CA3234149A1 - Use of an extract of part of a plant for stimulating plant defenses against viruses, and associated composition and methods - Google Patents

Abstract

The elicitor composition which stimulates plant or tree defences, reducing the effects of an attack by a virus, comprises an extract of at least one part of at least one of the following plants: rocket salads, including of the genera Eruca sativa, Diplotaxis, Erucastrum and Bunias, Cakile, plants of the genera Allium, mustard (Sinapis alba, Brassica nigra, Sinapis arvensis, Brassica juncea), wasabi (Eutrema japonicum), horseradish (Armoracia rusticana), watercress (Nasturtium officinale), plants of the species Brassica rapa, Brassica ruvo, Brassica napus, Raphanus sativus, Barbarea verna, Erysimum allionii, Erysimum cheiri, Tropaeolum majus L, Alliaria petiolata, Salvadora persica, Carica papaya and Brassica oleracea.

Description

USE OF A PLANT PART EXTRACT TO STIMULATE THE DEFENSES OF
PLANTS AGAINST VIRUSES, COMPOSITION AND ASSOCIATED METHODS
Technical field of the invention The present invention relates to a use of a plant extract special for stimulate the defenses of plants or trees against viruses, and composition and processes associates. The present invention thus aims to reduce the effects of an attack of viruses.
State of the art In their permanent struggle for survival, plants, like animals, have developed a diversified range of protection and defense systems, which allow them to resist infectious diseases and parasites (fungi, bacteria, viruses, etc.). Sometimes, protective systems pre-exist before any contact with the pathogen with the host but the more often, resistance (induced resistance) sets up after the meeting the plant with his attacker.
For a certain number of plants, there are defensive means which pre-exist before attack by a potential pathogen or pest. For example, the plants have morphological barriers. For example, it could be cuticles thickly covered with hydrophobic substances (wax, cutin, suberin, etc.) preventing agents pathogens to enter or to develop. We can observe the presence of thorns on the surfaces leaves and stems having a repellent power against animals (in particular herbivorous mammals), or still the closure of the stomata in the leaves and the lenticels of stems to avoid penetration of fungal spores, bacteria, etc.
Plants also produce biologically active secondary metabolites and inert precursors to protect against harmful organisms. These secondary metabolites are not necessary for an essential biological activity, such as growth and reproduction, but help the plant to adapt to its environment, which transform into active molecules during attack by a pest, pathogen or after injury.
In response to attack by a pathogen, the plant also deploys modifications metabolic induced in the plant which very often causes either a defense late and discreet or an active and rapidly induced resistance.
Late defense is often linked to receptors present in the membrane or the plant cell, capable of recognizing associated molecular signals to microbes or Pathogens. For example, cell wall fragments, chitin or patterns peptides from the flagella of bacteria (exogenous elicitor), proteins (endogenous elicitor) are part patterns, which, once perceived by the plant, can initiate a response basic immune (PAMP-Triggered Immunity or PTI), in order to limit invasion and delay the progress of the agent pathogenic.
On the other hand, the specific recognition of the attack of a pathogen inducing a rapid host response can also restrict or stop invasion and allows the defense to trigger very quickly. Recognition of foreign components in plants WO 2023/062025

2 PCs is coordinated by specialized receptors, which trigger the responses of host defense after detection of the pathogen. In this specific case of perception, the elicitor is coded in the pathogen by genes called avirulence genes (AVR). THE
receptors the plant then recognizing the product of the avirulence gene, are coded by genes of resistance (R) of the plant. The interaction between the gene product avirulence of the parasite and the gene Plant resistance is commonly referred to as a gene-for-gene relationship.
.
R genes have been isolated from various plant species (e.g. tomato, linen, rice, tobacco, Arabidospsis, sugar beet). Despite the important diversity of parasites to which they confer resistance (fungi, bacteria, viruses or even nematodes), their comparison reveals strong sequence homology as well as conservation of patterns structural:
The majority of proteins deduced from R genes isolated from plants share different structural elements characterized within protein domains involved in the defense mechanisms in yeast, Drosophila and even vertebrates.
By analogy, the Most R products would combine a receiving domain and a ensuring effector two major functions respectively: recognition of molecules elicitors by mechanisms of protein-protein interactions and direct activation or indirect signals of transduction.
Five main structural domains conserved within R genes have been distinguished:
1/ The majority of products deduced from R gene sequences cloned to date present in the C-terminal position an LRR domain (Leucine-Rich Repeats intra- or extra-cytoplasmic. The LRR domains correspond to the repetition of a motif of variable size including leucines. They would intervene in the mechanisms protein-protein interactions and proteinpolysaccharide 2/ The NBS (Nucleotide Binding Site) or NB domain, associated with the LRR domain (NBS-LRR), is widely distributed within cloned R genes. This domain, composed in particular different conserved kinase-type motifs, corresponds to a binding site and hydrolysis of nucleotide triphosphates ATP and GTP. It presents analogies with animal proteins potentially involved in apoptotic cell death phenomena, notably APAF-1 identified in humans and CED-4 identified in Caenorhabditis elegans. All these areas are grouped under the terminology NB-ARC (Van der Biezen and Jones, 1998).

3 and 4/ Two other areas, called TIR and CC (or even LZ), less frequent within the identified R genes, can be associated, in the Nterminal position, to NBS products-LRR. The TIR (Toll Interleukin Receptor) domain presents important homologies of sequences with the intracellular domains of isolated protein receptors Drosophila (Toll receptor) and humans (Interleukin-1 receptor). Based on these analogies, a role in the cell signaling cascade is assigned to the TIR domain. The domain CC (Coiled-Coil) or LZ (Leucine-Zipper) which can vary in size and position is known to play a role in the homodimerization or heterodimerization of proteins.

WO 2023/062025 3 PCs 5/ Finally, the Serine/Threonine kinase domain occurs alone (for example product of Pto gene) or, associated with an LRR domain (for example, product of the Xa21 gene). He would be involved in phosphorylation reactions during signaling cascades.
In the majority of cases, during an incompatible interaction, the very defense active, leading to a situation of resistance, a whole series of events is set in motion place at the end of the perception of the pathogen by the plant.
The entire metabolism of infected cells is called upon and made intervene numerous mechanisms whose function is to destroy and trap parasites : proteins G, ionic fluxes, notably Ca2-E, HE, KE and Cl-, activated forms of oxygen (or ROS
for Reactive Oxygen Species) and a whole cascade of phosphorylation/dephosphorylation proteins such as MAPKs (Mitogen-ActivatedProtein-Kinase).
Most often, this signal surge translates phenotypically into the death rapid pathogen-infected cells and lesion formation localized necrotic around the site of parasite penetration. This deterministic reaction genetics is called reaction hypersensitivity (or < HR) or apoptosis (programmed cell death).
The important metabolic modifications allowing the aggressor to be confined in sound penetration site are the result of the expression of numerous genes of defense at the level of infected area. Cell death (HR) also generates signals that activate the resistance locally acquired (LAR), in contact with lesions, and systemic resistance acquired (SAR), to distance from the site of infection.
During the establishment of the LAR, the defense responses are intense and All particularly localized at the level of the ring of cells surrounding the HR area. These responses include the synthesis of a broad spectrum of anti-microbial compounds such as PR proteins (Pathogenesis Related Protein), certain secondary metabolites of antibiotic properties, by example phytoalexins, as well as the accumulation of molecules involved in the ways of signaling, including ROS and various hormones including acid salicylic. These two levels local resistance (HR and LAR) are accompanied by established resistance to the scale of the plant, SAR, whose expression, later and less intense, can persist several weeks later the infection. The SAR keeps the plant in a state of wakefulness which allows it to resist not only to the original aggressor, but also to a wide range of other parasites able to intervene later.
The establishment of this systemic resistance, at the level of tissues not infected, depends on an elaborate intercellular communication network. Signals released by cells expressing HR diffuse and reach other cells which in turn trigger a response specific. Salicylic acid, ethylene, jasmonic acid or even systemin were recognized for several years as chemical messengers. These messengers alert them uninfected cells and direct their metabolism towards the establishment of defense responses, in particular, the strengthening of the cell wall, the stimulation of the pathways of metabolism secondary (enzymes of phenylpropanoid metabolism, of the biosynthesis of ethylene and jasmonic acid) and protein accumulation.

