CN102363750B - Insecticidal fungus and application thereof - Google Patents
Insecticidal fungus and application thereof Download PDFInfo
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- CN102363750B CN102363750B CN 201110326065 CN201110326065A CN102363750B CN 102363750 B CN102363750 B CN 102363750B CN 201110326065 CN201110326065 CN 201110326065 CN 201110326065 A CN201110326065 A CN 201110326065A CN 102363750 B CN102363750 B CN 102363750B
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Abstract
The invention relates to the field of biological control, especially to an insecticidal fungus and its application. The invention provides an insecticidal fungus Beauveria bassiana (Balsamo) Vuillemin strain BBNS-J9-16(Beauveria bassiana), the collection number of which is CGMCC No. 5288. The invention also provides an application of the Beauveria bassiana (Balsamo) Vuillemin strain for killing aphid, and provides a efficient composite pesticide, which contains the above Beauveria bassiana (Balsamo) Vuillemin strain BBNS-J9-16 thalline and/or its spore powder. The high-efficient composite pesticide provided by the invention is used to infect hosts by the utilization of the insect fungus which is environment friendly and compatible with the environment so as to control insects. The insecticidal fungus can be used to protect crops and minimize the use of chemical pesticides, and plays an active role in guaranteeing the agricultural product safety and agro-environmental safety.
Description
Technical field
The present invention relates to the biological control field, relate to particularly a kind of disinsection fungal and application thereof.
Background technology
Aphid has the advantages that individuality is little, growth is fast, breeding potential is high, resistance is strong, and is strong to environment-adapting ability, is distributed widely in the various agriculture and forestry plants in the world.It not only directly takes food the harm plant, goes back the infect indirectly plant virus, causes serious financial loss for the productions such as vegetables, grain and oil, fruit tree, gardens.Aphid Species on vegetables mainly contains the red aphid Myzus of peach persicae (Sulzer), radish aphid Lipaphis erysimi (Kaltenbach), brevicoryne brassicae (vegetable aphid) Brevicoryne brassicae (Linn.).Black peach aphid is national distribution, euryphagy, 50 sections more than 400 kind of plant be can endanger, Cruciferae, solanaceous vegetable (such as wild cabbage, Chinese cabbage, green vegetables, radish and eggplant, potato, tomato etc.) and Rosaceae fruit tree (peach, Lee, apricot, plum, cherry etc.) mainly comprised.Radish aphid is distributed in south China more, happiness food Chinese cabbage, radish, rape.Brevicoryne brassicae be northern sociales in Zhejiang, river, Fujian etc. be also on the books, and causes harm and increase the weight of, and especially has a liking for wild cabbage, Cauliflower.Aphid slows down the serious dehydration of plant, malnutrition, growth with piercing-sucking mouthparts feeding plant juice, tikka, leaf roll, withered so that dead occur.Aphid or various plants virus vector become the medium of virus disseminating between plant, the effect of often the Plant diseases phase being added fuel to the flames.The honeydew of simultaneously its secretion is placed on plant surface, is conducive to the propagation of fungi and brings out sooty mold and cause the infringement of plant.
The control of aphid mainly contains that gardening is removed, the physics trapping, chemistry is killed and the method such as biological restriction, although front two kinds of methods can significantly reduce aphid insect population quantity, but because the aphid reproduction speed is fast, under suitable humiture and food condition, can breed more than 100 times in 1 month, therefore also must in time use chemical pesticide in the planting process, be controlled such as interior absorption organic phosphates, pyrethroid, amino methyl ester class etc.Studies confirm that, prolonged and repeated use agricultural chemicals has made aphid that the resistance of Multiple Pesticides is increased severely, for example the melon in Distributions in Liaocheng of Shandong Province, Shouguang (cotton) aphid reaches 120,000 and 270,000 times unexpectedly to the resistance level of fenvalerate, and pyrethroid insecticides prevents that in the locality aphid is substantially invalid.Aphicide such as Aphox, Provado etc. are through in a few years using efficiently originally for some, and resistance also increases by tens times.Excessively use agricultural chemicals to cause that agricultural-food Chinese medicine is residual to exceed standard and Sichuan.
