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    • 专利摘要:
    Composition and procedure to increase the content of glucosinolates in adult plants of the Brassica genus. The present invention relates to a composition comprising methyl jasmonate and a polysiloxane polyether for increasing the content of glucosinolates in adult plants of the Brassica genus, for example, broccoli. By means of foliar applications of said composition in adult plants that have a developed cuticle, a significant increase in glucosinolate concentrations (mainly glucorrafanine and neoglucobrasicin) is achieved, in the foil of the plants, without degradation of their organoleptic characteristics. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2737454A1
    申请号:ES201830674
    申请日:2018-07-05
    公开日:2020-01-14
    发明作者:Alcaraz Micaela Carvajal；Fernandez Diego A Moreno；Ruiz Juan Jose Rios；Dura Javier Bernabeu；Sanchez Agatha Agudelo
    申请人:Sakata Seed Iberica S L U；Consejo Superior de Investigaciones Cientificas CSIC；
    IPC主号:
    专利说明:

    [0001]
    [0002] Composition and procedure to increase the content of glucosinolates in adult plants of the Brassica genus
    [0003]
    [0004] The present invention relates to a composition and a method for increasing the content of glucosinolates in adult plants of the Brassica genus , preferably those used in human nutrition, such as broccoli, by foliar application once the reproductive vegetative phase of the plant, without any degradation of the organoleptic characteristics of the parts of the plant intended for human consumption.
    [0005]
    [0006] BACKGROUND OF THE INVENTION
    [0007]
    [0008] Broccoli, a plant of the order Brassicales, is a rich source of bioactive compounds, among which glucosinolates, compounds of secondary metabolism with nitrogen and sulfur in its molecule, stand out, which are receiving great scientific attention for their implications in the physiology of plants (plant defense) and food quality and bioactivity (organoleptic quality and functionality). These compounds are found almost exclusively in the cruciferous (Order Capparales) and are hydrolyzed when they come into contact with the enzyme myrosinase (vegetable glucosidase) as well as by the action of glucosidases of the intestinal microbiota, giving rise to isothiocyanates, bioactive compounds, which they are involved in the detoxifying mechanisms of cells through different mechanisms of stimulation and inhibition of certain enzymes. Two of the glucosinolates that can be found in broccoli varieties ( Brassica oleracea var. Italica) are glucorrafanine and neoglucobrasicin. Hydrolyzed derivatives of these glucosinolates give rise to isothiocyanate sulforraphane (from glucorrafanine) and indole-3-carbinol (from glucobrasicin and its related compounds). Both have attracted the attention of very diverse research groups worldwide because they are involved in the antioxidant and protective properties against free radicals derived from the consumption of broccoli (Moreno, DA et al. (2008) Journal of the Science of Food and Agriculture 88, 1472-1481). Therefore, either fresh or as derived ingredients, broccoli offers the possibility of developing foods enriched in bioactive compounds that help reduce incidence of diseases, with the consequent importance in the agri-food industry (Dominguez-Perles, R. et al. (2012) Acta Horticulturae, 939, 159-163). In fact, it is recognized as "the vegetable with the highest nutritional value" in relation to the weight of the edible part.
    [0009]
    [0010] The aforementioned glucosinolates, also called "thioglycosides", are nitrogenous sulforated anionic secondary metabolites found exclusively in the Brassicaceae family (Verkerk, R. et al. (2010) Acta Horticulturae vol. 856, pp. 63-69). The basic skeleton of glucosinolates (PD-thioglucoside-N-hydroxysulfate) consists of a pD-thioglucose group, a sulfonated oxime and a side chain derived from the amino acids methionine, phenylalanine, or tryptophan (Figure 1). The chemical structure of glucosinolates can vary between species and between varieties within the same species (Dominguez-Perles, R. et al. (2010) Journal of Food Chemistry 75, 383-392). According to the side chain, glucosinolates can be broadly classified into: aliphatic, aromatic, benzoic, multiple glycosylated, with sulfur in their side chain and heterocyclic or indole (Halkier, BA et al. (2006). Annual Review of Plant Biology, 57 (1), 303-333). In the case of broccoli, the most representative are aliphatic, aromatic and indole (A).