WO 2023/062025

4 PCs The particular case of viruses:
A phytovirus, or plant virus, is a virus that attacks organisms plants.
These viruses have the particularity of penetrating the plant cell of their host in order to divert to their take advantage of the cell's mechanisms and allow them to reproduce.
Among the viruses infecting plants, those whose genome consists of a or several single-stranded RNA molecules of messenger (positive) polarity are the most important both by their number (85 to 90% of known viruses) and by their impact economic. Although presenting varied morphologies, these viruses have a very simple, particles unwrapped being made up of a single or a few (2 or 3) types of subunits of capsid protein (CP).
For example, plants use RNA interference, or RNAi, to defend against many viruses. During their replication phase, RNA viruses go through a stage Double-stranded RNA (dsRNA) which is recognized by the plant as a warning signal and allows the setting en route to its defense system which could lead to the elimination of virus. For the majority RNA viruses, an effective antiviral RNAi response mainly relies on the activity of the DICER-LIKE4 (DCL4) protein to produce from dsRNA the major part of RNA
Antiviral “interferents” (siRNA). These are associated with a protein ARGONAUT
(AGO), in order to guide it towards the viral RNA, which leads to its degradation.
Plant viruses have four essential characteristics compared to other agents pathogens:
1/ Unlike other pathogens, viral populations have a very high evolutionary potential. This rapid evolution leads to an appearance of viral variants capable of circumventing and adapting very quickly to resistance plant genetics.
2/ Plant viruses are incurable by chemical means once the illness has installed in a field. No effective cure exists for viruses plants. Plants Infected susceptible individuals will retain the virus in their tissues until they die.
3/ It is the host cells which are responsible for multiplying the viruses which parasitize. THE
Plant viruses can infect all parts of a plant. Only the meristems (cells not differentiated) buds escape their invasion. They are the most often the cause of generalized diseases.
4/ Viruses can only survive in a living plant. They are destroyed as soon as the plant dies. They also require external agents to propagate from a plant to a other.
Within the virus/plant interaction itself, very specific mechanisms are put in place, different from the metabolic events occurring in the other interactions plants/pathogens (Bacteria, fungi, etc.).
Thus, phytoviruses are major pathogens found in crops of the world entire, and will represent an increasingly serious threat. From Infectious diseases emerging, viruses alone represent almost half of pathogens involved, either WO 2023/062025

5 PCs 47% versus 30% for fungi, and only 16% for bacteria (Anderson et al., 2004).
In this context, nowadays, the deployment of plant varieties carrying of genes resistance against phytoviruses remains the most effective approach in the control of these viral pathogens, in order to use the existing plant defense machinery already implemented.
One of the major criteria for breeders remains to obtain varieties having a durable resistance, that is to say which remains effective for a long period when the variety is cultivated intensively in an environment conducive to the development of disease.
Presentation of the invention The present invention aims to remedy all or part of these drawbacks.
To this end, according to a first aspect, the present invention aims at elicitor composition according to claim 1.
The action of this composition circumvents viral resistance. The bypass of a resistance translates temporally into the emergence and propagation of a virulent variant to the scale of the cell then of the plant to finish at the plot level whole. The virulence of a pathogenic, according to the definition of plant pathologists, designates the component qualitative of pathogenicity, that is to say the capacity or not to infect a genotype bearer of resistance given. Evolutionary forces will be strongly involved in the appearance virulent variants, which do not naturally pre-exist in an agroecosystem. Variations genetics go therefore be linked to variations/mutations of AVR genes: The first step correspond to the appearance at the cellular level of a virulent variant capable of infecting a plant having a resistance gene in the case of a gene-for-gene type interaction. That means that he accumulates the necessary mutations in its avirulence factor to become virulent.
It has been demonstrated that the composition which is the subject of the invention, preferably in its form extracted from an arugula plant, particularly Eruca Sativa and diplotaxis tenuifolia, acts very early after its spraying, demonstrating that this composition mimics the gene relationship for gene. For example, thirty minutes after its spraying, the composition preferential generates a series of metabolic events which show that the mechanisms defense triggered resemble those described during a gene-for-gene relationship:
1/ The production of reactive forms of oxygen (FAO or ROS), within 30 minutes following foliar spraying on Arabidopsis thaliana. The production transient species or reactive forms of oxygen (ROS for reactive oxygen species) constitute one of the answers the earliest following gene-for-gene recognition of an agent pathogenic by the plant.
These activated forms of oxygen are produced in the first minutes, even a few hours, after the elicitation. They are essentially made up of (02), (OH) and (H202) The significant accumulation of these molecules, commonly called burst oxidative, is often correlated with the control of cell proliferation and death cellular, development plants and the induction of defense responses. In the case of the response to agents pathogenic, these reactive forms of oxygen play a role of both antimicrobial compounds, in cell wall strengthening and signaling. THE
potential changes WO 2023/062025

6 PCs redox are integrated by the cell, which results in the activation of genetic programs leading to the establishment of the HR.
2/ The release of intercellular signals (phytohormones such as acid salicylic, jasmonic acid, ethylene) to cells adjacent to sites of infection conditioning to a immunization and an increase in the resistance of the latter (LAR for Local Acquired Resistance) in peach trees infected with bacteriosis, and treated with preferential composition.
3/ immunization of the whole plant (SAR for Systemic Acquired Resistance ) And potentiation phenomenon during a second attack by a parasite when the plant has been previously treated with this composition.
4/ production of I3-1,3 glucanase (PR2 Proteins) from ten hours later spraying preferential composition.
This composition which is the subject of the invention is therefore capable of acting independently of the structure of Resistance and Avirulence gene products, allowing overcome all mutation of the virus.
In conclusion, while the interaction between plants and viruses is very particular and different from other plant/parasite interactions (no chemical solution possible), it is demonstrated that the composition which is the subject of the invention acts effectively against viruses, by acting in very early times by simulating a gene for gene interaction, this which allows the plant to overcome all the weaknesses of a plant vis-à-vis a virus (too rapid appearance of variants, virus replication, etc.).
Given the rapid evolution of viruses, this composition positions the plants in a state of resistance, by acting downstream of the interaction between the R genes and AVR, in order to short-circuit possible virus mutations. Thus, the object composition of the invention, has the capacity to trigger all the defense mechanisms appearing after a phenomenon of gene-for-gene recognition, both for viruses and others pathogens, notably bacteria and fungi.
The effectiveness of the present invention reduces the effects of viruses from plants, plants, or trees, so as to allow the plant, in particular a tree, to continue properly to grow despite this attack, by overcoming the consequences of this attack, that is to say say by allowing the plant or tree to develop despite the virus, by reducing or eliminating the impact of virus. The plant extract can also, in certain uses, allow for the plant to eradicate certain viruses. This use can be curative or preventive.
We note that the reduction of the effects of viruses on plants or trees attacked by these virus involves, in certain cases, the total or partial reduction of symptoms. In fact, the plants and trees have, when their defense system is functional (notably thanks to the stimulation obtained by the implementation of the invention), the capacity to defeat a virus. Use of the composition which is the subject of the present invention aims to stimulate what the plants already know how to do, but that they do not do in cases of sensitivity, because they do not do not recognize their aggressor. The decline in pathology thus appears in certain examples of the description.

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7 PCs In preferred embodiments, the extract is obtained from of at least one of the following plants: Rockets, including Eruca sativa, Diplotaxis, Erucastrum and Bunias, Cakile, broccoli, plants of the Allium genera (garlic, onion, shallot, leek and chives), cabbage common, cauliflower, Brussels sprouts, kale, mustard, wasabi, watercress fountain, horseradish, white cabbage, field cabbage (Brassica rapa), including pak choi, cabbage Chinese and turnip, kohlrabi, collard greens, red cabbage, Chinese broccoli, broccoli rabe, rapeseed, radish, siberian wallflower, wallflower, Indian watercress, garlic herb, pilu tree, papaya, especially its fruit.
An elicitor composition which is the subject of the invention may comprise a crude extract total obtained by grinding and extracting the plant, into a fraction enriched in active compound(s) of such total extract, or in one or more active compound(s) in mixture. Such a composition allows advantageously, in an effective quantity in a composition, to combat symptoms of the attack of a plant or tree mentioned above by a virus mentioned above.
In preferred embodiments, the extract is obtained from of at least one of the following plants: Rockets, including Eruca sativa, Diplotaxis, Erucastrum and Bunias, Cakile, broccoli, common cabbage, cauliflower, Brussels sprouts, kale, mustard, watercress fountain, white cabbage, field cabbage (Brassica rapa), including pak cabbage choi, Chinese cabbage and turnip, kohlrabi, collard greens, red cabbage, Chinese broccoli, broccoli rabe, radish, and siberian wallflower.
In preferred embodiments, the extract is obtained from of at least one of the following plants:
- Rockets, including Eruca sativa, Diplotaxis, Erucastrum and Bunias, - Cakile.
In preferred embodiments, the extract is obtained from of at least one plants of the species Brassica oleracea.
In preferred embodiments, the extract is obtained from of at least one plant not containing Methyl-isothiocyanate and/or Propenyl isothiocyanate.
The inventor noted that these isothiocyanates could have effects harmful to plant growth while reducing the effects of viruses.
In embodiments, the extract is obtained from at least one plant containing at least one butyl isothiocyanate. The inventor noted that these isothiocyanates have favorable effects on plant growth while reducing effects of viruses.
In embodiments, the extract is obtained from at least one plant containing 1,3-thiazepane-2-thione. The inventor noted that this compound which, due to its structure, is not an isothiocyanate has favorable effects on the plant growth all by reducing the effects of viruses.
The inventor discovered that these extracts possess at least one substance active which reduces the effects of viruses, the presence of their diffusion vector and/or their colonization by insects.
In embodiments, the extract of at least part of plants is an extract obtained from a crushed material of said plants, and:

WO 2023/062025

8 PCs - said extract of at least one part of plants comprises at least leaves of the said plants, preferably mainly leaves and flowers, and - the process for obtaining said liquid extract comprises the steps following:
a) a step of grinding said plants;
b) filtration of the ground material obtained; And c) recovery of the liquid extract obtained after filtration.
We mean here to designate by the term essentially includes by including at least 75%, preferably at least 95%, of leaves and flowers of said plants by weight, per dry example, relative to the total plant weight, before mixing with solvent aqueous.
In embodiments, the extract is obtained by a process comprising, moreover, a step of nebulizing the liquid extract and passing the extract nebulized liquid in a flow of hot air.
According to a second aspect, the present invention aims at a method of reduction effects viruses on plants, including trees, having one stage application of the composition elicitor object of the invention.
In embodiments, the method is applied to reducing effects of a attack from one of the following viruses:
- Beet yellows viruses, - Tobacco mosaic virus, - Cassava mosaic virus, - The Banana Bunchy Top Virus (BBTV), - Banana streak virus (BSV), - Barley yellow dwarf virus (JNO or BYDV), - Cucumber mosaic virus, - Sugarcane mosaic virus (SCMV), - Corn lethal necrosis disease viruses (M LN, MCMV, SCMV), - The Potyviridae family of viruses, - Sweet potato feathery mottle virus (Potyvirus), or SPFMV, - Sweet potato chlorotic stunt virus (Crinivirus), or SPCSV
and SPVD, - Sweet potato moderate mottle virus, or SPMMV, - Sweet potato latent virus, or SPLV, - Sweet potato chlorotic spot virus or SPCFV, - Sweet potato virus G, or SPVG, - Sweet potato leafroll virus, or SPLCV, - Tomato brown rough fruit virus (ToBRFV), - Tomato spotted wilt virus , TSWV), - Tomato mosaic virus (Tomato mosaic virus, ToMV), - Zucchini yellow mosaic virus viruses, or ZYMV), WO 2023/062025

9 PCs - the rose mosaic virus.
In embodiments, the method is applied to reducing effects of a attack of at least one of the beet yellows viruses.
In embodiments, the method is applied to reducing effects of a attack of at least one of the cucumber mosaic viruses.
In embodiments, application of the elicitor composition is a Foliar application on plants.
According to a third aspect, the invention aims at a use of the composition object elicitor of the invention to reduce, on plants, including trees, the effects of an attack from one of the viruses listed above.
In embodiments, the application to the plant or tree is made by foliar spraying, ground watering, drip, use in hydroponic crops, seed treatment and/or seed coating.
In embodiments, the application to the plant or tree is made with a dilution in water of the composition between 2 g/L and 2000 g/L expressed as grams of plants on which was carried out the extraction per liter of product applied.
In embodiments, the application to the plant or tree is made with a dilution in water of the composition between 5 g/L and 200 g/L expressed as grams of plants on which was carried out the extraction per liter of product applied.
In embodiments, said extract of at least part of plants is an extract liquid from said plants.
In embodiments, application to the plant or tree is produced by foliar spraying, soil watering, soil irrigation, drip, hydroponics cultivation, seed treatment and/or seed coating.
In embodiments, at least one active ingredient is obtained from leaves of said plants.
In embodiments, at least one active ingredient is obtained from of flowers say plants.
In embodiments, at least one active ingredient is obtained by grinding of au least part of the said plants.
In embodiments, at least one active ingredient is obtained by extraction watery.
In embodiments, at least one active ingredient is obtained, by extraction of oil, by solvent extraction, or by extraction of cakes or pasta.
In embodiments, the composition is formulated as a powder, soluble powder, wettable powder, granules, dispersible granules, or wettable granules or slow diffusion, to be diluted in water at the time of use.
In embodiments, the composition is formulated as a liquid, liquid soluble concentrate, emulsifiable concentrate, suspension concentrate, or ready to employment.

WO 2023/062025

10 PCs According to a fourth aspect, the present invention aims at a production process of a composition which is the subject of the invention, which comprises a grinding step of at least part of the called plants, to provide a crushed material, and a filtering step to extract solid parts of said crushed material and obtain a liquid.
The particular advantages, purposes and characteristics of this composition and of this process being similar to those of the composition which is the subject of the present invention, they are not remembered here.
According to a fifth aspect, the present invention aims at a simulation method physical of gene-for-gene interaction, a composition capable of being extracted by aqueous extraction of a given plant in the claimed lists, in particular of a plant of rocket, and particularly of the Eruca Sativa or Diplotaxis type or plants genetically modified to produce this composition.
Thus, the process and the composition which is the subject of the invention carry out an action on the plant within hours, or even less than an hour, after the interaction bothers gene with the virus or its variant, thus avoiding the problem of mutations, and providing an answer reinforcing immediately the resistance of treated plants against viruses, bacteria, fungi and other pathogens.
While the interaction between plants and viruses is very particular and different from other plant/parasite interactions, since there is no solution possible chemical, it is demonstrated that the composition which is the subject of the invention, particularly in its form extracted from plants rockets, and particularly Eruca Sativa or Diplotaxis Tenu ifolia, acts effectively against viruses, by acting in very early times by simulating a gene for gene interaction, which allows the plant to overcome all the weaknesses of a plant with respect to live with a virus (too rapid appearance of variants, replication of the virus, etc.) and, more generally, of a pathogenic.
The advantages, purposes and particular characteristics of other aspects of the invention, in particular those which are claimed, also apply to this process and this composition and to the use of this composition to simulate, in a plant whose resistors must be stimulate, a gene-for-gene interaction.
Brief description of the figures Other advantages, aims and characteristics of the present invention will emerge from the description which follows, made for explanatory purposes and in no way restrictive, with regard to the drawing annexed, in which:
Figure 1 represents, in the form of a flowchart, the steps of a mode of carrying out a process for producing and using a ground material, which is an example preferential production of the composition which is the subject of the invention, Figure 2 represents dates of evaluations of beet treatments, Figure 3 shows numbers of Myzus persicae insects, Figure 4 shows incidences of the pest Myzus persicae, WO 2023/062025

11 PCs Figure 5 shows numbers of Aphis fabae insects, Figure 6 represents incidences of the pest Aphis fabae Figure 7 represents assessments of areas infected by jaundice, Figure 8 represents a photograph of the untreated modality (control), Figure 9 represents a photograph of the modality treated with the product PP1, Figure 10 represents a photograph of the modality treated with the product PP1, and Figure 11 represents a descriptive table of the treatment methods of the first one demonstration.
Description of embodiments The elicitor composition stimulating the defenses of plants or trees reducing the effects of an attack by a virus comprises an extract of at least a part of at least one of the plants following: Rockets, including genera Eruca sativa, Diplotaxis, Erucastrum and Bunias, Cakile, plants of the genera Allium, mustard (Sinapis alba, Brassica nigra, Sinapis arvensis, Brassica juncea), wasabi (Eutrema japonicum), horseradish (Armoracia rusticana), watercress fountain (Nasturtium officinale), plants of the species Brassica rapa, Brassica ruvo, Brassica napus, Raphanus sativus, Barbarea verna, Erysimum allionii, Erysimum cheiri, Tropaeolum majus L, Alliaria petiolata, Salvadora persica, Carica papaya and Brassica oleracea.
The preferred embodiments and certain viruses are listed below.
above.
Before presenting the different aspects of the present invention, we describe, below, viruses on which the eliciting composition object of the test has been successfully tested.
the invention:
For the sake of clarity and conciseness, the demonstrations of the description who will follow do not cover all of the plant combinations listed above, but illustrate the effectiveness of the present invention in all these combinations.
First demonstration: Effect of the eliciting composition object of the invention against beet yellows virus.
Beetroot is the primary reservoir of yellows viruses. It is therefore important to remove all harvest residues (digging cords, beets trailing) because the Leaf regrowth can become sources of infection. he is also important to good manage weeds, in plots and at the edges of fields, because a certain number of species are hosts to vector aphids and sometimes also to aphid viruses.
jaundice.
To combat this virus, the following techniques are used:
Preventive control by using seeds treated with an insecticide systemic in coating (imidacloprid). This technique presents by far the best effectiveness results and afterglow. However, it is important to note that in 2018 the neonicotinoids have been banned.
2020 was the second year without a neonicotinoid (NN I) on seeds since 1993.
Imidacloprid was authorized as a seed treatment that year. It is a very insecticide effective, neurotoxic. As a result, the aphid only bites once and so there is no longer of contamination per round (INRAE, 2020).
In Europe, imidacloprid will no longer be approved from 7/31/2022, but some countries kept it or granted exemptions, when ANSES in France withdrawn all marketing authorizations WO 2023/062025

