CN110547303B

CN110547303B - Application of bacillus thuringiensis G033A in prevention and treatment of southern American tomato leaf miner

Info

Publication number: CN110547303B
Application number: CN201910970856.4A
Authority: CN
Inventors: 张�杰; 张桂芬; 束长龙; 张毅波; 耿丽丽; 刘万学; 宋福平
Original assignee: Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Current assignee: Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Priority date: 2019-10-14
Filing date: 2019-10-14
Publication date: 2021-07-16
Anticipated expiration: 2039-10-14
Also published as: CN110547303A

Abstract

The invention relates to application of bacillus thuringiensis G033A in preventing and controlling southern American tomato leaf miner. The invention discovers for the first time that Bt G033A has high toxicity to diamond back moth and potato beetle, and also has excellent insecticidal activity to the world pest Nanmei tomato Spodoptera frugiperda, particularly the insecticidal activity to the high age (3 age and/or 4 age) of the Nanmei tomato Spodoptera frugiperda is obviously better than that of the low age (1 age and/or 2 age). In addition, the pesticide has obvious effect taking speed higher than that of other biogenic pesticides such as chrysanthemum indicum-azadirachtin, pyrethrin, rotenone and the like, and is a pesticide with application prospect. Therefore, the invention provides the application of bacillus thuringiensis G033A (Bt G033A) in preventing and controlling southern American tomato leaf miner.

Description

Application of bacillus thuringiensis G033A in prevention and treatment of southern American tomato leaf miner

Technical Field

The invention relates to application of bacillus thuringiensis G033A in preventing and controlling southern American tomato leaf miner, in particular to application of bacillus thuringiensis G033A (Bt G033A) in preventing and controlling southern American tomato leaf miner larvae.

Background

The south America tomato leaf miner Gelechiideae is a world destructive pest newly introduced into China, mainly uses larvae to eat leaf flesh, fruit, top tip, tender bud, tender stem and side branch of the tomato, and can reduce the yield of the tomato by 80 to 100 percent when the tomato is seriously damaged. The south America tomato leaf miner is harmful and concealed, has generated serious resistance to common insecticides comprising organic phosphorus, pyrethroid, abamectin, anthranilamide and the like, and simultaneously generates resistance to various biological source agents comprising spinosad, azadirachtin, chitin synthesis inhibitor and the like.

Bacillus thuringiensis (Bt) is one of the most widely applied biopesticides in the world today. The Insecticidal activity of Bt derives mainly from its ability to produce Insecticidal Crystal Proteins (ICPs), i.e., delta-endotoxins, which are non-toxic to humans and animals and do not pollute the environment.

Disclosure of Invention

Bacillus thuringiensis (Bt) G033A belongs to the genus Bacillus, Bacillus thuringiensis species, the kurstaki subspecies. It is obtained by introducing a plasmid vector containing cry3Aa7 gene into wild strain G03 strain by electroporation. The insecticidal composition has high toxicity on important agricultural pests such as diamondback moth of the Lepidoptera and potato beetle of the Coleoptera, and the field control effects on the young diamondback moth and the potato beetle are 82.9 to 85.2 percent and 85.3 to 90.2 percent respectively. It has stomach toxicity effect on insects, low toxicity on fish, bird and bee, and high toxicity on silkworm; has no bioaccumulation and no obvious influence on the soil microbial community structure.

The invention discovers for the first time that Bt G033A has high toxicity to diamondback moth and potato beetle, and has excellent insecticidal activity to the world pest Nanmei tomato subdata moth, especially the higher age (3 and/or 4 age) insecticidal activity of the Nanmei tomato subdata moth is obviously better than the lower age (1 and/or 2 age) insecticidal activity. In addition, the pesticide has obvious effect taking speed higher than that of other biogenic pesticides such as chrysanthemum indicum-azadirachtin, pyrethrin, rotenone and the like, and is a pesticide with application prospect.

Therefore, the invention provides the application of bacillus thuringiensis G033A (Bt G033A) in preventing and controlling southern American tomato leaf miner.

In a specific embodiment, the south american tomato leaf miner is a larval stage south american tomato leaf miner.

In a specific embodiment, the south american tomato leaf miner is a larval stage south american tomato leaf miner of at least one of

age

1, 2, 3 and 4.

In one embodiment, the amount of G033A is 75 to 400G/acre.