    [0011]
    [0012] Table A. Classification of the most representative glucosinolates in broccoli according to their structure.
    [0013]
    [0014]
    [0015]
    [0016]
    [0017] Glucosinolates have diverse and important physiological properties and constitute a defense mechanism for the plant against herbivores and pathogens, which at high concentrations produce metabolic stress in invertebrate herbivores, affecting their growth.
    [0018] They have biological properties of interest for the prevention of some diseases in humans that consume them (Jeffery, EH et al. (2009). Phytochemistry Reviews, vol. 8, 283-298). In addition, they contribute to the maintenance of body tissues, increase resistance to infections, regulate the proper development of the nervous system and intervene in growth, and are beneficial for the elaboration of enzymes in the liver (Moreno, DA et al. (2008 ), Journal of the Science of Food and Agriculture , 88, 1472-1481; Jeffery, EH et al. (2009). Phytochemistry Reviews, vol. 8, 283-298).
    [0019]
    [0020] Eliciters or biostimulators are natural substances that induce physiological changes in the plant, activating its defense mechanisms. Plants respond to these factors by activating a series of mechanisms, similar to defense responses to pathogen infections or environmental stimuli, that affect plant metabolism and improve the synthesis of phytochemicals or secondary metabolites. The first biotic eliciters were described in the early 1970s (Keen, NT (1975) Science 187, 74 75). Since then, numerous publications have accumulated evidence of application of compounds that induce defense responses in intact plants such as broccoli (Ku KM et al. (2016) Int J Mol Sci. 15, 17 (7), 1135). The use of eliciters is also performed as a tool to improve phytochemical content in plants for food improvement purposes, applied alone or in combinations at selected time points of plant growth (for review see: Baenas, N. et al. (2014) Molecules, 19 (9), 13541-13563). Eliciters can be classified as biotic and abiotic compounds, and plant hormones (salicylic acid, jasmonates, etc.) can also be considered eliciters.
    [0021]
    [0022] Elicitation can be used as a pre-harvest or post-harvest treatment. Among the pre-harvest treatments, the preparation of seeds (“priming”) is also considered, consisting of hydrating the seeds in a solution with the elicitator to induce the defense response of the cells. Thus, for example, it has been observed that parsley seeds treated with jasmonate showed an increase in antioxidant systems and the induction of defense systems through the synthesis of phenylpropanoids (Conrath, U. et al. (2015) Annual Review of Phytopathology 53, 97-119). The eliciters can also be applied as a gas in a closed environment (such as MeJA), in liquid form to a hydroponic solution or by exogenous aerosols.
    [0023] Jasmonic acid (JA) and its methyl ester, methyl jasmonate (MeJA), are widely known to elicit a wide range of compounds by inducing the expression of plant genes for various biosynthetic pathways. These small signaling molecules, also defined as "hormones," are induced in cells in response to wounds or attack by pathogens in plants, and can, in turn, induce cellular responses at low concentrations distant from their site of synthesis.
    [0024]
    [0025] In postharvest practices, treatments with specific eliciters have been used to improve phytochemical content and food quality such as broccoli (Villarreal-García D. et al (2016) Front Plant Sci. 10; 7: 45). The results concluded that a simple and effective treatment with MeJA improved the content of individual glucosinolates and phenolic components in broccoli during storage, protecting it from contamination in wounds.
    [0026]
    [0027] Research in recent years has demonstrated the potential of the application of JA and MeJA in Brassicas to increase the concentration of bioactive compounds such as glucosinolates. Almost all articles are focused on applications and determinations in sprouts (Baenas, N. et al. (2014) Journal of Agricultural and Food Chemistry, 62, 1881-1889) and in leaves of young plants (Yi, GE et al. ( 2016) Molecules 24, 21 (10)), that is, in plants without developed cuticle, although some research with inflorescence application also appears (Kim, HS et al. (2001) J. Amer. Soc. Hort. Sci. 136 (4): 239-246).
