A method for breeding of phytoseiulus persimilis and application thereof
By using wheat bran and vermiculite substrate combined with specific feed mites, the problem of the unsuccessful artificial large-scale propagation of Amblyseius tamataf was solved, enabling the rapid reproduction of Amblyseius tamataf and effective control of micro-sized pests, thereby reducing environmental pollution.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- YUNNAN TOBACCO CO LTD KUNMING BRANCH
- Filing Date
- 2024-05-24
- Publication Date
- 2026-06-05
AI Technical Summary
The artificial propagation of *Amblyseius tamataf* has not been achieved on a large scale. Existing propagation methods suffer from problems such as high workload, high space occupation, low cost but easy mold growth, and low propagation efficiency.
Using wheat bran and vermiculite as a substrate, yeast is added to make feed, which is combined with feed mites such as carrion mites, sweet fruit mites and insectivorous anticlimactites, and is bred in a specific ratio and inoculation sequence. Under the breeding conditions, the artificial large-scale breeding of Amblyseius tamataf is achieved.
It significantly increases the population of Amblyseius tamatae, enabling large-scale artificial breeding with rapid proliferation, reducing pesticide use, and is environmentally friendly, controlling micro-sized pests on tobacco, flowers, fruits, and vegetables.
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Figure CN118415142B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biological control technology for agricultural pests, specifically relating to a method for breeding and applying the amblyseius tamataf mite. Background Technology
[0002] Amblyseius tamatavensis belongs to the genus Amblyseius of the family Phytoseiidae in the order Parasitiformes of the class Arachnida. It was named after the port of Tamatavensis in Madagascar where it was first discovered.
[0003] Records indicate that *Amblyseius tamataf* can prey on a variety of microscopic pests, including whiteflies, silver leaf whiteflies, thrips, red palm mites, and citrus rust mites, making it a superior predatory mite germplasm resource. Assessing the development potential of a predatory mite involves evaluating not only its predatory ability but also its potential for large-scale propagation to achieve pest control. Currently, relying solely on the natural reproduction of predatory mites in the field is insufficient for pest control; multiple artificial releases of large quantities of predatory mites are typically required to effectively control pests and mites.
[0004] Therefore, researching methods to achieve mass reproduction of predatory mites is particularly important. Currently, commonly used methods include natural prey reproduction, substitute prey reproduction, artificial feed reproduction, and pollen reproduction. Natural prey reproduction involves using natural prey, such as spider mites, to feed the predatory mites and achieve mass reproduction. Although this method results in rapid population growth and large population numbers, it requires cultivating plants to obtain a sufficient quantity of prey before introducing the predatory mites for reproduction. This process is labor-intensive, space-consuming, and technically demanding. Artificial feed reproduction uses chemically formulated diets with different nutritional components to enable successful reproduction of predatory mites. While some artificial feeds can promote the development and reproduction of predatory mites, problems such as low egg production and short lifespan, as well as the unresolved issue of mold growth in artificial feeds, have prevented their effective use. Therefore, artificial feed reproduction of predatory mites is rarely used currently. Pollen reproduction involves feeding the predatory mites with one or more types of pollen, allowing them to multiply rapidly. However, using pollen to feed predatory mites also presents challenges such as pollen mold growth and the limited variety of mites that can be reared. Alternative prey methods utilize storage mites like *Aleuroglyphus ovatus*, *Tyrophagus putrescentiae*, and *Carpoglyphus lactis* as substitute prey. The advantages of these storage mites are that their diet consists of wheat bran and rice husks, which are low-cost, widely available, require minimal space for reproduction, and have high reproductive efficiency. However, commercially developed predatory mites, such as *Neoseiulus cucumeris* and *Neoseiulus californicus*, utilize alternative prey for large-scale propagation. Currently, *Amblyseius tamataf* has not been successfully propagated on a large artificial scale. Summary of the Invention
[0005] This invention addresses the current problem of the lack of large-scale artificial propagation of *Amblyseius tamataf*, and provides a method for breeding *Amblyseius tamataf* and its application, enabling large-scale artificial propagation of *Amblyseius tamataf*.
[0006] The technical solution claimed by this invention is as follows:
[0007] A method for breeding the tarmataef amblyseius mite includes the following steps:
[0008] S1: The substrate is sterilized at 100°C and then cooled to room temperature; the substrate includes wheat bran and vermiculite;
[0009] S2: Mix the substrate obtained in S1 with wheat bran and vermiculite in a ratio of 1-10:1, then add water to adjust the moisture content to 13%-35%, and then add 10-100g of yeast to make feed.
