CN113519560B

CN113519560B - Plant extract formula of compound molluscicide and preparation method thereof

Info

Publication number: CN113519560B
Application number: CN202110499422.8A
Authority: CN
Inventors: 杨绪勤; 孙亚萍; 张又文; 高玉忠; 杨香璿; 宛雯; 周霞; 师静静; 蒋继宏
Original assignee: Jiangsu Normal University
Current assignee: An Feng
Priority date: 2021-09-10
Filing date: 2021-09-10
Publication date: 2022-09-06
Anticipated expiration: 2041-09-10
Also published as: CN113519560A

Abstract

A formula of a compound biological molluscicide containing plant extracts comprises the following components: 40% of the extract of the knee of the milch cow, 10% of the tea saponin which accounts for 80%, 8% of the gypsum powder, 1% of the attractant and 41% of the flour, the tea saponin and the plant extract of the milch cow knee are compounded, so that metaldehyde in the traditional pesticide can be replaced, and the dosage of the chemical pesticide in the field can be reduced; people avoid the possibility of causing diseases and even cancers under long-time contact, and serious people can also damage the genetic mechanism of organisms to cause gene mutation; the invention can reduce the potential harm of metaldehyde to the environment and human body; the compound tea saponin and the camu knee compound have an addition effect on mutual molluscicidal effect while the drug effect is obvious, the drug cost is reduced, the ecological environment is protected.

Description

Plant extract formula of compound molluscicide and preparation method thereof

Technical Field

The invention relates to the field of biochemistry, and relates to a biological pesticide formed by compounding tea saponin and a radix achyranthis bidentatae extract, which is used as a molluscacide for preventing and treating ampullaria gigas disasters. The compound biopesticide of the invention has stronger drug effect and low price, can reduce the pollution to the environment and has huge industrialization prospect.

Background

Radix seu caulis Mucunae Sessileyanae (radix Achyranthis bidentatae)Cyathula capitata MoqIs a perennial herb cyathula plant of the amaranthaceae family. Rough and strong root, grayish brown or brownish red, round and conical root strip, little distortion. The stem is upright, nearly quadrangular, and branched, and the stem and the branch are sparse to grow grey and soft hair. Grows under the mixed trees in the 2300m hillside with the elevation of 1700-. Niu xi is mainly produced in the area of Jinshajiang river, Sichuan, Yunnan and Tibet. Collected in autumn and winter, removed soil, ground stem and leaf and fibrous root, dried in the sun or baked. The root is short and thick, the long cylindrical cone shape, the thickness difference of the two ends is large, the head is big, the tail is small, and the twisting is frequent. The surface is dark yellow and dark brown, and has skin pores and longitudinal wrinkles. The body is hard and brittle and is easy to break, and the section is slightly brownish red. Strong smell, slightly sweet, bitter and astringent taste, slightly numb taste.

Pomacea canaliculata is also called thamnaeus and apple snail. After 1984, ampullaria gigas have been widely cultivated in the Guangdong. Due to over-cultivation, plus bad taste, it is released to the field. The ampullaria gigas has strong environment-adaptive viability and can propagate quickly, and can live through winter naturally in many areas and two generations occur in the year. 2, 2.5 million young snails are co-propagated by 1 female snail through two generations in 1 year, the fertility is extremely strong, and the potential hazard is huge. Thus rapidly spreading to rivers, lakes and fields; the food intake is large, the variety of food is large, the growth of grain crops, vegetables and aquatic crops can be damaged, and the food becomes harmful animals in places such as Guangdong, Guangxi, Fujian, Yunnan, Zhejiang, Shanghai, Jiangsu and the like. The ampullaria gigas can naturally live through the winter in the area of south of the Yangtze river, and two generations occur in one year. In Guangxi Luzhai county and other places, the occurrence density of the ampullaria gigas in the rice field is as high as 16.95/square meter; the damage rate of the rice plants is generally 7% -15% and can reach 64%. The ampullaria gigas not only harms rice, but also harms vegetables and other aquatic crops, and also harms various aquatic plants such as narcissus, orchid and the like, and becomes a big hazard in water areas in south China. In addition, the ampullaria gigas is an intermediate host of parasitic diseases which are commonly suffered by people and livestock, and is very easy to bring health problems to surrounding residents. It is estimated that the water areas (ponds, lakes, paddy fields, etc.) harmed by China in provinces and cities in south China are not less than 10 ten thousand hectares. At present, chemical pesticides are mainly used for preventing and treating mollusks, about 50 kinds of mollusks mainly comprise 6% metaldehyde, shell snail killer, 6% mineral granules, 45% fentin acetate, 45% Bailuidi and the like, and the organic molluscicides have a certain prevention effect on ampullaria gigas, but have strong stimulation effect on human skin, some organic molluscicides even cause teratogenesis and carcinogenesis, have high toxicity on aquatic organisms, damage to ecological balance in the nature to a certain extent, and have high prevention and treatment cost, so the molluscicides have great limitation in practical application.