In one embodiment, the amount of G033A is 100 to 150G/acre.

In one embodiment, the dosage form of G033A is a wettable powder.

In a specific embodiment, said application is the application of said G033A to tomato plants for the control of said southern american tomato leaf miner.

Drawings

Figure 1 shows the indoor ovicidal activity of bacillus thuringiensis kurstaki subspecies G033A against south american tomato leaf miner, the same lower case letters above the histogram (both a) indicating insignificant differences at the P <0.05 level between concentration treatments (one-way anova, least significant difference test).

FIG. 2 shows the indoor insecticidal activity of Bacillus thuringiensis subspecies Kurstaki G033A on Nanmei tomato leaf miner larvae. Wherein A is the treatment of 1 to 2 instar larvae; b, treating 3-instar larvae; c is treatment of 4-instar larvae. Lower case letters (at least one of a, b and c) after concentration indicate differential results at P <0.05 level between different concentrations (paired data t test).

FIG. 3 shows the field ovicidal activity of Bacillus thuringiensis subspecies Kurstaki G033A and 3 other commonly used biopesticides against Nanmei tomato leaf miner. Lower case letters above the histogram indicate the results of differential analysis (one-way analysis of variance, least significant difference test) at P <0.05 level between each agent treatment.

FIG. 4 shows the field insecticidal activity of Bacillus thuringiensis subspecies Kurstaki G033A and 3 other commonly used biopesticides on young larvae of Nanmei tomato leaf miner. Wherein A is the treatment of 1 st larva; and B is the treatment of 2-instar larvae. The lower case difference above the same day (day 5 and day 7) survey data (mean ± SE, n ═ 4) in fig. 4A and 4B represent the results of differential analysis at a P <0.05 level between treatments (one-way analysis of variance, least significant difference test).

Detailed Description

The present invention is further illustrated by the following examples, which are intended to be purely exemplary of the invention and are not to be construed as limiting the invention in any way.

The Bt G033A wettable powder is given by Wuhan Kenuo Biotechnology GmbH, and the content of active ingredients is 32000 IU/mg.

Example 1

Biological Activity assay of indoor eggs

The experiment started in late 4 th ten days and ended in the end of 5 months.

32000IU/mG Bt G033A wettable powder is diluted to 5 concentrations of 40G/L, 30G/L, 20G/L, 10G/L and 5G/L by water, and a total of 6 treatments are carried out by taking clear water as a blank Control (CK). Wherein g in concentration unit refers to the mass of the wettable powder.

The bioassay method adopts a leaf soaking method: the collected fresh tomato leaves are completely unfolded (the longest part and the widest part are about 8cm and 5cm), the leaf stalks are obliquely and quickly cut by a paper cutter to absorb absorbent cotton, the absorbent cotton is wrapped by a preservative film to keep fresh, then the leaves are completely soaked in the concentration for 10s by holding the leaf stalks wrapped with the absorbent cotton, then the leaves are naturally dried, and the leaves are placed in a round plastic insect-culturing box (the diameter is 9cm, and the height is 3 cm).

Bioassay of eggs: the method comprises the steps of collecting tomato leaves with primary-spawned light yellow green eggs from a linkage greenhouse for planting hair powder tomato varieties, obliquely and quickly cutting leaf stalks by using a paper cutter to absorb absorbent cotton, wrapping the absorbent cotton by using a preservative film to keep fresh, naturally airing the leaves with primary-spawned eggs after completely soaking the leaves with the primary-spawned eggs in the concentration for 10s by using the leaf stalks wrapped by the absorbent cotton in a handheld mode, putting the leaves into a circular plastic insect breeding box (the diameter is 9cm, and the height is 3cm), and putting the fresh egg-free tomato leaves soaked in the same concentration. 10 eggs were treated at each concentration and 5 batches were repeated. The environment temperature is 20-25 ℃, the humidity is 40-70%, and the natural illumination is carried out. Eggs were observed to hatch into larval numbers 1 time per day, and continued for 7 days at regular intervals until no more or all were hatched. The egg hatching inhibition rate at 7d was calculated, as well as the corrected egg hatching inhibition rate at 7 d. After the data are processed by Excel2007, the corrected hatching inhibition rate data obtained by the indoor bioactivity determination are subjected to statistical analysis by SPSS 24.0. The corrected hatching inhibition rates of different concentrations are compared by adopting a One-way analysis of variance method, and a difference significance test (One-way ANOVA, LSD test) is carried out by using an LSD minimum significant difference method, wherein the significance level P is less than 0.05. The results are shown in FIG. 1.