[0010] S3: Feeding predatory mites with one or more of the feed mites from S2, including carrion mites, sweet fruit mites, and insectivorous narrow mites; the predatory mite is Amblyseius tamataf.
[0011] Preferably, the disinfection time in S1 is 60 minutes.
[0012] In a specific embodiment of the present invention, in step S3, the predatory mites are raised in a constant temperature box.
[0013] Preferably, the rearing conditions for predatory mites in step S3 are: temperature 24-26℃, humidity 90-95%, and photoperiod L:D = 12h:12h.
[0014] Preferably, in step S3, when only one type of feed mite is inoculated, feed mites are inoculated first, and after the feed mites are inoculated, predatory mites are inoculated for rearing; when two types of feed mites are inoculated, the feed mites are inoculated twice. The first inoculation of feed mites: select one type of feed mite for inoculation, and after the inoculation is completed, predatory mites are inoculated for rearing; after a period of rearing, the second inoculation of feed mites is carried out: inoculate the other type of feed mite, and after the inoculation is completed, the rearing of predatory mites continues.
[0015] Preferably, the predatory mites are reared for 19-23 days; when inoculating two types of feed mites, one type of feed mite is inoculated for 9-11 days before the other type of feed mite is inoculated; the ratio of the number of the two types of feed mites inoculated is 1:1; and the ratio of the number of predatory mites to feed mites is 1:100-1000.
[0016] In some embodiments of the present invention, in step S3, the feed mites inoculated are sweet fruit mite and insectivorous narrow mite, and the inoculation order is: sweet fruit mite, insectivorous narrow mite; the feed mites inoculated are detritivorous mites and sweet fruit mite, and the inoculation order is: detritivorous mites, sweet fruit mite.
[0017] In some other embodiments of the present invention, the feed mites inoculated in step S3 are Tyromitra saprophytica and Acer cristatum, and the inoculation order is: Tyromitra saprophytica, Acer cristatum.
[0018] Preferably, the inoculation ratio of predatory mites to feed mites is 1:600.
[0019] This invention provides the application of the Tamataf amblyseius mite bred using the above-mentioned breeding method in the control of micro-sized pests on tobacco, flowers, fruits, and vegetables, wherein the micro-sized pests include: spider mites, thrips, and whiteflies.
[0020] Beneficial effects:
[0021] This invention provides a method for breeding *Amblyseius tamatae*. One or more predatory mites, including *Tyrophagus tamariscina*, *Drosophila melanogaster*, and *Angiophagus tamariscina*, are selected to rear *Amblyseius tamatae*. Experiments showed that after 21 days of rearing with different predatory mites, the number of *Amblyseius tamatae* mites significantly increased. In particular, when using a combination of *Tyrophagus tamariscina* and *Angiophagus tamariscina*, with the inoculation order being *Tyrophagus tamariscina* followed by *Angiophagus tamariscina*, and the ratio of predatory mites to predatory mites being 1:600, the number of *Amblyseius tamatae* mites after 21 days was 19.27 times the initial number. This method achieves artificial breeding of *Amblyseius tamatae*, solving the current problem of the lack of large-scale artificial propagation of *Amblyseius tamatae*.
[0022] The method provided by this invention for breeding the Tamataf amblyseius mite can control small pests such as spider mites, thrips, and whiteflies on tobacco, flowers, fruits, and vegetables, reducing the use of pesticides, making it green and environmentally friendly, and reducing environmental pollution. Attached Figure Description
[0023] Figure 1 This is a flowchart of the present invention. Detailed Implementation
[0024] The present invention will be further described below with reference to specific embodiments. Unless otherwise specified, the components described in the present invention are all readily available to those skilled in the art.
[0025] The *Amblyseius tamatae* mite described in this invention was first discovered by the applicant in Yunnan, China, and was collected from blueberry plants; the specimen is deposited at Guizhou University. The *Tyrophagus putrescentiae*, *Carpoglyphus lactis*, and *Thyreophagus entomophagus* mites used below were all commercially available.
[0026] The instruments, consumables and reagents used include: 1L plastic box, wheat bran, vermiculite, paintbrush, alcohol, petri dish, counter, stereoscope, and incubator.
