CN111909854B - Zero-valent iron-Beauveria bassiana nano-particles and preparation method and application thereof - Google Patents
Zero-valent iron-Beauveria bassiana nano-particles and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a zero-valent iron-Beauveria bassiana nanoparticle and a preparation method and application thereof. The invention provides a preparation method of zero-valent iron-Beauveria bassiana nanoparticles, which comprises the following steps: s1, inoculating a beauveria bassiana spore suspension into a fermentation culture solution, and performing shake culture for 48-72 hours to obtain mycelia; s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture for 48-72 hours to obtain a conidium filtrate; s3, adding zero-valent iron into the conidium filtrate obtained in the step S2, and incubating for 24-96 h to obtain the zero-valent iron-Beauveria bassiana nano particles. The nano particles have high purity, strong activity and simple preparation method, have obvious control effect on noctuidae pests such as prodenia litura and the like, effectively solve the problems of low insecticidal speed, long action cycle and the like of beauveria bassiana, are favorable for delaying the occurrence and development of drug resistance of the noctuidae pests, and have very strong application potential in biological control of the noctuidae pests.
Description
Technical Field
The invention belongs to the technical field of biological control. More particularly, relates to a zero-valent iron-Beauveria bassiana nanoparticle, a preparation method and an application thereof.
Background
Spodoptera litura (Fabricius) is a typical noctuidae pest, and is an important agricultural pest distributed worldwide, and Spodoptera litura larvae are used as large crops for vegetables and fields with as many as 290 families as 99 families, such as cabbage, melons, tobacco, peanut, cabbage, cotton, leek, soybean, sweet potato and the like. Because prodenia litura has the characteristics of overlapping generations, multiple generations within one year, wide food intake and intermittent outbreak, the prodenia litura can cause great loss in a short period as long as the condition of proper climate, sufficient food and proper environment is adopted. The most common method for preventing and controlling prodenia litura is to use chemical pesticide, but with the unreasonable use of chemical pesticide, the environmental pollution caused by the chemical pesticide is serious, and a serious 3R problem is generated. Therefore, there is an urgent need for green, environmentally friendly biopesticides to control prodenia litura.
Beauveria bassiana (Beauveria bassiana) is a broad-spectrum entomopathogenic fungus, and researches show that the Beauveria bassiana can infect more than 700 insects of 15 meshes and 149 families. Meanwhile, the beauveria bassiana is friendly to the environment and warm-blooded animals, easy to culture, cheap and easily available in raw materials, wide in insecticidal spectrum and strong in pathogenicity; therefore, beauveria bassiana is currently the most widely used entomopathogenic fungus in China. The prior patent with publication number CN110066739A discloses a Beauveria bassiana SB010 which can effectively prevent and treat common thrips. However, the method for controlling agricultural pests by using entomogenous fungi alone has the defects of slow effect, easy environmental influence and the like.
Nanotechnology is a new field, and research and application of nanotechnology is mainly in the fields of materials and preparation, microelectronics and computer technology, medicine and health, aerospace and aviation, environment and energy, biotechnology and agricultural products, and the like. The nano-particles have high-strength target characteristics, and from the agricultural perspective, the application of nano-technology, particularly the use of nano-materials, has great potential application value in the aspect of promoting the development of pest management methods. The nano zero-valent iron (nZVI) is zero-valent iron particles with the particle size of 1-100 nm, has extremely small particle size and large specific surface area generated by the particle size, further shows high oxidation-reduction potential, and can reduce various halogenated organic pollutants, high-valent heavy metal ions, inorganic pollutants (such as nitric acid/nitrite) and the like. At present, the nano zero-valent iron is usually used for environmental heavy metal remediation, pollutant degradation and the like, and no report of using the nano zero-valent iron for biological control is found yet.
Disclosure of Invention
The invention aims to overcome the defects of the existing method for preventing and controlling noctuidae pests and provides a zero-valent iron-Beauveria bassiana nano particle and a preparation method and application thereof.
The invention aims to provide a preparation method of zero-valent iron-Beauveria brucei nanoparticles.
The invention also aims to provide the zero-valent iron-Beauveria bassiana nano-particles prepared by the method.
The invention also provides application of the zero-valent iron-beauveria brucei nano particles in controlling noctuidae pests or preparing noctuidae pest control preparations.
