Fast and portable fungal infection diagnosis and measuring NKP and ph kit for soil and water

Abstract

This invention is a fast and portable kit that is suitable for farmers and also the people who want to participate in the cultivation of plant fields and greenhouses; because it has two important features. The kit consists of five sections. The first part is dedicated to examining fungal infection in the incoming water, soil, and roots of plants, which can be used up to three times, and each time takes one to ten days for outcomes to appear. The remaining parts of the kit are designed for measuring the amount of significant elements in the soil, such as pH, nitrogen, phosphorus, and potassium. Each element has its own color chart for comparison with the seemed results, this part of the kit displays the results instantly at the moment. This helps farmers to understand how much fertilizer they should use and also if there is any threat to the plants.

Description

Fast and Portable Fungal Infection Diagnosis and Measuring NKP and pH Kit for Soil and Water

[0001] Plants are crucial for meeting human demands, especially in agriculture, which is one of the main sources of food and nutrition. Maintaining the quality and health of agricultural products is a crucial aspect of farming, and farmers have to put in a considerable amount of effort to achieve it. One of the biggest challenges that they face is the risk of fungal infections that can adversely impact the soil, water, and roots of plants. Fungal infections can reduce crop yield, poor plant growth, and even complete crop failure. Moreover, some fungal infections can also be harmful to human health, making it all the more crucial to prevent and control them effectively. Therefore, farmers must adopt various strategies such as crop rotation, fungicides, and proper irrigation techniques to safeguard their crops from fungal infections. Another important places for plants to grow are greenhouses, which are under exclusive protection. The growth and development of plants depend on various factors, including the quality of the soil and water they receive. Soil provides plants with essential nutrients while water gives them the hydration they need to thrive. However, both of these elements can become contaminated, which can negatively impact the growth and health of plants. Soil contamination can occur from the use of pesticides, fertilizers, and other chemicals, while water contamination can be caused by pollution and industrial waste. Maintaining the quality of soil and water is crucial to ensure that plants can grow and flourish in a healthy environment.

[0002] The presenting kit is divided into two sections. The first section is designed to help identify possible fungi in water, soil, and roots. The second section of the kit provides farmers with a way to measure the levels of various elements in the soil, including pH, nitrogen, phosphorus, and potassium. Each element has its own column and color chart, making it easy for farmers to ensure the right levels before fertilizing and to maintain the health of their water and soil.

[0003] G16Y 10 / 05 - A01G 9 / 14 - Y10S 435 / 81

[0004] After searching through databases such as Patent Scope, Free Patents, and Google Patents, no patents were found containing both features of this kit. However, there are separate patents for each feature.

[0005] WO 2001 / 066790 A2

[0006] Rapid and Sensitive Plate Method for Detection of Aspergillus Fumigatus

[0007] In this invention, microorganisms are identified based on a specific enzyme activity. In this kit, for the detection of Aspergillus fumigatus, using an arabinopyranoside substrate consisting of a fluorogenic, chromogenic, or fluorescent substrate. The method of carrying out the process in this invention includes steps a) formation of fungus on the membrane filter, b) reproduction of the fungus, c) removal of the membrane filter, d) contact of the said membrane filter with a solution containing arabinopyranoside substrate along with membrane permeable, e) detection of the presence of a signal which indicates the positive microcolonies in the mentioned membrane filter using the positive microcolonies in the mentioned membrane filter ChemScan™.

[0008] The type of compounds used in this invention is different from the materials mentioned in the claimed kit, and the use of this invention is to identify a type of fungus called Aspergillus fumigatus, while in the claimed kit, it is used to detect the contamination of pathogenic fungi in general. It is not designed only for a specific type of phytopathogenic fungus.

[0009] US 5558996 A

[0010] Fungus extraction method, kit, and extraction solution

[0011] This invention is used to identify the fungus Pyricularia oryzae in rice, wheat, barley, rye, and corn or banana plants. In this invention, a solution that includes acids such as hydrochloric acid, sulfuric acid, nitric acid, and dilute organic acids with a molarity of 1 to 50 mM and a linear alcohol detergent with a concentration of 1% is mixed with the plant tissue and the mixture is shaken for half an hour. It is given to identify the fungus by immunoassay method.

[0012] The composition of this solution for identifying fungal infection is different from the claimed kit, and this invention identifies a type of fungus called Pyricularia oryzae, which is found in the species of rice plants, while in the product, different types of plant pathogenic fungi appear in the designed kit.

