• 专利附录
  • 专利说明
  • 权利要求
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    • 专利摘要:
    A synergistic composition comprising paraffinic oil and an aqueous spore suspension of entomopathogenic fungi. in which the entomopathogenic fungus is selected from the Beauveria bassiana Bb15 strain, Metaíhizium anisopliae, Paediomyces fumosoroseus or Akanthomyces lecanil VI51, preferably Beauveria bassiana Bb15 or Akanthomyces lecanil VI51, its elaboration procedure and its use for the phytosanitary treatment of agricultural pests caused by arthropods and/or fungi. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2830475A1
    申请号:ES201931071
    申请日:2019-12-03
    公开日:2021-06-03
    发明作者:Fernandez Marta Suarez；Llorca Luis Vicente Lopez
    申请人:Universidad de Alicante；
    IPC主号:
    专利说明:

    [0004] Field of the invention
    [0006] The present invention falls within the general field of agriculture and, in particular, relates to synergistic compositions of aqueous suspensions of entomopathogenic fungi spores with paraffinic oils for the control of arthropod and fungal pests.
    [0008] State of the art
    [0010] Most countries in the world are going through a period of demographic changes that are difficult to predict, studies estimate that the population of planet earth will exceed 2.9 billion inhabitants in 2050. If it is also taken into account that climate change will favor the spread of pests in plants due to the "tropicalization" of temperate agricultural areas in the near future, highlights the great importance of ensuring food security for the human population at this time, as well as improving efficiency of food production, which involves controlling these pests in the most efficient way possible. Following this line, it is also interesting to keep food safety as preserved as possible between the process of collection and delivery to people.
    [0012] Postharvest treatment is of utmost importance, since many microorganisms cause diseases in plants and vegetables once they are harvested. Phytopathogenic fungi are the causal agents of the greatest number of diseases both in the field and in the citrus storage and conservation stage, among which the greatest pathogenic fungus of lemons stands out, which is Penicillium digitatum, also known as green mold. . This fungus only has an impact on citrus fruits but generates enormous economic losses worldwide, since it has an enormous impact on the processing, obtaining and storage of citrus fruits. It has been seen that fungi such as Akanthomyces lecanii can slow down the incidence of phytopathogenic fungi on citrus fruits (Benhamou, Phytopathology, 2004).
    [0013] A common method to control pests and diseases of crops in the field is the application of fumigants and other phytosanitary products. However, these products have a negative impact on the environment and can leave toxic residues on crops. In addition, the emergence of resistance from pests and pathogens to this type of product is increasing and represents a serious phytosanitary problem in some areas of the world. The need to reduce the use of chemical synthesis phytosanitary products implies the development of new strategies for the sustainable management of pests and diseases. One of these strategies is biological control, which can be defined as the study and use of a parasitic or pathogenic organism for the regulation of pests.
    [0015] Tetranychus urticae Koch (mite: Tetranychidae), known as the red spider, is one of the most polyphagous herbivorous arthropods, it feeds on more than 1,100 plant species belonging to more than 140 families. The use of chemical pesticides is the most used tactic for the control of T. urticae, but this species quickly develops resistance to these compounds (Van Leeuwen et al., Insect Biochemistry and Molecular Biology, 2010). Other species such as the red mite ( Panonychus citri), oriental mite ( Euttetranychus orientalis), red louse ( Aonidiella aurantii) and white louse ( Aspidiotus nerii) cause pests in crops. To carry out the biological control of these pests, entomopathogenic fungi such as Beauveria bassiana, Metarhizium anisopliae or Paecilomyces fumosoroseus are used (Ullah and Lim, Journal of Economic Entomology, 2015; Shi and Feng, Biological control, 2004). These biological control agents are used in the field in flood strategies. Multiplication of the antagonist in the field after multiple applications ensures control of insect populations.
    [0017] The use of paraffinic oils together with spores of entomopathogenic fungi provides a means in which the conidia disperse in a better way and helps them remain in the fruit, plant or pathogen, so that the fungus can develop correctly and lead to perform its function.
    [0019] Patent application WO9500020 discloses a method of producing a composition of paraffinic oils with conidia of entomopathogenic fungi, however, the procedure described in this patent involves a problem due to spore breakage that occurs when the oil is added thereto.