    [0028]
    [0029] On the other hand, surfactants, also known as surfactants are substances that influence by means of surface tension on the contact surface between two phases and that can, for example, facilitate the penetration of molecules (biocides, fertilizers, eliciters, etc.). ..) in plants. The number of surfactants known in the state of the art is very high, and their activity and effect depend on many factors, such as the plant species, their physiological state (presence of cuticle or not) or environmental conditions. It may even cause negative effects on the color or shape of the edible parts of the plant, including the edible part of Broccoli ( Brassica oleracea ssp. Italica) when combined with methyl jasmonate (J Sci Food Agric 2014; 94: 2090- 2096), east aspect that supposes a depreciation or reduction of the production very negative for the producer.
    [0030]
    [0031] The object of the present invention is a composition that is based on the combination of an elicitator such as methyl jasmonate and a surfactant such as polyether polysiloxane, which can be used by foliar applications in adult plants of the Brassica genus that have a developed cuticle, preferably in those used in human food and more preferably in broccoli, achieving a significant increase in glucosinolate concentrations (mainly glucorrafanine and neoglucobrasicin) in florets, without degradation of their organoleptic characteristics. In addition, its application allows the appearance of a secondary metabolite that are identified for the first time in plants of the order Brassicales, specifically a feruloyl-indolglucosinolate (hereinafter, feruloyl-GLS).
    [0032]
    [0033] DESCRIPTION OF THE INVENTION
    [0034]
    [0035] In a first aspect, the present invention relates to a composition characterized in that it comprises methyl jasmonate (MeJA), a polysiloxane polyether, water and a C2-C4 alcohol, preferably ethanol.
    [0036]
    [0037] In a preferred embodiment, the C2-C4 alcohol is ethanol.
    [0038]
    [0039] In a more preferred embodiment the composition of the invention consists of methyl jasmonate (MeJA), a polysiloxane polyether, water and a C2-C4 alcohol, preferably ethanol.
    [0040]
    [0041] The polyether polysiloxane is a copolymer of polysiloxane modified with polyether, preferably with surfactant activity. In the present invention the polyether polysiloxane will also be referred to as modified polyether polysiloxane (PPM).
    [0042]
    [0043] Polyether polysiloxanes suitable for use in the present invention are known in the state of the art and are commercially available. For example, modified polyetherpolysiloxanes are available under the brand Break-Thru 9902 ™, Break-Thru 9903 ™, Break-Thru 5503 ™, Break-Thru 9907 ™ and Break-Thru 9908 ™.
    [0044] More preferably, the polysiloxane polyether is a non-ionic trisiloxane, for example, available under the brand name BREAK-THRU® S 233.
    [0045]
    [0046] The concentration of methyl jasmonate in the final composition is between 50 and 500 pmolar, more preferably the concentration of methyl jasmonate in the composition is 100 pmolar.
    [0047]
    [0048] In a preferred embodiment of the invention, the composition comprises
    [0049] - methyl jasmonate between 0.0015% and 0.0028% by volume,
    [0050] - polyether polysiloxane between 0.068% and 0.1272% by volume,
    [0051] - C2-C4 alcohol between 0.15 and 0.28% by volume, and
    [0052] - water until 100% of the volume is completed (that is: water between 99.7805 and 99.59% by volume).
    [0053]
    [0054] A more preferred embodiment of the composition of the invention consists of: 0.0022% by volume of MeJA, 0.2% by volume of ethanol, 0.1% by volume of polyether polysiloxane and water to a total volume of 100%.
    [0055]
    [0056] The use of MeJA as elicitator in combination with a polysiloxane polyether results in a significant increase in glucosinolate concentrations and the generation of a feruloyl-GLS metabolite in the plants to which it is applied, even using minimal amounts of the MeJA elicitator.
    [0057]
    [0058] Another aspect of the invention relates to a process for increasing the content of glucosinolates in an adult plant of the Brassica genus which comprises the following steps:
    [0059]
    [0060] a) make a first foliar application of the composition defined in the first aspect of the invention at the time of appearance of the floral button on the plant; b) perform a second foliar application of the composition defined in the first aspect of the invention five days after the first application;
    [0061] c) perform a third foliar application of the composition defined in the first aspect of the invention five days after the second application.
    [0062]
    [0063] This procedure produces increases in glucosinolate content both in the inflorescence and in the leaves without any reduction in the organoleptic or commercial properties, preferably production, color, compaction, shape and taste of the former.