[0013] US 5916744 A

[0014] Testing for infestation of rapeseed and other Cruciferae by the fungus blackleg infestation Leptosphaeria maculans

[0015] This invention is used to identify Leptosphaeria maculans fungus. In this invention, the polymerase chain reaction produces a reaction and the amplified DNA is separated from the mixture before detection. Each primer has about 20-25 holes. Also, the primers of the said set are derived from LMR1 DNA sequences. The amount of GC in primers is 50-60%. Tm degree. C. of primers is 55-80. The mentioned process is by placing the surface disinfected tissue in a minimal liquid fungal medium, shaking the culture for at least 3 days at low ture, and finally collecting the fungal mycelium from the medium by centrifugation using an organic solution and DNA precipitation.

[0016] The compositions and methods used in this invention are different from the claimed kit because this invention is used to identify the fungus Leptosphaeria maculans, while in the claimed kit, different species of plant pathogenic fungi are identified.

[0017] CN103650814

[0018] Method for measuring use rate of nitrogen balance nitrogen fertilizer of greenhouse pot experiment

[0019] The invention relates to a measuring method for nitrogen fertilizer utilization and discloses a method for measuring the use rate of nitrogen balance nitrogen fertilizer in a greenhouse pot experiment. The method comprises the following steps selecting representative surface soil; arranging the pot experiment, carrying out management, recording the grain yield and the straw yield when crops are ripe, and reserving samples for analysis; measuring the base soil total nitrogen content, the grain total nitrogen content, the total nitrogen content of soil in each pot after the crops are harvested. The calculation formula is REN[balance]=U / (FN+SN), wherein the REN[balance] represents the nitrogen fertilizer use rate calculated through a nitrogen balance method, the U represents harvest nitrogen elements, the FN represents fertilizer nitrogen elements, and the SN represents the storage capacity reduction quantity of nitrogen elements of soil of a soil body. The method is short in measuring time and accurate in measuring results, influence caused by the external environment on the method is low, the considered factors are comprehensive and reasonable, the measuring result is stable and reliable, fertilizer application can be well guided according to the recommended, amount calculated through the method, of applied fertilizer, and the method has high market popularization value and broad market prospect.

[0020] This invention is only applicable to the element of nitrogen, while the claimed kit covers phosphor, potassium, and pH. Also, this invention can be used only in greenhouses, but the kit is also able to be used in farms and houses.

[0021] United States Patent Application 20190101505

[0022] Field Measurement of Soil Element Concentration

[0023] In an embodiment, a system for measuring soil element concentration in a field in real-time is disclosed. The system comprises an extraction apparatus coupled to a mobility component configured to move the system in the agricultural field. The extraction apparatus is configured to receive a plurality of soil samples successively from a soil probe coupled to the mobility component when the mobility component is operating. The extraction apparatus contains an extractant solution that is a solvent of the soil samples. In addition, the extraction apparatus comprises a mixer that is configured to mix the soil samples with the extractant solution, thereby forming a solution mix. The system also comprises a chemical sensor coupled to the extraction apparatus, the chemical sensor configured to measure the concentration level of a soil element in the solution mix. Furthermore, the system comprises a processor coupled to the chemical sensor, the processor is configured to calculate a concentration level of the soil element in each of the plurality of soil samples after the soil sample is received by the extraction apparatus and before a successive soil sample is received by the extraction apparatus.

[0024] The only similarity between the mentioned invention and our claimed kit is that they both aim to measure the elements in soil, however, the kit only measures exclusive elements, and also their methods are different because the earlier patent has a specific mobile component but the kit has its spots on it for the sample to be put in.

[0025] The fast and portable kit is a combination of two smaller kits with different functions. The first part is about examining the possible contamination of fungus in water, soil, and roots. For this purpose, a PDA plate is prepared in the kit so that the samples can be cultivated in them, but only water can be directly put in the kit. The soil must be diluted in distilled water and the root must be kept in alcohol to be cleaned and then enter the plate. After that, the plate should be vacuumed and placed in a dark and low temperature environment. The results will appear in one to three days, also this plate is able to be used three times.

[0026] The other part of the kit includes four kits to measure the NKP and pH of the soil. The sample must be diluted in water and settled. Then it is placed in the kits. Since each element has its kit, they have specific reagents and chart colors too. The sample soil reacts to the reagents and obtains a color. Eventually, the shade of color will be compared with the chart color above the kits and the amount of each element in the sample soil will be recognized.