    [0021] In Samish et al, Veterinary Parasitology 2014, several compositions are described made from different types of oils with Metarhizium brunneum spores to control Rhipicephalus annulatus ticks. However, these compositions are not applicable for the control of mites and fungi pests, to avoid the appearance of post-harvest diseases.
    [0023] Following these considerations, it is necessary to develop compositions to apply on different types of crops in a preventive way when the disease has not yet manifested itself but the organism that produces these diseases is already in the tree.
    [0025] Description of the invention
    [0027] In the present specification the terms spores and conidia are used interchangeably.
    [0028] In the present specification, the term "post-harvest treatment" refers to the treatment of the crop with the synergistic composition disclosed in such a way that the organism responsible for the injury is controlled and thus the disease does not manifest itself when the fruit is picked and stored.
    [0030] The present invention relates to a synergistic composition comprising paraffinic oil and an aqueous suspension of entomopathogenic fungal spores.
    [0032] The paraffinic oil can have a concentration between 40% and 90% weight / volume (w / v) in water, preferably between 50% and 80% w / v and even more preferably between 60% and 72% before be mixed with the aqueous suspension of entomopathogenic fungi spores.
    [0034] The entomopathogenic fungus, the spores of which are used in the synergistic composition, can be selected from among the species belonging to the Cordycipitaceae and / or Clavicipitaceae families: Beauveria bassiana Bb15, Metarhizium anisopliae, Paecilomyces fumosoroseus or Akanthomyces lecanii Vl51, preferably Akanthomyces lecanii Vl51 or Akanthomyces bassiana Bb .
    [0036] The paraffinic oil is mixed with the aqueous suspension of spores of the fungus in a 1: 1 ratio and / or the entomopathogenic fungi spores are found in a concentration of between 5106 and 109 conidia / ml, preferably 5107 to 2.5109 con / ml still more preferably 5108 entomopathogenic fungus conidia per ml of paraffin oil and water.
    [0038] Optionally, the synergistic composition can further comprise at least one inert substance such as, for example: emulsifiers, solvents, conidia carrier compounds, aerosol propellants, fragrances and colorants. They can be present in a proportion, for example, between 0 and 10% by volume relative to the total volume of the composition. All these substances are conventional and known in the state of the art for this type of composition (Shinoda and Saito, Journal of Colloid and Interface Science, 1969).
    [0040] A further object of the present invention relates to a process for obtaining the synergistic composition. Said method comprises: using a substrate for the growth of entomopathogenic fungi, said substrate can be a cereal, preferably rice.
    [0042] The process for preparing the synergistic composition comprises:
    [0043] - filter an entomopathogenic fungus grown on a substrate through a mesh of pore size between 0.1mm to 0.5mm, preferably 0.2mm and
    [0044] - add the previous filtrate over a paraffinic oil.
    [0046] An amount between 200 and 1000 grams is used, preferably 500 grams of substrate. They are then added to a commercial bag for fungal growth. Sufficient amount of distilled water is added to said bag so that when shaken the substrate is sufficiently moistened, preferably 20% of distilled water is added on the total weight of the substrate.
    [0048] The bag is then sealed with a heat sealer and at least 2 autoclaving cycles are carried out at a temperature in the range between 90 ° C and 140 ° C, preferably 100 ° C and 130 ° C and more preferably 121 °. C for a period of time between 10 and 40 minutes, preferably 15 and 30 minutes and more preferably 20 minutes to obtain a sterile substrate on which only the desired strain of fungus grows.
    [0050] Previously, the selected species of fungus has been cultured axenically in Corn Meal Agar (CMA) plates and the conids are extracted after a growth period of 10-30 days, preferably 21 days and concentrations in distilled water of between 105 and 107 conidia / ml, preferably 106conidia / ml.
    [0052] Under sterile conditions, a corner of the bag with autoclaved substrate is cut, a volume of the conidia suspension of the desired fungus is inoculated of between 0.1 and 10 ml, preferably 0.5 and 5 ml of the conidia suspension of the fungus and even more preferably 1 ml of said suspension whose concentration is between 105 and 107 conidia / ml, preferably 106 conidia / ml and the bag is resealed. It is shaken well so that the inoculated conidia are distributed over the substrate and it is kept growing in a room in dark conditions, low humidity (for for example, using a commercial dehumidifier) and constant temperature between 15 ° C and 28 ° C, preferably 22 ° C for a period of time between seven days and 3 months, preferably 15 days and a month, and more preferably one month. The bags are shaken promptly during this period to promote the development of the fungus.