[0027] First Example: Breeding Method of *Amblyseius tamataf*
[0028] This set of embodiments provides a method for breeding *Amblyseius tamataf*, such as... Figure 1 As shown, it includes the following steps:
[0029] S1: The substrate is sterilized at 100°C for 60 minutes and then cooled to room temperature; the substrate includes wheat bran and vermiculite;
[0030] S2: Mix the substrate obtained in S1 with wheat bran and vermiculite in a ratio of 1-10:1, then add water to adjust the moisture content to 13%-35%, and then add 10-100g of yeast to make feed.
[0031] S3: Inoculate one or more of the feed mites from S2 into a constant temperature incubator at a temperature of 24-26℃, relative humidity of 90-95%, and a photoperiod L:D = 12h:12h for predatory mite rearing for 19-23 days. The feed mites include carrion mites, sweet fruit mites, and insectivorous anthracids; the predatory mite is *Amblyseius tamatae*. Specifically, when inoculating only one type of feed mite, the feed mites are inoculated first, followed by predatory mites. When inoculating two types of feed mites, the feed mites are inoculated twice. For the first inoculation, one type of feed mite is selected, followed by predatory mites. After 9-11 days of rearing, the other type of feed mite is inoculated, followed by predatory mite rearing. The ratio of predatory mites to feed mites is 1:100-1000.
[0032] In some embodiments, the feed mites inoculated are fruit mites and insectivorous angiomites, and the inoculation order is: fruit mites, insectivorous angiomites, wherein the number ratio of fruit mites to insectivorous angiomites is 1:1; the feed mites inoculated are detritivorous mites and fruit mites, and the inoculation order is: detritivorous mites, fruit mites, wherein the number ratio of detritivorous mites to fruit mites is 1:1.
[0033] In other embodiments, the feed mites inoculated are Tyromitra saprophytica and Angiomyotis diffusa, and the inoculation order is: Tyromitra saprophytica, Angiomyotis diffusa, wherein the ratio of Tyromitra saprophytica to Angiomyotis diffusa is 1:1; the inoculation ratio of the predatory mite (Amblyseius tamataf) to the feed mites (Tyromitra saprophytica + Angiomyotis diffusa) is 1:600.
[0034] Second set of examples: Application of *Amblyseius tamataf*
[0035] This set of embodiments provides the application of Amblyseius tamataf, which can be used to control micro-sized pests such as spider mites, thrips, and whiteflies on tobacco, flowers, fruits, and vegetables.
[0036] Experimental Example 1. Effects of different feeder mites on the rearing population of *Amblyseius tamatae*.
[0037] 1. Experimental Methods
[0038] (1) Substrate disinfection: Disinfect the substrate (wheat bran and vermiculite) at 100℃ for 60 minutes for later use;
[0039] (2) Making feed: After the substrate that has been disinfected in step (1) is cooled to room temperature, mix it with wheat bran and vermiculite in a ratio of 1:1, add water, adjust the substrate moisture content to 13-17%, add yeast at a ratio of 10g yeast / L substrate, mix well and make feed.
[0040] (3) Making feeding boxes: The feed obtained in step (2) is divided into 1L plastic boxes, each containing 500ml of feed to make feeding boxes for later use.
[0041] (4) A total of 6 treatments were set up in the experiment: T1, T2, T3, T4, T5 and T6. Different combinations of feed mites were selected for the 6 treatments (Table 1). Feed mites were inoculated for the first time in the feeding boxes prepared in step (3) according to the inoculation method in Table 1.
[0042] Table 1. Six different treatments and corresponding feed mite combinations, and the inoculation method for the first feed mite inoculation.
[0043] deal with feed mite combination vaccination method T1 putrefactive typhoid mite Introducing 2 million carrion mites T2 Sweet fruit mite Introducing 2 million sweet fruit mites T3 Insectivorous mites Introducing 2 million insectivorous anthracids T4 Tyrophaga putrificans + Insectivorous mites Introducing 1 million carrion mites T5 Sweet fruit mite + insectivorous narrow mite Introducing 1 million sweet fruit mites T6 Tyrophaga putrescent mite + sweet fruit mite Introducing 1 million carrion mites
[0044] (5) Inoculation with predatory mites: After the first inoculation with feed mites is completed, 2,000 Tamaratavus amblyseius mites are introduced into the 6 treatments corresponding to the feeding boxes set in step (4).