With the continuous development of social science and technology, biological pesticides are also continuously developed, which is not separated from the research of biological control and the current situation of chemical pesticide use flooding. The novel-technology biopesticide effectively protects national agricultural development and green organic products, reduces the damage and pollution degree to the environment while continuously enhancing the capability of preventing and controlling plant diseases and insect pests, reduces the problem of pesticide pollution to the environment to a certain extent, and protects the ecological environment. In order to ensure the stable grain yield and prevent and treat plant diseases and insect pests, farmers can spray a large amount of chemical pesticides into the field. Not only the ecological environment is destroyed, but also the fertility of the land is influenced, the production cost of the crops is increased, and the burden of farmers is caused. China is a vast agricultural country with broad farmers, the cultivated land area in China is 14.3 hundred million acres, and the rice planting area is stabilized at 4.5 hundred million acres. This provides a solid foundation for the development of biological pesticides in China. In european countries, the success of synthesizing or modifying plant-derived active substances as lead structures depends on two major advantages, namely, full automation of compound synthesis and automation of high-throughput screening technology. In China, a system for independently creating new pesticides is not really formed at present. Has wide market prospect.

People have recognized the importance of protecting the ecological environment, and the strategy of sustainable development becomes a consensus of people all over the world. The reduction of chemical fertilizer and pesticide is an effective measure which must be implemented in developing China. Although the biopesticides still have certain problems in the development process, many successful achievements are achieved, and the future development situation is great.

Disclosure of Invention

The current chemical pesticide molluscicide circulating in the market is a 6% metaldehyde preparation, although the molluscicide has obvious killing effect, serious environmental pollution and ecological damage can be caused after long-term use, and the molluscicide greatly harms human health. The molluscacide is prepared by mixing the most suitable proportion for killing the ampullaria gigas, which is obtained by taking the botanical biological pesticide as the main material and the gypsum powder, the attractant and the flour as the auxiliary materials, and the raw materials are easy to obtain and have lower cost. The invention ensures that crops have no chemical pesticide residue, and ensures the development of green agriculture.

The invention is a compound biological pesticide which takes plant-derived biological pesticides as main materials and can effectively kill ampullaria gigas, can replace 6% metaldehyde chemical pesticides circulating in the market to prevent and control the harm of ampullaria gigas, can achieve the effect of killing ampullaria gigas in a short time, and can promote the development of green pesticides. The combination ratios of the present invention will be provided below:

the invention comprises the following steps: the components are as follows by weight percent: 80% of tea saponin 20%, 8% of gypsum powder, 1% of attractant and 71% of flour, and fully stirring and mixing to obtain the 80% tea saponin wettable preparation.

The second invention is that: the components are as follows by weight percent: 60% of radix achyranthis bidentatae extract, 8% of gypsum powder, 1% of attractant and 31% of flour are added to 100% and fully stirred and mixed to prepare the wettable preparation of the radix achyranthis bidentatae extract.

The invention III: the weight percentage of each component is as follows: 80% of tea saponin, 40% of the extract of the knee of the Ma niu, 8% of gypsum powder, 1% of attractant and 41% of flour, which are complemented to 100%, are fully stirred and mixed to prepare the tea saponin-extract wettable preparation of the Ma niu.

According to the invention, the tea saponin and the plant extract of the camento knee are compounded, so that metaldehyde in the traditional pesticide can be replaced, and the dosage of the field chemical pesticide is reduced; people avoid the possibility of causing diseases and even cancers under long-time contact, and seriously damage the genetic mechanism of organisms to cause gene mutation; the invention can reduce the potential harm of metaldehyde to the environment and human body; the composition has the advantages that the pesticide effect is obvious, the pesticide cost is reduced, the ecological environment is protected, the compounded tea saponin and the camu knee have an addition effect on mutually killing the snails, the ratio of the tea saponin in the first invention is 20%, the ratio of the radix achyranthis bidentatae in the second invention is 60%, the two plant extracts in the third invention are compounded, the snail killing effect is more efficient, and the snail killing effect is the same as that of the chemical pesticide metaldehyde on the basis of saving raw materials.

Drawings

Fig. 1 is a line graph of mortality rate of ampullaria gigas in 168h under different concentrations of tea saponin, and CK is a clear water control group. As can be seen from the figure, the tea saponin has good molluscicidal effect under the concentration of 40mg/L, 60mg/L, 80mg/L and 100 mg/L.

