CN110066739B - Beauveria bassiana SB010 and application thereof in prevention and treatment of common thistle marigold - Google Patents
Beauveria bassiana SB010 and application thereof in prevention and treatment of common thistle marigold Download PDFInfo
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Abstract
The invention discloses an application of a beauveria bassiana strain SB010 in prevention and treatment of thistle horses, wherein the strain SB010 is preserved in Guangdong province microorganism strain preservation center in 2018, 4 months and 19 days, and the preservation number is GDMCC NO: 60359. the strain SB010 can effectively inhibit the common thrips population, is safe to people, livestock, plants and environment, can partially replace thrips chemical prevention pesticides, and has great application potential in biological prevention and control.
Description
Technical Field
The invention relates to the field of plant protection, in particular to the field of biological control, and especially relates to beauveria bassiana and application thereof in control of common thistle horses.
Background
Chemical pesticides play a great role in pest control, and the usage amount and yield of pesticides in China are the first place in the world for a long time. However, according to statistics, the effective utilization rate of global pesticides is less than 30%, and the annual chemical pesticide use area of China is 2.8 hundred million hm2Above, the application amount reaches 50-60 ten thousand tons, about 80% of chemical pesticide directly enters the environment (Liliang, Zhang Ying, Sun Hui Ying, etc.. pesticide hazard and green plant protection technology discussion [ J]Hubei plant protection, 2018, (4): 63-64). With the long-term use of a large amount of chemical pesticides, the problems of pest Resistance (Resistance), pesticide Residue (Residue), pest Resurgence (Resurgence) and environmental pollution are more and more serious. Therefore, changing the traditional control mode mainly using chemical pesticides, implementing the plant protection policy of 'prevention mainly and comprehensive control', insisting on the basis of agricultural control and biological control, and combining other control measures to carry out comprehensive control on pests has extremely important significance in sustainable healthy development (Yangyuan Hua, Dukai book, Shimingwang. biological control research progress of entomogenous fungi [ J]The proceedings of the Henan academy of science and technology, 2011,39(1): 34-37).
Thrips is a generic term for insects of the thrips family (Thripida) of the order Thysanoptera (thysannoptera). The thrips pests have tiny body types, are easy to hide, have strong fertility, short growth cycle and serious generation overlapping, and increase the difficulty for preventing and treating. Thrips are small in size or hidden in activity, are not easily perceived by people at the early stage of damage, and are often discovered after serious disasters occur. At present, the pest control mainly depends on chemical control, and due to the lack of ideal and efficient chemical agents for a long time, a large number of chemical agents are unreasonably used, so that various problems of pest drug resistance, natural enemy killing, pesticide residue, environmental pollution and the like are caused, and the quality and safety of products are threatened. In recent years, the spread and spread of thrips has been accelerated by global trade and climate warming, and thrips has now become an important agricultural and forestry pest worldwide. Therefore, the search for environment-friendly control strategies is extremely urgent today, and biological control has wide application prospects in the control work of thrips pests. In addition, biological control can ensure the quality safety of products in the production process and can effectively reduce the using amount of chemical agents, so far, researches on biological control measures for controlling thrips pests have been reported.
Beauveria bassiana belongs to Deutero mycotina, Moniliales, Moniliaceae and Beavinia, and is an important biocontrol strain. Beauveria bassiana is found on Melanopius mexicanus for the first time, and is applied to Western fimbristipula fimbriata as a biocontrol bacterium for the first time in the early 70 s, and the application succeeds, and attracts wide attention. For example, Huangpeng et al disclose that a Beauveria bassiana BB-T02 with strong infectivity on thrips ficuum has good infection lethal effect on thrips ficuum (CN 107384803A); liyingping et al (2013) separate Beauveria bassiana SZ-26 strain (China biological control report) with high efficiency on Frankliniella occidentalis from Asian corn borer cadavers in northeast of China; yuanhongyong et al (2011 and 2013) isolated from the larvae of the silvery looper to obtain beauveria bassiana MZ050724 strain with high virulence against Frankliniella occidentalis, and beauveria bassiana Bb 080717 strain and MZ050724 strain (university of southwest, environmental insect school newspaper, Chinese forest pest) with high virulence against Frankliniella occidentalis; two strains of beauveria bassiana (CN 104212724A and CN 104263655A) with good control effect on frankliniella occidentalis are separated from corn borers by Wanghaihong and the like; beauveria bassiana (CN 106135295A) which has good control effect on alfalfa thrips is found by daubing the soldier and the like (2016), and the like. Among them, the biocontrol effect of beauveria bassiana on thrips was most studied.