Still another object of the present invention is to provide a noctuidae pest control agent.
The above purpose of the invention is realized by the following technical scheme:
the invention firstly provides a preparation method of zero-valent iron-Beauveria bassiana nanoparticles, which comprises the following steps:
s1, inoculating a Beauveria bassiana spore suspension into a fermentation culture solution, and performing shake culture for 48-72 hours to obtain a mycelium;
s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture for 48-72 hours to obtain a conidium filtrate;
s3, adding zero-valent iron into the conidium filtrate obtained in the step S2, and incubating for 24-96 h to obtain the zero-valent iron-Beauveria bassiana nano particles.
Preferably, in step S1, the beauveria bassiana strain SB010 is deposited at the Guangdong province collection center for microbial cultures at 7/5/2018 with a deposit number of GDMCC No:60359, and the preservation address is No. 59 building No. 5 building of No. 100 Dastaring Medium furnature, guangzhou city.
Preferably, it isThe concentration of the beauveria brookfield spore suspension in the step S1 is 1 multiplied by 10 5 ～1×10 9 conidia/mL.
More preferably, the concentration of the beauveria brookfield spore suspension in step S1 is 1 × 10 7 ～1×10 9 conidia/mL.
Still more preferably, the concentration of the beauveria brookfield spore suspension in the step S1 is 1 × 10 8 conidia/mL.
Preferably, the zero-valent iron in step S3 is nano zero-valent iron powder.
Preferably, the particle size of the nano zero-valent iron powder is 60-80 nm.
More preferably, the particle size of the nano zero-valent iron powder is 70nm.
Preferably, the mass-to-volume ratio of the zero-valent iron to the conidium filtrate in the step S3 is 1g: 500-1000 mL.
More preferably, the mass-to-volume ratio of the zero-valent iron to the conidium filtrate in step S3 is 1g:500mL.
Preferably, the rotation speed of the shaking culture in the step S1 and the step S2 is 120-180 rpm/min.
More preferably, the rotation speed of the shaking culture in the step S1 and the step S2 is 180rpm/min.
Preferably, the temperature of the shaking culture in step S1 and step S2 is 24-26 ℃.
More preferably, the temperature of the shaking culture in step S1 and step S2 is 25 ℃.
Preferably, the shaking culture time of the step S1 and the step S2 is 60h.
Preferably, the incubation time in step S3 is 24-72 h.
More preferably, the incubation time of step S3 is 72h.
Preferably, the fermentation culture solution in step S1 is Sabouraud agar medium (SDA).
Preferably, the preparation method of the beauveria brookfield spore suspension in step S1 comprises the following steps: selecting mature spores of Beauveria bassiana (balsamo) Blume on a PDA (personal digital assistant) plate for activated culture at 24-26 ℃ for 7-10 days, adding distilled water to wash off the spores, and shaking forcefully for 3-5 min to obtain a Beauveria bassiana spore suspension.
According to the invention, a large number of researches and exploration experiments show that the zero-valent iron-beauveria bassiana nanoparticles prepared by the method have high pathogenicity on noctuidae pests, and the zero-valent iron and the beauveria bassiana can generate obvious synergistic interaction effect and have obvious control effect on the noctuidae pests; therefore, the zero-valent iron-Beauveria brucellosis nanoparticles prepared by the method and the application of the zero-valent iron-Beauveria brucellosis nanoparticles in controlling noctuidae pests or preparing noctuidae pest controlling preparations are also within the protection scope of the invention.
Preferably, the noctuidae pest is prodenia litura.
In addition, the invention also provides a noctuidae pest control preparation which comprises the zero-valent iron-beauveria bassiana nano-particles. The prevention and treatment preparation takes zero-valent iron-beauveria brookfield nanometer particles as main active ingredients, and auxiliary agents can be added according to needs to prepare the prevention and treatment preparation.
In a preferred embodiment of the present invention, the particle size of the zero-valent iron-Beauveria brucei nanoparticles is 270-320 nm.
The invention has the following beneficial effects:
the zero-valent iron-beauveria bassiana nano particles with high purity, strong activity, no pollution and no residue are successfully prepared, the zero-valent iron and the beauveria bassiana in the nano particles show obvious synergistic interaction, and the zero-valent iron-beauveria bassiana nano particles have obvious control effect on noctuidae pests such as prodenia litura and the like through long-term infection biological research and indoor bioassay.
