CN115369058A - Bacillus licheniformis ZLP-81 and its application - Google Patents

Abstract

The present invention relates to a Bacillus licheniformis ZLP‑81, the preservation number of which is CGMCC No. 25028. The strain has the functions of preventing diseases, killing insects and promoting plant growth. The invention also discloses the application of the bacterial strain in the preparation of biological control preparations, biological pesticides, soil restoration agents or biological fertilizers.

Description

Bacillus licheniformis ZLP-81 and its application

technical field

The invention relates to a bacillus licheniformis ZLP-81 and application thereof.

Background technique

Plant diseases and insect pests bring serious losses to agricultural production. Conventional chemical pesticide control not only kills a large number of natural enemies and beneficial microorganisms, destroys the ecological balance, but also causes a sharp increase in the resistance of pests, causing great harm to the environment and human health. For this reason, the research and development of safe, efficient and environmentally friendly products and alternative technologies for chemical fertilizers and pesticides has become a strategic requirement to ensure food security, food safety and environmental safety.

Bacillus licheniformis is a kind of aerobic, spore-forming rod-shaped bacteria, which has the characteristics of non-toxic and harmless to humans and animals, environmental friendliness, strong stress resistance, fast reproduction speed, etc., and can secrete a variety of active substances. Studies have shown that such microorganisms are against Rhizoctonia solani, Botrytis cinerea, Fusarium, Fusarium wilt, Early blight, Leaf mold, Black spot, Anthracnose, Downy mildew, Scab, Ringworm, Spot disease, etc. have a strong inhibitory effect, and can promote crop growth.

However, there is no relevant report about the bacterial strains that simultaneously have the functions of preventing diseases, killing insects and promoting plant growth.

Contents of the invention

The object of the present invention is to provide a kind of bacillus licheniformis ZLP-81 and application thereof that have disease prevention, insecticide and plant growth promotion functions simultaneously.

The present invention adopts following technical scheme:

A Bacillus licheniformis ZLP-81, the deposit number is CGMCC No.25028, and the depository unit is the General Microbiology Center of the China Committee for the Collection of Microorganisms, and the address is: Yard 3, No. 1 Beichen West Road, Chaoyang District, Beijing, China No. Institute of Microbiology, Chinese Academy of Sciences, date of preservation is June 9, 2022.

A kind of application of above-mentioned Bacillus licheniformis ZLP-81 in preventing and treating Lepidoptera pests, Blattaorder pests, Hymenoptera pests, Centipede pests, Hemiptera pests, Coleoptera pests or Orthoptera pests.

Further, the Lepidoptera pests include diamondback moth, beet armyworm, litura and cabbage caterpillar; the hemiptera pests include aphids, whitefly, and ichthyophthora; the coleoptera pests include grubs, day lily Insects or pseudodips; said Orthoptera pests include locusts, crickets, and mole crickets.

A kind of application of above-mentioned Bacillus licheniformis ZLP-81 in preventing and treating plant pathogenic bacteria, aquatic pathogenic bacteria and Staphylococcus aureus.

Further, the plant pathogens include: tomato leaf mold, tomato cinerea, pear black spot, fusarium, cotton wilt, cucumber cinerea, grape cinerea, cucumber anthracnose, cucumber downy mildew, Cucumber scab, Apple ringworm, Rhizoctonia solani, Corn leaf spot and Tomato early blight.

Further, the aquatic pathogenic bacteria include: Aeromonas hydrophila, Edwardsiella tarda and Aeromonas victoria.

A kind of application of above-mentioned bacillus licheniformis ZLP-81 in the preparation of biocontrol agents, biological pesticides, soil remediation agents or biological fertilizers.

Further, the biopesticide, biopesticide, biofertilizer, biological seed coating agent or soil remediation agent contains Bacillus licheniformis ZLP-81 thallus and/or fermentation broth; or the surface isolated from fermentation broth Actin, iturin, fenlin, or spermatin.

An insecticide, which comprises the following components in parts by weight: 23.45-76.3 parts of the original powder of Bacillus licheniformis ZLP-81 strain, 15.0-20.0 parts of binder, 1.0-1.5 parts of dispersant, and 2.5-3.0 parts of disintegrant 0.9-1.4 parts of wetting agent and 0.55-2.15 parts of synergist.

Further, the original powder of the Bacillus licheniformis ZLP-81 strain is prepared by the following method: adding 7-13% of β-cyclodextrin, 5-13% of CaCO ₃ and 1.5-5.5% MgSO ₄ are then spray-dried.