12 PCs for agricultural purposes. Two other NNIs were withdrawn at the end of 2019 (thiamethoxam, clothianidin). Alone thiamethoxam and Imidacloprid were used in our territory in seed coating beetroot.
Preventive control by the application, during sowing, of micro-insecticides long pellets persistence, adapted to the fight against aphids which vector jaundice.
Curative control using sprays based on aphicidal products.
As of today, the products available are as follows:
Teppeki (50% Flonicamide), registered brand, has been approved since 21 December 201 8 with the following conditions of use (authorization of on the market, or AMM, no. 2050046):
Approval dose of 0.14 kg/ha, One application per year from the six true leaf stage, Effective only on aphids (selective of auxiliaries), Possible mixture with herbicides, Remanence of at least two weeks, Add one liter of oil according to regulations.
Movento (Spirotetramate at 100 g/1), registered brand, has obtained marketing authorization derogatory 120 days for use in 2020 with conditions of use following:
Approval dose of 0.451/ha, Two applications per year (minimum interval of 14 days) from the stage two leaves until the beets are covered, Effective only on aphids (selective of auxiliaries), Do not use as a mixture because there is a risk of reduced effectiveness and Minimum persistence of two weeks.
However, the use of these two active substances in 2020 did not allow of sufficiently control aphid populations throughout the territory. These two substances do not constitute a sustainable solution. At this stage, none chemical solution or not chemical treatment does not come close in effectiveness to chemical treatments based on NNI
and does not allow cope with an exceptional situation (INRAE, 2020) Given the urgent need to find an effective solution to combat against the beet yellows, the entirely biosourced elicitor composition object of the invention is capable of significantly reducing, or even eliminating, the effects caused by viral diseases serious to plants and until now incurable. The toxicity tests of this composition have demonstrated that it is not toxic. A test was set up with the Ephydia company (marked registered, BPE approved company for good environmental practices) against the virus beet yellows, the results of which are presented below.
Material and methods Plant material: Beta vulgaris is a species of plants in the family of Amaranthaceae.

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13 PCs It is cultivated throughout the world for sugar production, and in a manner annex for make ethanol or baker's yeast from molasses made from residues from white sugar manufacturing.
Growing conditions Planting date: 03/25/2021 Depth: two meters Spacing between rows: 45 cm Row spacing: 18.5 cm Planting rate: 100,000 plants/ha Planting method: seed Test carried out by: Ephydia Width of the treated plot: 2.7 m Length of the treated plot: 9.25 m Surface area of the treated plot: 24,975 m2 Number of repetitions: four Soil texture: Loamy Test location:
Town / Municipality: Attics (80) Country: France Postcode: 80360 Description of treatments:
Control (untreated plot), Application of PP1, at 50% V/V, Application of PP1, at 50% V/V and Teppeki, at 0.14 kg/ha, Application of Teppeki, at 0.14 kg/ha.
In this first demonstration, extracts of eruca sativa and diplotaxis tenuifolia have been used. The results being very similar, these two extracts have been grouped under the term credits of PP1 below.
Description of treatment methods Type of application: Foliar application Table 1 in Figure 11: Description of treatment modalities (or Trt) Table 2: Date and stage of application of treatments ABCD
Appiwation date 1.1/05/21 05/24/21 06/01/21 06/14121 BBCH Stadium 12 16 18 32 39 Teppeki Product Description:
No AMM: 2050046 Date of first product authorization: 04/21/2005 Active substance: Flonicamide 500 g/kg WO 2023/062025

14 PCs Function: Insecticide Use: 150531 06 Industrial and fodder beets*Trt Part.Aer.*Aphids Date of authorization of use: 08/12/2021 Maximum application dose: 0.14 kg/ha Maximum number of applications: One Application stage: Minimum: BBCH 12 Maximum: BBCH 49 Deadlines before harvest: 60 days It is important to note that the Teppeki product is only authorized to be applied only one times on the cultivation of beet, however during the test the program received three applications of the Teppeki product and six applications of the Teppeki product alone for reference modality. So the results which will follow will be nuanced knowing that these levels efficiency with the reference are never achieved in the fields under real conditions.
Description of the evaluations carried out during the trial.
Aphid rating at each application and five days after each treatment:
Myzus persicae: green peach aphid Aphis fabae: black bean aphid Notation at the appearance of symptoms, during the summer and before harvest Gross weight harvest and saccharimetry Figure 2: Date of assessments.
Results and discussion.
In Figures 3 to 6, each set of four vertical bars represents, of left to right, the untreated control, the treatment with the object composition of the invention, the combination of this composition and Teppeki and Teppeki alone.
Evaluation of the number of aphids and the incidence of the pest ¨ Myzus Persicae.
Figure 3: Number of Myzus persicae insects.
Figure 4: Pest incidence - Myzus persicae.
Note: DA ¨ A means Day after treatment A (days after treatment treatment A), etc.
The green peach aphid, Myzus persicae, is the main vector of jaundice of the beet. It has very good transmission capabilities for viruses.
moderate jaundice (BChV
and BMYV) as well as severe jaundice (BYV).
The results illustrated in Figure 3 show that the PP1 treatment makes it possible to decrease statistically the number of aphids vectors of yellows (except for rating 31 DA-A, associated letter a) in comparison to the untreated modality. All the modalities treated are statistically identical up to 14 DA-A and notation 41 DA-A (letters associated b) and different from 21 DA-A to 35 DA-A of the modality treated with Teppeki in significant overdose.
In addition, the results illustrated in Figure 4 show that the PP1 treatment allows a reduction in incidence compared to the control except for the 10 DA- notations A, 21 DA-A and 31 DA-A. These results show that PP1 has an effect on the presence of the vector of jaundice of the beetroot- Myzus persicae.

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15 PCs Aphid presence values are directly correlated with the appearance of symptoms of jaundice. Therefore, the results obtained show that PP1 allows the reduction symptoms.
Assessment of the number of aphids and the incidence of the pest ¨ Aphis Fabae.
Figure 5: Number of Aphis fabae insects Figure 6: Pest incidence - Aphis fabae The black bean aphid, Aphis fabae, is a secondary vector of BYV
(jaundice serious), but does not transmit BChV or BMYV (jaundice virus moderate). Jaundice is not never transmitted to the offspring of contaminated aphids.
The results illustrated in Figure 5 show that the PP1 treatment makes it possible to decrease statistically the number of aphids vector of jaundice in comparison to the modality no processed, except at 41 DA-A (associated letters a). Furthermore, we can observe that from 31 DA-A, the PP1 product is statistically equivalent to the Teppeki reference, even applied in significant overdose (associated letters b).
In addition, the results illustrated in Figure 6 show that the PP1 treatment allows a reduction in incidence compared to the control except for notation 31 DA-A
(associated letters has ).
These results show that PP1 has an effect on the presence of the vector of jaundice Aphis fabae beet. In addition, the aphid presence values are directly correlated with the appearance of symptoms of jaundice. Therefore, these results show that PP1 allows reduction of symptoms.
Areas infected by beet yellows (figure 7). Figure 7 represents a evaluation of the area infected by jaundice depending on the treatments. There risky period begins as soon as the first aphids appear in the plots end of April beginning of May, or from the two-leaf stage until ground cover at the end of June. THE
latency time is usually two to four weeks but would be shorter for jaundice serious (one to two weeks) than for moderate jaundice (four to six weeks).
In the case of this trial, Myzus persicae aphids were detected on the test of 05/14/2021 to 06/24/2021 and Aphis fabae aphids were detected on the test of 06/09/2021 as of 06/24/2021. The first symptoms of the disease were observed on 3/09/2021 as shown in Figure 7. These results demonstrate that the plots begin to show symptoms of beet yellows and that the product PP1 is also effective than the reference TEPPEKI (which was applied six times while it is only applied once only once in condition normal) and helps reduce symptoms compared to the non-normal modality.
processed.
Thus, PP1 could prove to be much more effective than the reference product Teppeki in normal application condition. PP1 thus makes it possible to reduce the presence of aphids vectors but also to limit the appearance of symptoms.
Figure 8: Photograph of the untreated modality ¨ plot 401 Figure 9: Photograph of the modality treated with the product PP1 ¨ plot Figure 10: Photograph of the modality treated with the product PP1 ¨ plot WO 2023/062025