    [0064]
    [0065] In the present invention, the glucosinolates are preferably selected from glucorrafanine, glucobrasicin, methoxyglucobrasicin, neoglucobrasicin and combinations thereof, more preferably, neoglucobrasicin.
    [0066]
    [0067] Additionally, and surprisingly, by this method, at least one compound (secondary metabolite), preferably a feruloylindole glucosinolate, in which a phenolic acid and an indole glucosinolate (tentatively, neoglucobrasicin, NGB) are also obtained.
    [0068]
    [0069] As far as it is known, it is the first time that this secondary metabolite has been described in plants of the Brassicales order , derived from glucosinolate and phenolic compound, which can be a compound for mobilization of glucosinolates between organs of the plant. Additionally, and since the components of the secondary metabolite have separately beneficial effects on the health of the person ingesting them, it is expected that the combination of both will also retain that effect.
    [0070]
    [0071] The magnitude of the increase in the concentration of glucosinolates obtained with the application of the composition of the invention, using a very low concentration of the MeJA elicitator (preferably not exceeding 100 pmolar), is needed in other tests carried out with the same elicitator and collected. in the state of the art, of much higher concentrations to reach equal magnitude (see examples of the present invention).
    [0072]
    [0073] In a preferred embodiment of the invention, the plant of the Brassica genus is selected from the list comprising broccoli, cauliflower, cabbage, turnip greens, Brussels sprouts, kohlrabi, turnip, eelgrass, kale and rapeseed. Even more preferably, the plant is broccoli.
    [0074]
    [0075] In a preferred embodiment of the invention, an amount between 100 and 150 ml of the composition is applied in each of the foliar applications per plant each plant.
    [0076] Preferably, the foliar application of the composition is by spray.
    [0077]
    [0078] In a more preferred embodiment of the process of the invention:
    [0079] - The foliar application of the composition is by spray applying an amount between 100 and 150 ml of the composition per broccoli plant in each application.
    [0080] - The composition comprises 0.0022% by volume of MeJA, 0.2% by volume of ethanol, 0.1% by volume of polyether polysiloxane and water to a total volume of 100%.
    [0081]
    [0082] The process of the invention allows to obtain functional foods that, in addition to their usual nutritional effects, have biological compounds with added positive effect on one or several functions of the organism and which have beneficial effects on health, improving it or reducing the risk of suffering from diseases . All this because the synthesis and content of glucosinates and at least one secondary metabolite derived from glucosinolate and phenolic compound (feruloyl-GLS) in the different tissues of plants, including florets, are greatly increased concentrations of methyl jasmonate when the plants have developed their cuticle, without damaging them.
    [0083]
    [0084] A final aspect of the present invention relates to the use of the composition to improve defense mechanisms against herbivores and pathogens in a Brassicales plant .
    [0085]
    [0086] Until now, in the state of the art, the possibility of increasing the glucosinolates of plants of the Brassica genus has been described, such as broccoli by means of methyl jasmonate, alone or in combination with a surfactant, both in sprouts and in leaf and In inflorescence. In the present invention, methyl jasmonate in combination with a polyether polysiloxane is applied in a reduced concentration in leaves and achieves an increase in glucosinolates in leaves and in florets, as well as the generation of at least one secondary metabolite derived from glucosinolate and phenolic compound. , a feruloil-GLS, with only three applications. This increase is also observed in different varieties of broccoli. Additionally, the composition of the invention does not cause a reduction of the organoleptic properties as described in the publications included in the state of the art and demonstrates in the results included in table 4 of the present invention, using a combination of methyl jasmonate and a surfactant of different nature.
    [0087]
    [0088] In the present invention, the term "adult plant" refers to a plant that has finished its vegetative period passes to the reproductive period. Adult plants have a cuticle thicker than those found in the initial stages of the vegetative period. In fact, in the reproductive phase of the development of the plant, the cuticle of the leaf reaches its maximum thickness.The cuticle is a layer of lipid polymers impregnated with waxes that is present on the external surfaces of the primary organs of all vascular terrestrial plants The cuticle forms a homogeneous external covering whose function is to prevent water permeability to prevent evaporation from the cells of the epidermis.