For ensureing the farm crop high and stable yields, guarantee simultaneously agricultural product quality and safety, nuisanceless plant protection technology becomes study hotspot gradually in recent years, and the biology restriction method of aphid is more and more favored.More successful method mainly is that protection is with release aphidius gifuensis, vegetable aphid cocoon honeybee, (example: Finland was in the success of the test of protection ground in 1978 for food aphid cecidomyiia Aphidoletes aphidimyza at present, China 1986 introduces from Canada and discharges, quantity benefit evil is than being 1: 20, preventive effect reached 90% in 7-9 days) etc. natural enemy of aphid, improve the population density of parasitism or predation control Field Pests; Moreover be to use the insect pathogenic fungus such as aphid mould Entomophthora aphidis, Verticillium Verticillium lecanii (Britain commodity registration Vertalec), make the insect death of falling ill by the pathogen infection effect.Insect pathogenic fungus is to the vertebrates high safety, and is compatible with environmental friendliness.Its mechanism of action to host insect be Spore adhesion in insect cuticle, the insect cuticle degrading enzyme by the secretion obligate passes body wall, breeds after entering the insect body cavity, consume its nutrition, discharge toxin, disturb normal physiological function, the infected tissue organ makes the host fall ill and dead at last.Owing to need not through digestive tube, therefore to preventing that the piercing-sucking mouthparts aphid from having a clear superiority in.This research group is engaged in for many years insect pathogenic fungus research and uses, find that muscardine (Beauveria bassiana) has parasitism and lethal effect to aphid, the effect of different strains is renderd a service and is differed greatly, and needs could obtain by artificial screening the bacterial strain of high virulence.In the association area, former studies person also has the higher Strain of Beauveria bassiana of virulence that obtains the multiple aphids such as wheat aphid are become aphid by the indoor bioassay screening.Find after deliberation, muscardine is as other insect pathogenic fungus, existence is to the slower weakness of host's onset, taking 3-5 days just can reach deadly peak usually in the biological assay, relatively lag behind the reproduction speed of aphid and plant to the tolerance degree of aphid damage, thereby usually can not in time control insect pest, do not reach the desirable preventive effect of people's expectation.In order to improve the muscardine preventive effect, need to seek to improve the method for muscardine desinsection speed.
Summary of the invention
In order to address the above problem, the present inventor proposes and has finished the present invention.
The purpose of this invention is to provide a kind of fungi Strain of Beauveria bassiana BBNS-J9-16 that aphid is efficiently infected.
A further object of the present invention provides Strain of Beauveria bassiana BBNS-J9-16 to the insecticidal action of aphid.
A further object of the present invention provides the high efficiency composition sterilant of a kind of aphid, and it comprises the spore powder of above-mentioned Strain of Beauveria bassiana BBNS-J9-16.
A further object of the present invention provides above-mentioned high efficiency composition sterilant to the insecticidal action of aphid.
The invention provides a kind of high efficient strain beauveria bassiana BBNS-J9-16.The inventor respectively from the Inner Mongol, natural forest area, meadow or the farmland on the ground such as Anhui, Jiangxi, Guizhou collect soil and infect the worm corpse of muscardine illness, in the laboratory, carry out microorganism separation, purifying, preserve the beauveria bassiana bacterial classification of purifying in the PDAY test-tube culture medium, obtain altogether 68 bacterial strains.The inoculation of purifying on the PDAY plate culture medium, was cultivated 12-14 days in 25 ± 1 ℃ of incubators, collect conidial powder, carry out biological assay, according to the final mortality ratio of aphid and bombys batryticatus rate, preliminary screening obtains 4 bacterial strains with higher toxicity action.These 4 primary election bacterial strains are inoculated the aphid polypide again and from the sick worm again separation and purification and biology measure and select, finally obtain high efficient strain beauveria bassiana BBNS-J9-16 (Beauveria bassiana).This bacterial strain is stored in (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on September 23rd, 2011, Institute of Microorganism, Academia Sinica, 100101), its deposit number is: CGMCC No.5288.