[0027] Due to the increasing global population and growing food demands, the agricultural industry has undergone significant advancements. Additionally, excessive use of fertilizers can lead to contamination of soil and underground water due to leaching. Pathogenic fungi can contaminate various agricultural and greenhouse environments, including hydroponic and soil greenhouses, as well as apartment flowers.

[0028] Pollution in the farming field can come from various sources such as contaminated incoming water, human activity, the entry of polluted soil or seeds, and more. In addition, the incorrect use of agricultural fertilizers can lead to extra costs for farmers and lower crop productivity. On the other hand, excessive use of fertilizers can accumulate harmful chemicals in crops and negatively affect human health.

[0029] Given the climate of most regions in Iran and the country's water scarcity issues, farmers must be equipped with the necessary training and tools to manage the use of soil and water. This will enable them to effectively regulate the amount of fertilizer used, which is a critical aspect of proper farm management.

[0030] It is feasible to determine the nutrient levels of soil by sending soil samples to agricultural laboratories. Similarly, to investigate fungal contamination, soil, water, or root samples must be sent to relevant laboratories. However, the process of receiving test results is both time-consuming and costly.

[0031] Due to the high cost and time-consuming sort of soil and water testing, some farmers are hesitant to send samples regularly. As a result, contaminated areas may go undetected until the contamination has spread. This often leads to farmers fertilizing their crops based on experience rather than accurate measurements of soil nutrients.

[0032] This invention is a cost-effective and advanced analyzing kit that can accurately determine the acidity of soil and the levels of three essential elements needed by plants - nitrogen, phosphorus, and potassium. In addition, it can detect fungal and pathogenic contamination and determine the presence or absence of such contamination in water, soil, or plant roots, regardless of the type of fungus, all within 30 minutes. This technology helps determine the optimal level of fertilizer use based on the level of nutrients and soil acidity, thus minimizing fertilizer loss and leaching.

[0033] The claimed invention is an advanced kit that comprises two features. These features can identify fungal infections in water, soil, and plant roots. Additionally, the kit can measure the soil's acidity level and the presence of three vital elements - nitrogen, phosphorus, and potassium - required for plant growth. Many agricultural environments and greenhouses, such as hydroponic and soil-based ones, as well as apartment flowers, are prone to contamination by pathogenic fungi or other pathogens. As a result, most farmers send samples of water, soil, or plant roots for laboratory testing every three to six months to make sure that there is no fungal contamination present. Pollution can enter the agricultural environment through various means such as contaminated water, people, soil, and seeds. In the case of hydroponic greenhouses, the water entering the greenhouse must be continuously tested as they are more at risk of contamination.

[0034] Plant diseases are caused by various types of fungi, which add up to about 8000 species. Different species of fungi can cause diseases in plants such as Magnaporthe oryzae, Botrytis cinerea, Puccinia spp, Fusarium graminearum, Fusarium oxysporum, Blumeria graminis, Mycosphaerella graminicola, Colletotrichum spp, Ustilago maydis, and Melampsora lini. Fungi grow in various pH environments. Some thrive in acidic and some in alkaline conditions.

[0035] Each package of this product contains five kits, and each kit consists of five experimenting spots. A plate is included in the kit to detect the presence of fungal contamination, and it can be used for up to three tests. Additionally, there are four other kits included in the kit that can be used to test the levels of different elements in the soil. Each of these four kits can be used for one test.

[0036] To check for fungal contamination in water, soil, and plant roots, samples need to be taken. For water samples, take about 1 ml of the aqueous sample and pour it into the plate.

[0037] The prepared place for this purpose contains a PDA plate of which 2 percentages of plate constituents are 3% PDA, at least 0.001% antibacterial, and at least 0.01% lithium chloride and water to excess the mass.

[0038] Then, vacuum the plate with tape or parafilm and place it in a dark room for 1 to 3 days at a low temperature. This will allow any possible fungi to grow, and the presence or absence of fungal contamination can be determined.

[0039] If the sample is soil, it needs to be diluted with distilled water and then placed on the plate. After that, it should be vacuumed to ensure there's no exchange with the environment. In the case of a root sample, the sample must be placed in 70% alcohol to destroy bacterial strains. Once it is done, the root sample should be placed in distilled water and then transferred to the plate by sterile forceps. The kit should be covered by tape or parafilm and vacuumed again. Finally, the kit should be placed in a dark environment with a low temperature. After 1 to 3 days, if there's any fungus in the plate, it will grow and show up as a halo.

