CN114586582A - Method for killing root-knot nematodes by using methyl p-hydroxyphenylpropionate - Google Patents

Method for killing root-knot nematodes by using methyl p-hydroxyphenylpropionate Download PDF

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CN114586582A
CN114586582A CN202210161727.2A CN202210161727A CN114586582A CN 114586582 A CN114586582 A CN 114586582A CN 202210161727 A CN202210161727 A CN 202210161727A CN 114586582 A CN114586582 A CN 114586582A
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soil
root
methyl
hydroxyphenylpropionate
water
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Inventor
王甲辰
王学霞
李钰飞
刘建斌
张树彬
武战会
梁浩
陈延华
曹兵
倪小会
梁丽娜
李博文
高宏双
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Beijing Academy of Agriculture and Forestry Sciences
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Beijing Academy of Agriculture and Forestry Sciences
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G13/00—Protecting plants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
    • A01N37/38—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01M—CATCHING, TRAPPING OR SCARING OF ANIMALS; APPARATUS FOR THE DESTRUCTION OF NOXIOUS ANIMALS OR NOXIOUS PLANTS
    • A01M17/00—Apparatus for the destruction of vermin in soil or in foodstuffs
    • A01M17/002—Injection of toxic gases or fluids into the soil

Abstract

The invention discloses a method for killing root-knot nematodes by using methyl p-hydroxyphenylpropionate, and belongs to the technical field of biology. The method of the invention is to apply methyl p-hydroxyphenylpropionate to the soil. The invention utilizes the methyl p-hydroxyphenylpropionate with a certain concentration to kill or inhibit root-knot nematodes at any time and anywhere, even in all growth periods of the crops, thereby ensuring the healthy growth of the crops.

Description

Method for killing root-knot nematodes by using methyl p-hydroxyphenylpropionate
Technical Field
The invention relates to the technical field of biology, in particular to a method for killing root-knot nematodes by using methyl p-hydroxyphenylpropionate.
Background
Root-knot nematodes are a public nuisance in the field of planting, and can damage root systems of many economic crops such as cucumbers, tomatoes, beans and the like, and seriously affect the growth of the root systems and the absorption of nutrients and water of the crops. Once it occurs, the mild person has reduced labor, the severe person is almost no longer in harvest, and it is extremely difficult to completely kill the disease. Root-knot nematodes spread rapidly in plantations and can be transmitted by nursery soil, machinery, working tools or shoe sole clay.
Currently, treatment methods include chemical, physical, electrical, and biological methods. The chemical method mainly adopts linear phosphorus, miloer, Yishubao, fritillaria pallidiflora and the like, the agents are scattered on the soil, and the greenhouse is covered with a film after rotary tillage for about 30 days. The physical method is to cover the soil with a three-prevention cloth after rotary tillage, introduce water vapor into the soil, and kill soil organisms and microorganisms by using heat and simultaneously kill root-knot nematodes. The electrical method mainly passes a large current of 5A or more and a voltage of 900 v or more. The biological method is to utilize the combination of para hydroxybenzene methyl propionate (MHPP) secreted by sorghum and ammonium nitrogen fertilizer or organic fertilizer capable of mineralizing to produce ammonium ions to kill root-knot nematodes. These methods all have their own drawbacks, mainly expressed as: the chemical method has the problems of high reagent price, unwilling acceptance by users, needing professional operation, polluting soil, incomplete killing and the like; the physical method mainly adopts special equipment, has high cost, destroys the soil ecosystem and also cannot kill thoroughly. The ammonium ions have strong killing effect on the root-knot nematodes, such as lime nitrogen and liquid ammonia. Lime nitrogen has the defects of uneven spreading, great harm to people and the like, while liquid ammonia has the problems of high-pressure storage, difficult control of operator protection during application, difficult control of gas tank release amount, easy nitrogen source pollution caused by excessive soil nitrogen nutrients and the like, and the two measures have the defect that ammonium ions are quickly nitrified into nitrate nitrogen by soil nitrifying bacteria to lose the killing effect, so that the killing efficiency is not ideal. Although the patent disclosed at present utilizes the root system of special crops such as sorghum, peanut, rice and the like to secrete natural nitrification inhibitor, ammonium ions can be stored in soil for a long time to form a soil layer for ammonium ion long-term storage, so that the root-knot nematode can not tolerate the ammonium ions and die. However, the main problem of the technology is that the operation is complex, the accumulated high-concentration ammonium ions inhibit the growth of the next-stubble crops or the crops can be cultivated only by washing off excessive ammonia nitrogen, so that the soil application efficiency is reduced. In conclusion, although the method of combining chemical, physical, electric, lime nitrogen, liquid ammonia, biological nitrification inhibitor, ammonium nitrogen fertilizer and the like for killing root-knot nematodes has certain effect, a series of problems of soil pollution, high cost, incomplete killing, complex operation and the like exist.