The nucleotide sequence in the ITS1-5.8S-ITS2 rDNA zone of described bacterial strain is shown in SEQ ID No.1:
tccgtaggtg aacctgcgga gggatcatta ccgagttatc caactcccaa cccctgtgaa 60
ttataccttt aattgttgct tcggcgggac ttcgcgcccg ccggggaccc aaaccttctg 120
aattttttaa taagtatctt ctgagtggtt aaaaaaatga atcaaaactt tcaacaacgg 180
atctcttggt tctggcatcg atgaagaacg cagcgaaatg cgataagtaa tgtgaattgc 240
agaattcagt gaatcatcga atctttgaac gcacattgcg cccgtcagta ttctggcggg 300
catgcctgtt cgagcgtcat tacgcccctc aagtcccctg tggacttggt gttggggatc 360
ggcgaggctg gttttccagc acagccgtcc cttaaattaa ttggcggtct cgccgtggcc 420
ctcctctgcg cagtagtaaa acactcgcaa caggagcccg gcgcggtcca ctgccgtaaa 480
accccccaac tttttatagt tgacctcgaa tcaggtagga ctacccgctg aacttaagca 540
tatcaataag cggagga 557
The application that the present invention also provides above-mentioned Strain of Beauveria bassiana BBNS-J9-16 to be used for killing aphis, preferably this muscardine bacterial strain is used for killing the application of vegetable aphid and flowering plant aphid.
The present invention also provides a kind of high efficiency composition sterilant, wherein, comprises thalline and/or its spore powder of above-mentioned Strain of Beauveria bassiana BBNS-J9-16.
According to high efficiency composition sterilant of the present invention, wherein, comprise chemical insecticide.
According to high efficiency composition sterilant of the present invention, the described high efficiency composition sterilant of preferred every gram contains 0.4～2 * 10
9The Provado of spore and 1/300-1/600 gram, the desinsection working concentration is 1000 times of dilutions.
According to high efficiency composition sterilant of the present invention, the described high efficiency composition sterilant of preferred every gram contains 0.4～2 * 10
9The effective cypermethrin of spore and 1/220-1/260 gram, the desinsection working concentration is 1000 times of dilutions.
According to high efficiency composition sterilant of the present invention, the described high efficiency composition sterilant of preferred every gram contains 0.4～2 * 10
9The Rogor of spore and 1/25-1/37.5 gram, the desinsection working concentration is 1000 times of dilutions.
The application that the present invention also provides above-mentioned high efficiency composition sterilant to be used for killing aphis is preferred for the extremely application of radish aphid (Lipaphis erysimi).
The present invention screening has obtained a kind of Strain of Beauveria bassiana that aphid is efficiently infected, also screen simultaneously several can with the hypotoxicity agricultural chemicals of beauveria bassiana compatibility.These agricultural chemicals can effectively promote beauveria bassiana that attacked by aphids is renderd a service under the condition far below conventional amount used, significantly reduce the amount of application of agricultural chemicals.The high efficiency composition sterilant that forms thus, thus the entomomycete of utilization and environmental friendliness compatibility infects host's Control pests, realizes the protection farm crop while, and the use of minimizing chemical pesticide has a positive effect to ensureing agricultural product security and agro-environment safety.
Description of drawings
The micro-morphology of Fig. 1 bacterial strain BBNS-J9-16.
Fig. 2 CTAB method is extracted the gel figure of the DNA of beauveria bassiana BBNS-J9-16.