Disclosure of Invention
The invention aims to provide biocontrol bacteria with excellent biocontrol effect on thrips, particularly common thrips. In the research process, the beauveria bassiana strain SB010 with a certain effect on prodenia litura has a prominent poisoning effect on common thrips. The beauveria bassiana strain SB010 was identified in patent application 201810596222.2 (a pesticide combining matrine and beauveria bassiana and its application) proposed by the present applicant and was deposited in the guangdong province collection of microorganisms at 19 months 4 and 2018 with the deposit number GDMCC NO: 60359.
the invention accordingly provides the use of said strains for the control of thistle. Preferably, the thrips is common thrips.
In one embodiment, the strain is cultured to produce a spore solution, a spore-containing microbial inoculum. The spore concentration of the strain at the time of application was 107spore/mL to 109spore/mL, preferably 108spore/mL to 109spores/mL.
In addition, the bacterial strain can be combined with other medicaments to prevent and treat thrips, especially common thrips, and a better synergistic effect can be obtained.
The inventor finds that the beauveria bassiana strain SB010 with certain effect on prodenia litura has particularly outstanding effect on common thrips in screening (candidate strains also comprise a plurality of beauveria bassiana). Biological experiments show that the beauveria bassiana strain has 90 percent (10 percent) of control effect on the common thrips8spore/mL) whereas the typical effect of beauveria bassiana under the same conditions is only 36%; under the condition of strain fermentation liquor at the concentration of 0.4mg/ml, the mortality of SB010 to thrips at different treatment times is obviously higher than that of other strains, and the corrected mortality of 5 days reaches 93.3 percent and is far higher than that of other strains (see the experimental results and tables of specific examples). Therefore, the beauveria bassiana strain SB010 can effectively inhibit the common thrips population, is safe to people, livestock, plants and environment, can partially replace thrips chemical pesticide, and has biological control effectGreat application potential.
Detailed Description
The following description is given by way of development and specific embodiments of the present invention, and is not intended to limit the invention thereto.
Example (b): bioassay of strains
1 test insects
The common thrips population is collected from Chongcun cowpea field in Guangzhou, and is raised in laboratory for several generations under the conditions of 26 deg.C, 65% relative humidity and 12: 12 photoperiod L: D.
2 test strains
Some strains for selection are listed:
SB 009: beauveria bassiana (Beauveria bassiana) strain, which is preserved in Guangdong provincial microorganism culture Collection (GDMCC) and has the accession number of GDMCC NO:60588 (additionally proposed patent application 2019102239096).
SB 010: the strain Beauveria bracteata (Beauveria brongniartii) is preserved in Guangdong province microorganism culture collection center, and the preservation number is GDMCC NO: 60359 (patent application 201810596222.2 has additionally been filed, in which deposited information is provided).
SP 016: beauveria bassiana (Beauveria bassiana) which is preserved in the engineering research center of department of biological control education of southern China university of agriculture.
SP 535: isaria fumosorosea (Isaria fumosorosea) deposited at the center of engineering research of department of biological control and education at southern China university of agriculture.
SP 502: isaria fumosorosea (Isaria fumosorosea) deposited at the center of engineering research of department of biological control and education at southern China university of agriculture.
MZ 16: metarhizium anisopliae (Metarhizium anisopliae) is deposited in the center of engineering research of department of biological control and education of southern China university of agriculture.
3 Experimental methods
(1) Preparation of spore suspension
Culturing at 26 + -1 deg.C on PDA plate for 7 days, eluting conidia of entomogenous fungi with 0.05% Tween-80 sterile water, stirring with magnetic stirrer, and shaking at 180rpm and 25 deg.C in shaking tableCulturing for 25min, filtering with double-layer mirror paper, counting with blood counting plate, measuring the concentration of mother liquor, and making into 1 × 108spores/mL of spore suspension.
(2) Preparation of strain fermentation liquor
Each strain was prepared at 1X 10 in the above manner7spore/mL spore suspension, 5mL prepared spore suspension was inoculated into an Erlenmeyer flask containing 50mL SDA medium, incubated at 26 ℃ for 5 days with shaking at 180rpm, and the filtrate was collected by suction filtration on a Buchner funnel, assayed for protein concentration and diluted to 0.4 mg/mL.
