ADHESIVE COMPOSITION FOR JOINING FUNGUS SPORES TO THE SURFACE OF VEGETABLESEEDS IN A STABLE MANNER AND METHOD FOR QUANTIFYING THE ADHESION CAPACITY

Abstract

An adhesive composition for joining fungus spores to the surface of vegetable seeds in a stable manner, characterized in that it includes an aqueous solution of fermented maize liquor and/or soya protein hydrolysates. A method and to the use of the same for quantifying the adhesion capacity by means of an indirect measurement in which fluorescent microspheres joined to the surface of seeds are quantified by means of the fluorescence microscopy technique is also disclosed. This method allows an indirect assessment of the adhesion of fungus spores to the surface of seeds in a simple manner and consequently allows the quality of the adhesive composition of the present invention to be assessed.

Claims

1. A method for quantifying the adhesion capacity of fluorescent microspheres to the surface of vegetable seeds by means of an adhesive composition, characterized in that it comprises the following steps: a) taking an adhesive composition comprising an aqueous solution of corn steep liquor and/or soya protein hydrolysates, b) mixing the adhesive composition of step a) with silica or polyethylene fluorescent microspheres with a particle size of between 1-120 microns and continuing to stir, c) placing the vegetable seeds into contact with the mixture of step b), d) drying the seeds at room temperature, e) obtaining images of the fluorescent microspheres adhered to the seeds by fluorescence microscopy, exposing them to ultraviolet (UV) light corresponding to a wavelength of 365 nm, and f) counting the fluorescent microspheres adhered to the seeds based on the images obtained in step e).

2. The method according to claim 1, wherein the adhesive composition comprises from 3 to 30 ml of soya protein hydrolysate and/or 1 to 10 g of corn steep liquor in water to complete a final volume of 100 ml.

3. The method according to claim 1 wherein step c) is implemented by submerging the vegetable seeds in the mixture of step b) in a receptacle in a volumetric ratio (seeds:final volume) of 1:3, continuing to stir the mixture for at least 1 minute between 50 and 100 rpm, concluding with decantation and removing the excess liquid from the seeds.

4. The method according to claim 1, wherein in that step c) is carried out by spraying the mixture of step b) on the vegetable seeds in a homogenous manner using a proportion of 11 of mixture per 100 kg of seeds.

5. The method according to claim 1, wherein the microspheres have a density of 0.98 g/cm.sup.3 and a particle size range of 1-120 microns.

6. The method according to claim 1, wherein said vegetable seeds are cereal seeds.

7.-10. (canceled)

11. A method for catalyzing the adhesion of fungus spores characterized in that it comprises the addition of an adhesive composition comprising an aqueous solution of corn steep liquor and/or soya protein hydrolysates to the surface of vegetable seeds.

12. The method according to claim 7, wherein said adhesive composition comprises from 3 to 30 ml of soya protein hydrolysate and/or 1 to 10 g of corn steep liquor in water to complete a final volume of 100 ml.

13. The method according to claim 7, wherein said fungus spores are arbuscular mycorrhizae spores.

14. The method according to claim 9, wherein said arbuscular mycorrhizae spores are of the genera Rhizophagus intraradices and Funneliformis mosseae.

Description

EXEMPLARY EMBODIMENTS

[0071] The following examples are embodiments of the present invention, but are not limiting of the same.

Example 1. Obtaining the Adhesive Composition of the Present Invention

[0072] For the following example, three batches of 3 adhesive compositions have been made: [0073] Sample 1: 15 ml of soya protein hydrolysate and water to complete a final volume of 100 ml of composition. [0074] Sample 2: 4 grams of fermented maize liquor and water to complete a final volume of 100 ml. [0075] Sample 3: Control sample (A) consisting of sterile water.

[0076] All the samples were obtained by the following method:

[0077] a. -) taking a suitable quantity of fermented maize liquor and/or soya protein hydrolysate,

[0078] b. -) mixing with a volume of 50 ml of water,

[0079] c. -) stirring to complete homogenization, and

[0080] d. -) adding sterile water whilst stirring to a final volume of 100 ml of final adhesive composition.

Example 2. Method for Quantifying Fluorescent Microspheres Adhered to the Surface of the Seeds by Means of an Adhesive Composition Based on Soya Protein Hydrolysate

[0081] The following example represents a proof of concept for the efficacy of the adhesive composition corresponding to Sample 1 of example 1.