The gel figure of the beauveria bassiana BBNS-J9-16 rrna ITS fragment of Fig. 3 amplification.
Fig. 4 has shown the effect of beauveria bassiana BBNS-J9-16 to aphid, and a is healthy aphid, and b, c are susceptible aphid.
Beauveria bassiana BBNS-J9-16 (Beauveria bassiana) is stored in (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on September 23rd, 2011, Institute of Microorganism, Academia Sinica, 100101), its deposit number is: CGMCC No.5288.
Embodiment
The invention will be further described in conjunction with specific embodiments.
Separation, the purifying of embodiment 1 beauveria bassiana BBNS-J9-16 (Beauveria bassiana) bacterial strain
Respectively from the Inner Mongol, natural forest area, meadow or the farmland on the ground such as Anhui, Jiangxi, Guizhou collect soil and infect the worm corpse of muscardine illness, carries out in the laboratory that microorganism separates, purifying.
Separation and purification step are: 1) from soil separate targets microorganism: with 20 times, 100 times, 500 times in sterilized water dilution soil, be inoculated into respectively on the selective medium, cultivated 4-7 days under 25 ℃ of temperature, choose colony lift and obtain on the new selective medium plate, single bacterium colony was cultivated 7-10 days, the apparent and microscopic morphology check up and appraisal by bacterium colony, determine beauveria bassiana, transfer in the bacterium culture medium test tube, after cultivating 14 days under 25 ℃ of temperature, preserve in 4 ℃ of refrigerator cold-storages.2) from the bombys batryticatus separate targets microorganism of falling ill: with the thalline on the careful picking worm of the inoculating needle corpse, be inoculated on the selective medium, under 25 ℃ of temperature, cultivated 7 days, differentiate bacterium colony and transfer on the new selective medium plate single bacterium colony cultivation 7-10 days, the apparent and microscopic morphology check up and appraisal by bacterium colony, determine beauveria bassiana, transfer in the bacterium culture medium test tube, after cultivating 14 days under 25 ℃ of temperature, preserve in 4 ℃ of refrigerator cold-storages.
Preserve the beauveria bassiana bacterial classification of purifying in the PDAY test-tube culture medium, obtain altogether 68 bacterial strains.To be seeded on the PDAY plate culture medium, in 25 ± 1 ℃ of incubators, cultivate 12-14 days, collect conidial powder, carry out biological assay.Measuring method is: 1) the laboratory rearing aphid; 2) spore powder is mixed with quantitative certain density spore suspension, to the aphid body, every bacterial strain is processed 100 aphids to usefulness Potter spray tower with 1ml bacterium liquid even spraying, 3 repetitions; 3) aphid is placed under 25 ℃ of temperature to raise supported the dead quantity of observed and recorded aphid every day, 47 days) dead aphid is transferred in the clean culture dish, moisturizing 3-5 days, observe polypide and have or not the growth muscardine and become bombys batryticatus; 5) according to the final mortality ratio of aphid and bombys batryticatus rate, preliminary screening obtains 4 bacterial strains with higher toxicity action; 6) to this 4 primary election bacterial strain according to 1)-5) and step, do again further mensuration, obtain high efficient strain beauveria bassiana BBNS-J9-16 (Beauveria bassiana).This bacterial strain is stored in (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on September 23rd, 2011, Institute of Microorganism, Academia Sinica, 100101), its deposit number is: CGMCC No.5288.
The biological morphology feature of embodiment 2 bacterial strains
Preparation PDAY substratum, its component and compound method are boiled 15min for fresh potato 200g is added the about 1000mL of water, with continuing to employ liquid after the filtered through gauze, add glucose 20g, yeast extract powder 5g, agar 15g, heating is melted agar fully, quantitatively to 1000mL, divide and install in the triangular flask, place 121 ℃ of high pressure steam sterilizers to keep 20min, pour in the culture dish when cooling to about 60 ℃, make dull and stereotyped stand-by.
