Search

CN-122012303-A - W-5 strain for improving pesticide tolerance of natural enemy insects and preparation method thereof

CN122012303ACN 122012303 ACN122012303 ACN 122012303ACN-122012303-A

Abstract

The invention discloses a W-5 strain for improving the tolerance of natural enemy insect pesticides and a culture method thereof, wherein the W-5 strain can well grow in a minimum salt culture medium with lambda-efficient cyhalothrin as a unique carbon source, bacterial liquid OD 600 continuously rises after culture, and the degradation rate of the cyhalothrin pesticides reaches 98.24 percent. Further, after the yellow meal worm treated by the W-5 is fed to the lygus lucorum, the activities of two key detoxification enzymes, namely glutathione S-transferase and cytochrome P450 in the lygus lucorum can be remarkably improved. After continuous feeding of the strain for one week, W-5 successfully colonized the intestinal tract of the lygus lucorum and increased the survival rate of the lygus lucorum to 90.625% under lambda-cyhalothrin stress. Therefore, W-5 synergistically enhances the resistance of lygus lucorum to lambda-cyhalothrin through a dual mechanism of 'microbial direct degradation' and 'host detoxification metabolic activation'.

Inventors

  • XIAO YANSONG
  • YANG XUEYU
  • YU FAHUI
  • PENG JIANJIAN
  • PENG YUANYUAN
  • LI QI
  • XU XIAOXIA
  • LIU TING
  • CHEN NANA
  • LI HAO
  • XIE YIHENG
  • Huang Sunbin
  • HUANG CHENG
  • TAN WEN

Assignees

  • 湖南省烟草公司郴州市公司

Dates

Publication Date
20260512
Application Date
20251120

Claims (10)

  1. 1. A W-5 strain for improving pesticide tolerance of natural enemy insects is characterized by being classified and named Stapyhlococcus sp, wherein the preservation number is GDMCC No. 67176, the preservation date is 2025 and 10 months 29, the preservation unit is the microorganism strain preservation center in Guangdong province, and the preservation unit address is 100 in Xiuzhou martyr in Guangzhou province.
  2. 2. The W-5 strain for improving pesticide tolerance to natural enemy insects according to claim 1, wherein the 16S rDNA sequence of the W-5 strain is shown in SEQ ID No. 1.
  3. 3. Use of a W-5 strain according to any one of claims 1-2 for increasing the tolerance of a host to lambda-cyhalothrin.
  4. 4. The method for culturing a W-5 strain for improving pesticide tolerance of natural enemy insects according to any one of claims 1 to 2, comprising the steps of: (1) Sufficiently grinding intestinal tissues of natural enemy insects to obtain tissue homogenate, centrifuging, and sucking supernatant for later use; (2) Adding the supernatant obtained in the step (1) into a liquid culture medium, fully and uniformly mixing, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture to obtain bacterial liquid; (3) And (3) uniformly coating the bacterial liquid diluent obtained in the step (2) on a solid culture medium plate, culturing, and then picking single bacterial colony, and purifying and culturing to obtain the W-5 bacterial strain.
  5. 5. The method according to claim 4, wherein in the step (1), the natural enemy insect is a female adult stinkbug, and the centrifugation is a centrifugation at 3000 r/min at 5min.
  6. 6. The culture method according to claim 4, wherein in the step (2), the liquid medium is any one of LB liquid medium, EB liquid medium and NA liquid medium.
  7. 7. The method according to claim 4, wherein in the step (2), the culture broth is obtained by shaking culture of 12h in a shaking table at 37℃under 200 r/min; The dilution is to add sterile liquid culture medium or PBS into the obtained bacterial liquid for dilution, and the dilution treatment is repeated for a plurality of times, so as to sequentially obtain bacterial liquid with dilution degree of 10 -1 to 10 -9 .
  8. 8. The method according to claim 4, wherein in the step (3), 100. Mu.L of the bacterial liquid of 10 -5 to 10 -9 dilution is uniformly spread on the solid medium plate.
  9. 9. The culture method according to claim 4, wherein in the step (3), the solid medium plate is any one of LB solid medium plate, EB solid medium plate or NA solid medium plate.
  10. 10. The method according to claim 4, wherein in the step (3), the plate culture is carried out under conditions of 48 h in a 37℃incubator; the purification culture is that the purification is carried out once every 48h passage and the purification is carried out for 5 generations continuously.