Further, it also includes 0-52 parts of auxiliary materials.

Wherein, the binder is selected from soluble starch, carboxymethyl cellulose, and dextrin.

Wherein, the dispersant is selected from sodium lignosulfonate, sodium tripolyphosphate, sodium pyrophosphate.

Wherein, the disintegrating agent is selected from sodium sulfate, sodium lauryl sulfate, polysorbate 80.

Wherein, the wetting agent is selected from soybean lecithin, sulfonated oil, and acetylene glycol.

Among them, the synergists used are: siloxane copolymer and polyol mixture, polyether modified trisiloxane, ethoxylated polytrisiloxane and other organic silicon, synergist ether, butene di Detoxifying enzyme inhibitors such as diethyl phosphate and triphenyl phosphate, turpentine oil, tea saponin, natural tangerine peel essential oil, corn germ oil, rapeseed oil, soybean oil, green peel orange oil, pine oil, neem oil, castor oil, Peppermint oil, thyme oil, eucalyptus oil, sesame oil, olive oil and other vegetable oils, soybean lecithin nonionic surfactant mixture, polyaspartic acid, alkyl polyglucoside, d-limonene, honey, acetone, cinnamaldehyde, Any one or more of terpinen-4-ol, toosendanin, flicking fingers, beichuang, zozhanbao, and yikejia.

Further, it also includes 1.10-1.70 parts by weight of chemical pesticides, biological pesticides or plant pesticides.

Chemical insecticides include: alanine methyl ester, difenomethylcycline, saccharin, imported formazan, benzimidazole No. 44, melamine, thiram, imported blue powder, chlorantraniliprole, high-efficiency Cyhalothrin, spinosad, cypermethrin, imidacloprid, deltamethrin, gibberellic acid, indole acetic acid, indole butyric acid, adenine, hydroxyadenine, benzylaminopurine, 24-Epibrassinolide, 22,23,24-Epibrassinolide, 28-Epihomobrasinolide, 28-Homobrasinolide, 14-Hydroxybrassinosterol, Triacontanol, S-absorbin, ascorbic acid, furfuryl aminopurine, iron chlorin, allantoin, hypersensitivity protein, alternaria activator protein, amino oligosaccharins, lentinan, chitosan, dextran Sugar, oligosaccharides, β-lupine conglutinin polypeptide, cholecalciferol, milbemycin, sex attractant for Chilo borer, sex pheromone for Spodoptera litura, sex pheromone for Lygus green bug, sex pheromone for pear Any one or more of tropins and codling moth sex pheromones.

Biological insecticides include: Bacillus licheniformis, Bacillus brevius, Paenibacillus polymyxa, Bacillus thuringiensis, Bacillus methylotrophs, Bacillus marine, Bacillus firmus, Bacillus sphaericus, Bacillus cereus, Fluorescent false Bacillus monospora, Brevibacillus spp., Stenobacterium brevii, Bacillus licheniformis, Bacillus licheniformis, Rhodopseudomonas palustris, Rhodothrothia thiaphila, Beauveria bassiana, Beauveria bassiana, and Metarhizium anisopliae , Trichoderma harzianum, Trichoderma, Paecilomyces lilacinus, Verticillium chlamydia, ear mold, shell mold, shell mold, Pythium oligandrum, cockroach virus, polyhedrosis virus, granular virus, multocida Any one or more of mymycin, abamectin, and Microsporidia locusti.

Plant insecticides include: azadirachtin, matrine, veratrine, nicotine, rotenone, pyrethrins, pectin, eucalyptol, star anise oil, wolftoxin, triptolide, curcumol, Osthole, Eugenol, Emodin, Carvacrol, Berberine, Stenol, Tea Saponin, Spirocarb, Allicin, D-Limonene, Terpene Alcohol, Allyl Isothiocyanate , pentadecylphenolic acid, tridecylphenolic acid, and any one or more of meprofenone.

A bactericide comprises bacillus licheniformis ZLP-81 cell or fermentation liquid.

Further, the bactericide includes a fermentation broth of Bacillus licheniformis ZLP-81 and a carbendazim solution at a volume ratio of 500:1; the carbendazim solution is formed by mixing 1 g of carbendazim with 600-1000 mL of water .

A fungicide, comprising surfactant, iturin, fenchinin or spermatin isolated from the fermentation broth of Bacillus licheniformis ZLP-81.