Inoculate conidium at the PDAY plate culture medium, cultivate 2d, the visible bacterium colony of visual inspection for 25 ℃, the bacterium colony mycelia fine hair shape that is white in color continues to cultivate 3-4d, and visible bacterium colony enlarges growth, there is the oyster white Sporulation on the surface, after the spore maturation, and bacterium colony surface enrichment white aleurioconidium.
Drip the PDAY substratum of sterilization in the cell of matrix slide glass, the inoculation conidium is cultivated 3-4d for 25 ℃, observes in microscopically, and visible conidiophore and is born on the vegetative hyphae Dan Sheng; Conidiogenous cell clusters, and is ampuliform, and neck product spore axle is " Z " type and living the arrangement; The spore dispersion that easily comes off; Conidium is spherical, 2-3 μ m.As shown in Figure 1.
The rrna ITS region nucleotide sequence feature of embodiment 3 bacterial strains
Preparation LM substratum, its component and compound method add water to 1000mL for taking by weighing glucose 40g, peptone 5g, divide to be filled to triangular flask, place 121 ℃ of high pressure steam sterilizers to keep 20min, cool to room temperature for subsequent use.
The inoculation conidium is to above-mentioned substratum, and inoculum size is 1 * 10
5Spore/mL cultivates 48h on 25 ± 1 ℃, 200rpm shaking table, with staying mycelia behind the filter paper filtering, it is for subsequent use to extract moisture.
Extract genomic dna with the CTAB method.The DNA extraction operation is carried out routinely, obtains genomic dna, as shown in Figure 2.
Carry out pcr amplification with ITS1-5.8S-ITS2rDNA zone primer 15 '-TCCGTAGGTGAACCTGCGG and primer 25 '-TCCTCCGCTTATTGATATGC, the dna fragmentation that obtains increasing, as shown in Figure 3, fragment is carried out nucleotide sequence to be detected, obtain strain ribosome ITS nucleotide sequence, shown in SEQ ID No.1.
Embodiment 4 bacterial strains are to the action effect of vegetable aphid
The plantation Plantula Brassicae chinensis is raised radish aphid Lipaphis erysimi in the laboratory; It is 50,10,2,0.4,0.08 * 10 that the tween 80 aqueous solution with 0.05% is mixed with the series concentration gradient with the spore powder of bacterial strain BBNS-J9-16
6The spore suspension of spore/mL is got 1mL and is used Potter spray tower even spraying to the aphid body, and every bacterial strain is processed 100 aphids, 3 repetitions; Aphid placed under 25 ℃ of temperature to raise supported 7 days, the dead quantity of observed and recorded aphid every day is transferred to dead aphid in the clean culture dish, moisturizing 3-5 days, observes polypide and has or not the growth muscardine and become bombys batryticatus; According to the final mortality ratio of aphid and bombys batryticatus rate, estimate bacterial strain to the effect of aphid.The results are shown in Table 1 and Fig. 4, prove that bacterial strain can effectively cause the ill death of aphid.
Table 1 beauveria bassiana BBNS-J9-16 is to the toxicity action of vegetable aphid
Annotate:
1Be the contrast correction value, with " mean+SD " expression, suffix letter represents the significance of difference, represents without significant difference α=0.05 with same column comparison same letter;
2Be the bombys batryticatus ratio in the dead borer population.
Embodiment 5 bacterial strains are to the action effect of flowering plant aphid
In May, collect aphid from the Chinese rose blade; It is 1 * 10 that the tween 80 aqueous solution with 0.05% is mixed with the series concentration gradient
7The spore suspension of spore/mL is got 1mL and is used Potter spray tower even spraying to the aphid body, processes 2-3 at every turn and opens blade, about 100-130 aphid, 5 repetitions; Aphid placed under 25 ℃ of temperature to raise supported 7 days, the dead quantity of observed and recorded aphid every day is transferred to dead aphid in the clean culture dish, moisturizing 3-5 days, observes polypide and has or not the growth muscardine and become bombys batryticatus; According to the final mortality ratio of aphid and bombys batryticatus rate, estimate bacterial strain to the effect of aphid.The results are shown in Table 2, prove that bacterial strain can effectively cause the ill death of aphid.