16 PCs Conclusions of the first demonstration.
Data from this trial indicate that application of the herbal extract PP1 by foliar application helps reduce the presence of the two varieties of aphids vectors severe jaundice and moderate jaundice. These data are linked to the fact that PP1 allows you to reduce the symptoms of jaundice. Plants that have not been bitten by aphids continue their photosynthesis, have good vigor and continue their cycle of development.
Beet harvest depending on the conditions:
Witness: 0.8 Ton PP1: 1.2 Ton PP1 + Teppeki: 1.3 Ton Teppeki: 1.2 Ton Thus PP1 would prove to be a solution to the problem of jaundice and a solution of replacement of neonicotinoids. The PP1 product is capable of reducing significantly the incidence of beet yellows virus.
Second demonstration: Effect of PP1 against the mosaic virus cucumber.
This test is carried out in an area known for its contamination by the virus.
mosaic cucumber.
The control methods are generally as follows:
- Limit the presence of reservoir plants, - Immediately eliminate and destroy plants that show signs of symptoms, - Limit the presence of aphids, - Use clean tools when pruning and gardening work, - Choose healthy seeds or plants - Choose resistant cucumber varieties.
A trial is carried out to evaluate the effectiveness of PP1 against mosaic of the cucumber.
Plant material:
STYX variety, organic plants, Calendar: planting April 11, 2021, Harvest from May 23, 2021 to July 15, 2021, Device: Eight-meter tunnel, micro-perforated thermal opaque mulch, four rows double (distance 0.35 ni), Irrigation: one boom per crop row Modalities: Untreated controls (T) and plants treated with PP1 Four repetition block test Treatments: six treatments were carried out at a rate of 14 days.
Symptoms :
Chlorotic spots (more or less marked mosaic) appear on the youth leaves ; these can become deformed, embossed, or even dry out in serious cases. In the plots, we observe circular disease foci, which extend gradually.

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17 PCs An early attack causes complete dieback of young plants. THE
plants affected have reduced growth and a modified habit.
A plant infected with this virus remains a carrier of the virus until it dies.
Observations and measurements carried out: - Observations: Agronomic measurements of May 23 2021 to July 15, 2021:
Incidence represents the percentage of contaminated leaves or fruits.
Severity represents the percentage of surface area covered by symptoms of the disease.
Table 3: Incidence and severity on cucumber leaves ¨ May 23, 2021 Conditions Incidence Severity Oa Oa Witnesses PP1 Oa Oa The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 4: Incidence and severity on cucumber leaves ¨ July 15, 2021 Conditions Incidence Severity Witnesses 25a 30a PP1 5b 5b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 5: Incidence and severity on cucumbers (Fruits) ¨ July 15, 2021 Conditions Incidence Severity Witnesses 35a 40a PP1 10b 5b The severity and incidence were measured on 20 fruits taken in a manner random, for each modality. The analyzes were carried out according to Newman and Keuls. THE
different letters express significantly different results at the 5% threshold.
We observe, both for the leaves and for the fruits, the composition elicitor object of the invention (PP1) significantly reduces both the incidence and the severity of virus effects and significantly increases the quantity of beet harvested.
The results show the effectiveness of PP1 against the mosaic virus of cucumber.
Indeed, the notations on leaves and fruits show that the plants treated have significantly fewer symptoms than control plants.
Third demonstration: B1/ Broccoli extract (PP2) against the virus Mosaic cucumber (CMV). An extract of broccoli leaves is produced according to the protocol illustrated in figure 1, in liquid form, without concentration. This test is carried out in area known for its contamination by cucumber mosaic virus (presence of the vector).
No biocontrol solution is known to date to eradicate disease.

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18 PCs In this context, a trial is carried out to evaluate the effectiveness of PP2 against the virus of cucumber mosaic (CMV).
Symptoms :
Chlorotic spots (mosaic) appear on young leaves, which can deform, emboss, or even dry out in severe cases. An attack early causes complete dieback of young plants. Affected plants have a reduced growth and port changed. A plant infected with this virus remains a carrier of the virus until his death.
Material and method :
Cucumber variety: TYRIA, organic plants, not tolerant to the virus mosaic.
Calendar: planting April 8, 2020 Harvest: from May 28, 2020 to July 30, 2020 Device: Eight-meter tunnel, trellising on twine, two double rows (distance 0.35 m). Irrigation: one boom per crop row Modalities: Untreated controls (T) and treated plants (PP2) Layout: Block trial with four randomized repetitions Treatments: six treatments were carried out at a rate of 14 days.
Results :
Table 6: Incidence and severity on cucumber leaves ¨ May 30, 2020 Conditions Incidence Severity Oa Oa Witnesses PP2 Oa Oa The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 7: Incidence and severity on cucumber leaves ¨ July 20, 2020 Conditions Incidence Severity Witnesses 16a 10a PP2 9b 5b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 8: Incidence and severity on cucumbers (Fruits) ¨ July 20, 2020 Conditions Incidence Severity Witnesses 12a 15a PP2 5b 8b The severity and incidence were measured on 20 fruits taken in a manner random, for each modality. The analyzes were carried out according to Newman and Keuls. THE
different letters express significantly different results at the 5% threshold.

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19 PCs The results show the effectiveness of PP2 against the mosaic virus of cucumber.
Indeed, the notations on leaves and fruits show that the plants treated have significantly fewer symptoms than control plants.
Fourth demonstration: Brussels sprout extract (PP3) on virus mosaic of cucumber (CMV). An extract of Brussels sprout leaves is made according to the illustrated protocol in figure 1, in liquid form, without concentration.
This test is carried out in an area known for its contamination by the virus.
mosaic cucumber (presence of the vector).
No biocontrol solution is known to date to eradicate disease.
In this context, a trial is carried out to evaluate the effectiveness of PP3 against the virus of the cucumber mosaic.
Material and method :
Cucumber variety: TYRIA, organic plants, not tolerant to the virus mosaic.
Calendar: planting April 8, 2020 Harvest: from May 28, 2020 to July 30, 2020 Device: Eight-meter tunnel, trellising on twine, two double rows (distance 0.35 m). Irrigation: one boom per crop row Modalities: Untreated controls (T) and treated plants (PP3) Layout: Block trial with four randomized repetitions Treatments: six treatments were carried out at a rate of 14 days.
Results :
Table 9: Incidence and severity on cucumber leaves ¨ May 30, 2020 Conditions Incidence Severity Oa Oa Witnesses PP3 Oa Oa The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 10: Incidence and severity on cucumber leaves ¨ July 20, 2020 Conditions Incidence Severity Witnesses 27a 20a PP3 15b 10b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 11: Incidence and severity on cucumbers (Fruits) ¨ July 20, 2020 Conditions Incidence Severity Witnesses 32a 25a PP3 20b 15b WO 2023/062025