[0040] Each process related to fungal contamination takes a few days, but the other part of the kit displays the result quickly and instantly. The other four kits measure the concentration of nitrogen, phosphorus, potassium (NKP), and the level of soil acidity (pH). The farmers determine the amount of the mentioned parameters on the spot and quickly, and based on that, they begin the fertilization. This is done with the help of four reagents, which are:

[0041] 1- The pH reagent is composed of three reagents for its identification: 3% phenolphthalein, 6% methyl red, 4% bromothymol blue, and 87% calcium carbonate filler.

[0042] 2- Nitrogen identification reagent is obtained through the formation of colored diazonium salt. This reagent consists of a combination of multiple solid reagents, including 5% manganese sulfate, 8% sulfanilic acid, 2% from -1 naphthylmethylamine, and 85% calcium carbonate filler.

[0043] 3- Phosphorus identification reagent is obtained through the formation of a blue complex. This reagent consists of a combination of multiple solid reagents, including 5% sodium molybdate, 10% potassium persulfate, 7% citric acid, and 78% calcium carbonate filler.

[0044] 4- Potassium identification reagent is obtained through the formation of diazonium-colored salt. This reagent consists of a combination of multiple solid reagents, including 8% sodium cobalt nitrite, 2% naphthylamine, 1% citric acid, and 89% calcium carbonate filler.

[0045] These produced reagents are located in the kits separately, and each one has a special color assigned to it, according to each color, each substance can be distinguished. These colors are placed on the four parts in the form of color charts.

[0046] The section related to pH has a green tab that has four classifications from pale green to deep green.

[0047] The section related to nitrogen has a pink tab that has four classifications from pale pink to rich pink.

[0048] The section related to phosphorus has a blue tab that has four classifications from pale blue to deep blue.

[0049] The section related to potassium has an orange tab that has four classifications from pale orange to deep orange.

[0050] At the end of each of the four kits, there is a deepened spot where the soil sample can be put in. It is important to mix the soil with water before pouring it into these spots. After that, the mixture should be dissolved with the reagents provided in the kit. Each test will produce a specific color that can be compared to the color chart to determine the pH level or concentration of elements in the soil.

[0051] The function of these reagents is to alternate the color with the alternation of pH or the concentration of NKP elements, and the level of these parameters can be determined based on the intensity of the color created in comparison with the standard color chart. The pH indicator is yellow in acidic soil, but it is muddy green in loamy soil. In the nitrogen reagent, the color of the soil solution containing the reagent becomes pink, so pale pink indicates a low amount of nitrogen, medium pink indicates an average amount of nitrogen, and deep pink indicates a high level of nitrogen in the soil.

[0052] In the phosphorus reagent, the color of the soil solution containing the blue reagent is changed. Hence, pale blue indicates a low level of phosphorus in the soil, and as the intensity of the blue color increases, it indicates a higher level of phosphorus in the soil. The color of the soil solution containing potassium reagent is orange. Hence, a low color intensity indicates a low level of soil potassium and a high color intensity indicates a high level of soil potassium.

[0053] 1 - A simple and user-friendly method for all consumers, without the need for training or prior knowledge.

[0054] 2 - Obtaining the results quickly.

[0055] 3 - The cost is lower than the expenses of water, soil, and crop protection laboratories.

[0056] 4 - Examining on-site.

[0057] 5 - Being portable.

[0058] 6 - Realizing the results with bared eyes.

[0059] 7 - Containing the two features of examining soil and identifying fungus at the same time.Brief Description of Drawing

[0060] General function of the kit.

[0061] The function of the fungal infection examining side.

[0062] The function of pH and NKP measuring side.

[0063] overviews the two aspects of the portable kit which can detect fungal infections in water, soil, and roots, as well as measuring the levels of pH, phosphorus, nitrogen, and potassium in soil.

[0064] shows the kit is designed to examine fungal infections in water, soil, and roots. Before placing the sample root in the kit, it should be soaked in 70% alcohol. The soil needs to be diluted with distilled water before being added to the kit, while the water can be poured directly into it.