Fig. 2 is a line graph of mortality rates of ampullaria gigas over 168 hours at various concentrations of the extract of the knee of a marbled cattle, CK being the clear water control. As can be seen from the figure, the extract of the knee of the Ardisia mamillata has good molluscicidal effect under the concentration of 100mg/L, 200mg/L and 300 mg/L.

FIG. 3 is a comparison of the molluscicidal effect of tea saponin and camu knee extract wettable formulations with 6% metaldehyde GR over 21 days. CK: the effect of no pesticide application in the field is achieved; the effect of the 80% tea saponin wettable preparation and the sparrow knee extract wettable preparation is slightly lower than that of metaldehyde within 0-3 days, the effect of the 80% tea saponin wettable preparation is equivalent to that of the sparrow knee extract wettable preparation, and the effect of the 80% tea saponin wettable preparation is slightly better than that of the sparrow knee extract wettable preparation; the 80% tea saponin wettable preparation and the camu knee extract wettable preparation have obvious effect within 3-7 days and have equivalent effect with metaldehyde GR; the effect tends to be stable after 7 days, and the effect is good.

The experimental method comprises the following steps:

the invention adopts a method for testing toxicity in a laboratory, which comprises the following steps:

the amount of the required experimental material was calculated and weighed and dissolved in 2L of dechlorinated water. The experiments were repeated 3 times per treatment set up with the dechlorinated water as a control. Selecting 20-30 adult ampullaria gigas with 5-7 revolutions, placing the individual in an experimental container, adding vegetables such as water spinach and Chinese cabbage as food for the ampullaria gigas, changing the food every two days to keep the freshness of the food for the ampullaria gigas, and simultaneously covering a layer of gauze or net to prevent the ampullaria gigas from escaping. The temperature stability of the experimental region is strictly maintained so as to prevent the ampullaria gigas from being influenced by the large change of the early and late temperatures. And (5) observing the situation of the ampullaria gigas on time, and recording the death number of the ampullaria gigas.

The judgment standard of ampullaria gigas death in the invention is as follows:

(1) the soft body part at the mouth of the ampullaria gigas shell is stimulated by an iron wire or a writing brush to touch, the ampullaria gigas still alive can be quickly closed because of external stimulation, and if the ampullaria gigas exposed out of the soft body does not respond to the stimulation, the ampullaria gigas can be determined to be dead;

(2) the ampullaria gigas for tightly closing the snail shells can be made of iron wires or other hard substances, the ampullaria gigas shells are slightly picked up or pulled open, the still-living ampullaria gigas close the snail shells due to external stimulation, and dead individuals have no obvious response.

Detailed Description

Laboratory conditions: all experiments in the research are performed in key laboratories of medicinal plant biotechnology in Jiangsu province. The experimental department has an engineering technical center, a large number of advanced reaction instruments and detection equipment, provides a good hardware foundation for scientific research, and has conditions for developing and killing ampullaria gigas indoor experiments.

First, experimental material

Biological material: the ampullaria gigas are bred and provided by Xuzhou Nuote company, 5-7 ampullaria gigas individuals with high activity are selected indoors, 700-1000 ampullaria gigas are reserved in each round of experiment, and 3 times of experiments are repeated.

Experimental agents: the experimental medicament is the extract of the knee of the lepiota obtained by using an extraction and purification technology and 80 percent of tea saponin purchased in the market.

Preparing a radix achyranthis bidentatae extract: weighing 50g of plant powder, pouring into a 1L stainless steel pot, adding 500mL of distilled water, placing on an electric furnace, boiling with strong fire, keeping boiling with slow fire for 2.5-3.0h, filtering the solution with gauze, returning plant powder residue into the pot, adding 500mL of distilled water, boiling and extracting, repeating for 3 times, filtering and mixing the water solution obtained by boiling and extracting for 3 times, boiling and evaporating to dryness, cooling the extract, and placing in a refrigerator for later use.

Second, the experimental procedure

The invention comprises the following steps: the method comprises the following steps: the tea saponin and the components are prepared into wettable preparations according to the proportion, and the invention is not limited to the weight percentage. The formula is as follows: the 80% tea saponin wettable preparation comprises the following components in percentage by weight: 80% of tea saponin 20%, 8% of gypsum powder, 1% of attractant and 71% of flour, and fully stirring and mixing to obtain the 80% tea saponin wettable preparation. Step two: preparing experimental apparatus-transparent plastic basin, dividing into 7 groups, sequentially numbering as 80% tea saponin diluent reagent 12%, 14%, 16%, 18%, 20%, 22%, 24%. Blank without drug was used as control. 2L of dechlorinated water is poured into each group, 20-30 ampullaria gigas are added, vegetables such as water spinach and Chinese cabbage are added as food for the ampullaria gigas, the ampullaria gigas are placed on the net to prevent the ampullaria gigas from climbing out, the food is changed every 2 days, and 3 experiments are repeated. Application of the medicine: accurately sucking 1ml of each 80% tea saponin diluting reagent with different gradients by using a liquid-transferring gun, and sequentially adding the 80% tea saponin diluting reagents with different gradients according to the corresponding numbers. The contents of the tea saponin in different gradient 80% tea saponin dilution reagents are respectively as follows: 12%, 14%, 16%, 18%, 20%, 22%, 24%. Another group was prepared with 2L of dechlorinated water as a blank control. When the concentration of the reagent is changed, cross contamination or errors caused by drug residues and the like in the experiment are avoided.

