Cordyceps javanica FJDD-08 and application thereof

Abstract

The application discloses a Cordyceps javanica FJDD-08 and application thereof, and belongs to the technical field of biological control. The Cordyceps javanica FJDD-08 can parasitize in the body of a thrips pest, and effectively kills the thrips pest. In addition, the application provides a combined control method for the thrips pest, and the method applies the Cordyceps javanica, degradable armyworm glue and a novel bagging trinity control measure, kills the thrips pest in different growth periods and different spaces, and comprehensively and completely eliminates the pest from a soil source (entomogenous fungi), after emergence (degradable armyworm glue) and on crops (slow-release insecticide bagging), thereby providing technical support for the control of the thrips pest.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological control, in particular to a Cordyceps javanica FJDD-08 and application thereof. BACKGROUND

[0002] Thrips is the general term of Thysanoptera insects, and the body is tiny, which feeds on tender leaves, flowers and tender fruits. Thrips can harm more than 600 kinds of plants, and is an important pest of fruits, vegetables, flowers and other economic crops. Some thrips can also transmit plant viruses to cause indirect damage, spread plant viruses among crops, and ultimately cause devastating damage to crops.

[0003] Due to the fact that thrips are too small and spread in fruits, flowers and tender leaves, it is extremely difficult to control them. The overuse of chemical pesticides has caused serious resistance of thrips, especially the habit of thrips pupating in soil, which makes it more difficult to control thrips, and is not thorough. At present, for thrips adults, color plate trapping or plant trapping has certain trapping effect, but the damage to natural enemies is also large, and the above control measures have certain disadvantages.

[0004] Entomogenous fungi are a kind of parasitic fungi, which enter the insect body by invasion and absorb insect nutrition to finally cause the death of the host. Unlike chemical insecticides, it has the characteristics of multiple species, high safety and efficiency, long application period, benefit to natural enemies, not easy to produce resistance and can be quickly and massively produced, and is often used for biological control of pests. At present, Metarhizium and Beauveria are the most widely used for commercialization and application, but the pests controlled by them include cockroaches, termites, grubs, forest ticks and longicorn blood ticks, etc. There are few species of entomogenous fungi for thrips pests, and there is no effective killing method for thrips pests in different growth stages and different spaces.

[0005] Therefore, it is a technical problem to be solved by those skilled in the art to provide an entomogenous fungus for controlling thrips pests, and an effective killing method for thrips pests in different growth stages and different spaces. SUMMARY

[0006] In view of the above, the present application provides a Cordyceps javanica FJDD-08 and an application thereof, the Cordyceps javanica FJDD-08 can parasitize in a thrips pest, and effectively kill the thrips pest. In the combined prevention and control method of the thrips pest, the Cordyceps javanica, the degradable sticky rice glue and the new bagging are used in a trinity manner, the thrips pest in different growth periods and different spaces is killed, and the thrips pest is completely eliminated from the soil source (entomophaga), after the emergence (degradable sticky rice glue) and on the crops (slow-release insecticide bagging).

[0007] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0008] A Cordyceps javanica FJDD-08 is preserved in the China General Microbiological Culture Collection Center, the address is No. 1, Beichen West Road, Haidian District, Beijing, the preservation date is February 20, 2023, 100101, the preservation number is CGMCC No. 40502, and the taxonomic name is Cordyceps javanica.

[0009] Another object of the present application is to provide the application of the above-mentioned Cordyceps javanica FJDD-08 in the prevention and control of thrips pests.

[0010] Preferably, the thrips pest is one or more of flower thrips, palm thrips, western flower thrips, bean thrips, yellow thrips, tea yellow thrips and brown thrips.

[0011] Still another object of the present application is to provide a combined prevention and control method of thrips pests, comprising the following steps:

[0012] (1) the field is ploughed, and then the mixture of the above-mentioned Cordyceps javanica FJDD-08 is uniformly mixed with fine sand and scattered on the ground;

[0013] (2) a circle of degradable sticky rice glue is applied on the stem of the crops to prevent the hatched thrips from climbing to the upper part of the crops along the root stem;

[0014] (3) a paper bag with a slow-release insecticide is used to seal the fruit in the early fruiting stage to prevent the pests from feeding.

[0015] Preferably, the step (1) is specifically operated as follows:

[0016] For bare field, the field is ploughed, and the mixture of the above-mentioned Cordyceps javanica FJDD-08 and fine sand is evenly mixed and applied to the ground.

