commercialized in Japan—a tomato developed by Sanatech- Seed, a company originating from the University of Tsukuba. Interested consumers can buy the tomatoes online. And a number of gene-edited grape varieties have already been created using CRISPR technology.
Legal, ethical, environmental, and health issues Gene change for breeding and improvement has become technically feasible, but it raises a wide range of legal, ethical, environmental, and health issues. To reduce the ecological impact of genetically modified organisms, the Cartagena Act came into force internationally as early as 2003, and some 200 countries have now signed up to it. But there are also bioethical questions regarding genetic engineering, as well as concerns about the safety of gene-altered foods. Resistance is so great that there is a worldwide boycott of these products, strongly supported by environmental group Greenpeace, for example.1 On the other hand, improvements in genetic engineering, such as the advent of CRISPR, have brought a range of genetic technologies. Some may well achieve social acceptance, which would in turn have a major impact on agriculture. As mentioned above, radiation-induced mutation uses the natural mechanism of gene repair—a technique already accepted by many people. One of the CRISPR editing techniques can be said to be an update of the radiation technique. Based on genome information, CRISPR can precisely target a gene to cut it out, and it is then repaired naturally. By repeating this process, an error occurs and a change in the gene sequence is induced. This technique is called targeted mutagenesis. It is the ultimate shortening of natural mutation that may well be accepted by society soon. Sanatech-Seed used targeted mutagenesis for its edited tomatoes, which have already been approved by Japanese ministries for commercialization.2
There appears to be no real opposition
from consumers so far, though the Japan Organic Agriculture Research Association and others have expressed opposition.3
Knock-down and knock-out Gene-editing technologies vary widely. In mutagenesis, there are technologies called knock-down and knock-out. In both, a gene is cut out using a gene-editing tool such as CRISPR, and a mutation is introduced as part of the natural repair process. When the function of the gene is lost, it is knock-down, and when the gene is connected at both ends with the gene cut out, it is knock-out. For grapevines, Vivai Cooperativi Rauscedo (VCR) in Friuli,
Italy, collaborates with the Institute of Applied Genomics (IGA) of Udine, which uses knock-down to create edited grape varieties resistant to powdery mildew. Dr Elisa De Luca is the VCR director involved in this project, which she kindly explained to me. Powdery mildew is one of the most damaging vine diseases. When the pathogen adheres to a vine, the plant reacts and produces certain chemicals. The pathogen takes advantage of the vine’s response by detecting the chemicals and recognizing the vine to attack further. By knocking-down the gene responsible for the chemical production, the vine can avoid the pathogen attack. Vines resistant to powdery mildew can avoid damaging losses in quality and yield, and therefore loss of income for the grower. It also reduces the chemical spraying required to control the disease, which lowers production costs, as well as reducing environmental impact and health risks to neighboring residents. In 2023, Bordeaux experienced unprecedented powdery mildew
damage due to the wet growing season. According to the Chamber of Agriculture of the Gironde, 92% of monitored vineyards were infected, and more than 55% of grapes lost their commercial value due to powdery mildew. Similar damage was observed throughout the region.4
At Domaine Ducourt in the
Entre-Deux-Mers, to take only one example, a total of 18 chemical sprays were applied to organically grown vineyards in 2023, while in dry and warm years, only ten chemical sprays are needed. As for the food safety of gene-edited varieties, it should not
provoke a problem, at least in theory. The edited vines in Dr De Luca’s collaboration project are only non-functional in the relevant genes, so their other traits are no different from those of the original vines. That said, it is still necessary to grow them for at least ten years to monitor their food safety, quality, and response to diseases. And there is already a concern. This knock- down variety is only hiding from powdery mildew. Whether or not it will be found, sooner or later, remains to be seen.
Knock-in
The disadvantage of mutagenesis (knock-down and knock-out) is that it is inefficient (even if it is still dramatically quicker compared to conventional breeding methods). If one inserts the desired gene directly, it is possible to create the desired variety far more rapidly. This method is called gene knock-in. Powdery and downy mildews are fungal diseases originating from the Americas. Indigenous American vine species have the resistance genes. On the other hand, Vitis vinifera, the main species used in wine production today, does not have any of these resistance genes. Knocking the resistance genes of American vines into Vitis vinifera would provide comparable disease resistance. Four genes (Rpv3.1, Rpv3.2, Rpv10, and Rpv12) have already been identified for resistance to downy mildew, and three others (Run1, Ren3, and Ren9) for resistance to powdery mildew. These genes cause a hypersensitive response in grapevines when they detect the pathogen and resist it. The more resistance genes there are, the stronger the resistance.
The knock-in technique is similar to the knock-out in
practice: A gene-editing tool such as CRISPR is used to cut out the relevant gene in vinifera, and the resistance gene is added along with it. Though not related to vines, a video of gene editing is available on YouTube for readers who want to see how CRISPR editing works.5
The advantage of knock-in is that vines can directly combat the diseases through resistance genes. It is very effective and seems more reliable than indirect methods, such as that in Dr De Luca’s project. A knock-in grape variety has already been created at the CREA-VE Research Institute in Veneto, Italy. The institute is part of the Italian Ministry of Agriculture, and the relevant team is led by Dr Luca Nerva. CREA-VE is working on knocked-in varieties with the downy mildew resistance gene, Rpv3. Their disease resistance is very high. Hybrid PiWi varieties have disease-resistance genes. Let’s look at the impact of the knock-in variety. Domaine Ducourt, mentioned above, grows Sauvignac, a high-quality PiWi variety that produces a crisp white wine similar to Sauvignon Blanc. Sauvignac has two genes for resistance to powdery mildew (Ren3+Ren9) and two for resistance to downy mildew (Rpv3+Rpv12). In 2023, a total of 18 chemical applications were required for the vinifera varieties grown organically, while only
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