The experimental result shows that when the concentration of the 80% tea saponin diluting reagent is 20%, 22% and 24%, the reagent has good lethal effect within 48-96 hours, the death rate of ampullaria gigas reaches more than 92%, the effect is stable within 72-96 hours, the effect is more obvious when the concentration is higher, and the effect is basically the same as that of a 6% metaldehyde preparation. In consideration of the molluscicidal effect and the cost of raw materials, the wettable preparation is prepared by selecting the tea saponin and other components in a weight ratio of 2: 8. Step three: grouping: preparing experimental tools, namely transparent plastic basins, which are divided into 5 groups, and numbering is 20mg/L, 40mg/L, 60mg/L, 80mg/L and 100mg/L of 80% tea saponin wettable preparation in sequence; blank without drug was used as control. 2L of dechlorinated water is poured into each group, 20-30 ampullaria gigas are added, vegetables such as water spinach and Chinese cabbage are added as food for the ampullaria gigas, the ampullaria gigas are placed on the net to prevent the ampullaria gigas from climbing out, the food is changed every 2 days, and 3 experiments are repeated. Application of the medicine: accurately weighing 80% tea saponin wettable preparation with balance, applying medicinal liquid corresponding to the number, and taking another group with 2L of dechlorinated water as blank control. When the types and the concentration of the medicines are changed, deionized water is used for repeatedly cleaning, and the method can effectively avoid cross contamination or errors caused by medicine residue and other reasons in an experiment. And (3) observing and recording: the subjects were observed every 12 hours after administration for a week. The experimental observation includes that firstly, ampullaria gigas with obvious openings and death are picked out, for ampullaria gigas with shells tightly closed all the time, surface medicines can be selected to be washed off and then the ampullaria gigas is placed in clear water, and whether the ampullaria gigas dies or not is observed after a plurality of days, or the ampullaria gigas dies and lives are determined by a tapping method. And (5) recording the number of dead snails in time after observation, and recording a table.