Inhibition of egg hatching (number of unhatched eggs/total eggs) × 100% (1)

Corrected egg hatching inhibition rate ═ ((Bt G033A-treated egg hatching inhibition rate-blank control egg hatching inhibition rate)/(1-blank control egg hatching inhibition rate)) × 100% (2)

As can be seen from FIG. 1, after treatment, 7d, Bt G033A slightly differed in ovicidal activity against the leaf miner of Nanmei tomato at various concentrations, but the difference was not significant (F)_4,201.188, P0.346; one-way ANOVA, LSD test), and all are less than 25%, and thus ovicidal activity is judged to be low accordingly.

Example 2

Determination of the biological Activity of indoor larvae

The experiment started in late 4 th ten days and ended in the end of 5 months.

1-4 instar larvae of Nanmei tomato leaf miner are picked from a linkage greenhouse planted with a variety of hair powder tomatoes, and starved for 4 hours after indoor feeding of tomato leaves for 24 hours.

Preparing Bt G033A liquid medicine: 32000IU/mG Bt G033A wettable powder was diluted with water to 5 concentrations of 40G/L, 30G/L, 20G/L, 10G/L, 5G/L, with clear water Control (CK).

The bioassay method adopts a leaf soaking method: the collected fresh tomato leaves are completely unfolded (the longest part and the widest part are about 8cm and 5cm), the leaf stalks are obliquely and quickly cut by a paper cutter to absorb absorbent cotton, the absorbent cotton is wrapped by a preservative film to keep fresh, then the leaves are completely immersed in the liquid medicine with different concentrations for 10s by holding the leaf stalks wrapped with the absorbent cotton in hand, and then the leaves are naturally dried and put into a round plastic insect-culturing box (the diameter is 9cm, and the height is 3 cm). Larvae of 1 to 2 instars, larvae of 3 instars and larvae of 4 instars, which were starved for 4h, were inoculated onto tomato leaves in each insect box, treated 10 per concentration, per instar, and repeated for 5 batches. The environment temperature is 20-25 ℃, the humidity is 40-70%, and the natural illumination is carried out. The survival state of the larvae is observed for 1 time every day (24h), the larvae are continuously observed for 7d at regular time, and the larvae are judged to die when the soft brush is light and the touch bodies do not react and have no luster. The number of dead larvae and the number of surviving larvae were recorded and larval mortality was calculated and corrected accordingly. A One-way ANOVA (least significant Difference) method is adopted to compare whether the corrected larva mortality rate has significant difference among different concentrations, and a LSD minimum significant difference method is adopted to carry out the significance test (One-way ANOVA, LSD test), wherein the significance level P is less than 0.05. The results are shown in FIG. 2.

Larval mortality (number of dead larval/total larval) x 100% (3)

Corrected larval mortality ═ ((Bt G033A treatment larval mortality-blank control larval mortality)/(1-blank control larval mortality)) × 100% (4)

The results in fig. 2 show that different concentrations of Bt G033A treated tomato leaves have better poisoning activity against all instar larvae of south american tomato leaf miner.

As can be seen from FIG. 2A, after 96h (4d) of treatment of 1 to 2 instar larvae with Bt G033A at 5 to 40G/L, the corrected larval mortality rates for the remaining treatments, except for the low concentration of 5G/L, were all above 95%; all larvae were dead including a low concentration of 5g/L after 144h (6d) of treatment. The difference between the insecticidal effects of Bt G033A at concentrations of 10G/L (t 2.800, df 4, P0.049; paired data t test), 20G/L (t 2.841, df 4, P0.047), 30G/L (t 2.979, df 4, P0.041) and 40G/L (t 3.178, df 4, P0.034) was not significant; and the insecticidal effect under the four concentrations is obviously better than that under the concentration of 5 g/L.