Compared with the traditional fungal spore powder, the nano particle biological pesticide has the advantages of being not easily affected by the environment, fast in effect, high in efficiency, longer in survival time of beauveria brookfield spores in the environment, more stable and more environment-friendly, obviously improves the control effect and stability of beauveria brookfield spores on noctuidae pests, effectively solves the problems of slow pest killing speed, long action period and the like of beauveria brookfield spores, and the preparation method of the nano particle is simple, convenient and fast; therefore, the nano-particle has very strong application potential in biological control of noctuidae pests.
The nano particles prepared by the method have good control effect on noctuidae pests, can reduce the using amount of strains, has the characteristics of low toxicity and low drug resistance, meets the requirement of organic food production, has no pollution and residue on the environment, and is favorable for delaying the occurrence and development of drug resistance of the noctuidae pests.
Drawings
FIG. 1 is a UV-Vis spectrum of zero-valent iron-Beauveria brucei nanoparticles at different absorbance values.
FIG. 2 is an SEM image of a zero-valent iron-Beauveria brucei nanoparticle.
FIG. 3 is a graph showing the result of EDX spectroscopy of zero-valent iron-Beauveria bassiana nanoparticles.
FIG. 4 is an XRD pattern of a zero-valent iron-Beauveria brucei nanoparticle.
FIG. 5 is an FTIR spectrum of a zero valent iron-Beauveria brucei nanoparticle.
FIG. 6 is a graph showing the effect of zerovalent iron-Beauveria bassiana nanoparticles on the mortality rate of prodenia litura.
FIG. 7 is a graph of the effect of different concentrations of zerovalent iron-Beauveria bassiana nanoparticles on the relative growth rate of prodenia litura; wherein, different lower case letters indicate that the difference of the relative growth rate of the prodenia litura among the zero-valent iron-Beauveria bassiana nano-particles with different concentrations at the same time is obvious (P < 0.05).
FIG. 8 is a graph of the effect of varying concentrations of zerovalent iron-Beauveria brucei nanoparticles on the relative feeding rate of Spodoptera litura; wherein, different lower case letters indicate that the differences of the relative feeding rates of the prodenia litura among the zero-valent iron-beauveria brookfield nano-particles with different concentrations at the same time are obvious (P < 0.05).
FIG. 9 is a graph showing the effect of different concentrations of zerovalent iron-Beauveria brucei nanoparticles on the conversion rate of prodenia litura hairs; wherein, different lower case letters indicate that the conversion rate difference of prodenia litura hair between the zero-valent iron-beauveria bassiana nano-particles with different concentrations at the same time is obvious (P < 0.05).
FIG. 10 is a graph of the effect of different concentrations of zerovalent iron-Beauveria bassiana nanoparticles on pupation rate of Spodoptera litura; wherein, different lower case letters indicate that the pupation rate difference of prodenia litura among different concentrations of the zero-valent iron-beauveria brookfield nano-particles at the same time is obvious (P < 0.05).
FIG. 11 is a graph showing the effect of different concentrations of zerovalent iron-Beauveria bassiana nanoparticles on the emergence rate of prodenia litura; wherein, different lower case letters indicate that the difference of the emergence rates of the prodenia litura among the zero-valent iron-beauveria bassiana nano-particles with different concentrations at the same time is obvious (P < 0.05).
Detailed Description
The present invention will be further described with reference to the following specific examples, which are not intended to limit the invention in any manner. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
The beauveria bassiana strain SB010 used in the following examples was stored in the center of research on biological control engineering of the education department of agricultural university in south China. The strain is preserved in Guangdong province microbial strain preservation center in 2018, 5 and 7 months, and the preservation number is GDMCC No:60359, deposited at No. 59 building 5 of Ji-Tou 100 of Jieli Zhonglu, guangzhou (published in the prior patent with publication No. CN 110066739A).
Preparation of beauveria brookfield strain SB010 spore suspension: selecting mature spores of Beauveria bassiana strain SB010, performing activation culture on a PDA plate at 25 ℃ for 8.5d, adding distilled water to wash the spores, shaking forcefully for 4min to obtain a Beauveria bassiana strain SB010 spore suspension, and calculating the spore concentration by a blood counting method for later use.