[0017] For the existing crop field, the field is ploughed, and the mixture of the above-mentioned Cordyceps javanica FJDD-08 and fine sand is evenly mixed and applied to the ground.

[0018] Preferably, the mass ratio of the mixture of the Cordyceps javanica FJDD-08 to fine sand in step (1) is 1-4:5-15.

[0019] Preferably, the mixture of the Cordyceps javanica FJDD-08 in step (1) is composed of Cordyceps javanica FJDD-08 spore powder, wheat bran, talcum powder and soybean oil with a mass ratio of 2-5:40-100:2-5:7-15.

[0020] Preferably, the spore content of the Cordyceps javanica FJDD-08 spore powder is 3.5x10 8 6 / g, and the viable spore rate is 85%-100%.

[0021] Preferably, the height of the degradable armyworm glue applied in step (2) is 5-20 cm from the bottom of the crop roots.

[0022] Preferably, the depth of the field ploughing in step (1) is 3-5 cm.

[0023] According to the above technical solution, compared with the prior art, the present application has the following beneficial effects:

[0024] (1) The present application provides a Cordyceps javanica FJDD-08 and its application. The Cordyceps javanica FJDD-08 can parasitize in thrips pests, and the corrected mortality rate is 95.36%, which can effectively kill thrips pests.

[0025] (2) The combined control method of thrips pests provided by the present application applies the Cordyceps javanica, degradable armyworm glue and new type of bagging in a three-in-one control measure, which can kill thrips pests in different growth periods and different spaces, and can completely eliminate pests from the soil (entomogenous fungi), after emergence (degradable armyworm glue) and on crops (slow-release insecticide bagging). BRIEF DESCRIPTION OF DRAWINGS

[0026] In order to make the technical solutions in the embodiments of the present application or the prior art clearer, the accompanying drawings needed in the embodiments or prior art description will be briefly introduced below. Obviously, the accompanying drawings in the following description only represent some of the embodiments of the present application, and other drawings can be obtained by those of ordinary skill in the art without any creative effort based on the provided drawings.

[0027] Figure 1 The accompanying drawings are morphological characteristics and culture traits of Cordyceps javanica FJDD-08.

[0028] Note: A-B: FJDD-08 colony front view and back view; C-D: FJDD-08 mycelium conidiophore and conidium.

[0029] Figure 2 The accompanying drawings are phylogenetic tree of strain FJDD-08 based on 18S rDNA. DETAILED DESCRIPTION

[0030] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments only represent some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without any creative effort belong to the scope of protection of the present application.

[0031] Cordyceps javanica FJDD-08 involved in the following examples is preserved in the China General Microbiological Culture Collection Center, located at No. 1, Beichen West Road, Yard 3, Chaoyang District, Beijing, 100101, and the preservation date is February 20, 2023, and the preservation number is CGMCC No. 40502.

[0032] Example 1

[0033] The separation and identification of Cordyceps javanica FJDD-08 of the present application are as follows:

[0034] 1. Strain separation

[0035] Sample source: In June 2022, dead and dying insect corpses of various instars of thrips naturally infected with diseases were collected from the Biosafety Research Lab of the Institute of Plant Protection, Fujian Academy of Agricultural Sciences.

[0036] The separation steps are as follows:

[0037] (1) Soak the collected thrips mummies in 75% alcohol for 3s in a clean bench;

[0038] (2) Subsequently, rinse the sample 3 times with sterilized distilled water, place on a clean slide, and divide the cadaver into 3 pieces using a dissecting needle;

[0039] (3) Use an inoculation needle to inoculate the sample onto PDA solid medium, and repeat the above operation for other samples;

[0040] Culture conditions: place the culture dish in a constant temperature incubator for culture, culture temperature 27±1℃, humidity 70%±5% RH, light cycle 14L-10D, culture for 7 days;

[0041] (4) Pick the mycelium for 2 times of plate streaking separation to obtain pure culture colonies.

[0042] The strain isolated from the western flower thrips mummy is named FJDD-08, the sample name and isolation date are recorded, and is stored in a 4℃ refrigerator for standby.

[0043] PDA medium composition: potato dextrose agar medium (PDA) ordered from Beijing Coolab Technology Co., Ltd., containing potato powder, glucose, and agar. Weigh 46g of the medium, dissolve in 1L of distilled water, and sub-pack into triangular bottles for standby, sterilize at 121℃ for 30min.

[0044] Constant temperature incubator: model: PGX-250B; Ningbo Saifu Experimental Instrument Co., Ltd.