The experimental result shows (as shown in figure I), when the concentration of the 80% tea saponin wettable preparation is 40mg/L, 60mg/L, 80mg/L and 100mg/L, the formula has good lethal effect within 48-96 hours, the death rate of ampullaria gigas reaches more than 92%, the effect is stable within 72-96 hours, the effect is more obvious when the concentration is higher, the effect is basically the same as that of a 6% metaldehyde preparation, and the 100% molluscicidal effect is achieved after 144 hours. Therefore, the 80% tea saponin wettable preparation can replace 6% metaldehyde chemical pesticide which is popular in the market to prevent and control the harm of ampullaria gigas, the biological pesticide with the proportion can play a role in killing the ampullaria gigas in a short time, the toxic action of metaldehyde on the environment can be reduced, and the killing effect on the ampullaria gigas is ensured. The biological pesticide has the advantages of safe and convenient use, easily obtained raw materials for producing the biological pesticide, lower production cost than common chemical agents, easy acceptance by the masses of farmers when being put into the market, real prevention and control of the ampullaria gigas disasters, and consideration of human and animal protection and environment friendliness. The second invention is that: the method comprises the following steps: the radix genuniae extract and the components are prepared into wettable preparations according to the proportion, and the invention is not limited to the weight percentage. The formula is as follows: the weight percentage of each component of the radix achyranthis bidentatae extract wettable preparation is as follows: 60% of radix achyranthis bidentatae extract, 8% of gypsum powder, 1% of attractant and 26% of flour are complemented to 100% and fully stirred and mixed to prepare the wettable preparation of the radix achyranthis bidentatae extract. Step two: preparing experimental tools, namely transparent plastic basins, which are divided into 7 groups, and numbering is 20%, 30%, 40%, 50%, 60%, 70% and 80% of the radix seu folium Cochinchinensis extract diluting reagent in sequence. Blank without drug was used as control. 2L of dechlorinated water is poured into each group, 20-30 ampullaria gigas are added, vegetables such as water spinach and Chinese cabbage are added as food for the ampullaria gigas, the ampullaria gigas are placed on the net to prevent the ampullaria gigas from climbing out, the food is changed every 2 days, and 3 experiments are repeated. Application of the medicine: accurately sucking 1ml of radix Achyranthis bidentatae extract diluting reagent with different gradients by using a pipette gun, and sequentially adding radix Achyranthis bidentatae extract diluting reagents with different gradients according to the corresponding numbers. The contents of the different-gradient radix cambogiae extract diluted reagent type radix cambogiae extracts are respectively as follows: 20%, 30%, 40%, 50%, 60%, 70%, 80%. Another group was prepared with 2L of dechlorinated water as a blank control. When the concentration of the reagent is changed, cross contamination or errors caused by drug residues and the like in the experiment are avoided. The experimental result shows that the concentration of the radix achyranthis bidentatae extract diluted reagent is 60%, 70% and 80%, the lethal effect is good within 48-96 hours, the death rate of ampullaria gigas is more than 90%, the effect is stable within 72-96 hours, the effect is more obvious when the concentration is higher, and the effect is basically the same as that of a 6% metaldehyde preparation. Considering the snail killing effect and the cost of raw materials, the proportion of the extract of the knee of the Cannabis sativa to other components is 6: 4 to prepare a wettable formulation. Step three: grouping: preparing experimental tools, namely transparent plastic pots, which are divided into 4 groups, and sequentially numbering the wettable preparations of the 48% radix et caulis Entadae extract at 200mg/L, 300mg/L, 400mg/L and 500 mg/L; blank without drug was used as control. 2L of dechlorinated water is poured into each group, 20-30 ampullaria gigas are added, vegetables such as water spinach and Chinese cabbage are added as food for the ampullaria gigas, the ampullaria gigas are placed on the net to prevent the ampullaria gigas from climbing out, the food is changed every 2 days, and 3 experiments are repeated. Application of the medicine: accurately weighing wettable preparation of radix et caulis Opuntiae Dillenii extract with balance, applying medicinal liquid corresponding to the number, and collecting another group with 2L of dechlorinated water as blank control. When the types and the concentration of the medicines are changed, deionized water is used for repeatedly cleaning, and the method can effectively avoid cross contamination or errors caused by medicine residue and other reasons in an experiment. And (3) observation and recording: the subjects were observed every 12 hours after administration for a week. The experimental observation is that firstly, ampullaria gigas with obvious openings and death are picked out, and for the ampullaria gigas with the shells closed tightly all the time, the ampullaria gigas can be placed in clear water after surface medicines are washed out, and whether the ampullaria gigas die or not is observed after a plurality of days or a tapping method is selected to determine the death and the death of the ampullaria gigas. And (5) recording the number of dead snails in time after observation, and recording a table.