Preparation of a Sabouraud agar culture medium: 20g of glucose and 5g of peptone, distilled water is added to the mixture to be constant volume of 1L, and the mixture is sterilized at the high temperature of 121 ℃ for 15min.
Example 1 preparation of zero-valent iron-Beauveria brucei nanoparticles
A preparation method of zero-valent iron-Beauveria bassiana nanoparticles comprises the following steps:
s1, adjusting the concentration to be 1 multiplied by 10 8 Inoculating conidium/mL Beauveria bassiana strain SB010 spore suspension into fermentation culture solution SDA, and performing shake culture at rotation speed of 180rpm/min and temperature of 25 deg.C for 60h to obtain mycelium;
s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture for 60 hours at the rotation speed of 180rpm/min and the temperature of 25 ℃ to obtain a conidium filtrate;
and S3, adding nano zero-valent iron powder with the particle size of 70nm into the conidium filtrate obtained in the step S2, and incubating for 72 hours to obtain the zero-valent iron-Beauveria bassiana nano particles.
Wherein the mass-to-volume ratio of the nano zero-valent iron powder and the conidium filtrate in the step S3 is 1g:500mL.
Example 2 preparation of zero-valent iron-Beauveria bassiana nanoparticles
A preparation method of zero-valent iron-Beauveria bassiana nanoparticles comprises the following steps:
s1, adjusting the concentration to be 1 multiplied by 10 7 Inoculating conidium/mL Beauveria bassiana strain SB010 spore suspension into fermentation culture solution SDA, and performing shake culture at a rotation speed of 120rpm/min and a temperature of 26 ℃ for 48h to obtain mycelium;
s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture at the rotating speed of 120rpm/min and the temperature of 26 ℃ for 48 hours to obtain a conidium filtrate;
s3, adding nano zero-valent iron powder with the particle size of 60nm into the conidium filtrate obtained in the step S2, and incubating for 24h to obtain the zero-valent iron-Beauveria bassiana nano particles.
Wherein the mass-volume ratio of the nano zero-valent iron powder in the step S3 to the conidium filtrate is 1g:1000mL.
Example 3 preparation of zero-valent iron-Beauveria brucei nanoparticles
A preparation method of zero-valent iron-Beauveria bassiana nanoparticles comprises the following steps:
s1, adjusting the concentration to be 1 multiplied by 10 9 conidia/mL of Beauveria bassiana strain SB010 spore suspension is inoculated into fermentation culture solution SDA, and is subjected to shaking culture at the rotation speed of 160rpm/min and the temperature of 24 ℃ for 72hObtaining mycelium;
s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture at the rotating speed of 160rpm/min and the temperature of 24 ℃ for 72 hours to obtain a conidium filtrate;
s3, adding nano zero-valent iron powder with the particle size of 80nm into the conidium filtrate obtained in the step S2, and incubating for 36h to obtain the zero-valent iron-Beauveria bassiana nano particles.
Wherein the mass-volume ratio of the nano zero-valent iron powder in the step S3 to the conidium filtrate is 1g:800mL.
Example 4 preparation of zero-valent iron-Beauveria brucei nanoparticles
A preparation method of zero-valent iron-Beauveria bassiana nanoparticles comprises the following steps:
s1, adjusting the concentration to be 1 multiplied by 10 5 Inoculating conidia/mL Beauveria bassiana strain SB010 spore suspension into fermentation culture solution SDA, and performing shake culture at rotation speed of 140rpm/min and temperature of 25 deg.C for 60h to obtain mycelium;
s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture for 60 hours at the rotating speed of 140rpm/min and the temperature of 25 ℃ to obtain a conidium filtrate;
s3, adding nano zero-valent iron powder with the particle size of 75nm into the conidium filtrate obtained in the step S2, and incubating for 50h to obtain the zero-valent iron-Beauveria bassiana nano particles.
Wherein the mass-volume ratio of the nano zero-valent iron powder in the step S3 to the conidium filtrate is 1g:550mL.
