WO2010107121A1 - 新規土壌診断方法 - Google Patents
新規土壌診断方法 Download PDFInfo
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- WO2010107121A1 WO2010107121A1 PCT/JP2010/054892 JP2010054892W WO2010107121A1 WO 2010107121 A1 WO2010107121 A1 WO 2010107121A1 JP 2010054892 W JP2010054892 W JP 2010054892W WO 2010107121 A1 WO2010107121 A1 WO 2010107121A1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
- A01C21/007—Determining fertilization requirements
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/24—Earth materials
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- the present invention mainly relates to a novel soil diagnosis method using the number of soil bacteria and the circulation activity of substances as indices, and a soil quality control method and improvement method using the diagnosis method.
- Microorganisms play an important role in the conversion and circulation of substances in various environments including soil. For example, in order to convert nitrogen fertilizer into a form that is taken up by agricultural crops in farmland, it is necessary for microorganisms to “nitrify”.
- Non-Patent Documents 1 and 2 soil evaluation has been centered on analysis for chemical farming methods, which have been mainly determined by physicochemical properties such as the concentration and pH of various inorganic ions, and the activity of microorganisms has not been considered.
- the present invention provides a soil diagnosis method, a quality control method, and an improvement method in consideration of material circulation by soil microorganisms, and particularly provides a farmland quality diagnosis method, a management method, and an improvement method suitable for plant growth. This is the main issue.
- the present inventor has conducted extensive studies on soil evaluation and diagnosis methods reflecting the activity of soil microorganisms. As a result, it was found that by combining the soil bacteria count and the analysis related to nitrogen, phosphorus and potassium, it was possible to make an appropriate diagnosis of soil, and further studies were made to complete the present invention.
- the present invention relates to the following soil diagnosis method, quality control method, and improvement method.
- Item 1 Circulating activity index calculated using the following (I) to (III): (I) Ammonia reduction rate in the target soil, (II) Soil diagnostics using phytic acid production activity from phytic acid in target soil, (III) potassium production activity from compost in target soil, and (IV) soil bacteria count in soil Diagnosis method.
- Circulation activity index is With respect to the area of the equilateral triangle formed with the reference value of the ammonia reduction rate set in advance, the reference value of the phosphate generation activity from phytic acid and the reference value of the potassium generation activity from compost as the apex, (I) Ammonia reduction rate, (II) Phosphate production activity from phytic acid, and (III) Potassium production activity from compost placed on the line connecting the corresponding vertices from the center of gravity of the equilateral triangle Item 2.
- the diagnostic method according to Item 1 which is a ratio of the area of a triangle formed with the measurement points of the above as apexes.
- Item 3 A) Nitrogen circulation activity index calculated using the following (A-1) to (A-3): (A-1) Number of soil bacteria in the target soil, (A-2) Ammonia reduction rate in the target soil, and (A-3) Nitrite reduction rate in the target soil, B) Phosphorus circulating activity index calculated using (B-1) to (B-3) below: (B-1) Number of soil bacteria in the target soil, (B-2) Phosphate producing activity from phytic acid in the target soil, and (B-3) Phosphate producing activity from compost in the target soil, And C) an index of potassium circulation activity calculated using the following (C-1) to (C-3): (C-1) number of soil bacteria in the target soil, (C-2) A soil diagnosis method characterized in that soil diagnosis is performed using at least the potassium release rate in the target soil and (C-3) potassium generation activity from compost in the target soil.
- Nitrogen cycling activity index is With respect to the area of the equilateral triangle formed with the reference value of the soil bacteria count set in advance, the reference value of the ammonia reduction rate, and the reference value of the nitrite reduction rate as a vertex, The (A-1) soil bacteria count, (A-2) ammonia reduction rate, and (A-3) nitrite reduction rate placed on the line connecting the corresponding vertices from the center of gravity of the equilateral triangle Item 4.
- the diagnostic method according to Item 3 which is a ratio of the area of a triangle formed with a measurement point as a vertex.
- Phosphorus circulating activity index is With respect to the area of an equilateral triangle formed with the reference value of the preset number of soil bacteria, the reference value of the phosphate production activity from phytic acid, and the reference value of the phosphate production activity from compost as the apex, (B-1) the number of soil bacteria placed on the line segment connecting the corresponding vertices from the center of gravity of the equilateral triangle, (B-2) the activity of producing phosphate from phytic acid, and (B-3) Item 5.
- the diagnostic method according to item 3 or 4 which is a ratio of the area of a triangle formed with each measured value of phosphoric acid production activity from compost as a vertex.
- the potassium circulatory activity index is With respect to the area of the equilateral triangle formed with the reference point of the preset soil bacteria count, the reference value of the potassium release rate, and the reference value of the potassium production activity from compost as the apex, The (C-1) soil bacteria count, (C-2) potassium release rate, and (C-3) potassium production from the compost placed on the line connecting the corresponding vertices from the center of gravity of the equilateral triangle Item 6.
- the diagnostic method according to any one of Items 3 to 5, which is a ratio of an area of a triangle formed with each measured value of activity as a vertex.
- Item 7 A soil quality control method, wherein the soil quality is managed by performing the diagnostic method according to any one of Items 1 to 6 over time and analyzing the change in the index over time .
- Item 8 A method for improving soil, comprising performing the diagnosis method according to any one of Items 1 to 6 and performing a process for improving the index based on the obtained diagnosis result.
- Soil diagnostic method 1.1 Soil diagnosis method (1) In the soil diagnosis method (1) of the present invention, the circulating activity index calculated using the following (I) to (III): (I) Ammonia reduction rate in the target soil, The soil diagnosis is performed using (II) phosphoric acid producing activity from phytic acid in the target soil, (III) potassium producing activity from compost in the target soil, and (IV) the number of soil bacteria in the soil.
- the ammonia reduction rate in the target soil is a value indicating the reduction rate of the concentration of the ammonia compound administered to the target soil.
- the ammonia reduction rate can be calculated as a value obtained by the following formula when an ammonia compound is administered to the target soil.
- Ammonia reduction rate (%) [1- (N 1 ⁇ N 2 ) / N 1 ] ⁇ 100
- N 1 represents the amount of ammonia nitrogen on the day of administration of the ammonia compound.
- N 2 represents the amount of ammonia nitrogen after a certain period from the administration of the ammonia compound.
- the ammonia compound administration date means the administration date of the ammonia compound to the target soil.
- the amount of ammonia nitrogen on the day of administration of the ammonia compound can be expressed as the amount of ammonia nitrogen on day 0 of administration.
- “after a certain period of time after administration of the ammonia compound” means the day when a certain period has elapsed since the administration of the ammonia compound to the target soil.
- the amount of ammonia nitrogen after 3 days from the date of administration of the ammonia compound to the target soil can be expressed as the amount of ammonia nitrogen on the third day of administration.
- the length of the fixed period can be set as appropriate, but is preferably 3 to 7 days, particularly 3 days after administration. When the period is shorter than this, or when it is extremely long, the difference in activity becomes difficult to distinguish.
- ammonia reduction rate in the target soil is preferably a value obtained by the following formula:
- the amount of ammonia nitrogen means the amount of ammonia nitrogen (NH 4 + ) per unit dry weight of the target soil.
- the amount of ammonia nitrogen can be measured by the indophenol method, potassium chloride solution leaching method, high performance liquid chromatography and the like. More specifically, it can be measured by the ammonia nitrogen determination method in the examples.
- ammonia compound to be administered to the target soil is not particularly limited, and examples thereof include ammonium salts such as ammonium sulfate, ammonium chloride, ammonium nitrate, and ammonium carbonate. Of these, ammonium sulfate, which is a common agricultural fertilizer, is preferably used.
- the amount of ammonia compound to be administered to the target soil is not particularly limited, but considering the concentration of nitrogen-containing compounds in general field soil, it is 30 to 100 ⁇ g-N / g-dry per unit dry weight of the target soil.
- the ammonia reduction rate reflects the conversion efficiency from ammonia nitrogen to nitrite nitrogen.
- the larger the decrease rate the more the number of ammonia-oxidizing bacteria in the target soil or the activity per unit cell mass. It is thought to contain high ammonia oxidizing bacteria.
- the smaller the decrease rate the smaller the number of ammonia-oxidizing bacteria.
- the phosphate producing activity from phytic acid in the target soil is a value indicating the conversion activity of phytic acid administered to the target soil.
- the phosphate production activity from phytic acid in the target soil can be calculated as a value obtained by the following formula when phytic acid is administered to the target soil.
- Phosphate production activity from phytic acid [(P 3 ⁇ P 2 ) / P 1 ] ⁇ 100 (In the formula, P 1 represents the amount of phosphoric acid in phytic acid. P 2 represents the amount of water-soluble phosphoric acid on the day of phytic acid administration. P 3 represents the amount of water-soluble phosphoric acid after a certain period of time from phytic acid administration. To express.)
- the amount of phosphoric acid in phytic acid can be calculated from the dose of phytic acid based on the fact that 6 moles of phosphoric acid are contained in 1 mole of phytic acid.
- the phytic acid administration date means the phytic acid administration date for the target soil.
- the amount of water-soluble phosphate on the day of administration of phytic acid can be expressed as the amount of water-soluble phosphate on day 0 of administration.
- “after a certain period of time from phytic acid administration” means the day when a certain period has elapsed since phytic acid administration to the target soil.
- the water-soluble phosphoric acid after 3 days from the day of administration of phytic acid to the target soil can be expressed as the amount of water-soluble phosphoric acid on the third day of administration.