[0065] validates the kit measures the acidity, nitrogen, phosphorus, and potassium levels in soil. To use it, mix a sample of the soil with water until it settles. Then, place the resulting solution in special spots for each material. Each spot has its color chart, and the result is determined by comparing it with the corresponding color shades.Examples

[0066] To test for contamination in the water that is supplied to the greenhouses, we take a sample of 1 milliliter and put it in the testing kit. Alternatively, we can mix the soil sample with distilled water and add it to the PDA plate. After that, we cover the kit with tape or parafilm and store it in a dark place. Any possible infections will be visible within the first 24 hours. We start by cleaning the root sample to check for the presence of bacteria. To do this, we put the sample in 70% alcohol for two minutes and then transferred it to distilled water in the kit space using a pipette. Once the sample is in the kit, we need to seal the kit with tape or parafilm before leaving it in a dark place. After 3 days, we can check for contamination in all three samples. Any contamination will appear as a halo. If the result is unclear, we can add 3 drops of logos solution or 5 drops of salt water to make the halo more visible.

[0067] To measure the pH, we mix the soil with water and then allow the soil to settle for 5 minutes, and then pour it into its special spot to get the color and compare it with the standard color chart to match. Determine the color and pH of the soil.

[0068] To measure the concentration of soil elements, we mix the soil sample with water in a ratio of 1 to 5 and stir thoroughly for one minute. Then the mixture is allowed to settle in the soil. Then, with a dropper, we pour it into the special chamber of each element, and by comparing the color that appears with the standard color, we find out the concentration level of that element in the soil.

[0069] Soil and hydroponic greenhouses

[0070] Apartment plants and flowers

[0071] Cultivation Farms

Claims

A kit containing a plate for diagnosing the fungal contamination in soil, roots, and water by sampling with alcohol and distilled water without requiring specific knowledge.According to Claim No. 1, the fungal contamination diagnosis kit contains a PDA plate that allows for three cultivations.According to claim No. 2, 2 percentages of plate constituents are 3% PDA, at least 0.001% antibacterial, and at least 0.01% lithium chloride and water to excess the mass.According to claim No. 3, to perform the first test sample, the soil or plant root sample should be placed in 70% alcohol to eliminate bacterial contamination, then 3 different samples can be cultivated in three different parts of the plate.According to claim No. 4, to test the soil sample, it should be diluted with distilled water and then cultivated on the plate.According to claim No. 5, thisplate can also test the water used in agriculture and greenhouses, for this purpose, water can be put directly in the place of the plate to ensure that it is free from fungal contamination.According to claim No. 6, after planting any of the samples, including roots, soil, or water in the plate, the plate should be vacuumed and placed in a dark environment, preferably at a low temperature, for 1 to 3 days until the appearance of fungal contamination in form of halos.A combination of four kits for measuring the amount of NKP and pH elements in the soil can be used by sampling with water and comparing with the color chart in real-time without requiring specific knowledge.According to claim No. 8, this combination includes four specific kits for each element containing their reagents, which are placed in the kits.According to claim 9, the pH plate agent is a mixture of its three identification reagents, 3% phenolphthalein, 6% methyl red, 4% bromothymol blue, and 87% calcium carbonate filler.According to claim 10, the Nitrogen kit reagent is a mixture of several solid reagents, including 5% sodium molybdate, 10% potassium persulfate, 7% citric acid, and 78% calcium carbonate filler.According to claim 11, Phosphorous plate reagent is a solid multi-reactive compound, including 5% sodium molybdate, 10% potassium persulfate, 7% citric acid, and 78% calcium carbonate filler.According to claim 12, The potassium plate reagent is a mixture of several solid reagents, including 8% sodium cobalt nitrite, 2% naphthylamine, 1% citric acid, and 89% calcium carbonate filler.According to claim 13, the soil sample is mixed with water and after settling, it is entered into each kit to obtain a special color due to the reaction of these reagents.According to claim 14, the pH indicator color chart has four classifications: minor, low, medium, and high from pale green to deep green.According to claim 15, the nitrogen reagent color chart has four classifications: minor, low, medium, and high, from pale pink to rich pink.According to claim 16, the Phosphorus indicator color chart has four classifications: minor, low, medium, and high from pale blue to deep blue.According to claim 17, the potassium indicator color chart has four classifications: minor, low, and medium.According to claim 18, to determine the amount of each element in the soil mixture, the color obtained by placing it in the kit is compared to the color chart provided for each element.

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