[0045] (6) Incubator rearing: Place the rearing boxes (6 treatments) inoculated with feed mites and predatory mites in step (5) into an incubator for rearing. The temperature of the incubator is 24-26℃, the relative humidity is 90-95%, and the photoperiod is L:D = 12h:12h.
[0046] (7) Secondary inoculation with feed mites: For treatments T4, T5 and T6, when the tamataf ampulline mites were fed to the 10th day, 1 million insectivorous angiomites were introduced into treatment T4, 1 million insectivorous angiomites were introduced into treatment T5, and 1 million sweet fruit mites were introduced into treatment T6.
[0047] (8) Incubator rearing: After the second inoculation in step (7), the rearing boxes corresponding to treatments T4, T5 and T6 are placed in the incubator for continued rearing, and the rearing conditions remain unchanged.
[0048] (9) The number and growth rate of predatory mites fed with different feed mites were observed on day 21 after six different treatments.
[0049] (10) Detection of predatory mites: 1 ml of predatory mites was measured as a replicate point. The number of active mites other than eggs was counted under a stereomicroscope and recorded. Ten replicate points were taken for each treatment for detection. The average of the 10 points was multiplied by 500 to obtain the number of Amblyseius tamatae after 21 days of treatment.
[0050] (11) Calculation formula: Growth multiple = Number of Tamarataf amblyseius after 21 days / Number of Tamarataf amblyseius at the initial inoculation;
[0051] (12) Experimental replication: The above experiment was repeated 3 times.
[0052] Table 2. Changes in the population and growth rate of *Amblyseius tamata* under six different treatments.
[0053]
[0054] Note: The values in the table are mean ± standard error. Different lowercase letters in the same column indicate significant differences.
[0055] 2. Experimental Results
[0056] As shown in Table 2, after feeding *Amblyseius tamatae* with different feeder mites (6 treatments) for 21 days, the number of *Amblyseius tamatae* mites in treatment T4 was 36,066.67, which was 18.03 times the initial number. Treatment T3 was the second highest, with 27,800 mites, 13.90 times the initial number. Treatment T6 had the lowest number of mites, only 15,600, which was only 7.80 times the initial number. Therefore, treatment T4 (*Typhae carrionis* + *Streptococcus terrestris*) is the preferred feeder mite combination for *Amblyseius tamatae*.
[0057] Experimental Example 2. Effect of the inoculation ratio of predatory mites and feed mites (carrion mites + insectivorous stenosperms) on the rearing population of *Amblyseius tamatae*.
[0058] 1. Experimental Methods
[0059] (1) Substrate disinfection: Disinfect the substrate (wheat bran and vermiculite) at 100℃ for 60 minutes for later use;
[0060] (2) Making feed: After the substrate that has been disinfected in step (1) is cooled to room temperature, mix it with wheat bran and vermiculite in a ratio of 1:1, add water, adjust the substrate moisture content to 13-17%, add yeast at a ratio of 10g yeast / L substrate, mix well and make feed.
[0061] (3) Making feeding boxes: The feed obtained in step (2) is divided into 1L plastic boxes, each containing 500ml of feed to make feeding boxes for later use.
[0062] (4) The experiment set up three treatments: H1, H2 and H3. Each treatment corresponds to a breeding box obtained in step (3). The first inoculation of feed mites: H1 was inoculated with 300,000 mites of carrion in its corresponding breeding box, H2 was inoculated with 600,000 mites of carrion in its corresponding breeding box, and H3 was inoculated with 900,000 mites of carrion in its corresponding breeding box.
[0063] (5) Inoculation with predatory mites: After the first inoculation of feed mites for treatments H1, H2 and H3 is completed, 2000 Tamartaf amblyseius mites will be introduced into the corresponding breeding boxes for treatments H1, H2 and H3 respectively.
[0064] (6) Incubator rearing: Place the rearing boxes (treatments H1, H2, and H3) inoculated with feed mites and predatory mites into an incubator for 10 days. The rearing conditions are: incubator temperature 24-26℃, relative humidity 90-95%, and photoperiod L:D = 12h:12h.
[0065] (7) Second inoculation with feed mites: After step (6) is completed, 300,000 insectivorous mites are introduced into the breeding box corresponding to treatment H1, 600,000 insectivorous mites are introduced into the breeding box corresponding to treatment H2, and 900,000 insectivorous mites are introduced into the breeding box corresponding to treatment H3.