As can be seen in FIG. 2B, all larvae died after 120h (5d) treatment of 3 instar larvae with Bt G033A at 5 to 40G/L. The difference between the insecticidal effects of Bt G033A at concentrations of 10G/L (t-3.709, df-4, P-0.034), 20G/L (t-2.663, df-4, P-0.076), 30G/L (t-3.163, df-4, P-0.050) and 40G/L (t-4.019, df-4, P-0.028) was not significant; the insecticidal effect under the three concentrations of 10g/L, 30g/L and 40g/L is obviously better than that under the concentration of 5 g/L; however, the difference between the concentrations of 5g/L and 20g/L was not significant.

As can be seen from FIG. 2C, after treatment of 4 instar larvae with Bt G033A at 5 to 40G/L for 96h (4d), the corrected larval mortality rates for the remaining treatments, except for the low concentration of 5G/L, were all above 90%; after 120h (5d) of treatment, the corrected larva mortality rate including low concentration of 5g/L reaches more than 90%. The difference between the insecticidal effects of Bt G033A at concentrations of 5G/L and 10G/L was not significant; the difference between the insecticidal effects of Bt G033A at concentrations of 20G/L, 30G/L and 40G/L was not significant; but the insecticidal effect under the three concentrations of 10g/L, 30g/L and 40g/L is obviously better than that under the concentration of 5 g/L; and there are significant differences in concentrations between 5g/L (20g/L vs.5g/L: t: 3.303, df: 4, P: 0.030; 30g/L vs.5g/L: t: 2.964, df: 4, P: 0.041; 40g/L vs.5g/L: t: 3.114, df: 4, P: 0.036) and 10g/L (20g/L vs.10g/L: t 4.982, df: 4, P: 0.008; 30g/L vs.10g/L: t: 3.575, df: 4, P: 0.023; 40g/L vs.10g/L: t: 3.067, df: 0.034, P: 0.023; 40g/L vs.10g/L: t: 3.067, df: 0.037, P: 0.034, P: 0.023) and 20g/L (20 g/L).

After the data are collated by Excel2007, the corrected larva mortality data obtained by indoor bioactivity determination are statistically analyzed by SPSS 24.0 software, the corrected larva mortality range of 48h of each instar larva is 10% -90%, and based on the corrected larva mortality range, the lethal medium concentration (LC) of Bt G033A to 48h of Nanmei tomato leaf miner larva is calculated by SPSS 24.0 software₅₀) 95% Confidence Interval (CI) and correlation coefficient (r)²). The results are shown in Table 1.

TABLE 1 indoor virulence of Bacillus thuringiensis Kurstaki subspecies G033A against larvae of Spodoptera littoralis at different ages in south America

As can be seen from Table 1, LC of Bt G033A for 48h of Nanmei tomato leaf miner₅₀Respectively 14.63g/L of 4-instar larva, 15.59g/L of 3-instar larva and 23.17g/L of 1-2 instar larva, and the virulence effects are sequentially 4-instar larva>Larva of 3 years old>Larvae of 1 to 2 instars. The results show that the indoor toxicity of Bt G033A to 3-and 4-instar larvae of Nanmei tomato leaf miner is higher than that to 1-to 2-instar larvae.

In conclusion, when Bt G033A is used for controlling the larvae of the leaf miner of the Nanmei tomato, the preferable application concentration in the field is 10G/L, namely, 100 times of dilution.

Example 3

Test of field drug effect

The test field is Yunnan Yuxi. Yuxi is located in the middle of Yunnan province, the average annual temperature is 16.4-24.6 ℃, and the Yuxi has no severe cold in winter and no severe summer heat in summer.

The test field is an organic vegetable planting base, and the planted tomato crops mainly comprise two varieties, namely, hair powder and cherry tomatoes. Wherein, the field drug effect test takes the hair powder as a test host. Covering the ridge surface with black mulching film, and planting in a greenhouse, wherein the planting distance is about 50cm, and the row distance is about 70 cm; the greenhouse is arranged in a north-south direction, the width of the north-south direction is about 16m, the length of the east-west direction is about 50m, and the height is about 6 m.

The tomato planting and cultivation management process implements the national standard GBT1963.1-4-2005 organic vegetable production technical standard, and no chemical agent or chemical fertilizer is applied in the process; and (5) drip irrigation and watering at regular intervals. After the fruit is ripe, the fruit is mainly used for picking.