    [0054] The contents of the bag are then poured into containers with water, in a proportion of 500 grams of initial substrate per liter of water and shaken well to dislodge the spores from the substrate and keep them in an aqueous suspension. The concentration of spores obtained after this process can be between 107 and 1010 conidia / ml, preferably 108 to 5109 conidia / ml, even more preferably 109conidia / ml. It is filtered through a mesh of pore size between 0.1 mm to 0.5 mm, preferably 0.2 mm to separate the substrate from the aqueous suspension of conidia.
    [0056] The filtrate from the previous stage is added to a container with paraffinic oils to proceed to its application in the field in a 1: 1 ratio of paraffinic oil and aqueous spore suspension. The final spore concentration is between 5106 and 109 conidia / ml, preferably 5107 to 2.5108 with / ml still more preferably 5108 conidia / ml of paraffin oil and water.
    [0058] According to a further embodiment, in the synergistic composition according to the invention, the inert substance (s) may be present in a proportion, for example, between 0 and 10% by volume with respect to the total volume. of the composition.
    [0060] Inert substances can be selected from: emulsifiers, solvents, conidium carrier compounds, aerosol propellants, fragrances, colorants and combinations thereof, and would be added to the container with the paraffinic oils and the aqueous suspension of fungal spores previously mixed in a 1: 1 ratio.
    [0062] An additional object of the present invention is the use of the synergistic composition for the phytosanitary treatment of pests and diseases of agricultural crops, with the aim of reducing the presence of pests and diseases that cause serious losses in crops of enormous economic importance.
    [0064] The pest can be a fungus, preferably Fusarium spp., Penicillium italicum, Penicillium digitatum and Colletotrichum spp.
    [0066] The pest can be an arthropod, preferably a mite, more preferably an acariform, even more preferably: spider mite ( Tetranychus urticae Koch), mite red ( Panonychus citri), oriental mite ( Euttetranychus orientalis), red louse ( Aonidiella aurantii) and white louse ( Aspidiotus nerii), even more preferably red louse ( Aonidiella aurantii).
    [0068] Agricultural crops can be selected from: herbaceous and tree crops, within these citrus fruits are selected, preferably lemons.
    [0070] A particular embodiment of the invention refers to the use of the composition in which the cultivation is arboreal and consists of applying the composition in a volume of between 1 and 3 liters per tree, preferably 2L of the synergistic composition per tree.
    [0072] Since the concentration at which the fungus is obtained after growth in the initial substrate is between 107 and 1010 conidia / ml, preferably 108 to 5109 conidia / ml, even more preferably 109conidia / ml. and mixed with paraffinic oils in a 1: 1 ratio, the final concentration of fungus applied in the culture is between 5106 and 109 conidia / ml, preferably 5107 to 2.5109 con / ml, even more preferably 5108 conidia of entomopathogenic fungus for each ml of paraffinic oil and water on each tree.
    [0074] The synergistic composition can also contain at least one inert substance in a proportion, for example, between 0 and 10% by volume relative to the total volume of the composition.
    [0076] To reduce the plague in the crop, the application of the synergistic composition is carried out every 2-5 months, preferably every 4 months.
    [0078] The synergistic composition can be applied in liquid or spray form on the leaves of the crop.
    [0080] The synergistic composition described in the present invention represents an improvement over the disclosed compositions, since, by adding the spores suspended in water to the paraffinic oil, the possibility of spore breakage is reduced. Furthermore, the present application refers to the use in agriculture of the synergistic composition described for the control of mite and fungal pests, thus avoiding the appearance of post-harvest diseases.
    [0082] Brief description of the figures
    [0084] Figure 1 shows the location of the cultivation fields in which the experiments are carried out to develop the present technology. The course is located in the "Hacienda el Pino" farm, on the Carretera El Algar-Los Urrutias s / n 30368, in Cartagena, Murcia. The coordinates of the field in which it is applied are Latitude 37 ° 40 '13.61''N Longitude 0 ° 50' 54.42 '' W. a) Plot where strain Bb15 is found (section 15), b ) plots in which the treatments are applied (in the circle).
    [0086] Figure 2 shows the infective capacity of entomopathogenic fungi on red louse samples taken in the field 30 days after the application of their spores sprayed in paraffinic oil. a) Bb15-infected egg b) and c) Bb15 and Vl51-infected females respectively.