- the length of the fixed period can be set as appropriate, but is preferably 3 to 7 days, particularly 3 days after administration. When the period is shorter than this, or when it is extremely long, the difference in activity becomes difficult to distinguish.
- the phosphoric acid production activity from phytic acid in the target soil is preferably a value determined by the following formula:
- the amount of water-soluble phosphoric acid means the amount of water-soluble phosphoric acid per unit dry weight of the target soil.
- the amount of water-soluble phosphoric acid can be measured by the molybdenum blue method, high performance liquid chromatography or the like. More specifically, it can be measured according to the method for quantifying water-soluble phosphoric acid in the examples.
- the amount of phytic acid administered to the target soil is not particularly limited, but is about 0.5 to 5% (w / w), preferably about 1 to 2% (w / w) per unit dry weight of the target soil.
- the activity of producing phosphoric acid from phytic acid reflects the conversion efficiency from phytic acid to water-soluble phosphoric acid, and it is considered that the higher the activity, the easier it is to use phosphorus contained in the plant body. From this, it is considered that the greater the activity, the better the quality of the soil, and the more the amount of externally applied phosphorus fertilizer can be suppressed.
- the potassium production activity from compost in the target soil is a value indicating the conversion activity of potassium in the compost administered to the target soil to free potassium.
- the compost is the same as that described in the above phosphorus circulation index, and examples thereof include plant compost such as bark compost, livestock compost such as bark compost, cattle manure, and pig manure compost, and seaweed compost. These may be used alone or in combination of two or more.
- bark compost contains much potassium, more appropriate evaluation becomes possible.
- the total potassium (K 2 O) content of bark compost is 0.1% or more (dry matter).
- the administration form of compost is not limited, and compost mixed with culture soil may be used.
- the mixing ratio is about 10 to 50%, preferably about 25 to 35% by weight of compost with respect to the total amount of the cultured soil.
- the potassium production activity from compost in compost soil can be calculated as a value obtained by the following formula when compost is administered to the target soil.
- K 4 represents the potassium content in the compost.
- K 5 represents the potassium release amount on the compost administration day.
- K 6 represents the potassium release amount after a certain period from the compost administration.
- the potassium content in the compost can also be quantified according to a known method.
- a filtrate obtained by adding an aqueous ammonium acetate solution to compost and filtering is used as a potassium extract, and the obtained extract is used as an atomic absorption photometer.
- the date of compost administration means the date of administration of compost to the target soil.
- the amount of potassium released on the day of administration of compost can be expressed as the amount of potassium released on the 0th day of administration.
- “after a certain period of time from the administration of compost” means the day when a certain period has elapsed from the administration of compost to the target soil.
- the amount of released potassium after 3 days from the day when compost was administered to the target soil can be expressed as the amount of released potassium on the third day of administration.
- the length of the fixed period can be set as appropriate, but is preferably 3 to 7 days, particularly 3 days after administration. When the period is shorter than this, or when it is extremely long, the difference in activity becomes difficult to distinguish.
- the potassium generation activity from compost in the target soil is preferably a value obtained by the following formula:
- the amount of potassium released can be measured in the same manner as described later.
- the potassium generation activity from compost reflects the conversion efficiency of potassium in compost to free potassium, and it is considered that the greater the activity, the easier it is to use potassium in compost. From now on, it is considered that the greater the activity, the better the quality of the soil and the less the amount of potassium input from the outside.
- the number of soil bacteria represents the number of soil bacteria obtained based on the amount of DNA present per unit weight of sample collected from the target soil.
- the number can be expressed in units of the number (cells / g-soil or cells / g-sample) per unit weight of the target soil (or sample).
- the amount of DNA here refers to the amount of DNA present per unit weight of the sample collected from the target soil. More specifically, the total amount of DNA present per unit weight of the sample is shown regardless of the origin of the DNA.
- the number of soil bacteria can be determined by converting the amount of DNA present per unit weight of sample collected from the target soil by an appropriate technique.
- the correlation between the number of soil bacteria in the soil and the amount of DNA is obtained in advance, and the amount of DNA measured from the collected sample is collated with the correlation. be able to.
- the number of soil bacteria is determined by converting the amount of DNA per unit weight of a sample collected from the target soil according to the following formula.
- the sample collected from the target soil is the soil collected (sampled) from the target soil.
- the collection method is not particularly limited, and can be appropriately performed according to a known method.
- the collection conditions can be set as appropriate, from the viewpoint of appropriately judging the state of microorganisms in the target soil, it is preferable to collect the sample while avoiding the time when the target soil is not in a normal state due to rain or the like.
- the amount of DNA per unit weight of sample collected from the target soil can be measured by eluting the DNA present in the sample collected from the soil to be diagnosed and quantifying the amount of the DNA.
- the obtained sample should be kept at a low temperature (eg, about ⁇ 4 to ⁇ 80 degrees, preferably ⁇ 20 to ⁇ 80). Can be stored for about 1 day to 3 weeks.
- a low temperature eg, about ⁇ 4 to ⁇ 80 degrees, preferably ⁇ 20 to ⁇ 80.
- the method for eluting DNA from all the microorganisms contained in the sample is not particularly limited as long as the DNA is significantly decomposed or sheared and its quantification is adversely affected.
- one embodiment of the DNA elution method is a method of treating the sample with a DNA elution solution.
- Examples of the DNA elution solution used here include solutions generally used for eluting DNA from bacteria.
- an inhibitor of a DNA degrading enzyme such as EDTA or EGTA, a cationic surfactant, a solution containing an anionic surfactant and / or a buffer containing them, etc.
- the buffer can also contain a proteolytic enzyme such as proteinase K, thermolysin, and subtilisin.
- the blending ratio of each component can be appropriately set within a range that does not significantly impair DNA extraction.
- the elution conditions for DNA are not particularly limited.
- DNA can be eluted by adding 2 to 20 ml, preferably 5 to 15 ml, more preferably 8 to 12 ml of the above DNA extraction solution to 1 g of soil subjected to the elution treatment. .
- the elution temperature can be appropriately set according to the DNA elution solution to be used and the type of soil used for the elution treatment.
- the elution time varies depending on the type of DNA extraction solution to be used, the type of soil subjected to the elution treatment, the elution temperature, etc., and cannot be defined uniformly, but as an example, 0.1 to 4 hours, preferably 0.2 Up to 2 hours, more preferably 0.3 to 1 hour can be mentioned.
- the amount of DNA present in the target soil can be determined.
- the DNA quantification method is not particularly limited, and for example, the eluted DNA can be purified and recovered as necessary, and quantified by a known or conventional DNA quantification method.
- the DNA is stained with ethidium bromide and the fluorescence intensity of the DNA band on the gel is measured. The method of doing can be mentioned.
- a method of dissolving the DNA recovered by purification in a buffer solution and measuring the absorbance at 260 nm of the solution can be mentioned.
- the method for purifying DNA is not particularly limited, and can be performed according to a conventional method.
- Examples of the method include a step of adding a DNA precipitating agent such as isopropyl alcohol, ethanol, or polyethylene glycol to the DNA-containing layer obtained in the above step to precipitate the DNA and recovering the DNA.
- the DNA extraction efficiency in each sample should be measured in advance and corrected for each target sample based on the extraction efficiency. It is desirable to determine the amount of DNA.
- the DNA extraction efficiency means the ratio of the amount of DNA actually eluted and quantified from the sample to the amount of DNA contained in the sample collected from the target soil.
- the number of soil bacteria can be determined according to the method described above.
- the total amount of DNA derived from all bacteria present in the sample reflects the overall characteristics and status of the target soil. Therefore, the number of soil bacteria obtained based on the amount of DNA present per unit weight of sample collected from the target soil is an index for grasping the characteristics of the soil and the working conditions of the bacteria in the soil.
- the soil is suitable for plant growth If one of the following conditions is not satisfied, it is determined that the soil is not suitable for plant growth.
- a reference value for the activity of producing phosphate from phytic acid 1% (w / w) phytic acid per unit dry weight was administered to the target soil, and the amount of water-soluble phosphoric acid on day 0 and the water solubility on day 3
- the reference value can be used. If the measured value is 10% or more, preferably 30% or more, it can be evaluated that the soil has excellent activity of producing phosphoric acid from phytic acid.
- the standard value for the activity of producing potassium from compost is to administer 1% (w / w) compost per unit dry weight to the target soil and measure the amount of potassium released on day 0 and the amount of potassium released on day 3.
- the case where the value obtained by the above formula is 100% can be used as the reference value. If the measured value is 5% or more, preferably 20% or more, it can be evaluated that the soil has an excellent potassium production activity from compost.
- the method for calculating the circulatory activity index by combining the above (I), (II) and (III) is not particularly limited.
- the reference value of the ammonia reduction rate set in advance, the phosphate generation activity from phytic acid, (I) Ammonia reduction rate placed on the line segment connecting the corresponding vertex from the center of gravity of the equilateral triangle, with respect to the area of the equilateral triangle formed with the reference value and the standard value of potassium production activity from compost as the apex It is preferably calculated as a ratio of the area of a triangle formed with the measurement points of (II) phosphoric acid producing activity from phytic acid and (III) potassium producing activity from compost as vertices.
- the number of soil bacteria is 2 ⁇ 10 8 cells / g-soil or more, and the reference value is the top. If the ratio of the triangle formed with the measurement point with respect to the area of the equilateral triangle as a vertex is 10 or more, preferably 30 or more, it can be determined that the soil is suitable for the growth of the plant. It can be judged that the soil is not suitable for growth.