The beneficial effects of the present invention are:

The Bacillus licheniformis ZLP-81 of the present invention is a class of aerobic, spore-forming rod-shaped microorganisms, which has the characteristics of environmental friendliness, strong stress resistance, fast propagation speed, etc., and can secrete various active substances to promote plant growth.

The Bacillus licheniformis ZLP-81 of the present invention has stronger insecticidal activity, can effectively kill Lepidoptera pests such as diamondback moth, beet armyworm, Spodoptera litura, cabbage caterpillar, aphids, whitefly, spotted cicada etc. Pests of the order Hemiptera, pests of the order Coleoptera such as grubs, dayworms, and scorpion beetles, pests of the order Orthoptera such as locusts, crickets, and mole crickets, cockroaches of the order Blattata, ants of the order Hymenoptera, and centipedes of the order Centipede. At the same time, it has broad-spectrum antibacterial properties, and is effective against gray mold, sickle disease, fusarium wilt, early blight, leaf mold, black spot, anthracnose, downy mildew, scab, ring spot, small spot, etc. All plant pathogenic bacteria have obvious inhibitory effects, and have good application prospects in the development of functional microbial preparations such as insecticide or disease prevention.

Description of drawings

Figure 1 is a colony diagram of Bacillus licheniformis ZLP-81.

Figure 2 is a diagram of Bacillus licheniformis ZLP-81 strain.

Detailed ways

The technical solutions of the present invention will be described in detail below in conjunction with preferred embodiments. The following examples are only used to illustrate and explain the present invention, but not to limit the technical solution of the present invention.

Example 1 Isolation and Screening of Bacillus licheniformis ZLP-81

(1) Take soil samples: collect soil samples from Qamdo, Tibet, China, 50-100g each; put them into sterilized kraft paper bags, seal the bags, and record the sampling location, environment and date.

(2) Isolation and purification of Bacillus: the soil samples were separated by plate dilution method on PB medium. The specific operation is as follows: Accurately weigh 10 g of soil sample, add it into a 90 mL conical flask of 0.9% normal saline with glass beads, shake it on a shaker at 180 r/min for 30 min, and make the soil sample evenly distributed to make a soil suspension with a concentration of 10 ^-1 ; diluted in turn to concentrations of 10 ^-5 , 10 ^-6 , and 10 ^-7 . They were placed in a constant temperature water bath at 80°C for 30 minutes, and 100 μL of each dilution was taken to coat a PB plate, and the plate was incubated upside down in a constant temperature incubator at 30°C for 48 hours. Single colonies of different forms were picked and identified by spore staining. The bacillus was purified by streaking method and transferred to the slant of PB medium test tube. After culturing at 30°C for 48h, it was stored in a refrigerator at 4°C for later use.

(3) Screening of insecticidal and disease-preventing strains

Screening of insecticidal strains: Aphids are used as target organisms, after the purified microorganisms are fermented and cultured in liquid shake flasks, the fermentation broth is centrifuged at a speed of 12000r/min for 10min to remove bacteria, and the supernatant is filtered with a 0.22μm sterile filter membrane A sterile fermentation filtrate is prepared and the fermentation filtrate is diluted appropriately. Get the broad bean stem leaf of appropriate length and put into conical flask. Spray an appropriate amount of diluent on the live aphids and their stems and leaves and put them into conical flasks, cover the mouth of the flasks with gauze and fix them with rubber bands. After 2 days, the number of aphids was counted, and the strains with better insecticidal effect were screened out.

Screening of strains with disease-preventing effect: take cucumber gray mold as indicator bacteria, and use plate confrontation method for primary screening. In the aseptic operation room, the Botrytis cinerea pathogen growing on the PDA medium is made into d=5mm bacteria cake with a puncher, and then the bacteria cake is transferred to the center of the PDA medium plate, and each antagonistic bacteria obtained by the preliminary screening The aseptic filtrates were respectively connected to the place 2.5cm away from the pathogenic bacteria, and three repetitions were set up. At the same time, the plate with only the pathogenic bacteria cake was used as a control, and the constant temperature was cultivated at 26°C. Calculate the average inhibition rate. The strains with strong antagonistic effect on pathogenic bacteria were screened out. The strains with strong insecticidal and disease-resistant effects were frozen and preserved at -80 °C.

