A spherical torus of arthrographis and its use in preparing a highly effective fruit fly attractant

Abstract

The present application relates to a kind of spherical ring conidium and its application in the preparation of high-efficiency real fly attractant, the spherical ring conidium is (Torulaspora globose) F-HY017-Y1, the high-efficiency real fly attractant includes 20-40% of yeast source active ingredient, the yeast source active ingredient is the fermentation product of the above-mentioned spherical ring conidium.The present application is screened to obtain a kind of yeast, confirms that the fermentation product has high-efficiency attraction effect to real fly female and male, and the strain is isolated from the host of target pest feeding, and more specific;And in the preparation of high-efficiency real fly attractant, the present application optimizes the process, further enzymolysis is carried out after yeast fermentation, releases the core active ingredient, greatly improves the utilization of yeast fermentation product, and the attraction efficiency to real fly pest is higher.

Description

Technical Field

[0001] This invention relates to the field of pest control, specifically to a spherical Cyclospora yeast and its application in the preparation of highly effective fruit fly attractants. Background Technology

[0002] Fruit flies are an important quarantine pest in horticulture, primarily causing damage as larvae. Adults lay their eggs in the crevices between the fruit peel and flesh. After hatching, the larvae remain hidden inside the fruit, feeding extensively on the flesh before obvious symptoms such as premature yellowing and fruit drop appear. Their highly concealed damage makes them difficult to control, and ineffective control can lead to significant yield reductions or even total crop failure. Currently, the main methods for controlling fruit flies rely on chemical control and adult trapping. While chemical control is fast-acting, it is prone to pesticide contamination and pesticide resistance in pests. Therefore, adult trapping is an important method for the green control of fruit flies.

[0003] Currently, fruit fly attractants mainly consist of food baits or sex pheromones. While sex pheromones are effective at attracting fruit flies, they generally only attract males and are species-specific; one sex pheromone cannot be effective against multiple fruit fly species simultaneously. Food baits, on the other hand, attract multiple fruit fly species and can attract both males and females. Highly effective food baits are the best attractants for controlling fruit flies. However, there is a lack of highly effective fruit fly food baits in China, and imported food baits are expensive and have inconsistent and unsatisfactory attraction effects, hindering large-scale promotion. Therefore, developing highly effective and inexpensive fruit fly food baits is an urgent problem to be solved. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a spherical Cyclospora yeast and its application in the preparation of a highly efficient fruit fly attractant, in response to the above problems and requirements.

[0005] To solve the above technical problems, the present invention adopts the following technical solution:

[0006] A globose yeast, specifically Torulaspora globose F-HY017-Y1, was deposited at the China Center for Type Culture Collection on May 11, 2022, with the accession number CCTCC NO:M2022597.

[0007] The application of *Torulaspora globose* in the preparation of a highly effective fruit fly attractant is as follows: the fermentation product of *Torulaspora globose* F-HY017-Y1 is used as the effective active ingredient from the yeast source. The highly effective fruit fly attractant comprises the following components in the indicated mass fractions: 20-40% effective active ingredient from the yeast source, 4%-32% filler, 12%-40% disintegrant, 0%-4% lubricant, and 24%-48% preservative.

[0008] Furthermore, the disintegrant is any one or more of sodium carboxymethyl starch, crospovidone, citric acid, and sodium bicarbonate; the preservative is any one or more of sodium diacetate, potassium sorbate, calcium propionate, and methylparaben; the filler is any one or more of microcrystalline cellulose, glucose, dextrin, and soluble starch; and the lubricant is any one or more of magnesium stearate, calcium stearate, and talc.

[0009] Furthermore, the yeast fermentation product is a dry powder of enzymatically hydrolyzed protein, which is obtained by fermenting Torulaspora globose F-HY017-Y1, followed by enzymatic hydrolysis of the fermentation product, and then spray drying.

