cut the DNA strand – meaning that if you have a gene you do not want to be expressed anymore, you send that to CRISPR and it cuts it and that gene is not expressed anymore. You do not insert any foreign material, you just terminate the expression of endogenic genes.

For powdery mildew, there’s a gene in the plant that expresses a protein. Powdery mildew is a fungi so it walks around and it sees a plant with that protein – “ah, there’s that protein I like” – and it attacks the plant. If you edit that gene that expresses that protein and the protein is not expressed anymore, the powdery mildew does not see the protein so it does not attack the plant. That means the plant now is resistant to powdery mildew. That is exactly what we did. If you edit that gene and that protein will not be expressed, then cannabis will be become powdery mildew-resistant.

MV: Is this a gene that’s found in other plants that your team found in cannabis and then turned it off?

IM: Exactly. We looked at the genes that are responsible for that and we checked if those genes exist in cannabis and they do. And we patented it, meaning that anyone else who wants to silence a trait like powdery mildew through gene editing will have to come through us.

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MV: So you didn’t just patent silencing powdery mildew, you patented silencing any gene in cannabis?

IM: Not any gene. Genes that express agronomically important traits.

MV: It seems like the hardest part is finding these genes. Is that where the majority of the research is?

IM: That was actually the first step. Applying CRISPR in cannabis is extremely difficult. Whereas CRISPR works very easily in certain crops like tomatoes, it does not like to work on cannabis. We recruited a leading researcher in plant science from Israel’s leading plant research institute. She was doing gene editing of tomatoes and we recruited her to our company. We said, “We want you to do gene editing on cannabis.” And she said, “No problem, I’m going to take all my protocols and I am going to apply them to cannabis and in three to four months – that’s all it takes – we’re going to have an edited cannabis plant.” Well, three to four months pass, then a year passes, then two years pass and nada. And why is that? Because regeneration. I am not going to bore you with the technicalities, but one of the very key issues that is important in doing cannabis is regeneration. You do not put the CRISPR protein into a whole plant. You have to start with a group of cells. And those cells have to regenerate to a whole plant.

Cannabis does not like to be regenerated from cells. So we had to find a completely different method. We had to develop from scratch the basic research capabilities of applying CRISPR in cannabis, and it has taken us over two years and a lot of money.

MV: So you have created these stable seeds for powdery mildew-resistant plants. What’s the next step?

IM: It’s a work in process. We have stable seeds, and we also saw some initial promising results by editing on powdery mildew. We are now testing those edited plants for powdery mildew to see that they actually are resistant to powdery mildew. Nature is funny; sometimes it works by the book, sometimes it doesn’t. I cannot bring to the market something that I have not tested and say for a fact that it is powdery mildew-resistant. It took us over a year-and-a-half to develop protocols and capabilities for doing biological tests on powdery mildew. At this first stage we have 30 varieties. I assume that five to six will pass the test positively and move on to the commercial phase. And every four months we have additional varieties entering our testing regimes.

MV: When do you expect to get the first batch to market?

IM: Right now, we are sowing the seeds for the testing. That testing should take about four months. That means around the end of April or May we will be able to select from those 30 varieties that pass the test and we can start to do the commercial stage.

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