- Soil diagnosis method (2) The soil diagnosis method of the present invention is characterized by evaluating or diagnosing soil using at least (A) a nitrogen cycling activity index, (B) a phosphorus cycling activity index, and (C) a potassium cycling activity index.
- the nitrogen cycling activity index is an index for analyzing the relationship between conversion of nitrogen-containing compounds including nitrification and soil bacteria.
- the nitrogen circulation activity index in the present invention is: (A-1) Number of soil bacteria in the target soil, (A-2) A value calculated using the ammonia reduction rate in the target soil and (A-3) the nitrous acid reduction rate in the target soil.
- Organic nitrogen compounds added to the soil are decomposed into peptides, amino acids and the like, and then converted into ammonia nitrogen.
- ammonia nitrogen (NH 4 + ) is converted to nitrite nitrogen (NO 2 ⁇ ) and nitrate nitrogen (NO 3 ⁇ ) sequentially. Some denitrification occurs and is converted to nitrogen (N 2 ).
- the nitrous acid reduction rate in the target soil is a value indicating the reduction rate of the nitrite nitrogen (NO 2 ⁇ ) concentration administered to the target soil.
- the nitrous acid reduction rate can be calculated as a value obtained by the following formula when a nitrite compound is administered to the target soil.
- Nitrite reduction rate (%) [1- (N 3 ⁇ N 4 ) / N 3 ] ⁇ 100
- N 3 represents the amount of nitrite nitrogen on the day of administration of the nitrite compound.
- N 4 represents the amount of nitrite nitrogen after a certain period from administration of the nitrite compound.
- the nitrite compound administration date means the administration date of the nitrite compound to the target soil.
- the amount of nitrite nitrogen on the day of administration of the nitrite compound can be expressed as the amount of nitrite nitrogen on day 0 of administration.
- the term “after a certain period after administration of the nitrite compound” means the day when a certain period has elapsed since the administration of the nitrite compound to the target soil.
- the amount of nitrite nitrogen after 3 days from the day of administration of the nitrite compound to the target soil can be expressed as the amount of nitrite nitrogen on the third day of administration.
- the length of the fixed period can be set as appropriate, but is preferably 3 to 7 days, particularly 3 days after administration. When the period is shorter than this, or when it is extremely long, the difference in activity becomes difficult to distinguish.
- the nitrite reduction rate in the target soil is preferably a value obtained by the following formula:
- the amount of nitrite nitrogen means the amount of nitrite nitrogen (NO 2 ⁇ ) per unit dry weight of the target soil.
- the amount of nitrite nitrogen can be measured by the naphthylethylenediamine method, high performance liquid chromatography or the like. More specifically, it can be measured by the quantitative method of nitrite nitrogen in the examples.
- Nitrite reduction rate reflects the conversion efficiency from nitrite nitrogen to nitrate nitrogen. The greater the decrease rate, the more nitrite-oxidizing bacteria in the target soil or the amount per unit cell mass It is thought that nitrite oxidizing bacteria with high activity are contained. In addition, the smaller the decrease rate, the smaller the number of nitrite-oxidizing bacteria.
- the standard value of the number of soil bacteria 3.25 ⁇ 10 9 cells / g-soil, which is the average value of the number of soil bacteria in farmland soil, is used as the standard value. If the measured value is 10% or more, preferably 40% or more, it can be evaluated that the soil has an excellent number of soil bacteria.
- the standard value of the ammonia reduction rate 60 ⁇ g-N / g-dry soil of ammonium sulfate was administered to the target soil, and the amount of ammonia nitrogen on the 0th day and the amount of ammonia nitrogen on the 3rd day were measured.
- the case where the value obtained in step 100 is 100% can be used as the reference value.
- the measured value is 30% or more, preferably 60% or more, it can be evaluated that the soil has an excellent ammonia reduction rate. Since the concentration of nitrogen-containing compounds in general field soil is about 60 ⁇ g-N / g-dry oilsoil, if the reduction rate of this amount is 100%, it has necessary and sufficient ammonia conversion activity. It can be evaluated that it is doing.
- nitrite reduction rate 60 ⁇ g-N / g-dry soil potassium nitrite was administered to the target soil, and the amount of nitrite nitrogen on day 0 and the amount of nitrite nitrogen on day 3 were measured. The case where the value obtained by the above equation is 100% can be used as the reference value. If the measured value is 60% or more, preferably 90% or more, it can be evaluated that the soil has an excellent nitrite reduction rate. Since the concentration of nitrogen-containing compounds in general field soil is about 60 ⁇ g-N / g-dry soil, the necessary and sufficient nitrite conversion activity is obtained when the reduction rate of this amount is 100%. It can be evaluated as having.
- the method for calculating the nitrogen cycling activity index by combining the above (A-1), (A-2) and (A-3) is not particularly limited, but the preset reference value for the number of soil bacteria, ammonia reduction
- the (A-1) soil placed on the line segment connecting the corresponding vertex from the center of gravity of the equilateral triangle with respect to the area of the equilateral triangle formed with the reference value of the rate and the standard value of the nitrite reduction rate as the apex It is preferably calculated as a ratio of the area of a triangle formed with the measurement points of the number of bacteria, the (A-2) ammonia reduction rate, and the (A-3) nitrous acid reduction rate as vertices.
- the nitrogen circulation activity index can be calculated by the calculation method described in the examples.
- the phosphorous circulatory activity index is an activity to convert a phosphorus-containing organic compound into phosphoric acid, in other words, an activity to convert a phosphorous compound that cannot be used by plants into usable phosphoric acid. It is an index for analyzing the relationship between soil bacteria.
- the phosphorus circulating activity index in the present invention is: (B-1) Number of soil bacteria in the target soil, (B-2) Phosphate producing activity from phytic acid in the target soil, and (B-3) Phosphate producing activity from compost in the target soil, Is a value calculated using.
- plants absorb water-soluble phosphoric acid. For this reason, it is considered that phosphorus is easily absorbed by plants in soil with a large amount of water-soluble phosphoric acid in the soil.
- phytic acid and compost are considered to be particularly important as phosphorus compounds.
- Phytic acid is a substance for plants to store phosphorus, and is abundant in weeds and residues after harvesting crops. If the microorganisms in the target soil have a high activity of releasing phosphoric acid from phytin in these plants, it can be judged that the quality of the soil is high.
- compost is used as a means to replenish phosphorus that is deficient in the soil from the outside.
- compost contains phosphoric acid as a component of bark compost, not water-soluble phosphoric acid. If the activity of microorganisms in the target soil to generate phosphoric acid from compost is high, it can be determined that the quality of the soil is high.
- the phosphate production activity from compost in the target soil refers to the conversion activity of compost administered to the target soil into phosphoric acid, in other words, conversion of compost It is a value indicating the activity of decomposing to release water-soluble phosphoric acid.
- compost examples include plant compost such as bark compost, livestock compost such as chicken manure compost, cattle manure compost, and pig manure compost, and seaweed compost. These may be used alone or in combination of two or more.
- bark compost contains a lot of phosphoric acid in the form of phytic acid or the like, more appropriate evaluation becomes possible.
- the total phosphoric acid (P 2 O 5 ) content of bark compost is 0.5% or more (dry matter).
- the administration form of compost is not limited, and compost mixed with culture soil may be used.
- the mixing ratio is about 10 to 50%, preferably about 25 to 35% by weight of compost with respect to the total amount of the cultured soil.
- the phosphoric acid production activity from compost can be calculated as a value obtained by the following formula when compost is administered to the target soil.
- Phosphate production activity from compost (%) [(P 6 -P 5 ) / P 4 ] ⁇ 100 (In the formula, P 4 represents the amount of phosphoric acid in compost. P 5 represents the amount of water-soluble phosphoric acid on the day of compost administration. P 6 represents the amount of water-soluble phosphoric acid after a certain period of time from compost administration.)
- the amount of phosphoric acid in compost can be measured according to a known phosphoric acid content determination method. For example, after decomposing organic matter in compost with perchloric acid, it is extracted with 0.002N sulfuric acid and subjected to the molybdenum blue method. It can be obtained by quantifying total phosphoric acid.
- the date of compost administration means the date of administration of compost to the target soil.
- the amount of water-soluble phosphoric acid on the day on which compost is administered can be expressed as the amount of water-soluble phosphoric acid on day 0 of administration.
- “after a certain period of time from the administration of compost” means the day when a certain period has elapsed from the administration of compost to the target soil.
- the amount of water-soluble phosphoric acid after 3 days from the day when compost is administered to the target soil can be expressed as the amount of water-soluble phosphoric acid on the third day of administration.
- the length of the fixed period can be set as appropriate, but is preferably 3 to 7 days, particularly 3 days after administration. When the period is shorter than this, or when it is extremely long, the difference in activity becomes difficult to distinguish.
- the phosphoric acid production activity from compost in the target soil is preferably a value determined by the following formula:
- the amount of water-soluble phosphoric acid means the amount of water-soluble phosphoric acid per unit dry weight of the target soil, and can be measured by the same method as described above.
- the phosphoric acid production activity from compost reflects the conversion efficiency from compost to water-soluble phosphoric acid, and it is considered that the greater the activity, the easier it is to use phosphorus contained in the compost. From this, it is considered that the greater the activity, the better the quality of the soil and the lower the input of compost. Moreover, it is considered that the smaller the activity, the more difficult it is to use the phosphorus in the compost. From this, it is considered that the smaller the activity is, the lower the quality of the soil is, and it is necessary to increase the amount of compost input or to add phosphorus fertilizer.
- the standard value of the number of soil bacteria 3.25 ⁇ 10 9 cells / g-soil, which is the average value of the number of soil bacteria in farmland soil, is used as the standard value. If the measured value is 10% or more, preferably 40% or more, it can be evaluated that the soil has an excellent number of soil bacteria.
