CN110066739A - A kind of muscardine SB010 and its application in the common setose thistle Malaysia and China of prevention and treatment - Google Patents

Abstract

The present invention discloses the application of a Beauveria bassiana strain SB010 in the prevention and control of thrips, wherein the strain SB010 was deposited in the Guangdong Provincial Microbial Culture Collection Center on April 19, 2018, and its deposit number is GDMCC NO: 60359 . The strain SB010 can effectively inhibit the common thrips population, is safe to humans, animals, plants and the environment, can partially replace thrips chemical control pesticides, and has great application potential in biological control.

Description

A kind of Beauveria bassiana SB010 and its application in the control of common thrips

technical field

The invention relates to the field of plant protection, in particular to the field of biological control, in particular to Beauveria bassiana and its application in preventing and treating common thrips.

Background technique

Chemical pesticides play a huge role in pest control, and my country's pesticide usage and output have long ranked first in the world. However, according to statistics, the effective utilization rate of pesticides in the world is less than 30%. The annual area of chemical pesticides in China is more than 280 ^million hm2, and the application amount reaches 500,000-600,000 tons. About 80% of the chemical pesticides directly enter the environment (Li Liang, Zhang Zhanying). , Sun Huiying, et al. Discussion on pesticide hazards and green plant protection technology [J]. Hubei Plant Protection, 2018, (4): 63-64). With the long-term and large-scale use of chemical pesticides, the "3R" problems brought about by pests resistance (Resistance), pesticide residues (Residue) and pest resurgence (Resurgence) and environmental pollution problems are becoming more and more serious. Therefore, it is necessary to change the traditional control method based on the use of chemical pesticides, implement the plant protection policy of "prevention first, comprehensive control", adhere to agricultural control and biological control as the basis, and combine other control measures to comprehensively control pests. It is of great significance to continue the development of health care (Yang Yunhua, Du Kaishu, Shi Mingwang. Research progress in biological control of insect-borne fungi [J]. Journal of Henan University of Science and Technology, 2011,39(1):34-37).

Thrips is a collective name for insects in the family Thripida of the order Thysanoptera. The thrips pests are small in size, easy to hide, and have strong fecundity, short growth cycle, and serious overlapping of generations, which increases the difficulty of control. The thrips have small bodies or hidden activities, and are not easily noticed by people in the early stage of damage, and are often discovered after serious disasters are caused. At present, the control of such pests mainly relies on chemical control. Due to the lack of ideal and efficient chemical agents, a large number of chemical agents are used unreasonably, which leads to many problems such as pest resistance, killing natural enemies, pesticide residues, and environmental pollution. Product quality and safety are also threatened. In recent years, global trade and climate warming have accelerated the spread and dissemination of thrips, which have become an important agricultural and forestry pest worldwide. Therefore, it is extremely urgent to seek environmentally friendly control strategies today, and biological control has broad application prospects in the control of thrips pests. In addition, biological control can ensure the quality and safety of products in the production process, and can effectively reduce the use of chemical agents. So far, there have been many reports on biological control measures to control thrips pests.

Beauveria bassiana belongs to Deutero mycotina, Moniliales, moniliaceae, Beaveriaspp, and is an important biocontrol bacteria species. Beauveria bassiana was first discovered in the Mexican black locust (Melanopius mexicanus), and it was first used as a biocontrol bacteria in the western beetle in the early 1970s, and it was successful and attracted widespread attention. For example, Huang Peng et al. disclose a strain of Beauveria bassiana BB-T02 with strong infectivity to thrips in hot areas, which has a good lethal effect on thrips in hot areas (CN107384803A); Li Yin Ping Ping (2013) isolated Beauveria bassiana SZ-26 strain (Chinese Journal of Biological Control), which is highly effective against thrips occidentalis, isolated from Asian corn borer carcasses in Northeast China; Yuan Shengyong et al. (2011 and 2013) isolated Spodoptera Beauveria bassiana strain MZ050724, which is highly virulent to Thrips occidentalis, and Beauveria bassiana strains Bb 080717 and MZ050724, which are highly virulent to Thrips ficus, were isolated from larvae (Journal of Southwest University, Environmental Insects). Journal of Chinese Forest Diseases and Insects); Wang Haihong et al. isolated two strains of Beauveria bassiana (CN 104212724 A and CN 104263655 A) from corn borer with good control effect on Western flower thrips (CN 104212724 A and CN 104263655 A); Tu Xiongbing et al. (2016) found that A strain of Beauveria bassiana with good control effect on alfalfa thrips (CN 106135295 A), etc. It can be seen that the most studied is the biocontrol effect of Beauveria bassiana on thrips.