20 PCs The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
Conclusion :
The analyzes were carried out according to Newman and Keuls. Different letters express significantly different results at the 5% threshold. The results show the effectiveness of PP3 against cucumber mosaic virus. Indeed, the ratings on leaves and fruit show that treated plants have significantly fewer symptoms than control plants.
Fifth demonstration: Arugula extract (¶ PP1) on the virus disease tomato spotted wilt virus (TSWV).
Tomato spotted wilt virus (TSWV) is widely distributed in the world in temperate and subtropical zones where it has been on the rise since the early years 1980. Emerging in France since 1987, it has a wide range of hosts potential. He is transmitted by at least nine species of thrips.
Symptoms of Tomato spotted wilt virus virus, TSWV) can take on various appearances on the tomato foliage, such as deformations leaves with an apical curvature of the apex, blocking of vegetation, a mosaic more or less less contrasty, chlorotic spots and lesions becoming necrotic, chlorosis and a more or less marked bronze coloring of the blade or veins, accompanied rings, Small dark lesions becoming necrotic also visible on the petioles and stem, anthocyanization of the blade.
Fruits are also affected. They may be tanned and present wide Arabesques and more or less concentric chlorotic rings. Of the necrotic changes dry, cracks are sometimes visible. Early contaminations lead to a reduction in number and size of fruits; if they are late, the fruits grow normally but are poorly colored and more or less distorted.
No biocontrol solution is known to date to eradicate disease.
In this context, a trial is carried out to evaluate the effectiveness of the elicitor composition object of the invention against the tomato TSWV virus. The extracts from rocket leaves (Diplotaxis) are obtained according to the protocol illustrated in Figure 1 under their liquid form no concentrated (extracts called < PP1).
Material and method :
The experiment takes place in a 250m2 rigid greenhouse equipped with openings and side vents.
Experimental device: Type .................................................. ... complete blocks to four repetitions. Plot elemental of 10 plants.
Technical itinerary: Semi on December 30, 2021 for planting on the 26th January 2022.
Harvest over four months from the beginning of March to the end of June 2022.
Modalities: Control (untreated), PP1 (spray application foliar) Treatments: Foliar spraying Six applications of PP1, at a rate of 14 days.

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21 PCs Analysis method: Analysis of variance with a risk threshold of 5%. THE
ratings presenting the same letters are not significantly different.
Results :
Table 12: Incidence and severity on tomato leaves ¨ March 20, 2022 Conditions Incidence severity Witnesses 10a 5a PP1 10a la The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 13: Incidence and severity on tomatoes (Fruits) ¨ March 20, 2022 Conditions Incidence severity Oa Oa Witnesses PP1 Oa Oa The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
Table 14: Incidence and severity on tomato leaves ¨ April 25, 2022 Modalities Incidence severity Witnesses 35a 70a PP1 10b 30b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 15: Incidence and severity on tomatoes (Fruits) ¨ April 25, 2022 Modalities Incidence severity Witnesses 25a 55a PP1 5b 10b The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
Table 16: Incidence and severity on tomato leaves ¨ June 5, 2022 Modalities Incidence severity Witnesses 90a 80a PP1 10b 40b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 17: Incidence and severity on tomatoes (Fruits) ¨ June 5, 2022 Modalities Incidence severity Indicators 50a 60a WO 2023/062025

22 PCs PP1 10b 15b The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
The results show the effectiveness of PP1 against tomato virus disease (TSWV). Indeed, the notes on leaves and fruits show that the treated plants have significantly less than symptoms than control plants. The progression of the disease is very slowed down by PP1 foliar treatments.
Sixth demonstration: Arugula extract on the mosaic virus of the tomato (ToMV). Tomato mosaic virus (ToMV) is present on all continents. It is frequently found on tomatoes and peppers. He is grave as well in culture in the open field, only under cover. Although its incidence has decreased significantly with the variety broadcast of resistant tomatoes, the recent marketing of new varieties sensitive showed how the ToMV was always ready to attack sensitive plant material.
No biocontrol solution is known to date to eradicate disease.
In this context, a trial is carried out to evaluate the effectiveness of PP1 against the virus ToMV of the tomato.
The symptoms caused by the presence of this virus are very varied and are enough comparable. We can notice a slowdown in plant growth, as well as discoloration abnormalities that may appear on leaflets and leaves.
Other symptoms can still be expressed, on leaves, as a clarification of the veins, marbling, a mosaic of green or yellow areas with the blade which becomes embossed and edgy.
We can also see the fall of flowers. When the fruit arrives maturity, they are small in size and sometimes more or less bumpy. They also express yellow discolorations, sometimes in rings. These symptoms may be present on green fruits or ripe while the plant appears healthy. Late infections have no impact on the production.
Arugula leaf extracts (Diplotaxis) are obtained according to the protocol illustrated in figure 1 in their non-concentrated liquid form (extracts called PP1 by the following).
The experiment takes place in a greenhouse, above ground cultivation.
Experimental setup: Complete block type with four repetitions.
Plot elemental of 10 plants.
Technical itinerary: planting the buckets on February 5, 2020. Harvest on five months of beginning of March to end of July 2020.
Modalities: Controls (untreated), PP1 (foliar spray) Treatments: Foliar spraying - Six applications of PP1, alder .............. cadence of 14 days.
Analysis method: Analysis of variance with a risk threshold of 5%. THE
ratings presenting the same letters are not significantly different.
Results :
Table 18: Incidence and severity on tomato leaves ¨ March 5, 2020 WO 2023/062025

23 PCs Conditions Incidence severity Witnesses 5a 5a PP1 5a 5a The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 19: Incidence and severity on tomatoes (Fruits) ¨ March 5, 2020 Conditions Incidence severity Oa Oa Witnesses PP1 Oa Oa The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
Table 20: Incidence and severity on tomato leaves ¨June 30, 2020 Conditions Incidence severity Witnesses 55a 60a PP1 12b 20b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 21: Incidence and severity on tomatoes (Fruits) ¨ June 30, 2020 Conditions Incidence severity Witnesses 28a 30a PP1 8b 10b The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
The results show the effectiveness of PP1 against the mosaic virus of the tomato (ToMV). Indeed, the notations on leaves and fruits show that the treated plants have significantly fewer symptoms than control plants. The progression of the disease is very slowed down by foliar treatments of PP1.
Seventh demonstration: Arugula extract on yellow mosaic virus of the zucchini (Zucchini yellow mosaic virus, or ZYMV).
ZYMV is a potyvirus transmitted by aphids in a non-persistent manner.
It's one of the best examples of viruses emerging in plants. Insulated for first time in Italy then in France in the 70s, it spread in a few years in the whole world, in sometimes causing epidemics of exceptional severity. This recent spread and quickly in various types of cultivation (intensive, extensive, under cover, full field) and very varied ecosystems (temperate, tropical, Sahelian, island) is attested by the fact that the ZYMV causes very strong symptoms.

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24 PCs ZYMV is now reported on cucurbits in virtually all their areas of production in the world. However, its frequency can vary a lot depending on the region.
Encountered regularly in tropical or subtropical regions, its epidemics are more irregular in temperate countries such as France. A survey carried out by 2004 to 2008 in the main French production areas showed that ZYMV
was only present in 11% of 2660 samples analyzed, mainly on squash (23% of samples tested), zucchini (14%) and melon (8%), and to a lesser extent on cucumber (3%).
In the areas where this virus was detected, outbreaks were generally severe, with a strong impact on the yield. ZYMV causes very severe mosaic symptoms, yellowing, stunting and deformation of the foliage of virtually all cucurbits. He also causes discoloration and spectacular deformation of fruits which are then unmarketable. Early attacks can lead to loss total of harvests.
The experiment takes place in open fields.
Experimental setup: Complete block type with four repetitions Plot elemental of 10 plants.
Technical itinerary: planting the buckets on April 15, 2022. Harvest on the 30th June 2022.
Modalities: Controls (untreated), PP1 (foliar spray) Treatments: Foliar spraying - Six applications of PP1, at a rate of 14 days.
Analysis method: Analysis of variance with a risk threshold of 5%. THE
ratings presenting the same letters are not significantly different.
Results :
Table 22: Incidence and severity on zucchini leaves ¨ June 30, 2022 Modalities Incidence severity Witnesses 25a 15a PP1 8b 5b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
Table 23: Incidence and severity on zucchini (Fruits) ¨ June 30, 2022 Conditions Incidence severity Witnesses 20a 10a PP1 5b 5b The severity and incidence were measured on 20 fruits taken in a manner random, for each modality.
Results show effectiveness of PP1 against mosaic virus yellow of the zucchini ZYMV. Indeed, the notations on leaves and fruits show that the treated plants have significantly fewer symptoms than control plants. The progression of the disease is very slowed down by foliar treatments of PP1.