The experimental result shows (as figure II), 400mg/L and 500mg/L of the wettable preparation of the genu cambarbus extract is applied; the molluscacide can die every 12 hours, has good killing effect within 48-96 hours, has the death rate of the ampullaria gigas of more than 90 percent, has stable effect, has better molluscacidal effect when the concentration is higher, has basically the same effect as a 6 percent metaldehyde preparation, and reaches 100 percent after 96 hours, so the wettable preparation of the leprianum majus extract can replace a 6 percent metaldehyde chemical pesticide which is popular in the market to prevent and treat the harm of the ampullaria gigas, can play a role in killing the ampullaria gigas within a short time, can reduce the toxic action of the metaldehyde on the environment, and simultaneously ensures the killing effect on the ampullaria gigas. The invention has safe and convenient use, the hemp-cattle knee material for producing the biological pesticide is easy to obtain, the production cost is lower than that of common chemical agents, the hemp-cattle knee material is easy to be put into the market and is easy to be accepted by the masses of farmers, and the hemp-cattle knee material is a special-effect molluscicide which truly prevents and treats the occurrence of ampullaria gigas disasters, protects people and livestock and is environment-friendly. The invention III: the method comprises the following steps: after the tea saponin and the extract of the camu knee are extracted, a compound experiment is carried out: the tea saponin, the camu knee extract and the components are prepared into wettable preparations according to the proportion, and the invention is not limited to the weight percentage. The formula is as follows: the 80% tea saponin-sparrow knee extract wettable preparation comprises the following components in percentage by weight: 80% of tea saponin 10%, 40% of the extract of the knee of the Ma niu, 8% of gypsum powder, 1% of attractant and 41% of flour, which are complemented to 100%, are fully stirred and mixed to prepare the 80% tea saponin-extract wettable preparation of the Ma niu knee. Step two: grouping: preparing experimental tools, namely transparent plastic basins, which are divided into 5 groups, wherein 80% of tea saponin 20mg/L is respectively compounded with 40mg/L, 60mg/L, 80mg/L, 100mg/L and 120mg/L of the knee of the leprous cattle, and marking is carried out. In addition, 5 groups of transparent plastic basins are prepared, 12mg/L of tea saponin 80% extract wettable preparation is respectively compounded with 40mg/L, 60mg/L, 80mg/L, 100mg/L and 120mg/L of the knee of the numb, and marking is carried out; blank without drug was used as control. 2L of dechlorinated water is poured into each group, 20-30 ampullaria gigas are added, vegetables such as water spinach and Chinese cabbage are added as food for the ampullaria gigas, the ampullaria gigas are placed on the net to prevent the ampullaria gigas from climbing out, the food is changed every 2 days, and 3 experiments are repeated. Application of the medicine: accurately weighing 80% tea saponin-radix seu folium Jasmini Lanceolatae extract wettable preparation with a balance, applying medicinal liquid corresponding to the number, and taking another group with 2L of dechlorinated water as blank control. When the types and the concentration of the medicines are changed, deionized water is used for repeatedly cleaning, and the method can effectively avoid cross contamination or errors caused by medicine residues and the like in an experiment. And (3) observing and recording: the subjects were observed every 12 hours after administration for a week. The experimental observation is that firstly, ampullaria gigas with obvious openings and death are picked out, and for the ampullaria gigas with the shells closed tightly all the time, the ampullaria gigas can be placed in clear water after surface medicines are washed out, and whether the ampullaria gigas die or not is observed after a plurality of days or a tapping method is selected to determine the death and the death of the ampullaria gigas. And (5) recording the number of dead snails in time after observation, and recording a table. Experimental results show that the wettable preparation compounded by 80% of tea saponin 12mg/L and the extracts of the lepiota mauritiana 100mg/L and 120mg/L has a certain lethal effect within 48-96 hours, the death rates of the ampullaria gigas are all over 65%, the wettable preparation compounded by the tea saponin 80% of extracts 20mg/L and the extracts of the lepiota mauritiana 80mg/L, 100mg/L and 120mg/L has a good lethal effect within 48-96 hours, the death rates of the ampullaria gigas are all over 95%, the effect is stable within 72-96 hours, the higher the concentration is, the more obvious the effect is, and the effect is basically the same as that of the 6% metaldehyde preparation. Therefore, the invention selects 80% of tea saponin: extract of the knee of the Ardisia mamillata: other components = 1: 4: 5, the 80 percent wettable preparation of the tea saponin and the extract of the camu, and the 80 percent wettable preparation of the tea saponin and the extract of the camu can replace 6 percent metaldehyde chemical pesticide which is popular in the market to prevent and control the harm of the ampullaria gigas. In conclusion, 80% of tea saponin 10% and 40% of the camu knee extract wettable preparation, the biological pesticide compounded according to the proportion is economical, can achieve the effect of killing ampullaria gigas in a short time, can reduce the toxic action of metaldehyde on the environment, and simultaneously ensures the killing effect on the ampullaria gigas. The biological pesticide is safe and convenient to use, raw materials for producing the biological pesticide are easy to obtain, the production cost is lower than that of common chemical agents, and the biological pesticide is easy to be accepted by the masses of farmers when being put into the market.

Third, field experiment detection

Purpose of the test

3 molluscicidal reagents provided by key laboratories of medicinal plant biotechnology of Jiangsu province are definite and are used for preventing and treating the control effect and the safety of the paddy ampullaria gigas in the field. The test is carried out according to the pesticide field efficacy test rule (second) part 81: the national standard GB/T17980.81-2004 of the molluscicide for preventing and treating the ampullaria gigas.

2 conditions of the test

2.1 selection of test subjects, crops and varieties

Test subjects: pomacea canaliculata (Thunb.) TrevPomacea canaliculata

And (3) test crops: rice (Oryza sativa L.) with improved resistance to stress

2.2 environmental conditions

The test is carried out in a test base established in Guangxi agricultural academy of sciences, the crop variety is rice, and the period of node pulling and spike growing during the test refers to a period from the beginning of differentiation from node pulling and spike sprouting to the stage before spike emergence. The experimental field cultivation conditions (soil type, fertility and cultivation) are uniform and consistent, and the method accords with scientific agricultural practice (GAP). The planted crops are rice, and the rice grows well. No pesticide was used before the test.

3 test design and arrangement

3.1 Agents

3.1.1 test Agents

The 80% tea saponin wettable preparation pesticide and the radix achyranthis bidentatae extract wettable preparation pesticide are provided by plant biotechnology key laboratories for Jiangsu province medicine.