    [0088] Figure 3 shows the pathogenicity of entomopathogenic fungal spores used in the field on Gallería mellonella larvae.
    [0090] Figure 4 shows the compatibility of paraffinic oil with entomopathogenic fungal spores used in field trials. The growth of entomopathogenic fungi in culture media with paraffinic oils added at 1ppm (parts per million), 10ppm and 100ppm does not show significant differences (ANOVA multivariate analysis with p = 0.05) with the control (medium without paraffin oil). UFC, Colony Forming Units.
    [0092] Figure 5 shows that paraffinic oil does not affect the germination of the entomopathogenic fungi spores used. The germination of entomopathogenic fungi spores in paraffinic oil suspensions at concentrations of 1ppm, 10ppm and 100ppm of paraffinic oil in CMA culture medium does not show significant differences (ANOVA, p <0.05) with the control (aqueous medium without oil paraffinic).
    [0093] Figure 6 shows that the application of the synergistic composition in the field does not affect the weight (grams) of the fruits (lemons) collected throughout the year. "A" indicates the time of application of the composition.
    [0095] Figure 7 shows that the application of the synergistic composition in the field does not affect the size (millimeters) of the fruits (lemons) collected throughout the year. "A" indicates the time of application of the composition.
    [0097] Figure 8 shows the mean infestation of red louse for each lemon of the plots with the treatments (control and synergistic composition of entomopathogenic fungi spores Bb15 and Vl51) throughout the year. "A" indicates the time of application of the composition.
    [0099] Figure 9 shows the annual evolution of the lice infection by entomopathogenic fungi from plots treated with the synergistic compositions and the control. "A" indicates the time of application of the composition.
    [0101] Figure 10 shows the incidence of postharvest diseases in treated lemons in the field the synergistic compositions (and controls) one month after their collection. "A" indicates the time of application of the composition.
    [0103] Realization examples
    [0105] Entomopathogenic fungi
    [0106] A Beauveria bassiana strain (CECT 21157) isolated from the soil of the "El Pino" farm (Figure 1a, point number 15 of the plot) is selected, in which lemons are grown, which we call Bb15, and a strain of Akanthomyces lecanii Vl51 (CECT 21156) (Asensio et al., Spanish Journal of Agricultural Research 2003), a filamentous fungus that naturally infects mealybugs (a group to which the red louse belongs). The tests that give rise to the present document is the one shown in Figure 1b.
    [0108] The entomopathogenic fungi used are capable of infecting red louse samples from fields treated with paraffin oil suspensions of said fungi (Figure 2, photos taken with light microscope). These fungi, especially Bb15, have a high virulence on G. mellonella larvae (Figure 3). The pathogenicity tests are carried out following the protocols of Ricaño et al., 2013 in the Florida Entomologist journal.
    [0110] Paraffin oil toxicity study on selected strains of entomopathogenic fungi
    [0112] Under laboratory conditions, plates of CMA "Corn Meal Agar" culture medium are axenically prepared with paraffinic oils at 1ppm, 10ppm and 100ppm. To check its effect on the entomopathogenic fungi Bb15 and Vl51, the medium is inoculated with said fungi by adding a 5mm agar cylinder containing the fungus in the center of the plate and measuring its growth radius. At 14 days the growth of each strain is checked from the agar cylinder and at 5 days its germination on a plate (previously conidia of the fungus had been sown in a plate and after 5 days how many have germinated are counted and compared treatments with control). Figures 4 and 5 show that their interaction is completely viable, having similar values to the control. Therefore, the use of paraffinic oils is compatible with the application of entomopathogenic fungi in the field.
    [0113] Composition preparation example
    [0115] The growth of entomopathogenic fungi Vl51 and Bb15 is carried out using rice as substrate. 500g of rice are weighed and added to a mushroom growth bag (Unicorn Bag), 100ml of distilled water is added and shaken so that all the rice remains moistened. After this, the bag is sealed with a heat sealer and two autoclave cycles are carried out at 121 ° C for 20 minutes to obtain a sterile substrate on which only the desired strain of fungus grows.
    [0117] Conidia of fungi Bb15 and Vl51 are removed from 21-day-old CMA plates and concentrations of 106conidia / ml are prepared in sterile distilled water.