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25 PCs Eighth demonstration: Radish extract (PP4) on the virus rose bush mosaic.
Rose mosaic is a viral disease that affects rose bushes (Rosa sp.). She is due to several viruses of the Ilarvirus and Nepovirus genera which intervene separately or more often in combination, which has led some authors to speak of viral complex of rose bush mosaic. In certain cultivars, these viruses can cause variegation flowers. Other infected cultivars may remain asymptomatic.
The disease is not lethal to rose bushes, but infection has the effect of reduce the vigor of the plants and weaken them, so that they are more vulnerable to stress transplant or winter injury.
This disease causes various symptoms on the leaves: spots ring fingers, chlorotic lines, watermarks, leaf mottling, as well as patterns in yellow mosaic.
Signs of the disease include: bright yellow zigzag patterns on the leaves, arranged symmetrically in relation to the main vein; Tasks yolks to cream can be diffuse and draw a marbling; localized browning may recall a drying out of the leaves.
The radish extracts are obtained according to the protocol illustrated in Figure 1 in their form non-concentrated liquid (extracts called PP4 hereinafter).
The experiment takes place in a heated greenhouse.
Experimental device: Complete block type ................................... four repetitions. Plot elemental of 10 plants.
Technical itinerary: experiment carried out on ................................ rose bushes producing roses, aged four years. Six Applications, at a rate of 14 days.
Modalities: Controls (untreated), PP4 (foliar spraying) Analysis method: Analysis of variance with a risk threshold of 5%. THE
ratings presenting the same letters are not significantly different.
Results :
Table 24: Incidence and severity on leaves ¨ June 30, 2022 Conditions Incidence severity Witnesses 18a 20a PP4 5b 5b The severity and incidence were measured on 20 leaves taken in a manner random, for each modality.
The results show the effectiveness of the extract against the virus of rose bush mosaic. In Indeed, the notations on leaves and fruits show that the plants treated have significantly fewer symptoms than control plants. The progression of the disease is very slowed down by PP4 foliar treatments.
The composition which is the subject of the present invention, subsequently called CEI
(including notably PP1 to PP4) does not correspond to what the literature describes:
1/ The CEI composition is extracted from leaves, stems, flowers, seeds and/or roots, according to a mode WO 2023/062025

26 PCs preferential extraction, with or without added water, according to the process described next to Figure 1. When used in fields, the composition is preferably diluted in tanks (spray tanks) to be sprayed at foliar level (or other application methods described in the description of uses) 2/ The CEI composition, obtained under these extraction conditions, does not have none direct antimicrobial activity.
In the CEI production process, the leaves, stems, seeds, roots and/or flowers make the subject of an extraction of compounds, by a known technique, for example by pressing, by ultrasound, and/or by use of solvents, particularly oily or aqueous.
In embodiments of this method, parts of plants are crushed and strongly diluted in water. In embodiments of this method, plant parts are ground without adding water. Optionally, the filtered ground material is then formulated in the form of powder, by nebulization in an ascending current of hot dry air, preferably to a temperature below 60 'C. Optionally, the extract in liquid form is sterilized by passage at high temperature for a very short duration, using techniques known.
The elicitor composition which is the subject of the invention is used in particular, by application, for stimulate the defenses of plants or trees and reduce the effects of viruses, especially viruses from beet yellows and cucumber mosaic.
CEI acts by stimulating plant defenses, and allowing plants processed from defend themselves against these viruses.
We can define CEI as an elicitor, given that the molecules possessing the property of inducing a cascade of defense reactions within the plant against the agents Pathogens are called elicitors.
The demonstration of the eliciting activity of defense mechanisms is also demonstrated at several levels: Demonstration of the production of molecules defenses, such as jasmonic acid, salicylic acid, or even peroxidases, has was carried out after treatment by CEI, under conditions of infections on beet (Beta vulgaris subsp. vulgaris) CEI has the particularity of stimulating the defenses of plants, and allowing these of react effectively, even in the case of invasive viruses, which are difficult to combat.
As illustrated in Figure 1, in one embodiment, the method of manufacturing and use of the composition which is the subject of the present invention comprises a extraction step 105 of an extract of said plants.
In preferred embodiments, the extract is obtained from of at least one plant not containing Methyl-isothiocyanate and/or Propenyl isothiocyanate.
The inventor noted that these isothiocyanates could have effects harmful to plant growth while reducing the effects of viruses.
In embodiments, the extract is obtained from at least one plant containing at least one butyl isothiocyanate. The inventor noted that these isothiocyanates have favorable effects on plant growth while reducing effects of viruses.

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27 PCs In embodiments, the extract is obtained from at least one plant containing 1,3-thiazepane-2-thione. The inventor noted that this compound which, due to its structure, is not an isothiocyanate has favorable effects on the plant growth all by reducing the effects of viruses.
For example, this extraction is carried out according to the following procedure:
- During a grinding step 110, the leaves, roots, stems, seeds and/or the flowers of said plants are finely ground with running water, for fifteen minutes, in a suitable mixing device, in order to obtain a homogeneous ground material;
- During a filtration step 115, the ground material is filtered to separate the debris of organs operated and obtain a green liquid without residue (the filtrate), which is the basis of the composition object of the invention or constitutes it.
In a variant, no water is added before grinding the parts of source plants.
In a variant, at least one of the active ingredients of the ground material is obtained by extraction of oil. In a variant, at least one of the active ingredients of the material crushed is obtained by solvent extraction by mechanical or microwave extraction, or by oilcake extraction or pasta. In a variant, at least one of the active ingredients is obtained by extraction mechanical or microwave extraction.
Alternatively, the extraction step 105 includes a step of compressing the leaves, roots, stems, seeds or flowers of said plants and collecting the liquid extracted, by simple gravity or by centrifugation. Alternatively, a simple centrifugation is carried out work during the stage extraction 105, to extract the liquid from the parts of said plants used.
As explained in the description which follows, the inventor discovered that use of this composition has a significant effect on trees and plants above mentioned infected by the viruses mentioned above. The inventor also discovered that the elicitor composition object of the invention has biostimulating effects on plant growth processed without constituting, at the doses used, a fertilizer or feed the treated plants.
It is noted that the liquid composition obtained at the end of step 105 can be formulated to make it easier to use. For example, it can be used in powder form, soluble powder, wettable powder, granules, dispersible granules, or wettable granules or slow diffusion, to be diluted in water at the time of use, liquid, soluble concentrated liquid, emulsifiable concentrate, suspension concentrate, or ready to use, in depending on the formulation chosen and the intended use or infused on a substrate dispersed in the soil of cultivation. THE
formulations are made from the product of extraction step 105 according to techniques known to those skilled in the art.
Active fractions can potentially be purified, by some means that this or, to facilitate formulation. Different extraction steps can be added for improve its quality. The composition which is the subject of the invention can be diluted in water, depending on the required dose at the time of use.

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28 PCs During an optional step 120, volatile extracts of the extract obtained. By example, this extract is transformed into powder, for example by nebulization and passage from the extract nebulized in a flow of hot air, preferably ascending.
Regarding use, during step 125, the biostimulant is applied under whatever form it is (liquid formulation, powder, soluble powder, pellets, pellets dispersible, slow dispersing granules, and all formulations) according to the uses and formulation considered. The use of the biostimulant which is the subject of the invention is preferably carried out by Foliar application or foliar spray. Other ways of using the biostimulant object of the invention are soil watering, soil irrigation, drip, hydroponic crops, or even in seed treatment and/or seed coating.
Preferably, the leaves and flowers of said plants represent at least 75%, preferably at least 95%, of the parts of said plants on which are carried out extraction, percentage by dry weight, relative to the total weight of these plants.
The composition which is the subject of the invention can be used for an application single or to a cadence between one day and one hundred and twenty days, or continuously, or according to the key stages of plant development, or in accordance with good agricultural practices and calendar of treatments planned for each plant species. The composition of this invention can be mixed with other products (plant protection products, plant protection media) cultures and materials fertilizers, fertilizers, fertilizers, biocides, or any other product intended for agriculture).
The application doses and application rates are adapted to the uses and to plant models. The application doses are included, for example, between 0.001 g/L and 2000 g/L of extracted plants, preferably between 2 g/L and 2000 g/L
of extracted plants and, more preferably, between 5 g/L and 200 g/L of plants extracted, expressed in grams of plants on which the extraction was carried out per liter of product applied.
The doses per liter or per hectare can be adapted to the types of plants infected, at the level of infection and the level of symptoms caused by viruses. Doses and the cadences of treatments with the composition which is the subject of the invention will also be adapted to strategy preventive or curative action against these viruses.
Concerning the plants from which the extracts used in this article are taken invention, they are preferably freshly picked. Alternatively, plants or parties of interest are suitably dried, in a manner known to those skilled in the art. THE
grinding can be made with two crushers which are used with different speeds of blades. The first crushed material obtained in 10 minutes of grinding is then poured into the second grinder having a speed blade faster. The ground material is homogeneous, with no visible residue of part of leaves, stems, or flowers. The quantity of water added during grinding is between 0 and 200 mL of water, preferably between 20 and 150 mL of water, and even more preferably, between 50 and 120 mL
of water, at room temperature for 100 g of leaves, stem, root, flower or seed.
Two successive filtrations are carried out, with a filtration fabric in nylon (Dutcher, registered trademark) 1000 m then 500 m. Filtration is carried out at temperature ambient, without pressure. For the recovery of the filtrate which is active, depending on the quantity to spray, we WO 2023/062025