A natural nitrification inhibitor methyl p-hydroxyphenylpropionate (MHPP) secreted by sorghum roots is artificially synthesized at present, and is mainly used as a natural nitrification inhibitor to improve the utilization rate of nitrogen fertilizers and reduce the non-point source pollution of nitrogen. The latest research shows that the plant growth regulator can be used as a root system regulator to inhibit the growth of main roots and stimulate the generation of lateral roots, and is favorable for the absorption of plant roots to nutrients.
Disclosure of Invention
The invention provides a method for killing root-knot nematodes by utilizing methyl p-hydroxyphenylpropionate, which can remarkably inhibit the root-knot nematodes or has better killing effect on the root-knot nematodes after a certain concentration of methyl p-hydroxyphenylpropionate (MHPP) is used for acting for a period of time; and the concentration lower than or higher than the concentration range not only can not kill but also can promote the growth of the root-knot nematode; the invention utilizes the methyl p-hydroxyphenylpropionate with a certain concentration to kill or inhibit root-knot nematodes at any time and anywhere, even in all growth periods of the growth of crops, thereby ensuring the healthy growth of vegetable crops.
The invention provides a method for killing root-knot nematodes, which comprises the following steps: methyl p-hydroxyphenylpropionate is applied to the soil.
In the method, the concentration of the methyl p-hydroxyphenylpropionate after being applied to the soil is 0.001-500 mu g/L.
The concentration of the methyl p-hydroxyphenylpropionate after being applied to soil is preferably 0.01-300 mu g/L, and more preferably 0.01-200 mu g/L; furthermore, the concentration of the methyl p-hydroxyphenylpropionate after being applied to the soil is preferably 0.5-150 mug/L, 1-150 mug/L, 100 mug/L or 150 mug/L.
In the method, the using period of the methyl p-hydroxyphenylpropionate is the idle period, the early period or the middle and later periods of the growth of the crops.
In the method, the application method comprises the steps of firstly dissolving a small amount of water and directly spraying methyl p-hydroxyphenylpropionate solution on the ground surface, and then watering; or the like, or, alternatively,
and preparing a methyl p-hydroxyphenylpropionate solution, and irrigating at least one of water irrigation, drip irrigation, spray irrigation and small high-ridge irrigation.
In the method, the action time of the methyl p-hydroxyphenylpropionate after being applied to the soil is 5-21 days.
The invention has the following advantages:
(1) the method has the advantages of low consumption of methyl p-hydroxy-propionate, low price, simple use technology and no influence by the growth period, season and the like;
(2) MHPP is a natural sorghum secretion product, has no harm to a soil ecosystem and agricultural products, ensures food safety, promotes the absorption of plant nutrients and improves the yield.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention.
The experimental procedures in the following examples are all conventional ones unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The methyl p-hydroxyphenylpropionate (MHPP) used in the examples described below was obtained from Shanghai Allantin Biotech Co., Ltd.
The 4% formaldehyde is the volume percent concentration described in the examples below.
Example 1
1. Method for field operation of killing nematode
On the facility kidney bean soil with serious root knot nematode in southern garden of Dahancun, Zhou kou, Beijing, the idle period before the kidney bean planting (1 month, 25 months to 2 months and 4 days in 2020), the facility greenhouse area is about 800m2. The test shows that the soil moisture content is 18.5 percent (weight percentage), and the total moisture content of the soil plough layer (0-20cm) is 27.75 t. The economical irrigation quantity is 300m according to the facility of the garden3/hm2The actual irrigation is 300m3/hm2-27.75m3/hm2＝272.25m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.6g MHPP; processing 3, weighing 1.5g MHPP; treatment 4. weighing 3. mu.g MHPP, putting the weighed samples into a spraying pot with 15kg water, and after dissolving, uniformly spraying the samples onto 4 blocks of 100m2On the ground, then water irrigation quantity is calculated to be 2.72m3Respectively irrigating, respectively filling water with MHPP concentration of 0 mug/L (irrigation water), 200 mug/L, 500 mug/L and 0.001 mug/L in a soil layer of 0-20cm, and sampling after 10 days to test the killing rate of the root-knot nematodes.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weigh 60g of soil and put into a small sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the sieve with the diameter of 1mm is taken out (the soil sample is not used at this time), the water in the dish is poured into the sieve with the diameter of 500 meshes, the sieve is inclined at an angle of 45 degrees so as to be convenient for flushing and collecting the nematodes, and the nematodes are separated from the water and attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper liquid layer of the centrifuge tube is slowly sucked out, and about 5mL of liquid remains in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper (a small grid of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the visual field size of a microscope, so that counting is convenient, and finally the culture dish is placed under the microscope for observing and counting the root-knot nematodes.
