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Yeast-like symbiote and use thereof

A symbiotic bacteria and yeast technology, applied in the application, biocide, microorganism and other directions, can solve the problems such as the lack of accurate identification and characteristic research, the difficulty of in vitro culture of YLS, and achieve the effect of improving the control effect and reducing the use.

Inactive Publication Date: 2015-05-20
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because it is difficult for people to simulate the microenvironment in insects, the ex vivo culture of YLS is very difficult
Kusumi et al. (1979) obtained several strains of SBPH YLS in vitro, but no accurate identification and characterization of them were found.

Method used

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  • Yeast-like symbiote and use thereof
  • Yeast-like symbiote and use thereof
  • Yeast-like symbiote and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: ( Pichia anomala in vitro isolation and identification)

[0026] BPH were collected from paddy fields around Hangzhou and raised in an artificial climate box (temperature 26±1°C, relative humidity 80±5%, light time 16L:8D), and barley seedlings were subcultured for more than 50 generations. Take 10 newly emerged female SBPH, disinfect the surface with 75% alcohol 3 times, 3 minutes each time, rinse 3 times with sterile water, 3 minutes each time, dissect the abdominal fat body on the ultra-clean workbench and collect . The collected fat bodies were added to YPD, PDA, YM, and Candia Elective Agar medium respectively, spread evenly, placed in a constant temperature incubator at 28°C for 48 hours, observed and recorded the results. On the culture medium with bacteria growing, pick a single colony with an inoculation loop, add it to the corresponding liquid medium, and cultivate it at 28°C and 180r / min on a shaking table for 24h. Repeat 3 times.

[0027] ...

Embodiment 2

[0034] Example 2: DGGE ( Pichia anomala verification)

[0035] Take 100 newly emerged female adults, disinfect the surface with 75% alcohol three times, each time for 3 minutes, and rinse with sterile water three times for each time for 3 minutes. Abdominal fat body was dissected, disinfected with 75% alcohol, rinsed with sterile water, and then centrifuged with 30% percoll at 7000 r / min for 10 minutes to remove impurities and fat in the upper layer, and the final concentration of 75% percoll was used for the lower layer. 10000 r / min, ultracentrifugation for 20min, the flocs in the 75% layer are yeast-like symbionts, and the genome is extracted with the yeast genome DNA kit.

[0036]The first round of PCR: using the above-mentioned extracted genome as a template, amplify the full-length 18Sr DNA-5.8Sr DNA-ITS sequence, the amplification primers are universal primers NS1 / ITS4, and the amplification system and conditions are the same as above. Nested PCR: The PCR product is d...

Embodiment 3

[0045] Example 3: Pichia anomala Sensitivity analysis of different pesticides

[0046] Pesticides for testing in this experiment: The pesticides for testing in this experiment were all purchased from Bayer Hangzhou Crop Science Co., Ltd., including: Nadiwan water-dispersible granules (the mass concentration of active ingredients is 75%, of which trifloxystrobin accounts for 25%, tebuconazole Accounting for 50%), Antaisheng wettable powder (the mass concentration of the active ingredient propazeb is 70%), imidacloprid water-dispersible granules (the mass concentration of the active ingredient is 70%), fluopyram + propamocarb hydrochloride suspension agent (active ingredient 687.5 g / L), Shijiale 40% pyrimethanil suspension concentrate. What table 3 lists is under different concentrations of fungicides, the in vivo separation of SBPH P. anomala Growth on solid media. The inhibitory effect of Antaisheng is the most obvious, when the concentration reaches 400mg / L P. anomala ...

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Abstract

The invention discloses a yeast-like symbiote and use thereof. The yeast-like symbiote is obtained through being separated from a fat body of laodelphax striatellus by an in-vitro culture technology, is named Pichia anomala SP2014 through microbial taxonomic identification, has the collection number of CCTCC No: M 2014534, and is collected at the CCTCC (China Center for Type Culture Collection) at Oct. 17, 2014. Shown by research and analysis on the sensitivity of the yeast-like symbiote to different microbicides, two kinds of microbicides, namely Antracol and Nativo, with obvious inhibitory effects to P. anomala SP2014 are sprayed to wheat seedlings at the concentration of 800mg / L, the death rate reaches 46.7% and 63.3% respectively after the wheat seedlings are eaten by laodelphax striatellus, the death rate is remarkably higher than that of control, and the weight is remarkably lower than that of a control group. Proven by fluorescent quantitative detection results, the amount of yeast-like symbiotes in laodelphax striatellus bodies is remarkably reduced after the microbicides are used. The yeast-like symbiote can serve as an indication marker for monitoring vitality of laodelphax striatellus or be used for screening or producing pesticides for preventing and treating laodelphax striatellus.

Description

technical field [0001] The invention relates to the technical field of biological control, and relates to the isolation of a yeast-like symbiotic bacterium and the exploration of a new method for preventing and controlling the brown planthopper. technical background [0002] SBPH Laodelphax striatellus Belonging to the family Delphacidae of Hemiptera, it is an important agricultural pest. In addition to piercing and sucking damage to rice, barley, wheat and other crops, it also spreads rice stripe leaf blight and rice black-streaked dwarf disease. Viral diseases such as maize dwarf disease and rough dwarf disease have caused huge crop yield losses (Ding Jinhua and Su Jianya, 2001). The current control of SBPH is mainly based on the use of insecticides. However, due to the long-term and irrational use of insecticides in large quantities, SBPH has developed varying degrees of resistance to many insecticides (Wang Yanhua et al., 2010). Dosage of pesticides, thus forming a vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12Q1/04C12Q1/18A01N51/00A01N47/14A01P7/04A01P3/00C12R1/84A01N37/50A01N43/653
CPCA01N37/50A01N43/653A01N47/14A01N51/00C12Q1/04C12Q1/18C12N1/165C12R2001/84A01N2300/00
Inventor 马正俞晓平曹伟边亚琳许益鹏郝培应
Owner CHINA JILIANG UNIV
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