SUMMARY OF THE INVENTION

The object of the present invention is to provide biocontrol bacteria with excellent biocontrol effect on thrips, especially common thrips. During the research process, it was found that the Beauveria bassiana strain SB010, which had a certain effect on Spodoptera litura before, had a prominent poisoning effect on common thrips. Beauveria bassiana strain SB010 was identified in the applicant's patent application 201810596222.2 (a combination of matrine and Beauveria bassiana and its application), and published on April 19, 2018 It is preserved in the Guangdong Provincial Microbial Culture Collection Center, and its preservation number is GDMCC NO: 60359.

Accordingly, the present invention provides the application of the strain in controlling thrips. Preferably, the thrips are common thrips.

In one embodiment, the strain is cultured to prepare a spore liquid and a spore-containing inoculum. The spore concentration of the strain at the time of application is 10 ⁷ spores/mL to 10 ⁹ spores/mL, preferably 10 ⁸ spores/mL to 10 ⁹ spores/mL.

In addition, the strain can also be combined with other agents to control thrips, especially common thrips, and may obtain a better synergistic effect.

During screening (the candidate strains also include some Beauveria bassiana), the inventors found that the Beauveria bassiana strain SB010, which had a certain effect on Spodoptera litura before, had a particularly prominent effect on common thrips. The biological experiments show that the Beauveria bassiana strain of the present invention has a control effect of 90% on common thrips (experiment with 10 ⁸ spores/mL), while under the same conditions, the usual Beauveria bassiana effect is only 90%. 36%; under the condition of strain fermentation broth at a concentration of 0.4 mg/ml, the lethality of SB010 to thrips at different treatment times was significantly higher than that of other strains, and the corrected mortality rate on the 5th day was 93.3%, much higher than that of other strains. (See experimental results and tables in specific examples). Therefore, the Beauveria bassiana strain SB010 of the present invention can effectively inhibit the common thrips population, and is safe to humans, animals, plants and the environment, can partially replace thrips chemical control pesticides, and has great application in biological control potential.

Detailed ways

The following is an introduction through the research and development process and specific embodiments of the present invention, but does not constitute a limitation of the present invention.

Example: Bioassay of strains

1 Test insects

Common Thrips populations were collected from cowpea fields in Zhucun, Guangzhou, and were raised in the laboratory for many generations under the conditions of temperature 26°C, relative humidity 65%, and photoperiod L:D=12:12.

2 tested strains

List some strains for screening:

SB009: Beauveria bassiana bacteria, deposited in the Guangdong Provincial Microorganism Culture Collection Center (GDMCC), the deposit number is GDMCC NO: 60588 (another patent application 2019102239096 has been filed).

SB010: strain Beauveria brongniartii, deposited in the Guangdong Provincial Microbial Culture Collection Center, its deposit number is GDMCC NO: 60359 (another patent application 201810596222.2 has been filed, wherein the deposit information is provided).

SP016: Beauveria bassiana, deposited in the Engineering Research Center of the Ministry of Education for Biological Control, South China Agricultural University.

SP535: Isaria fumosorosea, preserved in the Engineering Research Center of the Ministry of Education for Biological Control, South China Agricultural University.

SP502: Isaria fumosorosea, preserved in the Engineering Research Center of the Ministry of Education for Biological Control, South China Agricultural University.

MZ16: Metarhizium anisopliae, preserved in the Engineering Research Center of the Ministry of Education for Biological Control, South China Agricultural University.