4. Results
The results are shown in Table 1. The best effect is treatment 2, the killing rate of root-knot nematodes is 85.63%, the concentration is not suitable for treatment 3 and treatment 4, and the killing rate is 27.05 and 12.96% respectively.
TABLE 1 Effect of different concentrations of MHPP on root-knot nematode killing
Figure BDA0003514253240000031
Figure BDA0003514253240000041
Example 2
1. Method for field operation of killing nematode
On the protected tomato soil with serious root-knot nematode in southern garden of Changzi, Yingzi, Yingcun, Beijing great Xing area, the idle period before tomato planting (1 month, 15-1 month and 20 days in 2021) is utilized, and the area of the greenhouse of the facility is about 800m2. Tests show that the water content of the soil is 19.2 percent (weight percentage) and the total water content of a soil plough layer (0-20cm) is 2880 t. The economical irrigation quantity is 300m according to the facility of the garden3/hm2The actual irrigation is 300m3/hm2-28.8m3/hm2＝271.2m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.0003g of MHPP; processing 3, weighing 1.5g MHPP; treatment 4. weighing 3. mu.g MHPP, putting the weighed samples into a spraying pot with 15kg water, and after dissolving, spraying the weighed samples onto 4 blocks of 100m2On the ground, then according to the calculated irrigation quantity of 2.71m3Respectively irrigating, respectively filling water with MHPP concentration of 0 mug/L (irrigation water), 0.1 mug/L, 500 mug/L and 0.001 mug/L in a soil layer of 0-20cm, and sampling 5 days later to test the killing rate of the root-knot nematodes.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weighing 60g of soil, and putting the soil into a small-size sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the 1mm sieve and the sieve is taken out (the soil sample is not used at this time), the water in the dish is poured into a 500-mesh sieve (the sieve is inclined at an angle of 45 degrees so as to be convenient for washing and collecting the nematodes), and the water and the nematodes are separated, and the nematodes can be attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper layer liquid in the centrifuge tube was slowly aspirated, and about 5mL of liquid remained in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper, a small grid of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the size of a microscope visual field, and counting is convenient. And finally, placing the solution under a microscope to observe and count the root-knot nematodes.
4. Results
The results are shown in Table 2. The best treatment was treatment 2, the root knot nematode kill rate was 83.01%, less suitable concentrations were treatment 3 and treatment 4, and the kill rates were 42.79% and 12.45%, respectively.
TABLE 2 killing effect of different concentrations of MHPP on root-knot nematodes
Figure BDA0003514253240000051
Example 3
1. Method for field operation of killing nematode
On the facility soil with serious cucumber root knot nematode in Changzi, river Yangchun garden in great Xingxing area of Beijing, the idle period before cucumber permanent planting (1 month, 22 days to 2 months, 11 days in 2021) is utilized, and the facility greenhouse area is about 800m2. Tests show that the water content of the soil is 17.25 percent (weight percentage), and the total water content of a plough layer (0-20cm) of the soil is 25.88 t. The economical irrigation quantity is 300m according to the facility of the garden3/hm2The actual irrigation is 300m3/hm2-28.8m3/hm2＝274.12m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.00003g MHPP; processing 3, weighing 1.5g MHPP; treatment 4. weighing 3 μ g (weighing 1g, using constant volume dilution method for preparing solution) MHPP, putting the weighed samples into a spray can with 15kg water, respectively, and after dissolving, spraying onto 4 blocks of 100m MHPP with the spray can2On the ground, then according to the calculated irrigation quantity of 2.74m3And (3) irrigating, wherein MHPP concentrations in a soil layer of 0-20cm are 0 mug/L (clear water), 0.01 mug/L, 500 mug/L and 0.001 mug/L respectively, and sampling is carried out after 21 days to test the killing rate of the root-knot nematodes.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weighing 60g of soil, and putting the soil into a small-size sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the sieve with the diameter of 1mm is taken out (the soil sample is not used at this time), the water in the dish is poured into the sieve with the diameter of 500 meshes, the sieve is inclined at an angle of 45 degrees so as to be convenient for flushing and collecting the nematodes, and the nematodes are separated from the water and attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper layer liquid in the centrifuge tube was slowly aspirated, and about 5mL of liquid remained in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper, a small grid of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the size of a microscope visual field, and counting is convenient. And finally, placing the solution under a microscope to observe and count the root-knot nematodes.