(4) Identification of insecticidal and disease-preventing strains

According to the experimental method in the "Common Bacteria System Identification Manual", morphological, physiological and biochemical identifications were carried out on the screened strains with strong insecticidal and disease control effects. The strain was cultured on a PB medium plate at 32±1°C for 14 to 16 hours, and it was observed that the colony numbered ZLP-81 was irregularly round, white in color, dull, and rough in surface, and the bacterium was rod-shaped ( As shown in Figures 1 to 2), the spores are oval. Its physiological and biochemical identification indicators are shown in Table 1.

Table 1 Physiological and biochemical characteristics of strain ZLP-81

Note: "+" is positive; "-" is negative.

Through morphological observation and physiological and biochemical experiments, it was determined that the strain numbered ZLP-81 belonged to the genus Bacillus. Then, molecular biology classification and identification were carried out, and 16S rDNA sequence analysis was used to extract the total genomic DNA of the strain as a template to amplify its 16S rDNA sequence. The general primer 27F/1492R was used to amplify the target fragment, the amplified product was detected by 1% agarose gel electrophoresis, and the gene sequence was carried out by bidirectional sequencing method. Sequencing results After BLAST sequence homology comparison of the GenBank database, the strain numbered ZLP-81 was identified as Bacillus licheniformis (Bacillus licheniformis), named Bacillus licheniformis ZLP-81.

Bacillus licheniformis ZLP-81 was sent to the General Microbiology Center of China Committee for the Collection of Microbial Cultures for preservation. The address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China, and the preservation number is CGMCC No.25028 , and the deposit date is June 9, 2022.

Embodiment 2 bacillus licheniformis ZLP-81 is to the killing effect of lepidoptera pest, cockroach, hymenoptera ants and centipede centipede larvae

Bacillus licheniformis ZLP-81 was fermented. Use NB liquid culture medium, formula is: beef extract 5g/L, peptone 10g/L, NaCl 5g/L, glucose 10g/L, pH7.0. According to the inoculum amount of 5%, it was inoculated into the culture medium and then placed in a shaking incubator at 180r/min and 32°C for 48h to obtain a fermentation broth.

The fermented broth of Bacillus licheniformis ZLP-81 was diluted to 10 times, 50 times, 100 times, and treated with clear water as contrast. The medicine soil was prepared, and the medicine soil was prepared with clear water as a control. And add an appropriate amount of food ingredients and mix well. Using the medicinal soil method, select Plutella xylostella xylostella, Spodoptera litura, Spodoptera litura and cabbage caterpillar, cockroaches of cockroaches, ants of Hymenoptera and centipede larvae of Centipede order with the same size of insect body, and the temperature is 25 ℃～28 ℃ environment for 24 hours. The number of test insects per treatment is 45, and each treatment has 3 parallels. After being inoculated, they were cultured at room temperature at room temperature, and the mortality rate was counted after 56 hours, as shown in Table 2.

Table 2 The killing effect of Bacillus licheniformis ZLP-81 on lepidoptera pests and cockroaches

Example 3 The insecticidal effect of Bacillus licheniformis ZLP-81 on Hemiptera pests such as aphids

Bacillus licheniformis ZLP-81 was fermented and cultured (same as Example 2).

Dilute the fermentation broth to 10 times, 50 times, 100 times, and use clear water as a control. The food used in the insecticidal experiment of bean aphid is young broad bean stem, the food used in the insecticidal experiment of cotton aphid is young cotton stem, the food used in the insecticidal experiment of cabbage aphid is young cabbage leaf, and the food used in the insecticidal experiment of whitefly is tomato leaf , the food used in the insecticidal experiment of Cicada variegata was Sophora japonica, and these five kinds of food were soaked in the above liquid for 10 minutes, then blotted dry with filter paper, took it out and put it in a jar lined with filter paper, and added 2 mL of deionized water Moisturize, then cover the jar with plastic wrap, pierce the hole, and add 1mL of water every 12h. For each concentration, 50 pests were tested, repeated 3 times, and 50 pests of similar age and size were selected and placed on the prepared petri dish to starve for 4 hours, and then respectively added the broad bean young stems, cotton young stems, cotton young stems, and Cabbage young leaves, tomato young leaves and Sophora japonica flowers were replaced every 12 hours, cultivated overnight in an incubator at 25°C, recorded the number of dead insects, and calculated the mortality of aphids under different dilutions.

Mortality rate = (number of dead insects ÷ number of tested insects) × 100%

Mortality determination: After the pests are treated overnight, touch the pests with the tip of a brush, and if the pests do not move, they are considered dead, and the number of dead pests is counted and recorded. As can be seen from the results of the implementation data in Table 3, Bacillus licheniformis ZLP-81 has a good control effect on Hemiptera insects.