[0082] The steps carried out for quantifying by fluorescence were the following:

[0083] a. -) taking Sample 1,

[0084] b. -) adding 36 mg of fluorescent microspheres and stirring to complete homogenization,

[0085] c. -) submerging the seeds to be treated in the mixture of the previous step, in a bottle with a lid in a volumetric proportion of 1:3 (volume of seeds:final volume), continuing to stir for at least 1 minute between 50 and 100 rpm and decanting the liquid and removing the excess and keeping the seeds,

[0086] d. -) drying the seeds at room temperature,

[0087] e. -) obtaining images by fluorescence microscopy, exposing the dry seeds to UV light, and

[0088] f. -) counting the microspheres adhered to the seeds based on the images obtained in the previous step.

[0089] At this point, in order to be able to assess the adhesion capacity, the seeds are placed in a container with a lid with a capacity equal to 300 ml and continued to be stirred for 15 minutes between 50 and 100 rpm with the aim of evaluating the adhesion force of the microspheres on the surface of the seed. Then, the seeds are removed from the container, they are dried at room temperature and images are again obtained by fluorescence microscopy, exposing the seeds to UV light in order to count the microspheres adhered to the seeds based on the images obtained in the previous step and thus to be able to calculate the percentage of microspheres joined to the seed after stirring by means of the following mathematical formula:

[0090] Microspheres of seeds after stirring100

[0091] Microspheres of seeds prior to stirring

[0092] Thus the percentage value of the microspheres adhered to the surface of the seeds is obtained after an aggressive stirring process.

Example 3. Method for Quantifying Fluorescent Microspheres Adhered to the Surface of the Seeds by Means of an Adhesive Composition Based on Fermented Maize Liquor

[0093] The following example represents a proof of concept of the efficacy of the adhesive composition corresponding to Sample 2 of example 1.

[0094] a. -) taking Sample 2,

[0095] b. -) adding 36 mg of fluorescent microspheres and stirring to complete homogenization,

[0096] c. -) spraying, by means of the spraying method, the mixture of step b) on the vegetable seeds in a homogenous manner using a proportion of 11 of mixture per 100 kg of seeds,

[0097] d. -) drying the seeds at room temperature,

[0098] e. -) obtaining images by fluorescence microscopy, exposing the dry seeds to UV light, and

[0099] f. -) counting the microspheres adhered to the seeds based on the images obtained in the previous step.

[0100] At this point, in order to be able to assess the adhesion capacity, the seeds are placed in a container with a lid with a capacity equal to 300 ml and continued to be stirred for 15 minutes between 50 and 100 rpm with the aim of evaluating the adhesion force of the microspheres on the surface of the seed. Then, the seeds are removed from the container and images are again obtained by fluorescence microscopy, exposing the seeds to UV light in order to count the microspheres adhered to the seeds based on the images obtained in the previous step and thus to be able to calculate the percentage of microspheres joined to the seed after stirring by means of the following mathematical formula

[0101] Microspheres of seeds after stirring100

[0102] Microspheres of seeds prior to stirring

[0103] Thus the percentage value of the microspheres adhered to the surface of the seeds is obtained after an aggressive stirring process.

Example 4. Comparative Example for Assessing the Adhesion Capacity of the Adhesive Composition on Cereal Seeds

[0104] In the experiments of examples 2 and 3, a total of 300 wheat, maize and soya seeds were used in each experiment and they were distributed in groups of 100 seeds per each class. Additionally, a control experiment was carried out following the same steps of examples 2 and 3, but substituting the adhesive composition for sterile water as a manner of control, that is to say using Sample 3 as the control sample. Each experiment was carried out three times.

[0105] Table 1 shows the number of fluorescent microspheres adhered to the surface of the seeds (averages and standard deviations) prior to and after washing the seeds with aggressive stirring. The percentages of microspheres joined to the surface of the seeds after aggressive agitation with respect to the number of microspheres joined to the surface of the seeds after normal agitation are shown in parentheses; that is to say, the percentages for retaining fluorescent microspheres after aggressive agitation are shown between parentheses.

TABLE-US-00001 TABLE 1 Wheat Maize Soya Prior to Prior to Prior to stirring After stirring stirring After stirring stirring After stirring A (control) 5.3 0.1.sup.a 2.0 0.1 (38) 29.3 1.0 18.9 1.3 (64) 26.3 1.2 20.7 1.3 (78) Sample 1 13.0 0.3 9.2 0.2 (71) 66.0 3.1 57.5 2.8 (87) 67.4 3.7 62.9 3.5 (93) Sample 2 12.5 0.4 9.7 0.3 (78) 65.8 4.4 57.6 2.6 (87) 68.4 2.9 64.8 3.0 (95)

[0106] After carrying out this experiment, it was observed that the adhesion capacity of the adhesive composition meant that the percentage of seeds maintaining the fluorescent microspheres adhered after the aggressive stirring process was significantly higher than in the control samples.