4. Results
The results are shown in Table 3. The best treatment was treatment 2, the root knot nematode kill rate was 84.96%, less suitable concentrations were treatment 3 and treatment 4, with kill rates of 25.17% and 29.96%, respectively.
TABLE 3 killing effect of different concentrations of MHPP against root-knot nematodes
Figure BDA0003514253240000061
Example 4
1. Method for field operation of killing nematode
In the protected tomato soil with serious root knot nematode in the garden of Beizhong Jinshan in Yangzhou village of Ni Jiazhuang village in Hebei Ligusticum urban area, the early stage of tomato fruit setting (3 months and 18 days in 2021) is utilized to make the protected tomato soilThe greenhouse area is about 600m2. The test shows that the soil moisture content is 20.34% (weight percentage), and the total moisture content of the soil plough layer (0-20cm) is 30.51 t. The water irrigation quantity is 300m according to the economical irrigation quantity of the garden facility3/hm2The actual irrigation is 300m3/hm2-30.51m3/hm2＝269.49m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.3g MHPP; processing 3, weighing 1.5g MHPP; processing 4. weighing 3 μ g MHPP, respectively putting the weighed samples into 15kg water containers, and respectively uniformly mixing the samples with the water consumption of 2.69t/100m of a drip irrigation system after the samples are dissolved2Drip irrigation respectively at 4 blocks of 100m2On the ground, soil solutions with MHPP concentrations of 0 mug/L (clear water), 100 mug/L, 500 mug/L and 0.001 mug/L are formed in a soil layer of 0-20cm, and sampling is carried out on 3-23 months in 2022 to test the killing rate of the root-knot nematodes.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weigh 60g of soil and put into a small sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the sieve with the diameter of 1mm is taken out (the soil sample is not used at this time), the water in the dish is poured into the sieve with the diameter of 500 meshes, the sieve is inclined at an angle of 45 degrees so as to be convenient for flushing and collecting the nematodes, and the nematodes are separated from the water and attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper layer liquid in the centrifuge tube was slowly aspirated, and about 5mL of liquid remained in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper, a small grid of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the size of a microscope visual field, and counting is convenient. And finally, placing the solution under a microscope to observe and count the root-knot nematodes.
4. The results are shown in Table 4. The best treatment was treatment 2 with a root knot nematode kill of 92.52%, less suitable concentrations for treatments 3 and 4, with kill rates of 39.86% and 19.11%, respectively.
TABLE 4 killing effect of different concentrations of MHPP on root-knot nematodes
Figure BDA0003514253240000071
Example 5
1. Method for field operation of killing nematode
In the protected cucumber soil with serious root knot nematode in Ni Hebei Ligusticum urban area, 4 months and 20 days in 2021 year, the protected greenhouse area is about 800m2. The test shows that the water content of the soil is 21.96 percent (weight percentage), and the total water content of the soil plough layer (0-20cm) is 30.51 t. The water irrigation quantity is 300m according to the economical irrigation quantity of the garden facility3/hm2The actual irrigation is 300m3/hm2-32.94m3/hm2＝267.06m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.45g MHPP; processing 3, weighing 1.5g MHPP; treatment 4. mu.g of MHPP were weighed, and the above weighed samples were placed in containers of 15kg of water, respectively, and after dissolution, the regions were 2.67m each3/1002The water quantity is uniformly sprayed on 4 blocks of 100m along with the water2On the ground where cucumbers grow, soil solutions with MHPP concentrations of 0 mug/L (irrigation water), 150 mug/L, 500 mug/L and 0.001 mug/L are formed in a soil layer of 0-20cm, and sampling is carried out to test the killing rate of root-knot nematodes at 30 months and 4 months in 2021.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weighing 60g of soil, and putting the soil into a small-size sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the sieve with the diameter of 1mm is taken out (the soil sample is not used at this time), the water in the dish is poured into the sieve with the diameter of 500 meshes, the sieve is inclined at an angle of 45 degrees so as to be convenient for flushing and collecting the nematodes, and the nematodes are separated from the water and attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper layer liquid in the centrifuge tube was slowly aspirated, and about 5mL of liquid remained in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper, a small grid with the size of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the size of a microscope visual field, and counting is convenient. And finally, placing the solution under a microscope to observe and count the root-knot nematodes.