[0045] 2. Strain identification

[0046] 2.1 Strain culture traits and morphological characteristics

[0047] After the strain FJDD-08 was inoculated on PDA medium and cultured at 27℃ for 7 days, the colony initially presented a point-like radial distribution, the front of the colony initially presented a white velvet-like appearance, the middle was slightly protruding, the edge presented an irregular sawtooth shape, and there was a circle of irregular wheel marks from the inside to the outside. The back of the colony had scattered brown pigment deposition groups (as shown in Figure 1 A and Figure 1 B figures, respectively). The conidial phialides were very short, and the conidia were spherical or short rod-shaped, about 1.0-2.2μm×2.0-3.8μm (as shown in Figure 1 C and Figure 1 D figures, respectively).

[0048] 2.2 Molecular identification

[0049] The rDNA-ITS sequence of strain FJDD-08 was amplified by using fungal universal primers ITS1 / ITS4, and the PCR product was purified and sent for testing. The results showed that the rDNA-ITS sequence of strain FJDD-08 was 568 bp in length, and the GenBank accession number was OQ704261. The DNA sequence was subjected to Blast comparison in NCBI, and the similarity between strain FJDD-08 and Cordyceps javanica (GenBank accession number MH532893.1) was as high as 99%. The DNA sequence of the strain with higher similarity was selected and downloaded, and a phylogenetic tree was constructed in MEGA11.0 software using the neighbor-joining method, as shown in FIG. 1. Figure 2 The results are shown in the accompanying drawings.

[0050] The results showed that strain FJDD-08 was in the same branch as Cordyceps javanica. Based on this, combined with the morphological identification results, it was determined that the strain was Cordyceps javanica.

[0051] Example 2

[0052] Laboratory pathogenicity test

[0053] The isolated strain FJDD-08 was cultured for 7 days, the colony was washed with 5 mL of sterilized 0.1% Tween-80, the conidial suspension was collected, the concentration of the spore suspension was calculated using a hemocytometer, and then a spore suspension with a concentration of 1×10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 spores / mL was prepared. The total number of western flower thrips adults in each test group was 30, and three replicates were set. After 3 days of inoculation, the mortality rate and LC 50 were calculated. The experimental results are shown in Table 1.

[0054] Table 1 Toxicity test results of FJDD-08 strain on thrips adults

[0055]

[0056] Analysis of results: when the concentration of FJDD-08 strain reached 1×10 8 spores / mL, the corrected mortality rate reached 95.36%, and the LC50 of strain FJDD-08 was calculated by the dose regression equation as 3.67×10 5 spores / mL.

[0057] Similarly, according to the preliminary test of the laboratory toxicity test, when the concentration of FJXH-08 strain reached 1×10 8The corrected mortality of flower thrips, palm thrips, bean thrips, yellow thrips, tea yellow thrips and brown thrips were all higher than 90% at 1×106 spores / mL.

[0058] The slow-release insecticide on the surface of the bag in Examples 3-7 was 5% emamectin benzoate produced by Shenzhen Nuopuxin Chemical Co., Ltd.

[0059] The degradable gypsy moth glue in Examples 3-7 was purchased from Quanzhou Lvuposen Biological Technology Co., Ltd.

[0060] The mango garden in Sesidi Village, Dongzhang Town, Fuqing City, Fujian Province was selected as the test site in Examples 3-7.

[0061] Example 3 (treatment 1)

[0062] (1) 15 g of Cordyceps javanica spore powder, 300 g of wheat bran, 15 g of talc powder, and 52.5 g of soybean oil were weighed. A small amount of wheat bran and talc powder were mixed to obtain a mixture, and the mixture was mixed with Cordyceps javanica spore powder, soybean oil, and the remaining wheat bran to obtain a mixture of Cordyceps javanica FJDD-08. Before application, it was mixed with 1912.5 g of fine sand, and after deep plowing 5 cm, it was uniformly applied to the ground.

[0063] (2) A circle of degradable gypsy moth glue was applied from the root to the top 10 cm of the crop stem to prevent thrips from crawling to the upper part of the crop along the root stem;

[0064] (3) At the initial stage of fruit setting, paper bags with 5% emamectin benzoate slow-release insecticide on the outside were used to seal the fruit to prevent pests from feeding.