[0010] Furthermore, the enzymatic hydrolysis method for yeast fermentation products includes the following steps: centrifuging the fermentation broth and collecting the precipitate, dissolving the precipitate in pure water to obtain a homogenate, adjusting the pH value to between 10 and 12, heating at a constant temperature of 40-55℃ for 3 hours, then adjusting the pH value to between 2 and 7, adding protease, and continuing to heat at a constant temperature of 40-55℃ for 10 hours to obtain a yeast enzymatic hydrolysate solution. The protease includes any one or more of papain, acidic protease, and neutral protease; the pH value of the homogenate is adjusted according to the usage conditions of different proteases.

[0011] Furthermore, the yeast fermentation method is as follows: after inoculating 5-7% of activated yeast strains onto the fermentation medium, culture it in an incubator for 24-48 hours.

[0012] Furthermore, the fermentation medium comprises the following components by mass fraction: 5%-7% carbon source, 0.5%-1.5% nitrogen source, and 0.3%-1% yeast extract, and also includes 0.03mol / L-0.07mol / L KCl; the pH value of the fermentation medium is 5-7, the culture temperature during the culture process is 25-28℃, and the volume of the fermentation medium is 30-100mL.

[0013] Furthermore, the carbon source includes any one or more of glucose, sucrose, corn starch saccharification products, potato starch saccharification products, and cassava starch saccharification products; the nitrogen source includes any one or more of polypeptone, fish meal, blood meal, and urea.

[0014] Furthermore, the method for spray drying the enzymatically hydrolyzed product is as follows: the enzymatically hydrolyzed protein solution is dried by passing it through a spray drying tower, wherein the inlet temperature of the spray drying tower is 180-220℃, the outlet temperature is 90-110℃, and the injection rate is 6-20ml / min.

[0015] The beneficial effects of this invention are as follows:

[0016] This invention screened a yeast strain (Torulaspora globose) F-HY017-Y1, confirming that its fermentation product has a highly effective attraction effect on both female and male fruit flies. The fermentation product was then used to prepare a highly effective fruit fly attractant. Furthermore, this strain, isolated from the host consumed by the target pest, exhibits stronger specificity. Existing techniques for producing yeast attractants involve directly using the fermentation broth after yeast fermentation. This invention optimizes the process by further enzymatically hydrolyzing the fermented product, releasing the core active ingredients and significantly improving the utilization rate of the yeast fermentation product, resulting in higher attraction efficiency for fruit flies.

[0017] The present invention will now be described in detail with reference to the embodiments. Detailed Implementation

[0018] The principles and features of the present invention are described below. The examples given are only for explaining the present invention and are not intended to limit the scope of the present invention.

[0019] Example 1

[0020] A yeast bait containing the following ingredients was prepared using a tablet press: 0.4 g yeast hydrolysate, 0.2 g dextrin, 0.2 g glucose, 0.3 g sodium carboxymethyl starch, 0.1 g crospovidone, 0.1 g magnesium stearate, 0.5 g sodium diacetate, and 0.2 g methylparaben. The control attractant was an imported commercial Candida albicans attractant.

[0021] Using common bottle-shaped traps as containers, two attractants or control attractants from this embodiment were placed in each trap, along with 200 ml of tap water. The traps containing only tap water served as negative controls.

[0022] The experiment was conducted in a simulated wild environment indoors at Huazhong Agricultural University. The indoors covered an area of ​​approximately 120 square meters and was planted with crops such as citrus, peaches, and pumpkins to simulate the wild environment. Every morning at 8:00 AM, 200 male and 200 female oriental fruit flies and 200 male and 200 female South Asian fruit flies were released into the indoors.

[0023] The bait containers were suspended 1 meter above the ground on tree branches in a location protected from direct sunlight, with at least 1.5 meters between each container. Each treatment was repeated 5 times. The number of fruit flies captured was counted the following day, and then every 2 days thereafter, for a total of 15 days. The experimental results are shown in Tables 1 and 2:

[0024] Table 1

[0025]

[0026] Table 2

[0027]

[0028] In experiments conducted in a net-filled chamber simulating a field environment, the attractant provided by this invention showed comparable effectiveness to imported Candida albicans attractants, indicating promising application prospects. Furthermore, it maintained its attractant ability until the end of the experiment, demonstrating a relatively long duration of effectiveness.