Taking the zero-valent iron-beauveria bassiana nanoparticles prepared in example 1 as an example, the following specific experimental methods and results are used for determining the performance characterization of the nanoparticles prepared in examples 1 to 4, the indoor toxicity of the nanoparticles on prodenia litura and the influence on the growth and development of the prodenia litura:
application example 1 Performance characterization of zero-valent iron-Beauveria brucei nanoparticles
1. Experimental method
Mixing the conidium filtrate of the Beauveria bassiana strain SB010 with nano zero-valent iron powder for reduction reaction. The absorbance values of the zero-valent iron-beauveria brookfield nanoparticles prepared in example 1 at 200nm, 300nm, 400nm, 500nm and 600nm were measured respectively, and uv-vis spectroscopy (uv-vis spectroscopy is an initial characterization step for analyzing the formation of FeNPs in an aqueous solution), scanning Electron Microscopy (SEM), EDX spectroscopy, X-ray diffraction (XRD) and fourier transform infrared spectroscopy (FTIR) were performed.
2. Results of the experiment
The reduction reaction results show that: during the preparation of the zero-valent iron-beauveria brookfield nanoparticles, the solution color changed from light black to dark black, indicating that the zero-valent iron-beauveria brookfield nanoparticles had formed.
The ultraviolet-visible spectrum of the zero-valent iron-Beauveria brucei nanoparticles at different absorbance values is shown in figure 1, and a characteristic surface plasma absorption band is observed at 300nm after incubation for 72h and is formed by Fe 0 Excitation spectra of the synthesized FeNPs were also observed.
SEM images of the zero-valent iron-Beauveria brucei nanoparticles are shown in FIG. 2, wherein (A) is an SEM image at 5 μm magnification, and (B) is an SEM image at 1 μm magnification; it can be seen that the nano zero-valent iron powder has obvious adhesion on the surface of conidium of the beauveria brookfield strain SB010, which indicates that the zero-valent iron-beauveria brookfield nanoparticle has been successfully synthesized.
The EDX spectrum analysis result of the zero-valent iron-beauveria broomrape nanoparticles is shown in fig. 3, and it can be seen that the characteristic peaks of zero-valent iron are 6.3KeV and 7.1KeV.
The XRD pattern of the zero-valent iron-Beauveria bassiana nanoparticle is shown in figure 4, the diffraction intensity is recorded at a 2 theta angle of 10 degrees to 90 degrees, and the XRD result shows four strong peaks with 2 theta values of 45.72 degrees, 64.47 degrees, 83.59 degrees and 84.05 degrees, which respectively correspond to 200 planes, 220 planes and 311 planes of the face-centered cubic structure of the nanoparticle; the target is a wavelength of
Indicating that the nanoparticle has a cubic structure. The XRD patterns of the zero-valent iron and other substances have no peak, which indicates that the zero-valent iron-Beauveria bassiana nano-particles have high purity.
The FTIR spectrum of the zero-valent iron-Beauveria brucei nano-particle is shown in FIG. 5, and it can be seen that the absorption peaks of the nano-particle are located at about 3394.52, 1629.51, 1384.14, 1047.25 and 575.95, and the area is 4000 to 400cm -1 Strong O-H stretching vibration, strong C = C stretching vibration, C-H bending vibration, C-O stretching vibration and strong C-C stretching vibration; indicating the possible presence of amino acid residues and proteins synthesized in the zero-valent iron-Beauveria bassiana nanoparticle.
From the results, the beauveria bassiana strain SB010 successfully prepares the high-purity zero-valent iron-beauveria bassiana nano-particles under the mediation effect of zero-valent iron.
Application example 2 indoor toxicity determination of zero-valent iron-Beauveria brucei nanoparticles to Spodoptera litura
1. Experimental method
(1) T1-T9 reagents (T1-T9 reagents and their spore concentrations are shown in Table 1) were formulated using the zero-valent iron-Beauveria brucei nanoparticles prepared in example 1, a suspension of spores of Beauveria brucei strain SB010, and 0.01% Tween 80.
TABLE 1 T1T 9 reagents and spore concentrations (ppm)
(2) Setting the reagents T1-T7 as a test group, the reagent T8 as a control group and the reagent T9 as a blank control group; 20 well-developed prodenia litura 2-instar larvae with similar states are selected from each group. Soaking the spodoptera litura larvae in the reagents T1-T9 for 30s respectively, taking out, and putting on a paper towel for airing. Air-dried and then placed into an insect breeding box with a mark, and the insect breeding box is placed in an illumination incubator at 25 ℃ and 12L. The fresh artificial feed is replaced regularly.
(3) The death status of prodenia litura was continuously observed for 7 days, and the number of deaths of prodenia litura was recorded day by day, and the corrected mortality, median lethal concentration (LC 50), and median lethal time (LT 50) at the maximum concentration of 500ppm were calculated.