Test hosts: the tomato is cultivated, and the variety is the hair powder. The cultivation method comprises planting in rectangular plastic pots (length, width and depth are about 55 × 30 × 25cm) with nutrient soil, and planting in late 5-month ten days, wherein each pot contains 2 plants and has a leaf period of 4-5 leaves. Before and during the test, no pesticide and chemical fertilizer are applied, and the growth vigor of the plants is basically consistent.

Cell design: in the linked greenhouse, the area is about 10m²Each treatment had 4 repeat cells, with a total of 20 cells, and guard rows (about 1.2m) between cells and around the test site. The tomato is in the seedling stage, the southern American tomato leaf miner is more serious (the quantity of single plant is about 20 per grain) before the test, and the tomato is in the egg and low-age larva (1 to 2 years old) generation stage.

Reagent to be tested: bt G033A wettable powder has an effective component content of 32000IU/mg, and the prevention and control dose range recommended by manufacturers for preventing and controlling pests such as diamondback moth or potato beetle is 75-400G/mu. In this test, a concentration of 150 g/acre was used, i.e. diluted 100 times and sprayed at a rate of 15L per acre. It is used as it is.

The concentration of the pyrethrum-azadirachtin (abbreviated as Pa in English, 2 percent of pyrethrum-azadirachtin soluble solution from Sphacelotheca yunnanensis Biotech, Limited liability company, (1.9:0.1) is 2 percent SL of the pyrethrum-azadirachtin, and the concentration is high in a pest control dosage range recommended by manufacturers, namely 180 g/mu, namely, 15L of diluted 83 times. It is used as it is.

The pyrethrin (Pc, 1.5% aqueous pyrethrin emulsion from Nnanbao Biotechnology Limited liability company) is diluted 83 times and sprayed at 15L/mu, with the concentration being 180 g/mu. It is used as it is.

Rotenone (abbreviated as Rt in english, 5% rotenone soluble solution available from yunnanbao biotechnology ltd) was used at a concentration of 180 g/mu, which is a high value recommended by manufacturers for the range of pest control dosage, that is, at a diluted 83-fold spray rate of 15L/mu. It is used as it is.

The spraying apparatus is a pneumatic sprayer, and the model is as follows: lontai Max 2.0L, purchased from Nanye household, Inc., Linhai, Taizhou, Zhejiang province.

Test time and method: the field test medication time is 6 months, 15 days afternoon and cloudy. The stem and leaf are sprayed uniformly at one time. 1d before application and 1, 2, 3, 5 and 7d after application, respectively adopting five-point sampling method, and 1 plant per point, and investigating the number of eggs and larvae (1-2 instars) of leaf miner of the Nanmei tomato on each tomato and the survival state thereof (determined by ultra-bright light zoom headlamp bottoming light observation). Eggs which did not hatch as larvae and did not have luster within 7d are marked as not hatching; the larvae with the dead larvae which do not respond to the strong light stimulation are marked as dead larvae; and calculating an egg hatching inhibition rate according to formula (1), correcting the egg hatching inhibition rate according to formula (2), correcting the larval mortality rate according to formula (3), and correcting the larval mortality rate according to formula (4). The field activity for south America tomato leaf miner eggs is shown in figure 3, and the field activity for south America tomato leaf miner low-age larvae is shown in figure 4.

As can be seen from FIG. 3, the corrected hatching inhibition rates of 7d and 4 medicaments on the eggs of the Spodoptera frugiperda after the spraying treatment are all less than 40 percent, but the differences are obvious (F)_3,12＝17.358,P<0.001). Wherein, the ovicidal activity of rotenone (Rt) is the highest (Rt vs.Pa: P ═ 0.004; Rt vs.Pc: P ═ 0.019; Rt vs.Bt: P ═ 0.004; Rt vs.Bt:. RTM. the ovicidal activity of the rotenone (Rt) is the highest<0.001), average 36.7%; secondly, pyrethrin-azadirachtin (Pa) and pyrethrin (Pc) (Pa vs. Bt: P ═ 0.004; Pc vs. Bt: P: (Pc/P:) (Pc/P: (Pc))<0.001), respectively 13.3% and 17.6%; bt G033A had a low ovicidal activity of only 1.7% (One-way ANOVA, LSD test).