3 Experimental methods

(1) Preparation of spore suspension

After culturing on a PDA plate at 26±1°C for 7 days, the fully sporulated entomogenous fungal conidia were eluted with 0.05% Tween-80 sterile water, stirred with a magnetic stirrer, and then placed on a shaker at 180rpm at 25°C. After shaking culture for 25 min, filter with double-layer lens paper, count with hemocytometer, measure the concentration of mother solution, and prepare spore suspension of 1×10 ⁸ spores/mL.

(2) Preparation of strain fermentation broth

The spore suspension of 1×10 ⁷ spores/mL of each strain was prepared by the above method, and 5 mL of the prepared spore suspension was inoculated into a conical flask containing 50 mL of SDA medium. The filtrate was collected by suction filtration, and its protein concentration was determined and diluted to 0.4 mg/ml.

(3) Toxicity determination of entomogenous fungi to common Thrips

Place the prepared spore suspension and fermentation broth in a flat-bottomed finger-shaped tube (15mm × 75mm), pour out the spore suspension after soaking for 2 hours, and then use the finger-shaped tube to air dry for use; ), immersed in the spore suspension, taken out for 30s, put into the treated finger-shaped tube after natural air-drying, and at the same time inserted 50 female adults of common thrips, sealed with cotton, placed in an artificial climate box, and spit at 0.05% Warm-80 sterile water was used as blank control, and each treatment was repeated 4 times. The mortality was recorded for 5 consecutive days.

(3) Data processing

SPSS 19.0 software was used for experimental treatment analysis, and one-way ANOVA was used to analyze the results, and Tukey's test was used to test the significance of differences.

4 Experimental results

The experimental results showed that the virulence of spore suspensions of different strains to common thrips was significantly different, and the strain SB010 was significantly higher than other strains (Table 1, Table 2); the same strain, different concentrations, different treatment time virulence differences exist , the mortality of female adults of common Thrips increased with the increase of concentration and treatment time. When the concentration was 1×10 ⁸ spores/mL (Table 1), the cumulative mortality of SB010 on the 5th day was 90%, a significant When the concentration is higher than 1×10 ⁷ spores/mL.

The results showed that the strain SB010 had a good lethal effect on common Thrips, and it belonged to a potential good biocontrol bacteria.

Table 1 Corrected mortality (%) of different strains to common Thrips

Note: After Tukey test, different lowercase letters in the same column indicate significant differences in pathogenicity among different strains (P<0.05)

Table 2 Corrected mortality (%) of different strains to common Thrips

Note: After Tukey test, different lowercase letters in the same column indicate significant differences in pathogenicity among different strains (P<0.05)

Further, the experimental results of measuring the virulence of strain fermentation broth to thrips showed that the lethality of SB010 to thrips at different treatment times was significantly higher than that of other strains (see Table 3), wherein at the concentration of 0.4 mg/ml, the The 5-day corrected mortality rate was 93.3%, much higher than other strains.

Table 3 Corrected mortality (%) of different strains of fermentation broth to common Thrips

Note: After Tukey's test, different lowercase letters in the same column indicate significant differences in virulence among different strains (P<0.05).

Claims (6)

1. The application of Beauveria bassiana strain SB010 in the prevention and control of thrips, wherein the strain SB010 was deposited in the Guangdong Provincial Microorganism Culture Collection Center on April 19, 2018, and its deposit number is GDMCC NO: 60359. 2. application as claimed in claim 1, is characterized in that, described thrips is common thrips. 3. The application according to claim 1, wherein the bacterial strain SB010 is cultivated to prepare spore liquid and a spore-containing inoculum. 4. The application of claim 3, wherein the spore concentration of strain SB010 is 10 ⁷ spores/mL to 10 ⁹ spores/mL during application. 5. The application of claim 4, wherein the spore concentration is ¹⁰ spores/mL to 10 ⁹ spores/mL. 6. The application according to any one of claims 1 to 5, wherein the bacterial strain is used in combination with other medicaments for controlling thrips.