3.1.2 control agents:

6% metaldehyde GR (CnDollo) supplied by Xuzhou Nott chemical Co.

3.1.3 dosage and numbering:

TABLE 1 test design of test agents

。

3.2 cell arrangement:

3.2.1 cell arrangements

。

3.2.2 cell area and repetition:

cell area: about 15 square meters (3 m x 5 m).

The number of repetitions: 3 replicates.

3.3 the application method comprises the following steps:

3.3.1 methods of use

And (6) broadcasting.

3.3.2 applicator devices

None.

3.3.3 application time and frequency

The test only applies the medicine once, and the application time is 9 months and 6 nightfall in 2019.

3.3.4 information on the agents for controlling other diseases and pests

The test field is special for the test, and no other insect and disease prevention medicament is used.

4 investigating, recording and measuring method

4.1 Meteorological and soil data

4.1.1 Meteorological data

Year 2019, month 9, day 6 (day of administration): the temperature is 25-35 ℃, the product is cloudy, and the southeast wind is 1-2 grade.

6 days in 2019 and 6-26 days in 9 months, and the whole test period is mainly cloudy or raining; the lowest temperature is 20 ℃, the highest temperature is 35 ℃, and the rainfall weather is more (see table 5 for details).

4.1.2 soil data

The sandy loam soil of the test field has good irrigation and drainage conditions, moderate fertility conditions and a very small amount of weeds.

4.2 investigation methods, time and frequency

4.2.1 investigation time and number of surveys

The population base of the worms was investigated before the drug, and the number of dead worms was investigated 3, 7, 14, and 21 days after the drug.

4.2.2 methods of investigation

The number of ampullaria gigas naturally occurring in each cell is investigated before pesticide application, and each cell is randomly checked for 3-4 m ² If the number of the hairs is less than 8 per square meter, the hairs are in the wildExternally collecting ampullaria gigas of the same age period and supplementing the ampullaria gigas to about 8 per square meter. Only the dead individuals on the soil surface, the plant base and the upper part are observed in the first 3 times of investigation after the medicine is applied, and the soil surface is not suitable to be turned over so as to prevent the medicine grains from being buried under the soil. And (3) removing dead ampullaria gigas individuals out of the field after recording the number of the dead ampullaria gigas in the investigation so as to prevent repeated recording in the next investigation, turning over the position 1-3 cm below the soil surface at the same time in the last investigation, and paying attention to the existence of living or dead individuals below a larger soil block.

Judging criteria of ampullaria gigas death: the soft body part at the mouth of the ampullaria gigas shell is stimulated by an iron wire or a writing brush to touch, the ampullaria gigas still alive can be quickly closed because of external stimulation, and if the ampullaria gigas exposed out of the soft body does not respond to the stimulation, the ampullaria gigas can be determined to be dead; the ampullaria gigas tightly closing the shells can be slightly picked up or pulled open by using iron wires or other hard objects, the still-living ampullaria gigas close the shells due to external stimulation, and dead individuals have no obvious response.

4.2.3 the calculation method of the drug effect:

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4.3 direct effects on crops

During the test period, no phytotoxicity phenomenon appears on the crops.

4.4 quality and yield of the product

The test was not evaluated for quality and yield.

4.5 Effect on other organisms

During the test period, no influence of the medicament on other pests and natural enemies is found.

Results and analysis

Test result of controlling rice ampullaria gigas by using 23 kinds of metaldehyde GR in table

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Note: the control (%) in the table above is the average of each repetition. The results were analyzed by the duncan's new double error (DMRT) method, with the same case letters after the data in the table indicating insignificant differences at the 1% and 5% levels.

5.1 evaluation of drug

The test results of the 3 kinds of metaldehyde GR for preventing and controlling the rice ampullaria gigas are shown in the table 2:

3 days after the pesticide is applied, the control effect of the test pesticide and the control pesticide on the paddy field is as follows in sequence: 6% metaldehyde GR >80% tea saponin pesticide > Masui niu Knoxia extract pesticide. The test results are analyzed by DMRT method, the results show that the control effect of 80% tea saponin pesticide is better than that of the camu knee extract pesticide, the two pesticide have obvious difference with 6% metaldehyde GR molluscicidal effect, and have obvious difference with blank control.

7 days after the pesticide is applied, the control effect of the test reagent and the control reagent on the rice ampullaria gigas is as follows in sequence: 6% metaldehyde GR >80% tea saponin pesticide > Masui niu Knoxia extract pesticide. The test results are analyzed by DMRT method, the results of 80% tea saponin pesticide and the pesticide of the extract of the radix camberidis cambogiae show that the control effect is equivalent and the difference is not obvious. The control effect of 80% tea saponin pesticide and the pesticide of the extract of the radix seu folium Jasmini Lanceolatae is equivalent to the control effect of the soul fighting Luo, and the difference is not obvious.