As can be seen from fig. 4, under field conditions, 4 kinds of bio-source medicines have no substantial control effect on 1 to 2-instar larvae of plutella xylostella of south america in the first 2 days after application, particularly, the pyrethrin-azadirachtin, pyrethrin and rotenone are still substantially free from control effect in the 3 rd day after application, while Bt G033A shows a certain insecticidal effect. The control effect of the 4 biological source medicaments is obviously improved along with the prolonging of time.

Specifically, as can be seen from FIG. 4A for 1 st instar larvae, Bt G033A showed the highest field control effect at 5d after whole plant spray treatment and the corrected mortality rate was 89.1% (Bt vs. Rt: P ═ 0.003; Bt vs. Pa: P:. Pa.)<0.001；Bt vs.Pc:P<0.001); secondly, rotenone (Rt) with the control effect of 60.2 percent (Rt vs. Pa: P ═ 0.006; Rt vs. Pc: P)<0.001); again chrysanthemum-nimbin (Pa), the control effect is 39.2% (Pa vs. Pc: P)<0.001); the control effect of the pyrethrin (Pc) is the worst and is only 14.9 percent; and the difference between 4 medicaments is obvious (F)_3,12＝53.580,P<0.001) (One-way ANOVA, LSD test). After the whole plant is sprayed, the control effect of 7d, Bt and Rt on 1-instar larvae is the best, and the control effect is respectively 100 percent and 90.4 percent (Bt vs<0.001；Bt vs.Pc:P<0.001；Rt vs.Pa:P<0.001；Rt vs.Pc:P<0.001) and the difference between the two is not obvious (Bt vs. rt: P ═ 0.119); pa, and a control effect of 62.5% (Pa vs. P: Pc)<0.001); the control effect of Pc is the worst, and is only 40.9%; and there is a significant difference between Bt and Rt and Pa, Pc (F)_3,12＝62.758,P<0.001)(One-way ANOVA,LSD test)。

As can be seen from FIG. 4B for 2-instar larvae, the field control effect of Bt G033A was the highest at 5d after the whole plant was sprayed, and reached 98.0% (Bt vs<0.001；Bt vs.Pc:P<0.001); secondly, Rt, the control effect is 59.9% (Rt vs.Pc: P ═ 0.002), but the difference between the two is not obvious (Bt vs.Rt: P ═ 0.098); pa again, control 38.9% (Pa vs. pc: P ═ 0.018), and no significant difference from Rt (Rt vs. Pa: P ═ 0.231); the control effect of Pc is the worst, and is only 16.5%. The field control effect difference of 7d and 4 medicaments on 2-instar larvae after whole plant spraying treatment is obvious (F)_3,1211.805, P0.001) (One-way ANOVA, LSD test). Wherein the control effects of Bt, Rt and Pa are respectively 100%, 94.6% and 78.0%, and although the control effects are different, the differences are not obvious (Bt vs. Rt: P ═ 0.695; Bt vs. Pa: P ═ 0.120; Rt vs. Pa: P ═ 0.227); the prevention effect of Pc is only 41.5%, andthe difference between other 3 medicaments is obvious (Bt vs. Pc: P)<0.001；Rt vs.Pc:P<0.001；Pa vs.Pc:P＝0.003)(One-way ANOVA,LSD test)。

In conclusion, Bt G033A showed faster field insecticidal activity than chloranthus japonicus-azadirachtin, pyrethrin and rotenone on 1 to 2 instar south american tomato leaf miner larvae, and overall insecticidal activity was also better.

While the invention has been described with reference to specific embodiments, those skilled in the art will appreciate that various changes can be made without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, and method to the essential scope and spirit of the present invention. All such modifications are intended to be included within the scope of the present invention as defined in the appended claims.

Claims

1. Application of bacillus thuringiensis G033A in preventing and controlling southern American tomato leaf miner, wherein the southern American tomato leaf miner is a larval stage southern American tomato leaf miner.

2. The use of claim 1, wherein the Nanume tomato leaf miner is a larval stage Nanume tomato leaf miner of at least one of age 1, 2, 3 and 4.

3. The use according to claim 1, wherein the amount of G033A is 75 to 400G/acre.

4. The use according to claim 3, wherein the amount of G033A is between 100 and 150G/acre.

5. The use according to any one of claims 1 to 4, wherein the dosage form of G033A is a wettable powder.

6. The use according to any one of claims 1 to 4, wherein said use is the application of said G033A to tomato plants for the control of said Nanmei tomato leaf miner.