2. Results of the experiment
The influence result of the zero-valent iron-beauveria bassiana nanoparticles on the mortality of the prodenia litura is shown in fig. 6, and it can be seen that the mortality of the 2-instar larvae of the prodenia litura also rises along with the increase of the concentration of the zero-valent iron-beauveria bassiana nanoparticles in different time intervals; wherein, on day 3, the killing effect of 100ppm, 200ppm and 500ppm medicaments on 2-instar larvae of prodenia litura is better than that of Beauveria bassiana strain SB010 spore suspension; on the 4 th to 7 th days, the killing effect of 200ppm and 500ppm medicaments on 2-instar larvae of prodenia litura is better than that of the Beauveria bassiana strain SB010 spore suspension.
The LC50 values of the zero-valent iron-beauveria brookfield nanoparticles treated with prodenia litura are shown in table 2, and the LT50 values of the zero-valent iron-beauveria brookfield nanoparticles treated with prodenia litura are shown in table 3, and it can be seen that the LC50 values of the nanoparticles after 3, 5 and 7 days treatment are 464.49ppm, 163.43ppm and 58.86ppm, respectively, for prodenia litura; LT50 values of the zerovalent iron-Beauveria brucei nanoparticles to Spodoptera litura at concentrations of 50ppm, 100ppm, 200ppm and 500ppm 7 days after treatment were 14.64, 8.55, 5.10 and 2.29, respectively.
The above results show that: the zero-valent iron-beauveria bassiana nanoparticles have a good effect on killing of prodenia litura, and the killing effect is more obvious along with the increase of time; the effect of 500ppm nanoparticles was significantly higher than 1000ppm beauveria brookfield strain SB010 spore suspension at 5-day and 7-day corrected mortality, and the lethal effect of 100ppm, 200ppm nanoparticles was also not significantly different from 1000ppm beauveria brookfield strain SB010 spore suspension. Therefore, the lethal effect of the zero-valent iron-Beauveria brucei nanoparticles on prodenia litura is remarkably stronger than that of Beauveria brucei strain SB010 spore suspension, and the zero-valent iron-Beauveria brucei nanoparticles have good development potential.
TABLE 2 LC50 values of zero-valent iron-Beauveria brucei nanoparticles treated Spodoptera litura
TABLE 3 LT50 values of zero-valent iron-Beauveria brucei nanoparticles for treating prodenia litura
Application example 3 influence of zero-valent iron-Beauveria bassiana nanoparticles on growth and development of prodenia litura
1. Experimental methods
The spodoptera litura larvae were cultured to 2 instars in an incubator, and the test solutions were prepared using the zero-valent iron-beauveria brookfield nanoparticles prepared in example 1, beauveria brookfield strain SB010 spore suspension, and 0.01% tween 80: the treatments at 6.25ppm, 12.5ppm, 25ppm, 50ppm, 100ppm, 200ppm, and 500ppm were defined as test groups, the 1000ppm Beauveria brinell strain SB010 spore suspension was defined as a control group, and the treatment at 0ppm was defined as a blank control group.
20 well-developed prodenia litura 2-instar larvae with similar states are selected from each group. And (3) respectively soaking the prodenia litura larvae in each group of test solution for 30s, taking out, and putting on a paper towel for airing. The 2 nd larvae were placed in a plastic round box with a diameter of 3cm and a height of 4cm and cultured in an incubator (25 ℃). The weight of 2-instar larvae which have molted for 12 hours is taken as an initial weight, an electronic balance with one ten-thousandth precision is used for weighing and recording the weight of each larva and the weight of feed, and the relative growth rate, the relative feeding rate and the wool conversion rate of the larva are calculated according to the following formulas. And recording pupation rate after pupation, and recording eclosion rate after eclosion.
Experimental data processing analysis was performed using one-way analysis of variance with SPSS 19.0 software for relative growth rate, relative feeding rate and hair transformation rate, and Tukey for significance of difference testing. The mapping was performed using Microsoft Excel 2010.
2. Results of the experiment
The influence results of the zerovalent iron-beauveria brookfield nanoparticles with different concentrations on the relative growth rate of the prodenia litura are shown in fig. 7, and it can be seen that the nanoparticles have certain influence on the growth and development of the prodenia litura, and compared with the control group, the effect of inhibiting the growth of the prodenia litura of the nanoparticles with the concentrations of 200ppm and 500ppm is obviously improved.