4. As a result, the
The results are shown in Table 5. The best treatment was treatment 2 with a root knot nematode kill of 92.41%, less suitable concentrations for treatment 3 and treatment 4 with kill rates of 39.86% and 37.98%, respectively.
TABLE 5 killing effect of different concentrations of MHPP against root-knot nematodes
Figure BDA0003514253240000081
Example 6
1. Method for field operation of killing nematode
Greenhouse cucumber soil with serious root knot nematode in northern garden of Daheijia Tuo village in Hebei LeTing countyOn the earth, the full bearing age of cucumber (2021 year, 4 months and 20 days) is utilized, and the greenhouse area of the facility is about 800m2. The test shows that the water content of the soil is 20.06 percent (weight percentage), and the total water content of the soil plough layer (0-20cm) is 30.09 t. The water irrigation quantity is 300m according to the economical irrigation quantity of the garden facility3/hm2The actual irrigation is 300m3/hm2-30.09m3/hm2＝269.91m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.003g MHPP; processing 3, weighing 1.5g MHPP; treatment 4. weighing 3. mu.g MHPP, placing the weighed samples into 15kg water containers respectively, and uniformly distributing the weighed samples to 100m after dissolution2In small high ridges, 4 blocks of 100m are irrigated in a common mode2Separately irrigate 2.70m on the ground3The MHPP formation concentration in a soil layer of 0-20cm is respectively 0 mug/L (irrigation water), 1 mug/L, 500 mug/L and 0.001 mug/L, and sampling is carried out on 5, 11 days in 2021 year to test the killing rate of the root-knot nematodes.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weighing 60g of soil, and putting the soil into a small-size sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the sieve with the diameter of 1mm is taken out (the soil sample is not used at this time), the water in the dish is poured into the sieve with the diameter of 500 meshes, the sieve is inclined at an angle of 45 degrees so as to be convenient for flushing and collecting the nematodes, and the nematodes are separated from the water and attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper layer liquid in the centrifuge tube was slowly aspirated, and about 5mL of liquid remained in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper, a small grid of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the size of a microscope visual field, and counting is convenient. And finally, placing the solution under a microscope to observe and count the root-knot nematodes.
4. Results
The results are shown in Table 6. The best treatment was treatment 2 with a root knot nematode kill of 93.17%, less suitable concentrations for treatments 3 and 4 of 52.77% and 31.18%, respectively.
TABLE 6 killing effect of different concentrations of MHPP against root-knot nematodes
Figure BDA0003514253240000091
Example 7
1. Method for field operation of killing nematode
In the loofah soil with serious root knot nematode in southern garden of great Han secondary village in township of Beijing Zhou kou shop, the full bearing period of loofah (2021 year, 9 months and 20 days) is utilized, and the greenhouse area of the facility is about 800m2. The test shows that the water content of the soil is 19.15 percent (weight percentage), and the total water content of the soil plough layer (0-20cm) is 28.73 t. The water irrigation quantity is 300m according to the economical irrigation quantity of the garden facility3/hm2The actual irrigation is 300m3/hm2-28.73m3/hm2＝271.27m3/hm2. Selecting 4 blocks of 100m each in greenhouse2The killing effect demonstration of different MHPP concentration treatments on root-knot nematodes is developed: an exemplary process is: processing 1, weighing 0g of MHPP; processing 2, weighing 0.0015g MHPP; processing 3, weighing 1.5g MHPP; treatment 4. weighing 3. mu.g MHPP, placing the weighed samples into 15kg water containers respectively, and uniformly distributing the weighed samples to 100m after dissolution2In the big bed, 4 blocks of 100m are irrigated in a common mode2Separately irrigate 2.71m on the ground3In a 0-20cm soil layer, the MHPP formation concentrations are 0 mug/L (water), 0.5 mug/L, 500 mug/L and 0.001 mug/L respectively, and the roots are sampled and tested from 10 months and 5 days in 2021 yearKill rate of the knot nematode.