(3) Determination of toxicity of entomogenous fungi to common thrips
Placing the prepared spore suspension and fermentation liquor in a flat-bottom finger-shaped tube (15mm multiplied by 75mm), soaking for 2h, pouring out the spore suspension, and naturally drying the finger-shaped tube for later use; cutting cowpea into segments (1cm, without holes at both ends), soaking in spore suspension, taking out for 30s, air drying, placing into treated finger-type tube, simultaneously inoculating 50 heads of female common thrips, sealing cotton, placing in artificial climate box, and repeating each treatment for 4 times with 0.05% Tween-80 sterile water as blank control. Mortality was recorded for 5 consecutive days.
(3) Data processing
SPSS 19.0 software is used for experimental treatment analysis, single-factor analysis of variance is used for analyzing each result, and Tukey is used for detecting the significance of difference.
4 results of the experiment
The experimental results show that the pathogenicity difference of spore suspensions of different strains to common thrips is obvious, and the strain SB010 is obviously higher than that of other strains (Table 1 and Table 2); the pathogenicity of the same strain at different concentrations and different treatment times is different, the death rate of the female common thrips increases along with the increase of the concentration and the treatment time, when the concentration is 1 multiplied by 108Cumulative mortality of SB010 at 5d was 90% at spore/mL (Table 1), significantly higher than the concentration 1X 107spore/mL.
The result shows that the strain SB010 has better lethal effect on common thrips, and belongs to potential excellent biocontrol bacteria.
TABLE 1 corrected mortality of different strains on common thrips (%)
Note that different lower case letters in the same column indicate significant pathogenicity difference among different strains through Tukey test (P < 0.05)
TABLE 2 corrected mortality of different strains on common thrips (%)
Note that different lower case letters in the same column indicate significant pathogenicity difference among different strains through Tukey test (P < 0.05)
Further, the results of experiments to determine the virulence of strain broth towards thrips showed that SB010 was significantly more lethal to thrips at different treatment times than the other strains (see table 3), with a corrected mortality rate of 93.3% at 0.4mg/ml at day 5, which is much higher than the other strains.
TABLE 3 corrected mortality of different strains of fermentation broth on common thrips (%)
Note that different lower case letters in the same column indicate significant pathogenicity difference between different strains (P < 0.05) by Tukey test.
Claims (5)
1. Beauveria bassiana (B.brucei)Beauveria bassiana) The application of the strain SB010 in prevention and treatment of common thistle horses is disclosed, wherein the strain SB010 is preserved in 19.4.2018 in Guangdong province microorganism strain preservation center with the preservation number GDMCC NO: 60359.
2. the use according to claim 1, wherein the strain SB010 is cultured to produce a spore liquid or a spore-containing microbial inoculum.
3. The use according to claim 2, wherein strain SB010 has a spore concentration of 10 at the time of administration7 spore/mL to 109 spores/mL.
4. The use of claim 3, wherein the spore concentration is 108 spore/mL to 109 spores/mL.
5. Use as claimed in any one of claims 1 to 4, wherein the strain is used in combination with other agents for the control of thrips.
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| CN111909854B (en) * | 2020-07-20 | 2023-02-03 | 华南农业大学 | Zero-valent iron-Beauveria bassiana nano-particles and preparation method and application thereof |
| CN112715577B (en) * | 2020-10-22 | 2021-12-28 | 华南农业大学 | Application of matrine and beauveria bassiana in preventing and treating common circium japonicum |
| CN114317291B (en) * | 2022-01-30 | 2023-05-12 | 华南农业大学 | Space breeding beauveria bassiana strain SCAUHT21 with high pathogenicity to common thrips and application thereof |
| CN114317292B (en) * | 2022-01-30 | 2023-05-12 | 华南农业大学 | Space breeding beauveria bassiana strain SCAUHT12 with high pathogenicity to common thrips and application thereof |
| CN114317294B (en) * | 2022-01-30 | 2024-03-26 | 华南农业大学 | High pathogenicity and high ultraviolet resistance space insect fungus strain SCAUHT56 for common thrips and application thereof |
| CN114317293B (en) * | 2022-01-30 | 2024-03-26 | 华南农业大学 | High pathogenicity and high ultraviolet resistance space insect fungus strain SCAUHT18 for common thrips and application thereof |
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Effective date of registration: 20211126 Address after: 512029 No. 5, Longling Road, Gantang Industrial Park, Wujiang District, Shaoguan City, Guangdong Province Patentee after: GUANGDONG GANJIALE BIOTECHNOLOGY CO.,LTD. Address before: No. 483, Wushan Town, Tianhe District, Guangzhou, Guangdong 510642 Patentee before: SOUTH CHINA AGRICULTURAL University |
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