    [0089]
    [0090] The term "floral button" refers to the primordium of the inflorescence that starts from the apical bud of the main stem. A stage in which the development of the inflorescence covered by small leaves begins for protection.
    [0091]
    [0092] The term "floreta", "floreta" or "pella" refers to the immature or developing inflorescence. It is the main organ of food consumption in the form of a compound corymbus, developed from the floral button. It consists of numerous floral primordia. held in floral stems or pedicels, which in turn are arranged on succulent peduncles.
    [0093]
    [0094] The term "organoleptic properties" refers to the properties that are measured through the senses. This sensory analysis is based on the basic parameters of color, shape, size, aroma, texture, taste and temperature. In the present invention, preferably They refer to color, compaction, roundness and flavor.
    [0095]
    [0096] The term "C2-C4 alcohol" refers to an organic alcohol with a carbon number ranging from two to four. For example: ethanol, propanol, butanol, tect-butanol, isobutanol.
    [0097]
    [0098] Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.
    [0099]
    [0100] BRIEF DESCRIPTION OF THE FIGURES
    [0101]
    [0102] FIG. 1: Basic structure of a glucosinolate.
    [0103]
    [0104] FIG. 2: Chemical structure of a feruloyl-indole-glucosinolate, in which the indolic glucosinolate is neoglucobrasicin.
    [0105]
    [0106] FIG. 3: Photograph showing the dispersion of a drop when methyl jasmonate is added with water, Triton X-100 and PPM (polyether polysiloxane modified).
    [0107]
    [0108] Fig. 4. Absorption spectrum (UV-Vis), with the maximum absorption peaks coinciding with a feruloil-GLS, the arrows indicate the maximum absorption peaks at 227 ± 3 nm (GLS) and 320 ± 16 nm (hydroxycinnamic acids like the ferulic).
    [0109]
    [0110] EXAMPLES
    [0111]
    [0112] The invention is illustrated below by tests carried out by the inventors, which show the effectiveness of the composition and process of the invention.
    [0113]
    [0114] Material and methods
    [0115]
    [0116] Field test
    [0117]
    [0118] Seedlings of three varieties of broccoli ( Brassica olerácea var. Italica) obtained from a seedbed where the seeds grew for 30 days were used. Specifically, the varieties were Ares, Parthenon and Marathón, all of them from the SAKATA company. The experiment was carried out during the autumn / winter period in an Experimental farm in the Mirador field in the municipality of San Javier, (Murcia) under a semi-arid Mediterranean climate. Average Daily temperature and relative humidity were calculated from measurements made every 10 minutes using a data logger. The humidity reached in the plot was 50/80% (day / night) and the air temperature ranged between 24/8 ° C day and night respectively.
    [0119]
    [0120] A total of 40 plants were planted for cultivation, 20 for each treatment. All seedlings were planted on the same day, under the same conditions and irrigated with fertirrigation [2 dS m-1 electrical conductivity (EC)] twice a day for 10 minutes during the first 45 days, going to 15 minutes during the following 45 days
    [0121]
    [0122] Once the reproductive phase was reached, this is when the floral button appears (approximately after 120 days from sowing the seeds in the seedbed) the treatments were applied using Composition 1 and Control Composition that were performed three times on the leaf. One at the time of the appearance of the floral button and the following at 5 and 10 days respectively after the first application. Each application was made by spray, applying 100-150 ml per plant. The leaves and inflorescences were collected for analysis once they reached their commercial size (approximately 18 days after the appearance of the flower bud) and frozen in liquid nitrogen and lyophilized immediately to avoid degradation of the compounds of interest until moment of determination of glucosinolates and other metabolites.
    [0123]
    [0124] During the application of the treatments the irrigation by fertirrigation was prolonged in the same conditions in which it was carried out before the application of the treatments.
    [0125]
    [0126] Composition 1: A solution was prepared containing 0.0022% by volume of MeJA (for a final concentration of 100 micromolar in the composition), 0.2% by volume of ethanol, 0.1% by volume of polyether polysiloxane (Break- Thru S 233 ®) and 99.6978% by volume of water.
    [0127]
    [0128] Control Composition: The control treatment consisted in the application of a solution containing 0.1% by volume of polysiloxane polyether and 99.9% by volume of water.