- a reference value for the activity of producing phosphate from phytic acid 1% (w / w) phytic acid per unit dry weight was administered to the target soil, and the amount of water-soluble phosphoric acid on day 0 and the water solubility on day 3
- the reference value can be used. If the measured value is 10% or more, preferably 30% or more, it can be evaluated that the soil has excellent activity of producing phosphoric acid from phytic acid.
- the reference value for the activity of producing phosphoric acid from compost is to administer 1% (w / w) compost per unit dry weight to the target soil, the amount of water-soluble phosphoric acid on day 0 and water-soluble phosphorus on day 3
- the reference value can be used. If the measured value is 10% or more, preferably 30% or more, it can be evaluated that the soil has excellent activity of producing phosphoric acid from compost.
- the method for calculating the phosphorus circulation activity index by combining the above (B-1), (B-2) and (B-3) is not particularly limited, but a preset reference value for the number of soil bacteria, phytic acid Is placed on a line segment connecting the corresponding vertices from the center of gravity of the equilateral triangle with respect to the area of the equilateral triangle formed with the reference value of the phosphate producing activity from compost and the reference value of the phosphate producing activity from compost as the vertex. Furthermore, the triangle formed with the measurement points of (B-1) soil bacteria count, (B-2) phosphate production activity from phytic acid, and (B-3) phosphate production activity from compost as vertexes It is preferable to calculate as a percentage of the area.
- the phosphorus circulating activity index can be calculated by the calculation method described in the examples.
- the potassium cycle activity index is an index for analyzing the relationship between conversion of potassium-containing compounds and soil bacteria.
- the potassium circulatory activity index in the present invention is: (C-1) number of soil bacteria in the target soil, (C-2) It is a value calculated using the potassium release rate in the target soil and (C-3) the potassium generation activity from compost in the target soil.
- compost is used as a means for supplying potassium deficient in the soil from the outside.
- a large amount of potassium is contained in the bodies of animals and plants in the compost.
- plants can use free potassium. Therefore, it is considered that the activity of converting potassium contained in compost into free potassium by microorganisms is an important factor.
- the potassium release rate in the target soil is a value indicating the amount of potassium per unit dry weight of the target soil.
- the potassium release rate in the target soil is a value calculated by the following formula.
- Potassium release rate (%) [(K 3 ⁇ K 2 ) / K 1 ] ⁇ 100 (In the formula, K 1 represents the potassium content in the target soil on the measurement start date. K 2 represents the potassium release amount on the measurement start date. K 3 represents the potassium release amount after a certain period from the measurement start date. .)
- the potassium content in the target soil can be quantified according to a known method.
- a filtrate obtained by adding an aqueous ammonium acetate solution to the soil and filtering is used as a potassium extract, and the obtained extract is subjected to atomic absorption spectrophotometry. It can be obtained by measuring the amount of potassium using a meter.
- Potassium liberation means the amount of potassium per unit dry weight of the target soil.
- Potassium liberation can be measured by atomic absorption spectrophotometry and ICP-MS. For example, it can be obtained by measuring a solution obtained by extracting potassium liberated from soil with distilled water using an atomic absorption photometer. Specifically, it can be measured by the method described in the quantitative determination method of potassium in soil by the atomic absorption photometer described in the examples.
- the potassium release rate is Value obtained by the following formula:
- the potassium release rate reflects the amount of potassium available to the plant, and the larger the value, the easier it is to use potassium in the soil. From this, it is considered that the larger the value, the better the quality of the soil, and the less the amount of potassium input from the outside.
- the smaller the value the more difficult it is to use potassium in the soil. From this, it is diagnosed that the smaller the value, the poorer the quality of the soil and the greater the amount of potassium input from the outside.
- the standard value of the number of soil bacteria 3.25 ⁇ 10 9 cells / g-soil, which is the average value of the number of soil bacteria in farmland soil, is used as the standard value. If the measured value is 10% or more, preferably 40% or more, it can be evaluated that the soil has an excellent number of soil bacteria.
- the activity when all potassium in the target soil is converted to free potassium in 3 days can be defined as 100%. If the measured value is 5% or more, preferably 10% or more, it can be evaluated that the soil has an excellent potassium release rate.
- the reference value for the activity of potassium production from compost is to administer 1% (w / w) compost per unit dry weight to the target soil and measure the amount of potassium released on day 0 and the amount of potassium released on day 3.
- the case where the value obtained by the above formula is 100% can be used as the reference value. If the measured value is 5% or more, preferably 20% or more, it can be evaluated that it has excellent potassium production activity from compost.
- the method for calculating the potassium circulatory activity index by combining the above (C-1), (C-2) and (C-3) is not particularly limited, but a preset reference value for the number of soil bacteria, potassium release With respect to the area of the equilateral triangle formed with the reference value of the amount and the reference value of the potassium production activity from compost as the apex, the (C-1 It is preferably calculated as the ratio of the area of a triangle formed with the measurement point of the number of soil bacteria, the (C-2) potassium liberation amount, and the (C-3) composting potassium production activity as the apex.
- the measurement point for the area of the equilateral triangle formed with the reference value as the vertex is formed as the vertex. If the ratio of triangles to be applied is 1 or more, preferably 5 or more, it can be determined that the soil has excellent potassium cycle activity, and if it is less than the above value, it can be determined that the soil has no excellent potassium cycle activity.
- the potassium cycle activity index can be calculated by the calculation method described in the examples.
- Target soil the kind of soil used as object is not specifically limited, For example, farmland, the soil after a bioremediation process, etc. are mentioned.
- the present invention is used as a farmland diagnostic method for diagnosing whether farmland is of a quality suitable for plant growth and whether the farmland needs to be improved for quality suitable for plant growth. Can do.
- the present invention can be used as a method for diagnosing purified soil for determining whether or not the material circulation activity of soil microorganisms is recovered in the soil after bioremediation treatment and can be used for ordinary applications.
- soil is diagnosed using the above circulating activity index, or the above (A) nitrogen circulating activity index, (B) phosphorus circulating activity index, and (C) potassium circulating activity index.
- the above-mentioned circulatory activity index and other indices other than the above (A) to (C) may be used.
- examples of other indicators include soil pH, electrical conductivity, dissolved oxygen concentration, particle size, or porosity. These can be measured according to a known method.
- an index related to carbon for example, an index related to carbon
- the total organic carbon content (TOC) may be used as an index.
- TOC total organic carbon content
- a carbon source as a biological component and an energy source for maintaining the activity are necessary, so the amount of carbon can also be an important factor. it is conceivable that.
- the method of using the above indicators (A) to (C) for diagnosis is not particularly limited, but for example, obtaining a comprehensive indicator as the sum or product of the indicators (A) to (C) or their calculated values. Can make a diagnosis.
- the soil with the larger sum of the indicators (A) to (C) has higher quality and can be diagnosed as soil suitable for plant growth.
- the indicators (A) to (C) are calculated as the ratio of the triangles formed with vertices as the measurement points relative to the area of the regular triangle formed with the reference values as vertices as described above, and the average of the ratios is calculated.
- the average is 10 or more, preferably 35 or more, it can be determined that the soil is suitable for plant growth, and if it is less than the above value, it can be determined that the soil is not suitable for plant growth.
- the index of (A) to (C) is calculated as a ratio of the triangle formed with the measurement point with respect to the area of the regular triangle formed with the reference value as the vertex as described above, and then with 100 as the vertex. If the ratio of the triangle formed with the above-mentioned ratio placed on the line connecting the corresponding vertices from the center of gravity of the regular triangle to the area of the regular triangle formed is 1 or more, preferably 5 or more, It can be determined that the soil is suitable for plant growth, and if it is less than the above value, it can be determined that the soil is not suitable for plant growth.
- each of the above-mentioned circulatory activity indicators or indicators (A) to (C) is diagnosed, which nitrogen, phosphorus or potassium circulatory system needs to be improved.
- a diagnosis may be made as to whether it is effective or the improvement of the state of soil microorganisms is effective.
- soil diagnosis can be performed as a comprehensive index combining the above indices (A) to (C) with other indices. Furthermore, diagnosis can be performed by examining the balance of the indicators (A) to (C) or by comparing with other indicators.
- Soil Quality Management Method According to the present invention, a method for managing soil quality using the diagnostic method of the present invention is provided.
- the above-described diagnostic method of the present invention is performed over time, and the circulation activity index, (A) nitrogen circulation activity index, (B) phosphorus circulation activity index, and (C) potassium
- the soil quality is managed by analyzing the time course of the circulation activity index.
- the analysis method of the change with time is not particularly limited, and can be appropriately performed according to a known method. For example, you may analyze using the value which further converted or calculated the parameter
- the analysis can be performed by combining the circulatory activity index and changes over time of other indices other than the above (A) to (C).
- the quality control method of the present invention not only the state of substances necessary for plant growth but also the state of microorganisms in the soil can be grasped. For this reason, according to the quality control method of the present invention, it is possible to grasp whether the ecosystem in the soil is well preserved through the growth of the plant and various substance circulation activities are functioning.
- Soil Improvement Method there is provided a method for improving the quality of soil using the diagnostic method of the present invention.
- the soil activity related to the circulation activity index (A) the nitrogen circulation activity index, (B) the phosphorus circulation activity index, and (C) the potassium circulation activity index.
- the soil is improved by obtaining a diagnosis result and performing processing according to the diagnosis content.