Table 2 beauveria bassiana BBNS-J9-16 is to the toxicity action of Chinese rose aphid
Embodiment 6 beauveria bassianas and Provado composite insecticide are to the toxic action of aphid
The test spore powder of beauveria bassiana BBNS-J9-16, content is 1 * 10
11Spore/g.Provado is that Zhenjiang, Jiangsu is built 10% wettable powder that Su Huagong company limited produces, and manufacturer recommends the field working concentration to be 2000-3000 times of dilution.4 kinds of composite insecticides are the spore powder of beauveria bassiana BBNS-J9-16 and the mixture of Provado, wherein recombiner A
1In every gram contain 2 * 10
9The Provado of spore and 1/300 gram, recombiner A
2In every gram contain 0.4 * 10
9The Provado of spore and 1/300 gram, recombiner A
3In every gram contain 2 * 10
9The Provado of spore and 1/600 gram, recombiner A
4In every gram contain 0.4 * 10
9The Provado of spore and 1/600 gram.
Test method: the plantation Plantula Brassicae chinensis is raised radish aphid Lipaphis erysimi in the laboratory; Solution or suspension of respectively processing below the preparation respectively: 1) take by weighing respectively 1g composite insecticide A
1, composite insecticide A
2, composite insecticide A
3With composite insecticide A
4, add water 99mL, again by dilution in 1: 9, obtain the suspension of their 1000 times of dilutions; 2) with 0.05% the tween 80 aqueous solution spore powder of bacterial strain BBNS-J9-16 being mixed with concentration is 0.4 * 10
6Spore/mL and 2 * 10
6The spore suspension of spore/mL namely is equal to respectively composite insecticide A
1And A
21000 times suspension in beauveria bassiana spore content; 3) take by weighing the Provado wettable powder of 1g 10%, add water 99mL, by successively dilution in 1: 2,1: 9,1: 9, obtain 30000 times diluent again, namely be equal to composite insecticide A
1And A
21000 times suspension in the content of Provado; By dilution in 1: 11 time, namely obtain 60000 times diluent again, namely be equal to composite insecticide A
3And A
41000 times suspension in the content of Provado.Get respectively the treatment solution 1mL for preparing, to the aphid body, process 100 aphids with Potter spray tower even spraying at every turn, 3 repetitions; Aphid placed under 25 ℃ of temperature raised 7 days, the dead quantity of observed and recorded aphid every day is transferred to dead aphid in the clean culture dish, moisturizing 3-5 days, observes polypide and has or not the growth muscardine and become bombys batryticatus; According to the final mortality ratio of aphid and bombys batryticatus rate, estimate bacterial strain to the effect of aphid.
Test-results sees Table 3, proves that beauveria bassiana and Provado composite insecticide energy Effective Raise is to poisoning mortality ratio and the rate of death of aphid.
Table 3 desinsection host and independent beauveria bassiana or Provado compare the virulence of aphid
Annotate:
1Be the contrast correction value, with " mean+SD " expression, suffix letter represents the significance of difference, represents without significant difference α=0.05 with same column comparison same letter;
2Be the bombys batryticatus ratio in the dead borer population."--" expression is not processed or is not existed.