Table 3 Insecticidal effect of Bacillus licheniformis ZLP-81 on Hemiptera insects such as aphids

The killing effect of embodiment 4 bacillus licheniformis ZLP-81 to coleopteran larvae

Bacillus licheniformis ZLP-81 was fermented and cultured (same as Example 2).

The fermented broth of Bacillus licheniformis ZLP-81 was diluted to 10 times, 50 times, 100 times, and treated with clear water as contrast. The medicine soil was prepared, and the medicine soil was prepared with clear water as a control. And add an appropriate amount of food ingredients and mix well. Using the medicine-soil method, select third-instar larvae of beetles, needleworms, beetle beetles and longhorn beetles with the same body size, and keep them in an environment with a temperature of 25°C to 28°C for 24 hours. The number of test insects in each treatment was 15, and each treatment had 3 parallels, repeated 3 times. After inoculation, they were cultured indoors at room temperature, and the mortality was counted after 56 hours, as shown in Table 4.

The killing effect of table 4 bacillus licheniformis ZLP-81 to coleoptera larvae such as grubs

Embodiment 5 Bacillus licheniformis ZLP-81 is to the insecticidal effect of orthoptera pests such as locust, cricket, mole cricket

Bacillus licheniformis ZLP-81 was fermented and cultured (same as Example 2).

Dilute the fermentation broth to 10 times, 50 times, 100 times, and use clear water as a control. The food used in the locust killing experiment was wheat, the food used in the cricket killing experiment was edamame, and the food used in the mole cricket killing experiment was cabbage leaves. After placing it in a jar lined with filter paper, add 2 mL of deionized water to moisturize it, then cover the jar with plastic wrap, pierce the holes, and add 1 mL of water every 12 hours. 30 test pests at each concentration, repeated 3 times, picked 30 pests with similar body weight and put them on the prepared petri dish to starve for 4 hours, then added the wheat, edamame and cabbage leaves treated with the above liquid respectively, Replace every 12 hours, cultivate overnight in a 25°C incubator, record the number of dead insects, and calculate the mortality of pests under different dilutions.

Mortality rate = (number of dead insects ÷ number of tested insects) × 100%

As can be seen from the results of the implementation data in Table 5, Bacillus licheniformis ZLP-81 has a good control effect on Orthoptera insects such as locusts, crickets and mole crickets.

Table 5 Control effect of Bacillus licheniformis ZLP-81 on Orthoptera insects such as locusts

The bacteriostatic spectrum determination of embodiment 6 bacillus licheniformis ZLP-81

Select common pathogens such as: tomato leaf mold, tomato cinerea, pear black spot, fusarium, cotton wilt, cucumber cinerea, grape cinerea, cucumber anthracnose, cucumber downy mildew, cucumber scab , Rhizoctonia solani, corn spot bacterium, tomato early blight and other plant pathogens; aquatic pathogens such as Aeromonas hydrophila, Edwardsiella tarda, Aeromonas victoria and other aquatic pathogens and Staphylococcus aureus was used as the target, and the antibacterial spectrum of the strain ZLP-81 was determined by the plate confrontation method to determine the inhibitory ability of the ZLP-81 strain to the growth of pathogenic bacteria.

According to the implementation result of table 6, bacterial strain ZLP-81 all has stronger antagonistic action to tested 18 kinds of pathogenic bacteria, wherein the inhibitory action to tomato leaf mold is the strongest, and the diameter of inhibition zone can reach 25.20mm. Explanation The fungicide with the fermentation broth of Bacillus licheniformis ZLP-81 as the active ingredient has good application value in agricultural production.

The inhibitory effect of table 6 Bacillus licheniformis ZLP-81 on different plant pathogenic bacteria

The preparation of embodiment 7 bacillus licheniformis ZLP-81 insecticide (water dispersible granule)

(1) Preparation of the original powder of Bacillus licheniformis ZLP-81 strain

NB liquid medium was used to cultivate Bacillus licheniformis ZLP-81, the formula was: beef extract 5g/L, peptone 10g/L, NaCl 5g/L, glucose 10g/L, pH7.0. According to the inoculum amount of 5%, it was inoculated in NB medium and placed in a shaking incubator at 180 r/min and 32°C for 48 hours. Spray-dry the cultured Bacillus licheniformis ZLP-81 fermentation broth, choose 11% β-cyclodextrin as the filler, 9% CaCO ₃ , 3% MgSO ₄ , the inlet temperature is 180°C, and set the feed liquid The temperature is 22°C at room temperature, the wind speed is 30m ³ /h, and the feed flow rate is 15mL/min. After spray drying, the original powder of Bacillus licheniformis ZLP-81 strain is obtained.