- the contents of the treatment include additional administration of nitrogen, phosphorus and / or potassium-containing fertilizers, administration of nutrients to activate the microorganisms in the soil, and the introduction of microorganisms having circulating activities of nitrogen, phosphorus and / or potassium. Administration is possible. For example, when the ammonia reduction rate is low, it is conceivable to administer ammonia oxidizing bacteria.
- the soil improvement method of the present invention there is an advantage that it is possible to determine which component of nitrogen, phosphorus, and potassium is necessary, and further, it is possible to determine the treatment content in consideration of the action of microorganisms in the soil.
- the plant cannot sufficiently use nitrogen. According to the present invention, it can be determined that treatment for increasing the activity of microorganisms and administration of microorganisms having such activities are effective for such situations. On the other hand, even if the amount of nitrogen in the soil is not sufficient and it is determined that external input of nitrogen is necessary, if the activity of the microorganism is known to be sufficient, the dosage of nitrogen can be adjusted and excess Administration can also be suppressed.
- the soil can be improved by effectively utilizing the function of the ecosystem in the soil, and efficient soil improvement and further efficient food production can be achieved. Can be possible.
- a soil diagnosis method reflecting the circulating activity in the soil, particularly a soil diagnosis method capable of judging the suitability for cultivation of agricultural products.
- the soil diagnosis method of the present invention also reflects the state of microorganisms in the soil that are closely involved in the material circulation, making it possible to accurately diagnose the quality of farmland according to the natural circulation system.
- the diagnostic method of the present invention can accurately diagnose the quality of soil suitable for farming methods that do not rely on chemical farming methods such as biomass.
- the diagnostic method of the present invention it is necessary to improve which circulatory system of nitrogen, phosphorus and potassium, which is important for plant growth, and for the improvement, addition of additional components is effective, or In addition, it is possible to determine the processing contents of whether the improvement of the state of soil microorganisms is effective.
- the quality control method of the present invention not only the state of substances necessary for plant growth but also the state of microorganisms in the soil can be grasped, and the ecosystem in the soil is well preserved through the growth of plants. It is also possible to grasp whether various substance circulation activities are functioning.
- the soil improvement method of the present invention it is possible to improve the soil by effectively using the function of the ecosystem in the soil, and it is possible to improve the efficiency of the soil and further improve the profitability of agricultural production. To.
- the present invention provides a soil quality diagnosis and improvement means based on the natural circulation function, and improves the profitability of farming methods in which the use of chemical substances such as organic farming methods has been reduced, and environmental conservation. This contributes to the establishment of a type agricultural production system.
- FIG. A is a drawing for soil No. 1 and B is a drawing for soil No. 2.
- ⁇ is an uninoculated strain
- ⁇ is a A strain administered
- ⁇ is a diagram showing the results when a B strain is administered.
- A Ammonium sulfate 4 ⁇ g-N / g-soil administration
- B Ammonium sulfate 40 ⁇ g-N / g-soil administration
- C Ammonium sulfate 400 ⁇ g-N / g-soil administration
- ⁇ Ammonia nitrogen
- ⁇ Nitrite nitrogen
- ⁇ Nitrate nitrogen It is drawing which shows the nitrogen circulation in the soil which administered potassium nitrite.
- A Potassium nitrite 6 ⁇ g-N / g-soil administration
- B Potassium nitrite 60 ⁇ g-N / g-soil administration
- C Potassium nitrite 600 ⁇ g-N / g-soil administration
- ⁇ Ammonia nitrogen
- ⁇ Nitrite nitrogen
- ⁇ Nitrate nitrogen It is drawing which shows the relationship between the amount of nitrification per day, and the number of soil bacteria.
- ⁇ Kakihata
- ⁇ Kaki Paddy Field
- ⁇ Error bars other than farmland are standard deviation
- nitrite nitrogen by naphthylethylenediamine method 1.0 ml of an inorganic nitrogen extract extracted from soil was dispensed into a 1.5 ml microtube, and 100 ⁇ l of the diazotizing agent shown in Table 3 was added and stirred. After leaving still at room temperature for 5 minutes, 100 microliters of coupling agents shown in Table 4 were added, and it left still again at room temperature for 20 minutes, and the light absorbency of 540 nm was measured. The amount of nitrite nitrogen (NO 2 -- N) was measured from a calibration curve prepared using a nitrite nitrogen standard solution.
- 700 ⁇ l of the aqueous layer was taken into a new microtube, 700 ⁇ l of chloroform / isoamyl alcohol (24: 1, v / v) was added and mixed, and then centrifuged at 16 ° C. and 13,000 rpm for 10 minutes. After centrifugation, 500 ⁇ l of the aqueous layer was taken into a new microtube, 300 ⁇ l of 2-propanol was added, gently mixed, and centrifuged at 16 ° C. and 13,000 rpm for 15 minutes. After centrifugation, the supernatant was removed, 500 ⁇ l of 70% (v / v) ethanol was added, and the mixture was centrifuged at 16 ° C.
- TE 10 1 buffer solution (pH 8.0) shown in Table 7 was added and dissolved well to obtain an eDNA solution.
- Distilled water was added to 2.0 g of agarose, 4.0 ml of 50 ⁇ TAE buffer (pH 8.0) shown in Table 8 and 20 ⁇ l of 0.1 mM ethidium bromide solution to make 200 ml, and a 1.0% agarose gel was prepared.
- a loading dye (Toyobo, Osaka) (1.0 ⁇ l) was mixed with eDNA solution (5.0 ⁇ l), and a total amount of 6.0 ⁇ l and a smart ladder (Nippon Gene, Toyama) containing a known amount of DNA were applied to an agarose gel. After electrophoresis at 100 V for 40 minutes, the agarose gel was irradiated with UV to confirm the DNA band. A smart ladder DNA band was analyzed using KODAK 1D Image Analysis software (KODAK, NY, USA), and a calibration curve of DNA amount against fluorescence intensity was prepared.
- the amount of DNA was determined from the fluorescence intensity of the DNA band of each sample DNA solution, and the amount of eDNA per 1.0 g of each soil was calculated.
- the number of soil bacteria was determined by a calibration curve for converting the amount of eDNA into the number of soil bacteria by DAPI staining.
- the ammonia reduction rate and nitrite reduction rate were calculated from the amount of inorganic nitrogen reduction.
- the ammonia reduction rate was calculated from the amount of ammonia nitrogen measured in the above 1b) and 1c) by the following formula:
- the reduction rate of nitrous acid was calculated by the following formula from the amount of nitrite nitrogen (NO 2 ⁇ ⁇ N) measured in the above b) and d).
- Fig. 1 shows the results of the ammonia reduction rate and nitrous acid reduction rate for each sample.
- the decrease rate of nitrous acid was almost 100% in all samples. However, the rate of ammonia reduction varied from sample to sample: 72.0% for the highest sample and 3.10% for the lowest sample. Moreover, since the decrease rate of ammonia was lower than the decrease rate of nitrous acid in all samples, it was considered that the reaction from ammonia to nitrous acid was rate-limiting in the nitrification reaction.
- the number of soil bacteria indicates the ratio of the number of soil bacteria in each sample, that is, the amount of bacteria when the average value of the number of soil bacteria in farmland soil is 3.25 ⁇ 10 9 cells / g-soil.
- ammonia reduction rate indicates the ratio of the ammonia reduction rate of each sample when the activity of reducing 100 ⁇ % of the ammonia compound of 60 ⁇ g-N / g-dry soil in 100 days is defined as 100.
- the nitrous acid reduction rate indicates the ratio of the nitrous acid reduction rate of each sample when the activity of reducing the nitrous acid compound of 60 ⁇ g-N / g-dry soil to 100% in 3 days is taken as 100.
- sample No. 2 has a relatively high ammonia reduction rate, nitrous acid reduction rate, and the amount of bacteria, so when organic nitrogen is added to the soil, it is quickly converted to ammonia, It is thought that it is oxidized to nitric acid.
- strains A and B Two types of autotrophic ammonia-oxidizing bacteria (strains A and B) were administered to the soil to examine whether nitrification was activated.
- the culture solution of strain A or strain B was concentrated by centrifugation and administered to sterilized soil (soil 1 and 2) to 1.0 ⁇ 10 7 cells / g-dry soil. Furthermore, the ammonia nitrogen was applied to the soil so as to be 60 ⁇ g-N / g dry soil and left to stand for 3 days, and the temporal change in the amount of inorganic nitrogen was analyzed. The results are shown in FIG. Fig. 3 shows changes in the amount of nitrite nitrogen and nitrate nitrogen over time.
- the amount of accumulated nitrite nitrogen and nitrate nitrogen increased when autotrophic ammonia-oxidizing bacteria were administered compared to when it was not.
- the phytic acid 1% (w / w) was added to the sampled soil, stirred well, and left at room temperature for 3 days. Weigh 2.0 g of this soil into a 50 ml centrifuge tube, extract the water-soluble phosphoric acid by the method described above, subject it to the molybdenum blue method, measure the amount of water-soluble phosphoric acid, The amount.
- the amount of phosphoric acid in phytic acid was calculated from the dose of phytic acid.
- the activity of producing phosphoric acid from phytic acid was calculated from the following formula.
- the phosphoric acid production activity from compost was calculated from the following formula.
- the number of soil bacteria indicates the ratio of the number of soil bacteria in each sample when the average value of 3.25 ⁇ 10 9 cells / g-soil of soil bacteria in farmland soil is 100.
- Phosphoric acid production activity from phytic acid is defined as 1% (w / w) of phosphoric acid in phytic acid converted to water-soluble phosphoric acid in 100 days. The ratio of the phosphate production activity from phytic acid of each sample is shown.