[0065] Example 4 (treatment 2)

[0066] (1) 15 g of Cordyceps javanica spore powder, 300 g of wheat bran, 15 g of talc powder, and 52.5 g of soybean oil were weighed. A small amount of wheat bran and talc powder were mixed to obtain a mixture, and the mixture was mixed with Cordyceps javanica spore powder, soybean oil, and the remaining wheat bran to obtain a mixture of Cordyceps javanica FJDD-08. Before application, it was mixed with 1912.5 g of fine sand, and after deep plowing 5 cm, it was uniformly applied to the ground.

[0067] Example 5 (treatment 3)

[0068] Only degradable gypsy moth glue was used to control thrips pests.

[0069] Example 6 (treatment 4)

[0070] Only bags containing 5% emamectin benzoate were used to control thrips pests.

[0071] Example 7 (Treatment 5)

[0072] The mango orchard without any treatment was used as the control group.

[0073] Each treatment had 4 replicates, 2 trees per replicate (5 treatments, 40 trees in total), and 5 points per tree were investigated. The field thrips occurrence was investigated before treatment and 3d, 7d, 15d, 30d after treatment. The field control effect results are shown in Table 2.

[0074] Table 2 Effect of different treatments on controlling mango thrips pests

[0075]

[0076] Results analysis: Treatment 1 (Java Cordyceps + degradable sticky glue + bag with slow-release insecticide) > Treatment 4 (bag with 5% emamectin benzoate) > Treatment 2 (Java Cordyceps) > Treatment 3 (degradable sticky glue) > Treatment 5 (control without any treatment). In each time period, the control effect of Treatment 1 was significantly higher than that of Treatments 2, 3 and 4, especially 3 days after treatment, the control effect of Treatment 1 was 1.32-3.78 times that of Treatments 2, 3 and 4.

[0077] The various embodiments in the specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0078] The above description of disclosed embodiments enables a person skilled in the art to implement or use the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to these embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A strain of Cordyceps javanica (Cordyceps javanica ) FJDD-08, characterized in that, It was preserved in China General Microbiological Culture Collection Center, the address is No. 1, Beichen West Road, Chaoyang District, Beijing, the preservation date is February 20, 2023, and the preservation number is CGMCC No. 40502.

2. The Cordyceps javanicus bacteria of claim 1, Cordyceps javanica ) Application of FJDD-08 in the prevention and treatment of western flower thrips.

3. A method for the integrated control of Frankliniella occidentalis, characterized by, The method comprises the following steps: (1) The field is turned over, and then the Cordyceps javanica fungus in claim 1 is applied to the field; Cordyceps javanica ) FJDD-08 is mixed with fine sand and spread on the ground;​ (2) A circle of degradable sticky insect glue is applied to the stem part of crops to prevent thrips from climbing to the upper part of crops along the rhizome part; (3) Paper bags with slow-release insecticides are used to seal the fruits in the early fruiting stage to prevent pests from feeding.

4. The method for combined control of Frankliniella occidentalis according to claim 3, characterized by, Step (1) is specifically operated as follows: For bare fields, turn the fields over and then apply the Cordyceps militaris described in claim 1. Cordyceps javanica Mix FJDD-08 with fine sand and spread it on the ground; for existing crop fields, till the area around the crops, and then apply the Cordyceps militaris described in claim 1. Cordyceps javanica Mix the FJDD-08 mixture with fine sand and spread it on the ground.

5. The method for combined prevention and control of Frankliniella occidentalis according to claim 3, characterized in that, The mixture of the step (1) said Java Cordyceps fungus Cordyceps javanica The mass ratio of the mixture of the Java Cordyceps fungus FJDD-08 to fine sand is 1-4:5-15.

6. The method for combined prevention and control of Frankliniella occidentalis according to claim 3, characterized in that, Step (1) describes the Java Cordyceps fungus ( Cordyceps javanica The mixture of FJDD-08 consists of *Cordyceps militaris* in a mass ratio of 2-5:40-100:2-5:7-15. Cordyceps javanica It is composed of FJDD-08 spore powder, wheat bran, talc powder and soybean oil.

7. The method for combined control of Frankliniella occidentalis according to claim 6, characterized in that, The said Java Cordyceps fungus Cordyceps javanica The spore content of the spore powder of FJDD-08 is 3.5×10 8 spores / g, and the viable spore rate is 85%-100%.

8. The method for combined prevention and control of Frankliniella occidentalis according to claim 3, characterized in that, The height of the degradable sticky insect glue applied in step (2) is 5-20 cm from the bottom of the crop roots.

9. The method for combined prevention and control of Frankliniella occidentalis according to claim 3, characterized in that, The depth of the field turning in step (1) is 3-5 cm.

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