Embodiment 7 beauveria bassianas and effective cypermethrin composite insecticide are to the toxic action of aphid
The test spore powder of beauveria bassiana BBNS-J9-16, content is 1 * 10
11Spore/g.Effective cypermethrin is 4.5% missible oil that Langfang, the Chinese Academy of Agricultural Sciences Institute of Plant Protection agricultural chemicals pilot plant is produced, and manufacturer recommends the field working concentration to be 1500-2000 times of dilution.4 kinds of composite insecticides are the spore powder of beauveria bassiana BBNS-J9-16 and the mixture of effective cypermethrin, wherein recombiner B
1In every gram contain 2 * 10
9The effective cypermethrin of spore and 1/220 gram, recombiner B
2In every gram contain 0.4 * 10
9The effective cypermethrin of spore and 1/220 gram, recombiner B
3In every gram contain 2 * 10
9The effective cypermethrin of spore and 1/260 gram, recombiner B
4In every gram contain 0.4 * 10
9The effective cypermethrin of spore and 1/260 gram.
The plantation Plantula Brassicae chinensis is raised radish aphid Lipaphis erysimi in the laboratory; Solution or suspension of respectively processing below the preparation respectively: 1) take by weighing respectively 1g composite insecticide B
1, composite insecticide B
2, composite insecticide B
3With composite insecticide B
4, add water 99mL, again by dilution in 1: 9, obtain their 1000 times diluent; 2) with 0.05% the tween 80 aqueous solution spore powder of bacterial strain BBNS-J9-16 being mixed with concentration gradient is 0.4 * 10
6Spore/mL and 2 * 10
6The spore suspension of spore/mL namely is equal to respectively composite insecticide B
1And B
21000 times of diluents in beauveria bassiana spore content; 3) measure the r of 1mL 4.5%, add water 98mL, by successively dilution in 1: 9,1: 9, obtain 9900 times diluent again, namely be equal to composite insecticide B
1And B
21000 times of diluents in the content of effective cypermethrin; By 11: 2 thin ups 1 time, namely obtain 11700 times diluent again, namely be equal to composite insecticide B
3And B
41000 times suspension in the content of effective cypermethrin.Get respectively the treatment solution 1mL for preparing, to the aphid body, process 100 aphids with Potter spray tower even spraying at every turn, 3 repetitions; Aphid placed under 25 ℃ of temperature to raise supported 7 days, the dead quantity of observed and recorded aphid every day is transferred to dead aphid in the clean culture dish, moisturizing 3-5 days, observes polypide and has or not the growth muscardine and become bombys batryticatus; According to the final mortality ratio of aphid and bombys batryticatus rate, estimate bacterial strain to the effect of aphid.
The results are shown in Table 4, prove that beauveria bassiana and effective cypermethrin recombiner energy Effective Raise is to poisoning mortality ratio and the rate of death of aphid.
Table 4 desinsection host and independent beauveria bassiana or effective cypermethrin compare the virulence of aphid
Annotate:
1Be the contrast correction value, with " mean+SD " expression, suffix letter represents the significance of difference, represents without significant difference α=0.05 with same column comparison same letter;
2Be the bombys batryticatus ratio in the dead borer population."--" expression is not processed or is not existed
Embodiment 8 beauveria bassianas and Rogor composite insecticide are to the toxic action of aphid
The test spore powder of beauveria bassiana BBNS-J9-16, content is 1 * 10
11Spore/g.Agricultural chemicals Rogor is 40% missible oil that Jiangsu Tenglong Biological Pharmaceutical Co., Ltd. produces, and manufacturer recommends the field working concentration to be 800-2000 times of dilution.4 kinds of composite insecticides are the spore powder of beauveria bassiana BBNS-J9-16 and the mixture of Rogor, wherein recombiner C
1In every gram contain 2 * 10
9The Rogor of spore and 1/25 gram, recombiner C
2In every gram contain 0.4 * 10
9The Rogor of spore and 1/25 gram, recombiner C
3In every gram contain 2 * 10
9The Rogor of spore and 1/37.5 gram, recombiner C
4In every gram contain 0.4 * 10
9The Rogor of spore and 1/37.5 gram.