- the phosphoric acid production activity from compost is 1% (w / w) of phosphoric acid in compost in 3 days when all are converted to water-soluble phosphoric acid in 100 days.
- the ratio of the phosphate production activity from the compost of a sample is shown.
- the soil of sample No. 12 has a high phosphorus cycle activity index, and can be evaluated as a soil in which plant phosphate absorption is easy to be performed.
- the soil potassium cycle was analyzed using 10 samples (No. 11 to 20) of soils with different application and fertilization conditions.
- the amount of potassium released was quantified in the same manner as the amount of potassium released on the third day.
- the potassium release rate was calculated by the following formula.
- the potassium content in the compost was measured in the same manner except that compost was used instead of the soil.
- the potassium production activity from compost was calculated according to the following formula.
- the number of soil bacteria indicates the ratio of the number of soil bacteria in each sample when the average value of 3.25 ⁇ 10 9 cells / g-soil of soil bacteria in farmland soil is 100.
- the potassium release rate indicates the potassium release rate of each sample when the activity when all the potassium in the soil is converted to free potassium during 3 days is defined as 100.
- the potassium generation activity from compost is 1% (w / w) of compost from each sample when the activity in converting all the potassium in compost to free potassium in 100 days is 100. The ratio of the potassium production activity is shown.
- the soil of sample No. 11 has a high potassium cycle activity index, and can be evaluated as a soil in which plant potassium absorption is easily performed.
- Table 16 shows the diagnostic values from the area of the triangle with the ammonia reduction rate, the phosphatic acid-forming activity from phytic acid, and the potassium-generating activity from compost as vertices. From this result, it was predicted that the higher the comprehensive diagnosis result of the soil, the better the plant growth.
- FIG. 6 shows that when ammonium sulfate was added at 4, 40 ⁇ g / g-soil, almost all ammonia nitrogen was reduced and nitrate nitrogen was accumulated on the fourth day. When administered at 400 ⁇ g / g-soil, the difference in the amount of ammonia nitrogen decreased was smaller than that at the start.
- ammonia nitrogen contained in various farmland was measured, it was in the range of 0-100 ⁇ g-N / g-soil in almost all soils. Therefore, the amount of ammonia nitrogen administered as a substrate on the basis of the following is shown. Were determined.
- the substrate input is 40-60 ⁇ g / g-soil based on the amount of ammonia nitrogen contained in general soil. I thought that it should be done in the range. In the end, it was decided to input so as to obtain 60 ⁇ g / g-soil that was easy to calculate.
- nitrite nitrogen was hardly contained, but nitrate nitrogen was contained in 0-100 ⁇ g-N / g-soil at the same level as ammonia nitrogen. From these facts, the amount of nitrite nitrogen to be administered as a substrate was determined on the basis of the following.
- FIG. 8 shows the amount of nitrification per day.
- the number of soil bacteria was quantified by the eDNA analysis method as in 1f.
- the amount of nitrification indicates the total amount of nitrite nitrogen and nitrate nitrogen measured after administration to soil so that ammonia nitrogen becomes 60 ⁇ g-N / g dry soil and left to stand for 1 day.
- the nitrification reaction did not proceed when the number of microorganisms was 200 million / g or less.
- the lower limit ammonia reduction rate of the comprehensive diagnostic value, the phosphate production activity from phytin, and the potassium production activity from compost are 30, 10, 5 or less, it is not judged that the soil is excellent.
- the ratio of the area of the triangle formed with the lower limit value placed on the line segment connecting the corresponding vertex from the center of gravity of the regular triangle to the area of the regular triangle formed with the reference value as the vertex is 1.7 points. Become. Therefore, if less than this point, it was judged that good plant growth could not be expected.
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Abstract
Description
(I)対象土壌におけるアンモニア減少率、
(II)対象土壌におけるフィチン酸からのリン酸生成活性、及び
(III)対象土壌における堆肥からのカリウム生成活性
並びに(IV)土壌における土壌バクテリア数
を用いて土壌診断を行うことを特徴とする土壌の診断方法。
予め設定されたアンモニア減少率の基準値、フィチン酸からのリン酸生成活性の基準値及び堆肥からのカリウム生成活性の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(I) アンモニア減少率、前記(II) フィチン酸からのリン酸生成活性、及び前記(III) 堆肥からのカリウム生成活性の測定点を頂点として形成される三角形の面積の割合である、項1に記載の診断方法。
(A-1)対象土壌における土壌バクテリア数、
(A-2)対象土壌におけるアンモニア減少率、及び
(A-3)対象土壌における亜硝酸減少率、
B)下記(B-1)~(B-3)を用いて算出されるリン循環活性指標:
(B-1)対象土壌における土壌バクテリア数、
(B-2)対象土壌におけるフィチン酸からのリン酸生成活性、及び
(B-3)対象土壌における堆肥からのリン酸生成活性、
並びに、C)下記(C-1)~(C-3)を用いて算出されるカリウム循環活性指標:
(C-1)対象土壌における土壌バクテリア数、
(C-2)対象土壌におけるカリウム遊離率、及び
(C-3)対象土壌における堆肥からのカリウム生成活性
を少なくとも用いて土壌診断を行うことを特徴とする土壌の診断方法。
予め設定された土壌バクテリア数の基準値、アンモニア減少率の基準値、及び亜硝酸減少率の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(A-1)土壌バクテリア数、前記(A-2)アンモニア減少率、及び前記(A-3)亜硝酸減少率の測定点を頂点として形成される三角形の面積の割合である、項3に記載の診断方法。