(2) The production process of pesticides

The preparation method of formula 1: with 26.7% Bacillus licheniformis ZLP-81 bacterium powder and 52.5% adjuvant (calcium carbonate), 15% binder soluble starch, 1.3% dispersant sodium lignosulfonate, 2.9% The disintegrating agent sodium sulfate, 0.8% wetting agent soybean lecithin and 0.8% synergist soybean oil are fully mixed, pulverized and mixed evenly by jet mill, then added distilled water and kneaded into blocks, and then processed by granulator Extrude and granulate, place the obtained sample in an oven for drying at 45-50°C, and pass through a 40-mesh vibrating sieve to obtain Bacillus licheniformis ZLP-81 insecticide.

The preparation method of formula 2: with the former powder of 24% bacillus licheniformis ZLP-81 bacterial strain and 51.5% adjuvant (calcium carbonate), 17.2% binder carboxymethyl cellulose, 1.6% dispersant sodium tripolyphosphate , 2.3% disintegrant sodium lauryl sulfate, 1.5% wetting agent sulfonated oil and 1.9% synergist polyether modified trisiloxane are fully mixed, pulverized by jet mill and then mixed evenly, Then add distilled water and knead into a block, then extrude and granulate through a granulator, place the obtained sample in an oven for drying at 45-50°C, and pass through a 40-mesh vibrating sieve to obtain Bacillus licheniformis ZLP-81 insecticide.

The preparation method of formula 3: the former powder of 25.35% Bacillus licheniformis ZLP-81 bacterial strain and 50.5% adjuvant (calcium carbonate), 18.5% binder dextrin, 1% dispersant sodium pyrophosphate, 2.5% Disintegrating agent polysorbate 80, 1.2% wetting agent acetylene glycol and 0.95% synergist ethoxylated polytrisiloxane are fully mixed, pulverized and mixed by jet mill, and then added distilled water to knead The block is synthesized, and then extruded and granulated by a granulator. The obtained sample is placed in an oven for drying at 45-50° C., and passed through a 40-mesh vibrating sieve to obtain Bacillus licheniformis ZLP-81 insecticide.

Formula 1, formula 2 and formula 3 had corrected insecticidal rates of 73%, 74% and 74% against diamondback moth; corrected insecticidal rates of bean aphid were 66%, 66% and 67%; Rates were 65%, 66% and 68%.

The preparation method of embodiment 8 bacillus licheniformis ZLP-81 bactericide (powder)

The fermentation broth of Bacillus licheniformis ZLP-81 (same as Example 2) and carbendazim solution (1g carbendazim is dissolved in 800mL water) are evenly mixed according to the volume ratio of 500:1, then spray-dried, after spray-dried A powder of Bacillus licheniformis ZLP-81 fungicide was obtained.

The growth-promoting effect of embodiment 9 bacillus licheniformis ZLP-81 bactericide

Select cucumber as the test crop, indoor potted cucumber seedlings 20 strains/pot, establish 3 parallel, treatment group uses Bacillus licheniformis ZLP-81 bactericide (embodiment 8), control group does not use any growth-promoting products. Then randomly select 10 28-day-old cucumber seedlings, measure the plant height of the seedlings with a ruler, and measure the stem thickness with a vernier caliper. Greening at 105°C for 15 minutes, drying at 75°C to constant weight, measuring the dry mass of shoots and underground parts, measuring the chlorophyll content with ethanol-acetone extraction method, and measuring photosynthetic parameters with a portable photosynthetic measurement system. From Table 7, it can be seen that the fungicide Bacillus licheniformis ZLP-81 has an effect on the plant height, stem diameter, fresh weight of shoots per plant, dry weight of shoots per plant, leaf area, fresh weight of roots per plant, and root weight per plant of cucumber seedlings. Dry mass, total root volume per plant, total chlorophyll and net photosynthetic rate increased by 3.68%, 10.99%, 18.687%, 15.41%, 0.49%, 77.33%, 8.51%, 119.57%, 63.54% and 89.03% compared with CK , and except for plant height, leaf area and root dry weight per plant, all reached the level of significant difference. It shows that the Bacillus licheniformis ZLP-81 fungicide of the present invention has a significant growth-promoting effect on cucumber seedlings.