    [0119] Under sterile conditions, a corner of the bag is cut with autoclaved rice, inoculated with 1 ml of the conidia suspension of the desired fungus and resealed to maintain sterility within it. It is shaken well so that the inoculated conidia are distributed over the rice and it is kept growing in a room in conditions of darkness, low humidity (for example, using a commercial dehumidifier) and a constant temperature of 22 ° C for a month, the bags being shaken punctually every week to promote the development of the fungus.
    [0121] After one month, the bags are transported to the place of application, opened and the content is dumped into containers with water (the content of one bag is added for each liter of water, at this time the concentration of conidia is of 109 conidia / ml), and shake well to loosen the spores from the rice grains and keep them in an aqueous suspension. Filter through a 0.2 mm pore size mesh to separate the rice grains from the aqueous suspension of conidia. It is added to a tank with paraffinic oils (Volck verano, Agrodan SA, 72% emulsifiable concentrate, registration No. 12033) in a 1: 1 ratio, obtaining a concentration of 5108 conidia per ml of paraffinic oil and water to proceed with its application in field.
    [0123] Effect of treatments on the development of lemons
    [0125] In Figures 6 and 7 it is observed that the application of the synergistic compositions of entomopathogenic fungi spores with paraffinic oils does not show significant differences with the controls (treated with paraffinic oil) in the development of the fruit on the tree throughout the year ( Tables 1 and 2).
    [0127] Table 1 shows the statistical data (Mean, SD = Standard Deviation and N = Sample size of lemons collected each month) for the weight data (in grams) of the lemons over a year (from January 2018 to January 2019) in the two plots treated with the synergistic composition formed from spores of entomopathogenic fungi (Bb15 and Vl51 respectively) and paraffinic oil and the plot treated with paraffinic oil (Control). Data showing significant differences with respect to the control treatment are marked in bold (multivariate statistical analysis ANOVA with p = 0.05).
    [0129] Table 2 shows the statistical data (Mean, SD = Standard Deviation and N = Sample size) for the length data (in mm) of the lemons throughout a year (from January 2018 to January 2019) in the two plots treated with the synergistic composition formed from entomopathogenic fungal spores (Bb15 and Vl51, 109 with / ml of final concentration) and paraffinic oil and the plot treated with paraffinic oil (Control). Data showing significant differences with respect to the control treatment are marked in bold (multivariate statistical analysis ANOVA with p = 0.05).
    [0131] Therefore, treatments with the synergistic composition comprising paraffinic oil with entomopathogenic fungal spores do not harm the harvest.
    [0132]
    [0133]
    [0134] Effect of treatments with the synergistic composition on the infestation of lemons by red louse
    [0136] Based on what is observed in Figure 8, it can be seen that, after a tree harvest cycle, treatment with entomopathogenic fungi and paraffinic oils significantly reduces the amount of infestation by red louse in the fruit (Table 3, observe preferably in September 2018 to January 2019).
    [0138] Figure 9 shows the red lice infected by entomopathogenic fungi collected from lemons of the tree in each of the months. It is observed that, after each application of 2L of spore suspension in liquid medium with paraffinic oils per tree (5108 conidia / ml), applying spray applications (applications carried out in February, June and October), the number of infected red lice by these fungi increases. Infection by Vl51 increases when temperatures are lower and the environment is more humid, and more favorable to Bb15 as more applications are made over time, despite the fact that in the summer months the temperature is higher (average of 26.1 ° C in July 2018 compared to 11.8 ° C in January 2018) or the environment is drier (average of 56% relative humidity in July 2018 compared to a relative humidity of 64% in January 2018) (Source: State Meteorological Agency. Territorial Meteorological Center of Murcia. Webs: www.econet.carm.es and www.meteomurcia.es, visited in October 2019).
    [0140] Table 3 shows the statistical data (Mean, SD = Standard Deviation and N = Sample size of lemons collected each month) for the data on infestation by red louse of lemons throughout a year (from January 2018 to January 2018). 2019) in the two plots treated with the synergistic composition formed from entomopathogenic fungi spores (Bb15 and Vl51) and paraffinic oil and the plot treated with paraffinic oil (Control). Data showing significant differences with respect to the control treatment are marked in bold.