    [0129] Greenhouse test
    [0130]
    [0131] In the culture chamber, seeds of the broccoli variety ( Brassica olerácea var. Italica) Parthenon were grown, which grew for 30 days. Subsequently they were allowed to grow in the greenhouse for 75 days until the inflorescences reached commercial size. In this case a foliar application was made by spray of between 100 and 150 ml per plant of the treatments indicated below: one at the time of the appearance of the floral button and the following at 5 and 10 days respectively after the first application .
    [0132]
    [0133] Control composition: consisting of a solution containing 0.2% by volume of ethanol and 99.8% by volume of water.
    [0134]
    [0135] Surfactant composition 1 (Triton X-100): consisting of a solution containing 0.1% by volume of the Triton x-100 surfactant and 99.9% by volume of water.
    [0136]
    [0137] Surfactant composition 2 (PPM): consisting of a solution containing 0.1% by volume of the modified polyether polysiloxane surfactant and 99.9% by volume of water.
    [0138]
    [0139] Elicitator composition (MeJA): consisting of a solution containing 0.0022% by volume of MeJA, 0.2% by volume of ethanol and 99.7978% by volume of water.
    [0140]
    [0141] Composition 2 (MeJA Triton x-100): consisting of a solution containing 0.0022% by volume of MeJA, 0.2% by volume of ethanol, 0.1% by volume of Triton X-100 and 99 , 70% by volume of water.
    [0142]
    [0143] Composition 1 (MeJA PPM): A solution was prepared containing 0.0022% by volume of MeJA (for a final concentration 100 micro molar in the composition), 0.2% by volume of ethanol, 0.1% by volume of polyether polysiloxane and 99.7968% by volume of water.
    [0144]
    [0145] Subsequently the plants and leaves were harvested and the inflorescences were frozen in liquid nitrogen and lyophilized immediately to avoid degradations in the compounds of interest until the moment of the determination of glucosinolates and metabolites of interest.
    [0146]
    [0147] Determination of glucosinolates and a feruloyl-GLS
    [0148]
    [0149] The analyzes were performed on sheet and inflorescence at the time of commercial size collection. The method used for the extraction of glucosinolates was based on the method developed by the research and public domain group currently: Dominguez-Perles et al. (2010) Journal of Food Science 75, 383-392. Quantitative analysis was performed by HPLC-DAD. The lyophilized and ground samples (100 mg) were extracted with 1 ml of 70% MeOH in a hot bath at 70 ° C for 30 minutes, stirring every 5 minutes in vortex, to optimize the extraction. The reaction was then stopped in an ice bath and the samples were centrifuged to precipitate the insoluble material, for 15 minutes, at 10,000 g and at 4 ° C. The supernatant was collected and the methanol was removed on a rotary evaporator with a thermostated bath at 38 ° C. The dry residue that was obtained was redissolved in ultrapure water and filtered (00.22 pm of PVDF) for further analysis.
    [0150]
    [0151] Each sample (20 pl) was analyzed on a high-precision liquid chromatograph (HPLC) equipped with a pump for the distribution of binary eluents, in-line degasifier, photodiode detector (DAD) autosampler. The separation of the compounds was carried out on a C18 column of 250x4 mm and 0.4um pore diameter, in reverse phase liquid chromatography (RP-HPLC) mode. The identification of glucosinolates was carried out using the methodology already described by the research group and publicly available for multipurpose analysis of intact glucosinolates (Martínez-Sánchez et al. 2006 Postharvest Biol. Technol. 42, 86-97; Dominguez-Perles et al. (2010) Journal of Food Sci. 75, 383-392), based on the retention times of the respective analytical peaks, their maximum absorption spectra in the UV-Vis range and the comparison with commercially available external standards, synigrine for glucosinolates aliphatic and glucobrasicin for indole glucosinolates (Baenas et al. (2012) J. Agric. Food Chemistry, 60, 11409 11420). Detection was performed at 227 nm and the concentration of glucosinolates was expressed in pmol / g dry weight of plant tissue.