2. Root-knot nematode sampling and separating method
On each treatment field, the area was divided into 3 areas (three replicates), 5 drill soil samples were randomly taken from 0-20cm soil layer in each area using a root drill, stirred for 15 minutes, and mixed uniformly. Then the mixture is sieved by a 2mm sieve and stirred and mixed evenly again. Weighing 60g of soil, and putting the soil into a small-size sieve with the aperture of 1mm and a matched small disc (the diameter of the sieve is larger). Adding water capable of submerging soil surface from the periphery, refrigerating the refrigerated cabinet for 48 hours, and supplementing water in time. After 48 hours, the soil sample in the sieve with the diameter of 1mm is taken out (the soil sample is not used at this time), the water in the dish is poured into the sieve with the diameter of 500 meshes, the sieve is inclined at an angle of 45 degrees so as to be convenient for flushing and collecting the nematodes, and the nematodes are separated from the water and attached to the surface of the sieve. Washing the plate with a mild water flow to remove residual nematodes, sieving, washing several times, pouring the water in the sieve into a 50mL centrifuge tube, standing for settling overnight (12 hours or more), sucking out the supernatant next day, adding about 5mL of the remaining liquid in the tube, and adding 20mL of 4% (v/v) formaldehyde for fixation.
3. Microscopic examination of root-knot nematodes
The upper layer liquid in the centrifuge tube was slowly aspirated, and about 5mL of liquid remained in the tube. The 5mL of liquid is dripped into a culture dish prepared in advance by a dropper, a small grid of 5 multiplied by 5mm is drawn at the bottom of the culture dish, the size of the small grid is basically consistent with the size of a microscope visual field, and counting is convenient. And finally, placing the solution under a microscope to observe and count the root-knot nematodes.
4. Results
The results are shown in Table 6. The best treatment was treatment 2 with a root knot nematode kill of 88.03%, less suitable concentrations for treatments 3 and 4 were 34.47% and 30.77%, respectively.
TABLE 7 killing effect of different concentrations of MHPP against root-knot nematodes
Figure BDA0003514253240000101

Claims (9)

1. A method for killing root-knot nematodes, comprising the steps of: methyl p-hydroxyphenylpropionate is applied to the soil.
2. The method of claim 1, wherein: the concentration of the methyl p-hydroxyphenylpropionate after being applied to soil is 0.001-500 mu g/L.
3. The method of claim 2, wherein: the concentration of the methyl p-hydroxyphenylpropionate after being applied to soil is 0.01-300 mu g/L.
4. The method of claim 3, wherein: the concentration of the methyl p-hydroxyphenylpropionate after being applied to soil is 0.01-200 mu g/L.
5. The method of claim 4, wherein: the concentration of the methyl p-hydroxyphenylpropionate after being applied to soil is 0.5-150 mu g/L.
6. The method of claim 5, wherein: the concentration of the methyl p-hydroxyphenylpropionate applied to the soil is 1-150 mu g/L.
7. The method according to any one of claims 1-6, wherein: the using period of the methyl p-hydroxyphenylpropionate is the idle period, the early period or the middle and later period of the growth of crops.
8. The method according to any one of claims 1-7, wherein: the application method comprises the steps of firstly dissolving a small amount of water and directly spraying methyl p-hydroxybenzoate solution on the ground surface, and then watering; or the like, or, alternatively,
and preparing a methyl p-hydroxyphenylpropionate solution, and irrigating at least one of water irrigation, drip irrigation, spray irrigation and small high-ridge irrigation.
9. The method according to any one of claims 1-8, wherein: the action time of the methyl p-hydroxyphenylpropionate after being applied to the soil is 5-21 days.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108605931A (en) * 2018-04-20 2018-10-02 中国热带农业科学院热带生物技术研究所 Application of four kinds of compounds in preparing nematicidal drug
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WO2019012377A1 (en) * 2017-07-10 2019-01-17 Basf Se Mixtures comprising an insecticide and a nitrification inhibitor such as 2-(3,4-dimethyl-1h-pyrazol-1-yl)succinic acid (dmpsa) or 3,4-dimethyl pyrazolium glycolate (dmpg)
CN108605931A (en) * 2018-04-20 2018-10-02 中国热带农业科学院热带生物技术研究所 Application of four kinds of compounds in preparing nematicidal drug
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