The results of the effect of the zerovalent iron-beauveria bassiana nanoparticles with different concentrations on the relative feeding rate of the prodenia litura are shown in fig. 8, and it can be seen that the nanoparticles have a certain effect on the feeding of the prodenia litura, and the relative feeding rates of the prodenia litura treated by the nanoparticles with concentrations of 200ppm and 500ppm are significantly reduced compared with the control group.
The influence results of the zerovalent iron-beauveria bassiana nanoparticles with different concentrations on the conversion rate of the prodenia litura hairs are shown in fig. 9, and it can be seen that the nanoparticles have certain influence on the conversion rate of the prodenia litura hairs, and the conversion rates of the prodenia litura hairs treated by the nanoparticles with the concentrations of 200ppm and 500ppm are 0.35% and 0.09%, which are significantly lower than those of a control group.
The influence results of the zerovalent iron-beauveria bassiana nanoparticles with different concentrations on the pupation rate of prodenia litura are shown in fig. 10, and it can be seen that the nanoparticles have a certain influence on the pupation rate of the prodenia litura, and the pupation rate of the prodenia litura treated by the nanoparticles with the concentration of 500ppm is 30%, which is significantly lower than that of a control group.
The effect results of the zerovalent iron-beauveria brookfield nanoparticles with different concentrations on the eclosion rate of the prodenia litura are shown in fig. 11, and it can be seen that the eclosion rates of the prodenia litura treated by the nanoparticles with concentrations of 50ppm, 100ppm, 200ppm and 500ppm are respectively 55%, 48%, 35% and 0%, which are significantly lower than those of the control group.
The above results show that: the zero-valent iron-Beauveria bassiana nanoparticles can obviously inhibit the relative growth rate, the relative feeding rate and the hair conversion rate of the prodenia litura (presumably because the prodenia litura enters the prepupation stage and is inhibited in growth), and the pupation rate and the emergence rate of the prodenia litura; compared with Beauveria bassiana strain SB010 spore suspension, the zero-valent iron-Beauveria bassiana nanoparticles can inhibit and delay the growth and development of prodenia litura, and the zero-valent iron-Beauveria bassiana nanoparticles with the concentration as high as 500ppm can significantly inhibit the growth and development of prodenia litura and reduce the quality of the prodenia litura.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such modifications are intended to be included in the scope of the present invention.
Claims (9)
1. A preparation method of zero-valent iron-Beauveria bassiana nanoparticles is characterized by comprising the following steps:
s1, inoculating a beauveria bassiana spore suspension into a fermentation culture solution, and performing shake culture for 48-72 hours to obtain mycelia;
s2, adding the mycelium obtained in the step S1 into distilled water, and performing shaking culture for 48-72 hours to obtain a conidium filtrate;
s3, adding zero-valent iron into the conidium filtrate obtained in the step S2, and incubating for 24-96 h to obtain the zero-valent iron-Beauveria bassiana nano particles;
the beauveria bassiana strain SB010 in the step S1 is preserved in Guangdong province microorganism strain preservation center in 2018, 5 and 7 days, and the preservation number is GDMCC No:60359.
2. the method according to claim 1, wherein the concentration of the Beauveria bassiana spore suspension in step S1 is 1 x 10 5 ～1×10 9 conidia/mL.
3. The method according to claim 1, wherein the zero-valent iron in step S3 is nano zero-valent iron powder.
4. The method according to claim 1, wherein the mass-to-volume ratio of the zero-valent iron to the conidium filtrate in step S3 is 1:500 to 1000.
5. The method according to claim 1, wherein the rotation speed of the shaking culture in the steps S1 and S2 is 120 to 180rpm/min, and the temperature of the shaking culture is 24 to 26 ℃.
6. The zero-valent iron-Beauveria brucei nanoparticles prepared by the method of any one of claims 1-5.
7. Use of the zero-valent iron-beauveria brookfield nanoparticles of claim 6 for controlling noctuidae pests or in the preparation of a noctuidae pest control formulation.
8. The use according to claim 7, wherein the noctuidae pest is prodenia litura.
9. A noctuidae pest control formulation comprising the zero-valent iron-beauveria brookfield nanoparticle of claim 6.
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