予め設定された土壌バクテリア数の基準値、フィチン酸からのリン酸生成活性の基準値、及び堆肥からのリン酸生成活性の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(B-1)土壌バクテリア数、前記(B-2)フィチン酸からのリン酸生成活性、及び前記(B-3)堆肥からのリン酸生成活性の各測定値を頂点として形成される三角形の面積の割合である、項3又は4に記載の診断方法。
予め設定された土壌バクテリア数の基準点、カリウム遊離率の基準値、及び、堆肥からのカリウム生成活性の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(C-1)土壌バクテリア数、前記(C-2)カリウム遊離率、及び前記(C-3)堆肥からのカリウム生成活性の各測定値を頂点として形成される三角形の面積の割合である、項3~5のいずれかに記載の診断方法。
1.1.土壌診断方法(1)
本発明の土壌の診断方法(1)は、下記(I)~(III)を用いて算出される循環活性指標:
(I)対象土壌におけるアンモニア減少率、
(II)対象土壌におけるフィチン酸からのリン酸生成活性、及び
(III)対象土壌における堆肥からのカリウム生成活性
並びに(IV)土壌における土壌バクテリア数
を用いて土壌診断を行うことを特徴とする。
本発明において、対象土壌におけるアンモニア減少率とは、対象土壌に投与したアンモニア化合物濃度の減少割合を示す値である。
(式中、N1はアンモニア化合物投与日のアンモニア態窒素量を表す。N2はアンモニア化合物投与から一定期間後のアンモニア態窒素量を表す。)
アンモニア化合物投与日とは、対象土壌に対するアンモニア化合物の投与日を意味する。アンモニア化合物投与日のアンモニア態窒素量は、投与0日目のアンモニア態窒素量と表すことができる。
本発明において、対象土壌におけるフィチン酸からのリン酸生成活性とは、対象土壌に投与したフィチン酸の変換活性を示す値である。
(式中、P1はフィチン酸中のリン酸量を表す。P2はフィチン酸投与日の水溶性リン酸量を表す。P3はフィチン酸投与から一定期間後の水溶性リン酸量を表す。)
本発明において、対象土壌における堆肥からのカリウム生成活性とは、対象土壌に投与した堆肥中のカリウムの遊離カリウムへの変換活性を示す値である。
(式中、K4は堆肥中のカリウム含有量を表す。K5は堆肥投与日におけるカリウム遊離量を表す。K6は堆肥投与から一定期間後のカリウム遊離量を表す。)
本発明において、土壌バクテリア数とは、対象土壌から採取した試料単位重量当たりに存在するDNA量に基づいて求められる土壌バクテリア数を表す。
対象土壌から採取された試料とは、上記対象土壌から採取(サンプリング)される土壌のことである。採取方法は特に限定されず、適宜公知の方法に従って行うことができる。
前記(I)、(II)、(III)、及び(IV)は、土壌中の窒素、リン酸、カリウム循環においていずれも重要な因子であり、これらを組み合わせて解析することが、適切な診断のために重要である。
本発明の土壌診断方法は、少なくとも(A)窒素循環活性指標、(B)リン循環活性指標、及び(C)カリウム循環活性指標を用いて、土壌の評価乃至診断を行うことを特徴とする。
本発明において、窒素循環活性指標とは、硝化を含む窒素含有化合物の変換と土壌バクテリアの関係を解析するための指標である。
(A-1)対象土壌における土壌バクテリア数、
(A-2)対象土壌におけるアンモニア減少率、及び
(A-3)対象土壌における亜硝酸減少率
を用いて算出される値である。
対象土壌における土壌バクテリア数については、前記(IV)循環活性指標に記載したとおりである。
対象土壌におけるアンモニア減少率については、前記(I)循環活性指標に記載したとおりである。
本発明において、対象土壌における亜硝酸減少率とは、対象土壌に投与した亜硝酸態窒素(NO2 -)濃度の減少割合を示す値である。
(式中、N3は亜硝酸化合物投与日の亜硝酸態窒素量を表す。N4は亜硝酸化合物投与から一定期間後の亜硝酸態窒素量を表す。)
亜硝酸化合物投与日とは、対象土壌に対する亜硝酸化合物の投与日を意味する。亜硝酸化合物投与日の亜硝酸態窒素量は、投与0日目の亜硝酸態窒素量と表すことができる。
前記(A-1)、(A-2)、及び(A-3)は、土壌中の窒素循環においていずれも重要な因子であり、これらを組み合わせて解析することが、適切な診断のために重要である。
本発明において、リン循環活性指標とは、リン含有有機化合物からリン酸への変換活性、別言すると、植物が利用できないリン化合物を利用可能なリン酸に変換する活性、と土壌バクテリアの関係を解析するための指標である。
(B-1)対象土壌における土壌バクテリア数、
(B-2)対象土壌におけるフィチン酸からのリン酸生成活性、及び
(B-3)対象土壌における堆肥からのリン酸生成活性、
を用いて算出される値である。
対象土壌における土壌バクテリア数については、前記(IV)循環活性指標に記載したとおりである。
対象土壌におけるフィチン酸からのリン酸生成活性については、前記(II)循環活性指標に記載したとおりである。
本発明において、対象土壌における堆肥からのリン酸生成活性とは、対象土壌に投与した堆肥のリン酸への変換活性、換言すると、堆肥を変換・分解して水溶性リン酸を遊離させる活性を示す値である。
(式中、P4は堆肥中のリン酸量を表す。P5は堆肥投与日における水溶性リン酸量を表す。P6は堆肥投与から一定期間後の水溶性リン酸量を表す。)
前記(B-1)、(B-2)、及び(B-3)は、土壌中のリン循環においていずれも重要な因子であり、これらを組み合わせて解析することが、適切な土壌診断のために重要である。
本発明において、カリウム循環活性指標とは、カリウム含有化合物の変換と土壌バクテリアの関係を解析するための指標である。
(C-1)対象土壌における土壌バクテリア数、
(C-2)対象土壌におけるカリウム遊離率、及び
(C-3)対象土壌における堆肥からのカリウム生成活性
を用いて算出される値である。
対象土壌における土壌バクテリア数については、前記(IV)循環活性指標に記載したとおりである。
本発明において、対象土壌におけるカリウム遊離率とは、対象土壌の単位乾燥重量当たりのカリウム量を示す値である。
(式中、K1は測定開始日における対象土壌中のカリウム含有量を表す。K2は測定開始日におけるカリウム遊離量を表す。K3は測定開始日から一定期間後のカリウム遊離量を表す。)
下記式で求められる値:
対象土壌における堆肥からのカリウム生成活性については、前記(III)循環活性指標に記載したとおりである。
前記(C-1)、(C-2)、及び(C-3)は、土壌中のカリウム循環においていずれも重要な因子であり、これらを組み合わせて解析することが、適切な土壌診断のために重要である。
本発明において、対象となる土壌の種類は、特に限定されないが、例えば、農地や、バイオレメディエーション処理後の土壌等が挙げられる。
本発明においては、上記循環活性指標、又は、上記(A)窒素循環活性指標、(B)リン循環活性指標、(C)カリウム循環活性指標を用いて、土壌の診断を行う。
本発明によれば、上記本発明の診断方法を利用して、土壌の品質を管理する方法が提供される。
本発明によれば、上記本発明の診断方法を利用して、土壌の品質を改善する方法が提供される。
(1-1)実験方法
1a)硝化能の評価
土壌10 gをガラスシャーレに量り取り、110℃で2時間乾燥後、重量減少量から含水率を算出した。2 mmメッシュのふるいにかけた乾燥重量15 gの土壌を50 ml容UMサンプル瓶に入れ、硫酸アンモニウム水溶液(0.080 mM)もしくは亜硝酸カリウム水溶液(0.16 mM)をそれぞれ60 μg-N/g-dry soilとなるように添加した。土壌をよくかき混ぜた後、25℃、含水率一定で3日間静置した。
50 ml容遠心チューブに土壌サンプル2.0 gと1 M塩化カリウム水溶液20 mlを加え懸濁し、100 rpmで1時間振とうした。振とう後、10,000 rpmで5分間遠心分離し、その上清を無機態窒素抽出液とした。
土壌から抽出した無機態窒素抽出液1.0 mlを2.0 ml容マイクロチューブに分注し、表1に示す次亜塩素酸ナトリウム溶液500 μlを加えて撹拌し、室温で5分間静置した。静置後、表2に示すフェノール・ニトロプルシッドナトリウム溶液500 μlを加えて撹拌し、30℃で60分間静置した。静置後、640 nmの吸光度を測定した。吸光度測定時にアンモニア態窒素標準液を用いて検量線を作成し、得られた関係式を用いてアンモニア態窒素量(NH4 +-N)を測定した。
土壌から抽出した無機態窒素抽出液1.0 mlを1.5 ml容マイクロチューブに分注し、表3に示すジアゾ化剤100 μlを加えて撹拌した。室温で5分間静置した後、表4に示すカップリング剤100 μlを加えて再び室温で20分間静置し、540 nmの吸光度を測定した。亜硝酸態窒素標準液を用いて作成した検量線から亜硝酸態窒素量(NO2 --N)を測定した。
土壌から抽出した無機態窒素抽出液800 μlと、表5に示すブルシン・スルファニル酸溶液400 μlを試験管に分注し、硫酸溶液(硫酸:水 = 20:3) 4.0 mlを加えて撹拌した。冷暗所で40分間静置後、410 nmの吸光度を測定した。吸光度測定時に硝酸態窒素標準液を用いて検量線を作成し、得られた関係式を用いて硝酸態窒素量(NO3 --N)を測定した。
50 ml容遠沈管に土壌1.0 gを量り取り、表6に示すDNA抽出緩衝液(pH 8.0)を8.0 ml、20 %(w/v)ドデシル硫酸ナトリウム溶液を1.0 ml加え、1,500 rpm、室温で20分間撹拌した。撹拌後、50 ml容遠沈管から滅菌済み1.5 mlマイクロチューブに1.5 ml分取し、16℃、8,000 rpmで10分間遠心分離した。水層を新たなマイクロチューブに700 μl分取し、クロロホルム・イソアミルアルコール(24:1、v/v)を700 μl加えて混和した後、16℃、13,000 rpmで10分遠心分離した。遠心分離後、水層を新たなマイクロチューブに500 μl分取し、2-プロパノールを300 μl加えて緩やかに混和し、16℃、13,000 rpmで15分遠心分離した。遠心分離後、上清を除去し、70 %(v/v)エタノールを500 μl加え16℃、13,000 rpmで5分遠心分離した。遠心分離後、上清を除去しアスピレーターで30分間減圧乾燥させた。これに表7に示すTE 10:1緩衝液(pH 8.0)を50 μl加えよく溶解させ、これをeDNA溶液とした。アガロース2.0 g、表8に示す50×TAE緩衝液(pH 8.0)4.0 ml及び0.1 mMエチジウムブロマイド溶液20 μlに蒸留水を加えて200 mlとし、1.0 %アガロースゲルを作製した。eDNA溶液5.0 μlにローディングダイ(東洋紡、大阪)1.0 μlを混合し、全量6.0 μl、既知量のDNAを含むスマートラダー(ニッポンジーン、富山)1.5 μlをアガロースゲルにアプライした。これを100 Vで40分間電気泳動を行った後アガロースゲルにUV照射し、DNAバンドを確認した。KODAK 1D Image Analysis software(KODAK、NY、USA)を用いてスマートラダーのDNAバンドを解析し、蛍光強度に対するDNA量の検量線を作成した。この検量線を用いて、各サンプルDNA溶液のDNAバンドの蛍光強度からDNA量を求め、各土壌1.0 g当たりのeDNA量を算出した。eDNA量をDAPI染色による土壌バクテリア数に換算する検量線によって土壌バクテリア数を求めた。定量したeDNA量を関係式
Y = 1.7 × 108 X(R2 = 0.96)[Y;土壌バクテリア数(cells/g-soil)、X;eDNA量(μg/g-soil)]を用いて土壌バクテリア数を算出した。