29 PCs adapts the dilution (dose per hectare). Depending on usage, between 5 g of plants extracted per liter of spray mixture and 2000 g of extracted plants per liter of spray mixture spray, as described next to the examples.
The inventor found that the filtrate obtained can be kept for at least six days.
in a can room temperature, without losing its activity of stimulating the defenses of plants and trees.
The extract of at least part of said plants can thus be an extract liquid obtained from from a ground material of said plants, and:
- said extract of at least one part of plants comprises at least leaves of the said plants, preferably mainly leaves, and - the process for obtaining said liquid extract comprises the steps following:
a) a step of grinding said plants in an aqueous medium;
b) filtration of the ground material obtained; And c) recovery of the liquid extract obtained after filtration.
Concerning the formulation in the form of powder, granules, granules dispersible, or slow diffusion granules, a drying temperature is used, and, in ways of realization, coating of particles with other natural molecules (preferably very hydrophilic) which allow very good dissolution in water. THE
formulations are classic formulations in agriculture, particularly for products phytosanitary products, intended for be transported and stored in the form of powder, etc... and be, just before application, diluted in water. The present invention relates to the use of a composition elicitor comprising a plant extract obtained as explained above to stimulate the defenses of plants or trees and reduce the effects of viruses on these plants.
In embodiments, the elicitor composition which is the subject of the invention includes, plus, at least one of the following substances, obtained by synthesis or by extraction from plants, in particular the plants mentioned above:
- 1,3-thiazepane-2-thione, and/or - a brassinosteroid.

Claims (16)

30 1. Eliciting composition stimulating the defenses of plants or trees reducing the effects of attack of a virus, characterized in that it comprises an extract of at least part of at least one of the following plants:
- Rockets, including genera Eruca sativa, Diplotaxis, Erucastrum and Bunias, - Caki le, - plants of the Brassica oleracea species, - plants of the Allium genera, - mustard (Sinapis alba, Brassica nigra, Sinapis arvensis, Brassica juncea), - wasabi (Eutrema japonicum), - horseradish (Armoracia rusticana), - watercress (Nasturtium officinale), - plants of the Brassica rapa species, - Plants of the Brassica ruvo species, - Plants of the Brassica napus species, - Plants of the Raphanus sativus species, - Plants of the species Barbarea verna, - Plants of the species Erysimum allionll, - Plants of the species Erysimum cheiri, - Plants of the species Tropaeolum majus L, - Plants of the Alliaria petiolata species, - Plants of the Salvadora persica species, - Plants of the Carica papaya species. 2. Eliciting composition stimulating the defenses of plants or trees according to claim 1, in which the extract is obtained from at least one of the plants following: Rockets, including Eruca sativa, Diplotaxis, Erucastrum and Bunias, Cakile, broccoli, plants of the genus Allium (garlic, onion, shallot, leek and chives), common cabbage, cauliflower, cabbage from Brussels, cabbage curly, mustard, wasabi, watercress, horseradish, white cabbage, cabbage country (Brassica rapa), including cabbage pak chcii, Chinese cabbage and turnip, kohlrabi, collard greens, red cabbage, broccoli Chinese, broccoli rabe, rapeseed, radish, Siberian wallflower, wallflower, watercress indian, garlic herb, tree of pilu, papaya, especially its fruit. 3. Eliciting composition stimulating the defenses of plants or trees according to one of claims 1 or 2, in which the extract is obtained from at least one of the plants following: Rockets, including Eruca sativa, Diplotaxis, Erucastrum and Bunias, Cakile, broccoli, common cabbage, cauliflower, Brussels sprouts, kale, mustard, watercress fountain, cabbage white, field cabbage (Brassica rapa), including pak choi, cabbage Chinese and turnip, cabbage rabe, collard greens, red cabbage, Chinese broccoli, broccoli rabe, radishes, and siberian wallflower. 4. Stirnulant elicitor composition of the defenses of plants or trees according to one of claims 1 to 3, in which the extract is obtained from at least one of the plants following:
- Rockets, including Eruca sativa, Diplotaxis, Erucastrum and Bunias, - Caki it. 5. Stirnulant elicitor composition for the defenses of plants or trees according to one of claims 1 to 3, in which the extract is obtained from at least one of the plants the species Brassica oleracea. 6. Eliciting composition stimulating the defenses of plants or trees according to one of claims 1 to 5, in which the extract is obtained from at least a plant not containing no Methyl-isothiocyanate and/or Propenyl isothiocyanate. 7. Eliciting composition stimulating the defenses of plants or trees according to one of claims 1 to 6, in which the extract is obtained from at least a plant containing minus one butyl isothiocyanate. 8. Stirnulant elicitor composition of the defenses of plants or trees according to one of claims 1 to 7, in which the extract is obtained from at least a plant containing 1,3-thiazepane-2-thione. 9. Eliciting composition stimulating the defenses of plants or trees according to one of claims 1 to 8, wherein said extract of at least part of plants is an extract obtained from a crushed material of said plants, and:
- said extract of at least one part of plants comprises at least leaves of said plants, preferably mainly leaves and flowers, and - the process for obtaining said liquid extract comprises the following steps :
a) a step of grinding said plants;
b) filtration of the ground material obtained; And c) recovery of the liquid extract obtained after filtration. 10. Eliciting composition according to claim 9, the extract is obtained by a process comprising, in addition, a step of nebulization of the liquid extract and passage of nebulized liquid extract in a flow of hot air. 11. Process for reducing the effects of viruses on plants, including trees, having a step of applying the eliciting composition according to one of the claims 1 to 10. 12. Method according to claim 11, for reducing the effects of an attack of one of the viruses following:
- Beet yellows viruses, - Tobacco mosaic virus, - Cassava mosaic virus, - The Banana Bunchy Top Virus (BBTV), - Banana streak virus (BSV), - Barley yellow dwarf virus (JNO or BYDV), - Cucumber mosaic virus, - Sugarcane mosaic virus (SCMV), - Corn lethal necrosis disease viruses (MLN, MCMV, SCMV), - The Potyviridae family of viruses, - Sweet potato feathery mottle virus (Potyvirus), or SPFMV, - Sweet potato chlorotic stunt virus (Crinivirus), or SPCSV
and SPVD, - Sweet potato moderate mottle virus, or SPMMV, - Sweet potato latent virus, or SPLV, - Sweet potato chlorotic spot virus or SPCFV, - Sweet potato virus G, or SPVG, - Sweet potato leafroll virus, or SPLCV, - Tomato brown rough fruit virus (ToBRFV), - Tomato spotted wilt virus , TSWV), - Tomato mosaic virus (Tomato mosaic virus, ToMV), - Zucchini yellow mosaic virus virus, or ZYMV), - the rose mosaic virus. 13. Method according to claim 11, for reducing the effects of an attack of at least one of beet yellows virus. 14. Method according to claim 11, for reducing the effects of an attack of at least one of cucumber mosaic virus. 15. Method according to one of claims 11 to 14, in which the application of the composition elicitor is a foliar application on plants. 16. Use of the eliciting composition according to one of claims 1 to 10, to reduce, on plants, including trees, from the effects of an attack by a virus following:
- Beet yellows viruses, - Tobacco mosaic virus, - Cassava mosaic virus, - The Banana Bunchy Top Virus (BBTV), - Banana streak virus (BSV), - Barley yellow dwarf virus (JNO or BYDV), - Cucumber mosaic virus, - Sugarcane mosaic virus (SCMV), - Corn lethal necrosis disease viruses (MLN, MCMV, SCMV), - The Potyviridae family of viruses, - Sweet potato feathery mottle virus (Potyvirus), or SPFMV, - Sweet potato chlorotic stunt virus (Crinivirus), or SPCSV
and SPVD, - Sweet potato moderate mottle virus, or SPMMV, - Sweet potato latent virus, or SPLV, - Sweet potato chlorotic spot virus or SPCFV, - Sweet potato virus G, or SPVG, - Sweet potato leafroll virus, or SPLCV, - Tomato brown rough fruit virus (ToBRFV), - Tomato spotted wilt virus , TSWV), - Tomato mosaic virus (Tomato mosaic virus, ToMV), - Zucchini yellow mosaic virus virus, or ZYMV), - Rose mosaic virus.