    [0152]
    [0153] The type spectrum of feruloyl-GLS (see Figures 2 and 4) in which a phenolic acid and an indole glucosinolate (tentatively, neoglucobrasicin, NGB) are joined determined by HPLC-DAD-ESI-MS / MS fragmentation, observing the parental ion of 724 (MH, m / z, 100%) and the resulting fragments of 499 m / z (100%), 259 m / z (10.2% , glucosinolate indicator) and 193 m / z (100%, feruloyl derivative indicator).
    [0154]
    [0155] The results of the application of the treatments in the two tests carried out are shown below.
    [0156]
    [0157] Field test
    [0158]
    [0159] Table 1. Concentration of glucosinolates and a feruloyl-GLS in leaves of different varieties of Brassica olerácea var. Italica) registered in the field trial using different treatments.
    [0160]
    [0161] Table 2. Concentration of glucosinolates and a feruloyl-GLS in inflorescences of different varieties of Brassica olerácea var. Italica) registered in the field trial using different treatments.
    [0162]
    [0163] In tables 1 and 2 it can be seen that the foliar application of the composition of the invention (Composition 1) in adult plants that have reached the reproduction phase and have developed cuticle causes, for plants of different cultivars of the Ares, Parthenon varieties and Marathón, an evident effect on the increase in the concentration of individual glucosinolates (GRA: Glucorrafanina, MGB: Metoxiglucobrasicina and NGB: Neoglucobrasicina), both in leaf and in inflorescence, and the appearance of a feruloyl-GLS, also in the inflorescences and in elicited leaves.
    [0164]
    [0165] Additionally, no decrease in the organoleptic properties of the inflorescences obtained in this test was recorded.
    [0166]
    [0167] Greenhouse test
    [0168]
    [0169] Table 3. Concentration of glucosinolates in leaves and inflorescences of the Parthenon variety in the greenhouse test using different treatments
    [0170]
    [0171]
    [0172]
    [0173]
    [0174]
    [0175]
    [0176]
    [0177] GR: Glucorrafanine
    [0178] GB: Glucobrasicin
    [0179] MGB: Methoxyglucobrasicin
    [0180] NGB: Neoglucobrasicin
    [0181] F-GSL: feruloil-GLS
    [0182] Tr: concentration in trace quantities
    [0183] . Dry weight
    [0184]
    [0185] As can be seen in Table 3, the application of Composition 1 (MeJA PPM), achieves increases in the concentration of glucosinolates substantially higher than the rest of treatments applied in inflorescences and leaves and, in addition, it is the only one that achieves the appearance of a feruloil-GLS. Composition 2 (MeJA Triton x-100) that also favors the increase of glucosinolates although not of a feruloyl-GLS, however, achieves a much smaller increase (specifically with composition 1 of 116%, and with composition 2 of 44 %).
    [0186]
    [0187] Table 4.- Results of the percentage of commercial inflorescences of the Parthenon variety in the greenhouse test
    [0188]
    [0189]
    [0190]
    [0191]
    [0192] As can be seen in Table 4, the application of Composition 1 (MeJA PPM) does not produce any degradation of the inflorescences, considering as degradation the irregular shape or the alteration of the color, which does occur ostensibly when another is used. surfactant
    [0193]
    [0194] Drop dispersion
    [0195]
    [0196] The dispersion of the drop of Compositions 1 and 2 and that of Elicitator Composition (MeJA) and was determined under a binocular (1.6x) (Figure 3). It can be seen that no dispersion is observed in the MeJA drop, there is more dispersion in the MeJA drop containing the Triton x-100 surfactant and a total dispersion in the drop containing the PPM surfactant.
    [0197] The calculation of the drop dispersion area indicates that Triton x-100 disperses 4 times more than water and PPM disperses 41 times more than water and 10 times more than Triton x-100 (see Figure 3) .
    [0198] 5
    权利要求:
    Claims (17)
    [1]
    1. Composition characterized in that it comprises methyl jasmonate (MeJA), a polysiloxane polyether, water and a C2-C4 alcohol.
    [2]
    2. Composition according to claim 1, characterized in that it consists of methyl jasmonate (MeJA), a polysiloxane polyether, water and a C2-C4 alcohol.