Table 7 The effect of Bacillus licheniformis ZLP-81 fungicide on the growth promotion of cucumber seedlings

Note: The same letter in the same row means P>0.05, the difference is not significant, and different letters mean P<0.05, the difference is significant.

The preparation of embodiment 10 bacillus licheniformis ZLP-81 insecticide (comprising other insecticides)

According to mass ratio (seeing table 8), make the bacillus licheniformis ZLP-81 insecticide formula that comprises chemical insecticide, biological insecticide, plant insecticide. Wherein the bacterial powder is the former powder of the Bacillus licheniformis ZLP-81 bacterial strain that the step (1) of embodiment 7 obtains.

Preparation method: mix the original powder of Bacillus licheniformis ZLP-81 strain with auxiliary materials, then fully mix with binder, dispersant, disintegrant, wetting agent, insecticide and synergist, and pulverize by jet mill Mix again, then add distilled water and knead into a block, then extrude and granulate through a granulator, place the obtained sample in an oven for drying at 45-50°C, pass through a 40-mesh vibrating sieve to obtain Bacillus licheniformis ZLP-81 insecticidal agent.

The auxiliary material used is calcium carbonate.

The binder used is: (1) soluble starch, (2) sodium carboxymethylcellulose, (3) dextrin.

The dispersants used are: (A) sodium lignosulfonate, (B) sodium tripolyphosphate, (C) sodium pyrophosphate.

The disintegrants used were: (a) sodium sulfate, (b) sodium lauryl sulfate, (e) polysorbate 80.

The wetting agents used were: (α) soybean lecithin, (β) sulfonated oil, (γ) acetylenic diol.

The synergists used are: (1) siloxane copolymer, (2) polyether modified trisiloxane, (3) ethoxy modified polytrisiloxane and other silicones, (4) synergistic Ether, (5) diethyl maleate, (6) detoxifying enzyme inhibitors such as triphenyl phosphate, (7) turpentine, (8) tea saponin, (9) natural tangerine peel essential oil, (10) Corn Germ Oil, (11) Rapeseed Oil, (12) Soybean Oil, (13) Green Peel Oil, (14) Pine Oil, (15) Neem Oil, (16) Castor Oil, (17) Peppermint Oil, (18) ) thyme oil, (19) eucalyptus oil, (20) sesame oil, (21) olive oil and other vegetable oils, (22) soybean lecithin nonionic surfactant mixture, (23) polyaspartic acid, (24) Alkyl polyglucoside, (25) d-limonene, (26) honey, (27) acetone, (28) cinnamaldehyde, (29) terpinen-4-ol, (30) toosendanin, (31) snap finger Any one or two or more of Jianjian, (32) Double Chuang, (33) Zhanzhanbao, (34) Yikejia.

Chemical insecticides include: (1) Alanine methyl ester, (2) Difenomethylcycline, (3) Sukling, (4) Imported methyltrop, (5) Benzimidazole No. 44, (6) Melanmethamine, (7) Thiam, (8) Imported Blue Powder, (9) Chlorantraniliprole, (10) Lambda-cyhalothrin, (11) Spinosyn, (12) Cervical Cypermethrin, (13) imidacloprid, (14) deltamethrin, (15) gibberellic acid, (16) indole acetic acid, (17) indole butyric acid, (18) adenine, (19) hydroxyene Adenine, (20) benzylaminopurine, (21) 24-epibrassinolide, (22) 22,23,24-epibrassinolide, (23) 28-epihomobrasinolide, ( 24) 28-homobrassinolide, (25) 14-hydroxybrassinosterol, (26) triacontanol, (27) S-abetin, (28) ascorbic acid, (29) furfurylaminopurine, ( 30) Iron chlorin, (31) Allantoin, (32) Hypersensitive protein, (33) Alternaria tennatus activating protein, (34) Amino oligosaccharides, (35) Lentinan, (36) Chitosan, (37) dextran, (38) oligosaccharin, (39) β-lupine conglutinin polypeptide, (40) cholecalciferol, (41) milbemycin, (42) Including the sex attractant of Chilo suppressalis, (43) Sex pheromone of Spodoptera litura, (44) Sex pheromone of Lygus spp. Any one or two or more of them.