[0029] The experiment used two different fruit flies, and the results showed that the attractant provided by this invention had a good effect on both the Oriental fruit fly and the South Asian fruit fly, and had an attraction effect on both male and female flies.

[0030] Example 2

[0031] A yeast bait containing the following ingredients was prepared using a tablet press: 0.2 g yeast hydrolysate, 0.2 g microcrystalline cellulose, 0.2 g soluble starch, 0.2 g sodium carboxymethyl starch, 0.1 g crospovidone, 0.3 g citric acid, 0.1 g sodium bicarbonate, 0.1 g magnesium stearate, 0.2 g potassium sorbate, and 0.4 g calcium propionate, totaling 2.0 g. The control attractant was an imported commercial Candida albicans attractant.

[0032] Using common bottle-shaped traps as containers, two attractants prepared in Example 2 or commercial Candida attractants were placed in each trap, followed by 200 ml of tap water. The negative control did not contain any attractants, only tap water.

[0033] The experiment was conducted at the orange orchard base on the campus of Huazhong Agricultural University. The attractant cans were suspended 1 meter above the ground on tree branches in areas not exposed to direct sunlight, with at least 2 meters between each can. Each treatment was repeated 5 times. The number of fruit flies captured in each treatment was counted the following day, and then every 5 days thereafter, for a total of 30 days. The results are shown in Table 3.

[0034] Table 3

[0035]

[0036] At the Huazhong Agricultural University base, the attractant provided in Example 1 of this invention also exhibited a similar attraction effect to the positive control, indicating that this invention can effectively attract oriental fruit flies in the wild.

[0037] In this experiment, neither Example 2 nor the positive control attracted South Asian fruit flies. This is presumably because there are currently no South Asian fruit flies in the base, rather than because the attractant is ineffective against South Asian fruit flies.

[0038] Example 3

[0039] A yeast bait containing the following ingredients was prepared using a tablet press: 0.6 g yeast hydrolysate, 0.2 g microcrystalline cellulose, 0.2 g sodium carboxymethyl starch, 0.3 g citric acid, 0.1 g sodium bicarbonate, 0.1 g magnesium stearate, 0.3 g sodium diacetate, and 0.2 g calcium propionate. The control attractant was an imported commercial Candida albicans attractant.

[0040] Using common bottle-shaped traps as containers, two attractants prepared in Example 3 or commercial Candida attractants were placed in each trap, followed by 200 ml of tap water. The negative control did not contain any attractants, only tap water.

[0041] The experiment was conducted in a mango orchard in Yazhou District, Sanya City, Hainan Province (109°10'7"E, 18°21'25"N). The attractant containers were suspended 1 meter above the ground on tree branches in areas without direct sunlight, with at least 3 meters between each container. Each treatment was repeated 5 times. The number of fruit flies captured in each treatment was counted the following day, and then every 5 days thereafter, for a total of 30 days. The results are shown in Tables 4 and 5.

[0042] Table 4

[0043]

[0044] Table 5

[0045]

[0046] In experiments conducted in Sanya, Hainan, Example 3 and the positive control provided by this invention showed similar attraction effects against both the oriental fruit fly and the South Asian fruit fly, indicating that the attractant provided by this invention can be used for the control of the oriental fruit fly and the South Asian fruit fly.

[0047] The inconsistent number of *Bactrocera dorsalis* and *Fructus auricularia* attracted in Example 3 and the positive control in this experiment may be due to the inconsistent population sizes of *Bactrocera dorsalis* and *Fructus auricularia* at the experimental site, rather than to the different attraction abilities of the attractant of this invention to *Bactrocera dorsalis* and *Fructus auricularia*.

[0048] Therefore, the yeast-derived fruit fly food attractant provided by this invention has good field applicability and sustainability, and has a good effect on the control of target pests. Moreover, compared with imported baits of this type, it is inexpensive and efficient, and has good application prospects.