[0066] (8) Continue to raise in constant temperature box: After the second inoculation of feed mites in step (7), the corresponding breeding boxes for treatments H1, H2 and H3 are put into the constant temperature box for raising again. The raising conditions are the same, namely: the temperature of the constant temperature box is 24-26℃, the relative humidity is 90-95%, and the photoperiod L:D = 12h:12h. On the 21st day of raising, observe the number and growth rate of predatory mites raised under different inoculation ratios of treatments H1, H2 and H3.
[0067] (9) Detection of predatory mites: 1 ml of predatory mites was measured as a replicate point. The number of active mites other than eggs was counted under a stereomicroscope and recorded. Ten replicate points were taken for each treatment for detection. The average of the 10 points was multiplied by 500 to obtain the number of Amblyseius tamatae in the rearing box after 21 days.
[0068] (10) Calculation formula: Growth multiple = Number of Tamarataf amblyseius after 21 days / Number of Tamarataf amblyseius at the initial inoculation;
[0069] (11) Experimental replication: The above experiment was repeated 3 times.
[0070] Table 3. Number and growth rate of Amblyseius tamatae at different inoculation ratios (treatments H1, H2, H3)
[0071]
[0072] Note: The values in the table are mean ± standard error. Different lowercase letters in the same column indicate significant differences.
[0073] 2. Experimental Results
[0074] As shown in Table 3, when the initial inoculation number of *Amblyseius tamatae* was 2000, the highest increase in *Amblyseius tamatae* population was observed in treatment H2 (a combination of 600,000 *Tyromitra saliva* and 600,000 *Angiomyophyte mites*), reaching 38533.33 individuals, which was 19.27 times the initial inoculation number. Therefore, treatment H2, with an inoculation ratio of *Amblyseius tamatae* to (*Tyromitra saliva* + *Angiomyophyte mites*) of 1:600, is the optimal inoculation ratio for rearing *Amblyseius tamatae*.
[0075] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.
Claims
1. A method for breeding *Amblyseius tamataf*, characterized in that, Includes the following steps: S1: The substrate is sterilized at 100°C and then cooled to room temperature; the substrate includes wheat bran and vermiculite; S2: Mix the substrate obtained in S1 with wheat bran and vermiculite in a ratio of 1-10:1, then add water to adjust the moisture content to 13%-35%, and then add 10-100g of yeast to make feed. S3: Predatory mites are raised by inoculating one or more of the feed mites from S2, including *Tetranychus saprophyticus* and *Angiomyxobolus tamarata*; the predatory mite is *Amblyseius tamata*. When inoculating two types of feed mites, the feed mites are inoculated twice. The first inoculation of feed mites involves selecting one type of feed mite for inoculation, followed by inoculation of predatory mites. After a period of rearing, the second inoculation of feed mites is performed: inoculating the other type of feed mite, followed by rearing of predatory mites. The feed mites inoculated are *Tetranychus saprophyticus* and *Angiomyxobolus tamarata*, and the inoculation order is: *Tetranychus saprophyticus*, *Angiomyxobolus tamarata*. The ratio of the number of predatory mites to feed mites is 1:
600.
2. The method for breeding *Amblyseius tamatae* according to claim 1, characterized in that, The disinfection time in S1 is 60 minutes.
3. The method for breeding *Amblyseius tamatae* according to claim 1 or 2, characterized in that, In step S3, the predatory mites are raised in a constant temperature chamber.
4. The method for breeding *Amblyseius tamatae* according to claim 3, characterized in that, The rearing conditions for predatory mites in step S3 are: temperature 24-26℃, humidity 90-95%, and photoperiod L:D = 12h:12h.
5. The method for breeding *Amblyseius tamatae* according to claim 1, characterized in that, The predatory mites are reared for 19-23 days; when inoculating with two types of feed mites, one type of feed mite should be inoculated for 9-11 days before the other type of feed mite is inoculated; the ratio of the two types of feed mites inoculated is 1:
1.
6. The application of *Amblyseius tamatae* mites bred using any of the breeding methods described in claims 4-5 in the control of micro- and small-scale pests on tobacco, flowers, fruits, and vegetables, wherein the micro- and small-scale pests include: Spider mites, thrips, whiteflies.