Biopesticides include: (1) Bacillus subtilis, (2) Stenobacter brevis, (3) Paenibacillus polymyxa, (4) Bacillus thuringiensis, (5) Methylotrophic Bacillus, (6) Marine Bacillus, (7) Bacillus firmus, (8) Bacillus sphaericus, (9) Bacillus cereus, (10) Pseudomonas fluorescens, (11) Bacillus sporogenes, (12) Stenobacter brevis, (13) Bacillus licheniformis, (14) Bacillus amyloliquefaciens, (15) Rhodopseudomonas palustris, (16) Rhodobacter thiaphila, (17) Beauveria bassiana, (18) Beauveria bassiana , (19) Metarhizium anisopliae, (20) Trichoderma harzianum, (21) Trichoderma, (22) Paecilomyces lilacinus, (23) Verticillium chlamydia, (24) Auricle, (25) Shield Shell mold, (26) Shell mold, (27) Pythium oligandrum, (28) Cockroach virus, (29) Polyhedrosis virus, (30) Granular virus, (31) Spinosad, (32) Any one or two or more of abamectin and (33) Microsporidia locusti.

Plant insecticides include: (1) azadirachtin, (2) matrine, (3) veratrine, (4) nicotine, (5) rotenone, (6) pyrethrins, (7) bitter vine (8) eucalyptol, (9) star anise oil, (10) wolftoxin, (11) triptolide, (12) curcumol, (13) osthole, (14) eugenol, (15) Emodin, (16) Carvacrol, (17) Berberine, (18) Sterol, (19) Tea Saponin, (20) Spirocarb, (21) Allicin, (22) ) d-limonene, (23) terpene alcohol, (24) allyl isothiocyanate, (25) pentadecyl phenolic acid, (26) tridecyl phenolic acid, (27) cyproterone Any 1 or 2 or more types.

Table 8 Bacillus licheniformis ZLP-81 insecticide formula

The above-described embodiments and methods are the best implementations of the present invention. On the premise of not departing from the technical principles of the present invention, some changes, modifications, substitutions, and combinations can also be made, all of which are included in the protection scope of the present invention. Inside.

Claims (10)

1. A Bacillus licheniformis ZLP-81, characterized in that the preservation number is CGMCC No. 25028. 2. a kind of bacillus licheniformis ZLP-81 as claimed in claim 1 is controlling lepidoptera pest, cockroach pest, hymenoptera pest, centipede pest, hemiptera pest, coleoptera pest or orthoptera Application in pests. 3. an application of bacillus licheniformis ZLP-81 as claimed in claim 1 in the prevention and treatment of plant pathogens, aquatic pathogens and staphylococcus aureus. 4. application according to claim 3, is characterized in that, described phytopathogen comprises: tomato leaf mold, tomato cinerea, pear black spot fungus, fusarium, cotton fusarium wilt, cucumber cinerea, grape ash Mildew, Cucumber Anthracnose, Cucumber Downy Mildew, Cucumber Scab, Apple Ringworm, Rhizoctonia solani, Corn Leaf Spot and Tomato Early Blight. 5 . The application according to claim 3 , wherein the aquatic pathogens include: Aeromonas hydrophila, Edwardsiella tarda and Aeromonas victoria. 6. The application of a Bacillus licheniformis ZLP-81 as claimed in claim 1 in the preparation of biocontrol agents, biological pesticides, soil remediation agents or biological fertilizers. 7. An insecticide, characterized in that it comprises the following components by weight: 23.45 to 76.3 parts of former powder of Bacillus licheniformis ZLP-81 strain, 15.0 to 20.0 parts of binder, 1.0 to 1.5 parts of dispersant, 2.5~3.0 parts of disintegrant, 0.9~1.4 parts of wetting agent and 0.55~2.15 parts of synergist. 8. the insecticide according to claim 7, is characterized in that, the former powder of described Bacillus licheniformis ZLP-81 bacterial strain is prepared by following method: in the fermented liquid of Bacillus licheniformis ZLP-81, add fermented liquid quality 7～ 13% of β-cyclodextrin, 5~13% of CaCO ₃ and 1.5~5.5% of MgSO ₄ for spray drying. 9. The insecticide according to claim 8, further comprising 1.10-1.70 parts by weight of chemical insecticides, biological insecticides or plant insecticides. 10. A bactericide, characterized in that it comprises Bacillus licheniformis ZLP-81 thallus, fermentation broth, or surfactant, iturin, fenneline or spermatin isolated from fermentation broth.

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