    [3]
    3. Composition according to any of claims 1 or 2, characterized in that:
    - methyl jasmonate represents between 0.0015% and 0.0028% of its total volume;
    - Polysiloxane polyether represents between 0.068% and 0.1272% of its total volume;
    - C2-C4 alcohol represents between 0.15% and 0.28% of its total volume; and - water represents the rest of the volume of the composition.
    [4]
    4. Composition according to any one of claims 1 to 3, characterized in that the C2-C4 alcohol is ethanol.
    [5]
    5. Composition according to any of claims 2 to 4, characterized in that it consists of 0.0022% by volume of MeJA, 0.2% by volume of ethanol, 0.1% by volume of polyether polysiloxane and the rest water up to a total volume of 100%.
    [6]
    6. Composition according to any one of claims 1 to 5, wherein the polysiloxane polyether is a non-ionic trisiloxane.
    [7]
    7. Composition according to any one of claims 1 to 6 wherein the concentration of methyl jasmonate is between 50 and 500 pmolar.
    [8]
    8. Composition according to claim 7, wherein the concentration of methyl jasmonate is 100 pmolar.
    [9]
    9. Procedure to increase the content of glucosinolates in a plant adult of the Brassica genre that includes the following stages:
    a) performing a first foliar application of the composition defined in any of claims 1 to 8, at the time of the appearance of the leaf button on the plant;
    b) perform a second foliar application of the composition defined in any of claims 1 to 8, five days after the first application;
    c) performing a third foliar application of the composition defined in any of claims 1 to 8, five days after the second application.
    [10]
    10. A method according to claim 9, wherein the glucosinolates are selected from the list comprising glucorrafanine, glucobrasicin, methoxyglucobrasicin, neoglucobrasicin and combinations thereof.
    [11]
    11. Method according to any of claims 9 to 10, wherein the plant of the Brassica genus is selected from broccoli, cauliflower, cabbage, turnip greens, Brussels sprouts, kohlrabi, turnip, kohlrabi, kale and rapeseed.
    [12]
    12. Method according to claim 11, wherein the plant is broccoli.
    [13]
    13. Method according to any of claims 9 to 12 wherein an amount of between 100 and 150 ml of the composition is applied in each of the applications.
    [14]
    14. Method according to any of claims 9 to 13 wherein the foliar application of the composition is by spray.
    [15]
    15. Method according to any of claims 9 to 14, wherein:
    - The composition comprises 0.0022% by volume of MeJA, 0.2% by volume of ethanol, 0.1% by volume of polyether polysiloxane and water to a total volume of 100%.
    [16]
    16. Use of the composition according to any of claims 1 to 8, to improve defense mechanisms against herbivores and pathogens in a Brassicales plant .
    [17]
    17. Use of the composition according to any of claims 1 to 8 to obtain a functional food.
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    同族专利:
    公开号 | 公开日
    ES2737454B2|2022-02-21|
    WO2020008089A1|2020-01-09|
    BR112021000085A2|2021-04-20|
    US20210282344A1|2021-09-16|
    EP3831198A1|2021-06-09|
    EP3831198A4|2022-02-23|
    引用文献:
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    优先权:
    申请号 | 申请日 | 专利标题
    ES201830674A|ES2737454B2|2018-07-05|2018-07-05|COMPOSITION AND PROCEDURE TO INCREASE THE CONTENT OF GLUCOSINOLATES IN ADULT PLANTS OF THE GENUS BRASSICA|ES201830674A| ES2737454B2|2018-07-05|2018-07-05|COMPOSITION AND PROCEDURE TO INCREASE THE CONTENT OF GLUCOSINOLATES IN ADULT PLANTS OF THE GENUS BRASSICA|
    EP19830689.6A| EP3831198A4|2018-07-05|2019-06-28|Composition and method for increasing the content of glucosinolates in adult plants of the genus brassica|
    BR112021000085-2A| BR112021000085A2|2018-07-05|2019-06-28|composition and method to increase the content of glucosinolates in adult plants of the genus brassica|
    US17/257,743| US20210282344A1|2018-07-05|2019-06-28|Composition and method for increasing the content of glucosinolates in adult plants of the genus brassica|
    PCT/ES2019/070457| WO2020008089A1|2018-07-05|2019-06-28|Composition and method for increasing the content of glucosinolates in adult plants of the genus brassica|
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