14 days after the pesticide is applied, the control effect of the test reagent and the control reagent on the rice ampullaria gigas is as follows in sequence: 6% metaldehyde GR >80% tea saponin pesticide > Masui niu Knoxia extract pesticide. The test results are analyzed by DMRT method, the results of 80% tea saponin pesticide and the pesticide of the extract of the radix camberidis cambogiae show that the control effect is equivalent and the difference is not obvious. The control effect of 80% tea saponin pesticide and the pesticide of the camu knee extract is equivalent to the control effect of the control medicament of the soul fighting Luo, and the difference is not obvious.

After 21 days, the control effect of the test agent and the control agent on the rice ampullaria gigas is as follows in sequence: 6% metaldehyde GR >80% tea saponin pesticide > Masui niu Knoxia extract pesticide. The test results are analyzed and tested by a DMRT method, and the results of 80% of tea saponin pesticide and the pesticide of the extract of the radix camberiae show that the control effect is equivalent and the difference is not obvious. The control effect of 80% tea saponin pesticide and the pesticide of the extract of the radix seu folium Jasmini Lanceolatae is equivalent to the control effect of the soul fighting Luo, and the difference is not obvious.

In a word, the insecticidal rates of the 2 biological pesticides reach more than 80% from the seventh day, the effect is slightly lower than the molluscicidal effect of 6% metaldehyde GR, the difference is small, and if the cost is not calculated, the traditional 6% metaldehyde (positive control) can be replaced.

5.2 safety

The molluscicidal granule for test has no phytotoxicity symptom to rice, and has no uncomfortable feeling after the pesticide is applied by a pesticide applicator.

6 small knot

According to the field test result of the next half year in 2019, 2 test agents have good control effects on the ampullaria gigas, the control effects are all more than 85% in 21 days after the application, and the control effects are as follows: the insecticidal rates of the 2 biological pesticides are all more than 80% from the seventh day, the effect is slightly lower than the molluscicidal effect of 6% metaldehyde GR, the difference is small, and the 2 organisms can replace the traditional 6% metaldehyde (positive control).

Compared with the test result of the first half year, the control effect of the second half year is very obvious, and the control effect is mainly influenced by the following 2 factors:

1. temperature: according to the weather condition record, the high temperature is continuously kept for the first half year. During the next half year test period, the daily average temperature is 22.5-35.5 ℃, the temperature is proper, and the performance is active.

2. Humidity: can find out by the weather condition table of table 5, use gust rain and cloudy weather as the owner during whole experiment, the rainwater is more, and the earth's surface is moist, is favorable to the medicament to spread on the earth's surface, does not need the sprinkling irrigation crop simultaneously, has avoided the medicament to be covered by soil.

Therefore, the results of comprehensive field tests prove that 2 molluscicides provided by Xuzhou Nuote chemical industry have good control effects on the ampullaria gigas of the rice, and the control effects are as follows in sequence under the condition of the same dosage: 6% metaldehyde GR80% tea saponin pesticide > Stauntonia chinensis extract pesticide, the difference is not big, 2 kinds of organisms can replace the traditional 6% metaldehyde. When the medicine is applied, attention should be paid to: 1) selecting to apply the pesticide under the active climate condition of ampullaria gigas; 2) the pesticide is applied after rain, at the evening with large dew or in the climate with large humidity as much as possible, so that the prevention and treatment effect of the pesticide is improved. Meanwhile, the pesticide is alternately used with other effective medicaments to slow down the generation of the drug resistance of pests.

Claims

1. A formula of a compound botanical extract biological molluscicidal pesticide is characterized in that: 40% of a knee extract of the milch cow, 10% of tea saponin 80%, 8% of gypsum powder, 1% of an attractant and 41% of flour; preparing a radix achyranthis bidentatae extract: weighing 50g of plant powder, pouring into a 1L stainless steel pot, adding 500mL of distilled water, boiling on an electric furnace with strong fire, keeping boiling on slow fire for 2.5-3.0h, filtering the solution with gauze, returning plant powder residues into the pot, adding 500mL of distilled water, boiling and extracting, repeating for 3 times, filtering the aqueous solution obtained by boiling and extracting for 3 times, boiling and evaporating to dryness, cooling the extract, and putting into a refrigerator for later use.

2. A method of preparing the formulation of the compounded botanical extract molluscicidal pesticide of claim 1, wherein the formulation comprises: weighing the tea saponin, the camu knee extract, the gypsum powder, the attractant and the flour according to a preset weight proportion, and putting the weighed materials into a stirring kettle in sequence to be fully stirred and uniformly mixed.