Description

W-5 strain for improving pesticide tolerance of natural enemy insects and preparation method thereof Technical Field The invention belongs to the technical field of strains, and particularly relates to a W-5 strain for improving pesticide tolerance of natural enemy insects and a preparation method thereof. Background The lygus lucorum is an indispensable "biological guard" in the agricultural ecosystem. The novel pest control system provides sustainable and pollution-free pest control service for agricultural production through the strong predation capability. Studies have shown that single line bugs (especially the elderly nymphs and adults) can predate a large number of pests throughout the growth and development. One adult can predate tens or even hundreds of aphids a day. However, lygus lucorum is highly susceptible to a broad spectrum of chemical pesticides such as organic phosphorus and pyrethroids. Therefore, when medication is required, selective pesticides such as certain insect growth regulators, microbial pesticides, etc. must be preferred and avoid application during peak natural enemy activities. Disclosure of Invention In order to solve the defects in the prior art, the invention provides a W-5 strain for improving pesticide tolerance of natural enemy insects, and a construction method and application thereof. The staphylococcus saprophyticus W-5 can cooperatively enhance the resistance of the lygus lucorum to lambda-high-efficiency cyhalothrin through a double mechanism of direct degradation of microorganisms and detoxification and metabolic activation of hosts. In order to solve the technical problems, the invention provides the following technical scheme: A W-5 strain for improving the tolerance of natural enemy insect pesticides is classified and named Stapyhlococcus sp.W-5, the preservation number is GDMCC No. 67176, the preservation date is 2025, 10 months and 29 days, the preservation unit is the microorganism strain preservation center of Guangdong province, and the preservation unit address is 100 roads in Xiuzhou martyr of Guangzhou province. Preferably, the 16S rDNA sequence of the W-5 strain is shown as SEQ ID No. 1. The W-5 strain for improving the pesticide tolerance of natural enemy insects is applied to improving the tolerance of hosts to lambda-cyhalothrin. The culture method of the W-5 strain for improving the pesticide tolerance of the natural enemy insects comprises the following steps: (1) Sufficiently grinding intestinal tissues of natural enemy insects to obtain tissue homogenate, centrifuging, and sucking supernatant for later use; (2) Adding the supernatant obtained in the step (1) into a liquid culture medium, fully and uniformly mixing, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture to obtain bacterial liquid; (3) And (3) uniformly coating the bacterial liquid diluent obtained in the step (2) on a solid culture medium plate for plate culture, picking single bacterial colonies with different forms, and purifying and culturing by adopting a plate streaking method to obtain the W-5 bacterial strain. In the step (1), the natural enemy insect is a female imago of a lygus lucorum; the centrifugation treatment is centrifugation at 3000 r/min for 5 min. In the step (2), the liquid culture medium is any one of LB liquid culture medium, EB liquid culture medium or NA liquid culture medium. In the step (2), placing the strain in a constant-temperature shaking table at 37 ℃ for shaking culture at 200 r/min for 12 h to obtain the culture bacterial liquid; The dilution is specifically that a sterile liquid culture medium or PBS is added into the obtained bacterial liquid for dilution, the dilution treatment is repeated for a plurality of times, and bacterial liquid gradient dilution liquid of 10 -1 to 10 -9 is obtained in sequence. In the step (3), 100 mu L of bacterial liquid with the dilution of 10 -5 to 10 -9 is respectively taken and uniformly coated on a solid culture medium plate. In the step (3), the solid culture medium plate is any one of an LB solid culture medium plate, an EB solid culture medium plate or an NA solid culture medium plate. In the step (3), the plate culture condition is that the plate is placed in a 37 ℃ constant temperature incubator for culturing 48 h; the purification culture is that the purification is carried out once every 48h passage and the purification is carried out for 5 generations continuously. Compared with the prior art, the invention has the following beneficial effects: (1) The invention provides a W-5 strain for improving the pesticide tolerance of natural enemy insects, which can well grow in a minimum salt culture medium taking 50 mg/L lambda-efficient cyhalothrin as a unique carbon source, and after 72-h of the culture medium, bacterial liquid OD 600 continuously rises, and the degradation rate of the cyhalothrin pesticide reaches 98.24%. Further, after the yellow meal worm treated by th