    [0141]
    [0142] The control treatment begins, in January 2018, with an average value of 9.7 red lice per lemon. In the month of April the number of lice per fruit decreases by more than half, but in the following months it returns to the initial values. In August, only green lemons are harvested, so the value of lice in the control treatment in this month is close to 0, since lice tend to infect yellow lemons, and the ideal time of year for the development of the lice coincides with the ripening of the fruit. As of August, the control shows an increase in the amount of red lice found per lemon, until reaching in January 2019 with an average value of 27.4 red lice per fruit, so that in one year the average number of lice per lemon in this treatment will have increased by 2.5 times compared to the initial values of January 2018.
    [0144] The field treated with Bb15 had a higher infestation by red lice in January 2018 than the rest of the treatments (24.8 lice on average per lemon). A progressive decrease in insects is observed in the field from January to April, the month in which the fruits of the plot are harvested. In May, the average value of lice per fruit increases compared to the previous month. In this month an application of the fungus is carried out, so that in the months after August, a progressive reduction in the average number of insects per fruit is observed again, until it is practically zero in August. In the month of September the infestation increases and in the following months it decreases and, subsequently, it remains stable, with values of 8 to 10 lice per fruit. A reduction in the number of red lice of approximately 70% is achieved compared to the control group in January 2019: 8.7 vs 27.4, see table 3.
    [0146] The field treated with Vl51 began in January 2018 with an average of 11.9 red lice per fruit. As in the other treatments, a reduction in lice infestation is observed in the months of February to April. In May, as in the treatment with Bb15, the number of insects in the plot increases, but, after the application of the fungus, there is a progressive decrease until August. From September to November there is a slight decrease in the appearance of lice per fruit. Achieving a reduction in the number of red lice of approximately 60% compared to the control group in January 2019: 11.7 vs 27.4, see table 3.
    [0148] Finally, comparing the data from the last months of the year on the number of red lice per fruit between the control and those treated, it is observed that the treatments present significant differences with respect to the control. Therefore, it is confirmed that treatments with compositions of paraffinic oils and fungal spores Entomopathogens in the field reduce pest infestation and disease occurrence.
    [0150] Effect of entomopathogenic fungi on postharvest fungal infection
    [0152] In the lemons collected in the control plot, the presence of postharvest fungi (mainly Penicillium digitatum) decreases slightly from January to April from 14% to 8%, Figure 10, then increases with two maximums in the months of May (40% of infected lemons) and July (65% of infected lemons), in August the percentage of infection decreases to 16%, but then increases progressively until January, ending the year with 45% of infected lemons. After one year the infection of the lemons by P. digitatum in the control group has gone from being 14% to 45%, it has tripled.
    [0154] Lemons treated with the synergistic composition of Bb15 spores and paraffinic oils show a lower incidence of postharvest fungal infection than the rest of the treatments. The percentage of infection fluctuates slightly in the months of January to September, remaining at values between 10% and 30%. In October, the maximum number of infected lemons is 38%, but the infection decreases in the following months until reaching 17% in January 2019. After a year the infection has gone from being 10% in January 2018 to 17% in January 2019.
    [0156] In the lemons corresponding to the fields in which the treatment with Vl is applied, a reduction in the infection of lemons is observed in the months of January to March. In April the infection of lemons increases to 20% and remains stable until September. In October there is a new increase in infection of around 35% which, finally, decreases to 25% in January 2019. In one year the infection of lemons from this field has been reduced by half, by 50% to 25%.
    [0158] These data confirm that treatments with Bb and, above all, Vl protect lemons in the field from postharvest infections by P. digitatum. All this information is contained in Figure 10.
    权利要求:
    Claims (18)
    [1]
    1. A synergistic composition comprising paraffinic oil and an aqueous spore suspension of entomopathogenic fungi.
    [2]
    2. A synergistic composition according to the preceding claim, in which the paraffinic oil has a concentration between 40% and 90% w / v in water, preferably between 50% and 80% w / v and even more preferably between 60% % and 72% before mixing with the aqueous suspension of entomopathogenic fungi spores.
    [3]
    3. A synergistic composition according to the preceding claim, in which the entomopathogenic fungus, whose spores are used in the synergistic composition, can be selected from the species belonging to the Cordycipitaceae and / or Clavicipitaceae families: Beauveria bassiana Bb15, Metarhizium anisopliae, Paecilomyces fumosomseus or Akanthomyces lecanii Vl51, preferably Beauveria bassiana Bb15 or Akanthomyces lecanii Vl51.