用途や施肥状況が異なる土壌10サンプル(No.1~10)を用いて、無機態窒素量として、上記1b)~1d)に従ってアンモニア態窒素量と亜硝酸態窒素量を測定し、土壌の硝化活性を解析した。各サンプルにおける静置0日目と3日目における無機態窒素量及びその減少量を表9に示す。
物質循環では土壌バクテリアが密接に関与していると考えられる。このため、各サンプルにおける土壌バクテリア数を前記1fの方法で解析した。また、農地土壌における土壌バクテリア数のデータベースの平均値となる3.25×109 cells/g-soilを100として、測定した土壌バクテリア数の相対値(以下、バクテリア量ともいう)を算出した。各サンプルにおける土壌バクテリア数とバクテリア量を表10に示す。
得られた土壌バクテリア数、アンモニア減少率、亜硝酸減少率の3項目に基づき、土壌における窒素循環活性を評価するために、図2に示すチャートを作成した。
硝化の活性、即ち、窒素循環活性が低い土壌には、アンモニアを酸化する微生物の投与が効果的であると考えられる。そこで、アンモニア酸化細菌の投与によって硝化が活性化するかを解析した。
用途や施肥状況が異なる土壌10サンプル(No.11~20)を用いて、土壌のリン循環を解析した。
2a)土壌バクテリア数の解析
土壌バクテリア数は、前記1f)と同様に、eDNA解析法により定量した。
250 ml容UMサンプル瓶に土壌サンプル100 gを入れ、よく撹拌した。この土壌2.0 gを50ml容遠心チューブに量り取り、蒸留水20 ml加え、100 rpmで60分間振とうした。10,000 rpm、5分間遠心分離し、その上清を水溶性リン酸抽出液としてモリブデンブルー法に供した。水溶性リン酸抽出液1.0 mlを1.5 ml容マイクロチューブに分注し、表12に示すモリブデンブルー ストック溶液:0.41 M L(+)-アスコルビン酸水溶液 = 5:1の混合溶液100 μlを加えて撹拌後、30℃で30分静置した。静置後、720 nmにおける吸光度を測定し、リン酸標準液を用いて作成した検量線から、土壌中の水溶性リン酸を定量し、0日目の水溶性リン酸量とした。
250 ml容UMサンプル瓶に土壌サンプル100 gを入れ、よく撹拌した。この土壌2.0 gを50ml容遠心チューブに量り取り、蒸留水20 ml加え、100 rpmで60分間振とうした。10,000 rpm、5分間遠心分離後、抽出した水溶性リン酸をモリブデンブルー法に供し、リン酸量を定量して、0日目の水溶性リン酸量とした。
土壌バクテリア数、フィチン酸からのリン酸生成活性、および堆肥からのリン酸生成活性の3項目を用いて、図4に示すチャートを作成した。
用途や施肥状況が異なる土壌10サンプル(No.11~20)を用いて、土壌のカリウム循環を解析した。
3a)土壌バクテリア数の解析
土壌バクテリア数は、前記1fと同様に、eDNA解析法により定量した。
土壌3.0 gを50 ml容三角フラスコに量り取り、0.5 M硝酸40 mlを加えて60分間スターラーで撹拌した。撹拌後、ろ過し、ろ液をカリウム抽出液とした。この抽出液を原子吸光光度計(Z-2300、日立ハイテクノロジーズ、東京)を用いて測定した。測定条件は、燃料ガスとしてアセチレンを、助燃ガスとして圧縮空気を用い、圧力は共に0.5 MPaで測定した。カリウム標準液を用いて作成した検量線から、土壌中のカリウム遊離量を定量し、測定開始日(0日目)のカリウム遊離量とした。
250 ml容UMサンプル瓶に土壌サンプル100 gを入れ、よく撹拌した。土壌3.0 gを50 ml容三角フラスコに量り取り、蒸留水40 mlを加えて60分間スターラーで撹拌した。撹拌後、ろ過し、ろ液をカリウム抽出液とした。この抽出液を、前記3b)と同様にして、原子吸光光度計に供し、カリウム量を定量して、0日目のカリウム遊離量とした。
土壌バクテリア数、カリウム遊離率、およびカリウムからのリン酸生成活性の3項目を用いて、図5に示すチャートを作成した。
窒素循環活性、リン循環活性、及びカリウム循環活性に基づく土壌の総合診断と、植物の生長の関係を以下の方法で解析した。土壌としては、用途や施肥状況が異なる土壌10サンプル(No.11~20)を用いた。
4a)土壌バクテリア数の解析
土壌バクテリア数は、前記1fと同様に、eDNA解析法により定量した。
窒素循環活性は1項と同様にして解析した。
リン酸循環活性は2項と同様にして解析した。
カリウム循環活性は3項と同様にして解析した。
育苗ポットに各土壌サンプルを入れ、各ウェルにコマツナの種子10個を播種した。これを25℃、6,000ルクスで1週間生育させた。1/5,000 aポットに赤玉土を敷き詰め、土壌サンプル約1 kg加えて、ほぼ同じ大きさの苗を1ポット当たり3本移植した。これを25℃、6,000ルクスでさらに3週間生育させ、コマツナの地上部の生重量を測定した。なお、土壌1サンプルにつき3ポットで試験を行い、その平均値で評価した。
各土壌サンプルについて得られた指標の和から、土壌の総合診断を行った。即ち、各指標の和を3で割った値を算出し、得られた値が大きいほど土壌の品質に優れ、値が小さいほど土壌の品質が十分でないと診断した。
種々の土壌における硝化活性とバクテリア数との関係を調べた。図8に一日当たりの硝化量を示す。土壌バクテリア数は、前記1fと同様に、eDNA解析法により定量した。硝化量は、アンモニア態窒素が60 μg-N/g dry soilとなるように土壌に投与して1日間静置後に測定した亜硝酸態窒素量及び硝酸態窒素量の合計量を示している。その結果、微生物数が2億個/g以下になると、硝化反応が進まないことが明らかとなった。
アンモニア減少率、フィチンからのリン酸生成活性、及び堆肥からのカリウム生成活性について、それぞれ30、10、5以下であれば優れた土壌であると判断されない。基準値を頂点として形成される正三角形の面積に対する、正三角形の重心から対応する頂点を結ぶ線分上に置かれたこれらの下限値を頂点として形成される三角形の面積の割合は1.7点となる。そのため、この点未満であれば、良好な植物生長が期待出来ないと判断した。
従来技術では土壌中に含まれる栄養素の量やpH、CEC(塩基置換容量)などを測定し、農作物の生産に役立ててきたが、これらの値では堆肥などの有機物を多く含む肥料を添加した時の肥効が正確に把握できない。
Claims (8)
- 下記(I)~(III)を用いて算出される循環活性指標:
(I)対象土壌におけるアンモニア減少率、
(II)対象土壌におけるフィチン酸からのリン酸生成活性、及び
(III)対象土壌における堆肥からのカリウム生成活性
並びに(IV)土壌における土壌バクテリア数
を用いて土壌診断を行うことを特徴とする土壌の診断方法。 - 循環活性指標が、
予め設定されたアンモニア減少率の基準値、フィチン酸からのリン酸生成活性の基準値及び堆肥からのカリウム生成活性の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(I) アンモニア減少率、前記(II) フィチン酸からのリン酸生成活性、及び前記(III) 堆肥からのカリウム生成活性の測定点を頂点として形成される三角形の面積の割合である、請求項1に記載の診断方法。 - A)下記(A-1)~(A-3)を用いて算出される窒素循環活性指標:
(A-1)対象土壌における土壌バクテリア数、
(A-2)対象土壌におけるアンモニア減少率、及び
(A-3)対象土壌における亜硝酸減少率、
B)下記(B-1)~(B-3)を用いて算出されるリン循環活性指標:
(B-1)対象土壌における土壌バクテリア数、
(B-2)対象土壌におけるフィチン酸からのリン酸生成活性、及び
(B-3)対象土壌における堆肥からのリン酸生成活性、
並びに、C)下記(C-1)~(C-3)を用いて算出されるカリウム循環活性指標:
(C-1)対象土壌における土壌バクテリア数、
(C-2)対象土壌におけるカリウム遊離率、及び
(C-3)対象土壌における堆肥からのカリウム生成活性
を少なくとも用いて土壌診断を行うことを特徴とする土壌の診断方法。 - 窒素循環活性指標が、
予め設定された土壌バクテリア数の基準値、アンモニア減少率の基準値、及び亜硝酸減少率の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(A-1)土壌バクテリア数、前記(A-2)アンモニア減少率、及び前記(A-3)亜硝酸減少率の測定点を頂点として形成される三角形の面積の割合である、請求項3に記載の診断方法。 - リン循環活性指標が、
予め設定された土壌バクテリア数の基準値、フィチン酸からのリン酸生成活性の基準値、及び堆肥からのリン酸生成活性の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(B-1)土壌バクテリア数、前記(B-2)フィチン酸からのリン酸生成活性、及び前記(B-3)堆肥からのリン酸生成活性の各測定値を頂点として形成される三角形の面積の割合である、請求項3又は4に記載の診断方法。 - カリウム循環活性指標が、
予め設定された土壌バクテリア数の基準点、カリウム遊離率の基準値、及び、堆肥からのカリウム生成活性の基準値を頂点として形成される正三角形の面積に対する、
前記正三角形の重心から対応する頂点を結ぶ線分上に置かれた前記(C-1)土壌バクテリア数、前記(C-2)カリウム遊離率、及び前記(C-3)堆肥からのカリウム生成活性の各測定値を頂点として形成される三角形の面積の割合である、請求項3又は4に記載の診断方法。 - 請求項1又は3に記載の診断方法を経時的に行って、前記指標の経時変化を解析することにより、土壌の品質を管理することを特徴とする、土壌の品質管理方法。
- 請求項1又は3に記載の診断方法を行って、得られた診断結果に基づき、前記指標を改善するための処理を行うことを特徴とする土壌の改善方法。
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JP2011504900A JP5578525B2 (ja) | 2009-03-19 | 2010-03-19 | 新規土壌診断方法 |
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US8682584B2 (en) * | 2011-08-19 | 2014-03-25 | Brookside Laboratories, Inc. | Nitrogen potential index |
FR2997961B1 (fr) * | 2012-11-12 | 2014-12-05 | Polyor Sarl | Diagnostic de l'etat microbiologique de sols en fonction de la resilience de populations bacteriennes qu'ils contiennent |
FR3072854B1 (fr) * | 2017-11-02 | 2019-11-29 | Sarl Polyor | Methode pour la formation d’un indicateur elementaire de l’efficacite agronomique d’azotobacteries en sols arables |
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CN109064039B (zh) * | 2018-06-04 | 2022-02-11 | 北京捷西农业科技有限责任公司 | 一种农田土壤健康评价方法 |
CN111912802B (zh) * | 2020-08-25 | 2023-05-26 | 潍坊市生态环境局寿光分局 | 一种分光光度法检测亚硝酸盐氮的方法 |
CN113020232B (zh) * | 2021-03-09 | 2022-03-15 | 农业农村部环境保护科研监测所 | 一种受污染耕地综合治理及动态调控方法 |
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