[0049] The above description provides examples of the preferred embodiments of the present invention. Parts not detailed herein are common knowledge to those skilled in the art. The scope of protection of the present invention is determined by the claims. Any equivalent modifications based on the technical teachings of the present invention are also within the scope of protection of the present invention.

Claims

1. A type of spherical Cyclospora yeast, characterized in that, Specifically, Cyclosporum spheroidum ( Torulaspora globosa F-HY017-Y1, isolated from a host damaged by fruit flies, was deposited at the China Center for Type Culture Collection on May 11, 2022, with the accession number CCTCC NO: M2022597.

2. The application of the enzymatically hydrolyzed protein obtained from the fermentation product of *Cyclospora spheroidosa* as described in claim 1 in the preparation of a highly efficient fruit fly attractant, characterized in that... The fruit flies are *Bactrocera dorsalis* and *Fructus aurantiacus*. The enzymatic hydrolysis method of the yeast fermentation product includes the following steps: centrifuging the fermentation broth and collecting the precipitate, dissolving the precipitate in pure water to obtain a homogenate, adjusting the pH to between 10 and 12, heating at a constant temperature of 40-55°C for 3 hours, then adjusting the pH to between 2 and 7, adding protease, and continuing to heat at a constant temperature of 40-55°C for 10 hours to obtain the yeast enzymatic hydrolysate solution. The protease includes any one or more of papain, acidic protease, and neutral protease. The pH of the homogenate is adjusted according to the usage conditions of different proteases.

3. The application according to claim 2, characterized in that, The specific application method is to use *Cyclospora spheroidosa* (… Torulaspora globosa The enzymatically hydrolyzed protein of F-HY017-Y1 is used as an effective active ingredient derived from yeast. The highly efficient fruit fly attractant includes the following components in the following mass fractions: 20-40% effective active ingredient derived from yeast, 4%-32% filler, 12%-40% disintegrant, 0%-4% lubricant and 24%-48% preservative.

4. The application according to claim 3, characterized in that, The disintegrant is any one or more of sodium carboxymethyl starch, crospovidone, citric acid, and sodium bicarbonate; the preservative is any one or more of sodium diacetate, potassium sorbate, calcium propionate, and methylparaben; the filler is any one or more of microcrystalline cellulose, glucose, dextrin, and soluble starch; and the lubricant is any one or more of magnesium stearate, calcium stearate, and talc.

5. The application according to claim 2, characterized in that, The enzymatic hydrolysis of the protein is achieved through the processing of *Cyclospora spheroidosa* (…). Torulaspora globosa F-HY017-Y1 was fermented to obtain fermentation products, which were then enzymatically hydrolyzed and spray-dried.

6. The application according to claim 5, characterized in that, The fermentation method for yeast is as follows: after inoculating 5-7% of activated yeast culture medium on the fermentation medium, culture it in an incubator for 24-48 hours.

7. The application according to claim 6, characterized in that, The fermentation medium comprises the following components by mass fraction: 5%-7% carbon source, 0.5%-1.5% nitrogen source, and 0.3%-1% yeast extract, and also includes 0.03mol / L-0.07mol / L KCl; the pH of the fermentation medium is 5-7, the culture temperature during the culture process is 25-28℃, and the volume of the fermentation medium is 30-100 mL.

8. The application according to claim 7, characterized in that, The carbon source includes any one or more of glucose, sucrose, corn starch saccharification products, potato starch saccharification products, and cassava starch saccharification products; the nitrogen source includes any one or more of polypeptone, fish meal, blood meal, and urea.

9. The application according to claim 5, characterized in that, The method for spray drying the enzymatically hydrolyzed product is as follows: the enzymatically hydrolyzed protein solution is dried by passing it through a spray drying tower, wherein the inlet temperature of the spray drying tower is 180-220℃, the outlet temperature is 90-110℃, and the injection rate is 6-20 ml / min.

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