The plantation Plantula Brassicae chinensis is raised radish aphid Lipaphis erysimi in the laboratory.Solution or suspension of respectively processing below the preparation respectively: 1) take by weighing respectively 1g composite insecticide C
1, composite insecticide C
2, composite insecticide C
3With composite insecticide C
4, add water 99mL, again by dilution in 1: 9, obtain the diluent of their 1000 times of dilutions; 2) with 0.05% the tween 80 aqueous solution spore powder of bacterial strain BBNS-J9-16 being mixed with concentration gradient is 0.4 * 10
6Spore/mL and 2 * 10
6The spore suspension of spore/mL namely is equal to respectively composite insecticide C
1And C
21000 times suspension in beauveria bassiana spore content; 3) measure the dimethoate emulsifiable concentrate of 1mL 40%, add water 99mL, by successively dilution in 1: 9,1: 9, obtain 10000 times diluent again, namely be equal to composite insecticide C
1And C
21000 times of diluents in the content of Rogor; By 2: 1 thin ups 1 time, namely obtain 15000 times diluent again, namely be equal to composite insecticide C
3And C
41000 times suspension in the content of Rogor.Get respectively the treatment solution 1mL for preparing, to the aphid body, process 100 aphids with Potter spray tower even spraying at every turn, 3 repetitions; Aphid placed under 25 ℃ of temperature to raise supported 7 days, the dead quantity of observed and recorded aphid every day is transferred to dead aphid in the clean culture dish, moisturizing 3-5 days, observes polypide and has or not the growth muscardine and become bombys batryticatus; According to the final mortality ratio of aphid and bombys batryticatus rate, estimate bacterial strain to the effect of aphid.
The results are shown in Table 5, prove that beauveria bassiana and Rogor recombiner energy Effective Raise is to poisoning mortality ratio and the rate of death of aphid.
Table 5 desinsection host and independent beauveria bassiana or Rogor compare the virulence of aphid
Annotate:
1Be the contrast correction value, with " mean+SD " expression, suffix letter represents the significance of difference, represents without significant difference α=0.05 with same column comparison same letter;
2Be the bombys batryticatus ratio in the dead borer population."--" expression is not processed or is not existed.
Claims (4)
1. a disinsection fungal beauveria bassiana (Beauveria bassiana) BBNS-J9-16, its deposit number is: CGMCC No.5288.
2. the described Strain of Beauveria bassiana BBNS-J9-16 of claim 1 is used for the application of killing aphis, and described aphid is vegetable aphid or flowering plant aphid.
3. a high efficiency composition sterilant is characterized in that, comprises thalline and/or its spore powder of Strain of Beauveria bassiana BBNS-J9-16 claimed in claim 1,
Wherein,
The described high efficiency composition sterilant of every gram contains 0.4 ~ 2 * 10
9The Provado of individual spore and 1/300-1/600 gram;
The described high efficiency composition sterilant of every gram contains 0.4 ~ 2 * 10
9The effective cypermethrin of individual spore and 1/220-1/260 gram; Perhaps
The described high efficiency composition sterilant of every gram contains 0.4 ~ 2 * 10
9The Rogor of individual spore and 1/25-1/37.5 gram.
4. the described high efficiency composition sterilant of claim 3 is used for the application of killing aphis, and described aphid is vegetable aphid or flowering plant aphid.
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| CN111727978A (en) * | 2020-07-22 | 2020-10-02 | 周辛平 | Method for preparing fungus biological insecticide by utilizing hermetia illucens |
| CN112522112B (en) * | 2020-12-07 | 2021-11-05 | 中国农业科学院植物保护研究所 | Fungus strain for preventing and treating alfalfa aphids and application thereof |
| CN117598324A (en) * | 2023-11-21 | 2024-02-27 | 西南大学 | Beauveria bassiana-based pesticide for controlling Bactrocera dorsalis and testing method thereof |
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