    [4]
    4. A synergistic composition according to the preceding claim, in which the paraffinic oil is mixed with the spores of the fungus in a 1: 1 ratio and / or the spores of entomopathogenic fungi are found in a concentration of between 5106 and 109 conidia / ml , preferably 5107 to 2.5109 with / ml even more preferably 5108 conidia of entomopathogenic fungus for each ml of paraffinic oil and water.
    [5]
    5. A synergistic composition according to the preceding claim, comprising at least one inert substance selected from emulsifiers, solvents, conidial carrier compounds, aerosol propellants, fragrances and colorants and combinations thereof.
    [6]
    6. A synergistic composition according to the preceding claim, in which the inert substance is present in a proportion of between 0 and 10% by volume with respect to the total volume of the composition.
    [7]
    7. Process for preparing a synergistic composition according to the preceding claim, comprising:
    - filter an entomopathogenic fungus grown on a substrate through a mesh of pore size between 0.1mm to 0.5mm, preferably 0.2mm and
    - add the previous filtrate over a paraffinic oil.
    [8]
    8. Process for preparing a synergistic composition according to the preceding claim, which comprises the addition of an inert substance in a proportion comprised between 0 and 10% by volume with respect to the total volume of the composition.
    [9]
    9. Process for preparing a synergistic composition according to the preceding claim, previously comprising:
    - prepare the substrate for the growth of the fungus, said substrate is a cereal, preferably rice, by adding distilled water and at least 2 autoclaving cycles and
    - Add an inoculum of the fungus to the substrate and grow for a period of time between 15 days and a month.
    [10]
    10. Use of a synergistic composition according to any one of claims 1 to 9, for the phytosanitary treatment of pests and diseases of agricultural crops.
    [11]
    11. Use of a synergistic composition according to the preceding claim, in which the pest is a fungus, preferably Fusarium spp., Penicillium italicum, Penicillium digitatum and Colletotrichum spp.
    [12]
    12. Use of a synergistic composition according to claim 10, in which the pest can be an arthropod, preferably a mite, more preferably an acariform, even more preferably: red spider ( Tetranychus urticae Koch), red mite ( Panonychus citri), oriental mite ( Euttetranychus orientalis), red louse ( Aonidiella aurantii) and white louse ( Aspidiotus nerii), even more preferably red louse ( Aonidiella aurantii).
    [13]
    13. Use of the synergistic composition according to one of claims 10 to 12, wherein the application of the synergistic composition on the culture is carried out every 2-5 months, preferably every 4 months.
    [14]
    14. Use of the synergistic composition according to one of claims 10 to 13, wherein the synergistic composition is applied in liquid or spray form on the culture.
    [15]
    15. Use of a synergistic composition according to one of claims 10 to 14, in which the crop is arboreal and the composition is applied in a volume of between 1 and 3 liters per tree, preferably 2L.
    [16]
    16. Use of a synergistic composition according to one of claims 10 to 15, in which the applied composition has a concentration of between 5106 and 109 conidia / ml, preferably 5107 to 2.5109 conidia / ml even more preferably 5108 conidia of fungus entomopathogenic per ml of paraffin oil and water.
    [17]
    17. Use of a synergistic composition according to one of claims 10 to 16, in which the applied composition comprises at least one inert substance in a proportion between 0 and 10% by volume relative to the total volume of the composition.
    [18]
    18. Use of a synergistic composition according to one of claims 10 to 17, in which the agricultural crops are selected from: herbaceous and tree crops, within these, citrus, preferably lemon trees are selected.
    类似技术:
    公开号 | 公开日 | 专利标题
    Shah et al.2003|Entomopathogenic fungi as biological control agents
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    同族专利:
    公开号 | 公开日
    ES2830475B2|2022-01-19|
    WO2021111028A1|2021-06-10|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    ES2257213A1|2004-04-23|2006-07-16|Intrachem Bio Italia S.P.A.|Plant protection by liquid biological agent employs Beauveria bassiana to inhibit insect egg deposition on the plants|
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    US20180325120A1|2017-05-10|2018-11-15|The United States Of America, As Represented By The Secretary Of Agriculture|Compositions and methods to reduce the population of wheat-stem sawfly and hessian fly|
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    PCT/ES2020/070763| WO2021111028A1|2019-12-03|2020-12-02|Compositions of entomopathogenic fungi and paraffinic oils for controlling post-harvest diseases and pests|
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