CN104099277B - A kind of bacillus thuringiensis for preventing and treating Bradysia odoriphaga and application thereof - Google Patents
A kind of bacillus thuringiensis for preventing and treating Bradysia odoriphaga and application thereof Download PDFInfo
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Abstract
The invention discloses a kind of bacillus thuringiensis (Bacillus thuringiensis) the bacterial strain JQ23 for preventing and treating Bradysia odoriphaga belonging to field of biological control, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 24th, 2014, and deposit number is CGMCC No.9375.The invention also discloses a kind of microbial insecticide containing JQ23 and preparation method thereof.Bacillus thuringiensis JQ23 of the present invention has the highest virulence to olaiphage larva in Chinese Chive, and the preventing and treating for Bradysia odoriphaga provides a kind of effective method；Secondly, bacillus thuringiensis of the present invention belongs to natural product, nontoxic to people and animals, environmentally safe；Additionally, microbial insecticide preparation method of the present invention is simple, easy to use.
Description
Technical field
The invention belongs to field of biological control, be specifically related to a kind of Su Yun gold spore bar for preventing and treating Bradysia odoriphaga
Bacterium, and the application that this bacillus thuringiensis is in vegetable-crop pest-insect prevents and treats.
Background technology
Bradysia odoriphaga (Bradysia odoriphaga) has another name called fragrant-flowered garlic maggot, yellow foot fungus gnat, belongs to Diptera, eye Sciaridae,
The liliaceous vegetables such as main harm Folium Allii tuberosi, Herba Alii fistulosi, Bulbus Allii Cepae, Herba Alii fistulosi, Bulbus Allii, the most also endanger Caulis et Folium Lactucae sativae, green vegetable, Herba Apii graveolentis etc., are Herba Alii fistulosis
One of primary pest of Bulbus Allii class vegetable, the most all has distribution.The larva of Bradysia odoriphaga lives in upper soll layer, group
Collect at the bulb of Folium Allii tuberosi underground part and tender stem's damage.Newly hatched larvae is first caused harm the upper of Folium Allii tuberosi leaf sheath base and bulb
End；Mainly cause harm in spring, autumn tender stem, cause rhizome to rot.Folium Allii tuberosi aerial growth of being injured is thin and delicate, under yellow leaf is wilted
Hang down, the last withered and yellow death of fragrant-flowered garlic leaf.During summer temperature height, larva moves down, Folium Allii tuberosi bulb of causing harm, and causes whole bulb to rot,
Time serious, whole pier Folium Allii tuberosi is withered.For the prevention and controls of Bradysia odoriphaga mainly have physical control (sweet-and-sour liquid, ozone etc.) and
Chemical prevention.Owing to physical control method exists complex operation, the prevention effect shortcoming such as big affected by environment, production is mainly adopted
Prevent and treat with chemical means such as pesticide pouring roots, but owing to the breeding of fragrant-flowered garlic maggot is fast, disguised by force, difficulty of prevention and cure is big, cause vegetable grower
High-toxic pesticide lack of standardization or excessive, even Misuse disabling is prevented and treated, and causes pesticide residues to exceed standard, pollutes environment, danger
Evil human health and murder the problems such as useful birds, on the other hand, long-term a large amount of use chemical pesticides strengthen pest resistance to insecticide,
Pollute environment, kill birds, cause the rampant once again vicious cycle of insect, it is impossible to reach the effective control to pest population, with
Shi Zaocheng Folium Allii tuberosi agriculture is residual to exceed standard, and makes consumer health be on the hazard.Poison Folium Allii tuberosi event takes place frequently, such as in recent years: in April, 2004 is " fragrant
River poison Folium Allii tuberosi is gone to the capital event ", in April, 2010 " Qingdao poison Folium Allii tuberosi " and in March, 2011 Nanyang, henan " poison Folium Allii tuberosi event ", continuous print
Poison Folium Allii tuberosi exposure event so that fragrant-flowered garlic complexion changed " is talked " by consumer, has had a strong impact on the economic interests of vegetable grower and the food peace of consumer
Entirely.
Biological control due to have specificity strong, to people and animals' nonhazardous, do not exist and develop immunity to drugs and environmental pollution etc.
Problem, is increasingly subject to the favor of people.In order to reduce the chemical pesticide pollution to environment and the threat to people's health, it is right to screen
Fragrant-flowered garlic maggot has highly pathogenic Biocontrol microorganism to be effective ways and the important measures preventing and treating fragrant-flowered garlic maggot.Bacillus thuringiensis
(Bacillus thuringiensis is called for short Bt) is a kind of Gram-positive separated in soil or dead insects body
Antibacterial, this type of bacterium can produce one or more to Lepidoptera, coleoptera, Diptera, Homoptera, film wing at sporulation period
The various pests such as mesh, Orthoptera, Isoptera, Trichoptera or Thysanoptera have the parasporal crystal protein of insecticidal activity
(Insecticidal Crystal Proteins is called for short ICPs).This insecticidal crystal protein is the most single-minded to target pest,
To person poultry harmless, free from environmental pollution, thus bacillus thuringiensis is widely used in the Biological control of insect.
The research utilizing Bacillus thuringiensis Prevention fragrant-flowered garlic maggot is relatively fewer, such as the effect of Bacillus thuringiensis Prevention fragrant-flowered garlic maggot
Fruit up to about 75% (Cheng Xuemei etc. plant protection technology and popularization.04 phase in 1998) or 84.85% (solution dimension star .2006, Central China
Agriculture university's Master's thesis).
From the foregoing, Bacillus thuringiensis Prevention Bradysia odoriphaga is utilized to have applications well prospect.
Summary of the invention
Present invention aim at providing a kind of bacillus thuringiensis for preventing and treating Bradysia odoriphaga.This bacterial strain is to double
Wing mesh eye Sciaridae insect Bradysia odoriphaga has high poisoning ability.
Another object of the present invention is to provide a kind of microbial insecticide.
The present invention the 3rd purpose is to provide mentioned microorganism method for producing insecticide.
The present invention the 4th purpose is to provide the purposes of above-mentioned bacillus thuringiensis.
Realize technical scheme as follows:
One bacillus thuringiensis of the present invention (Bacillus thuringiensis) bacterial strain JQ23, this bacterial strain in
On June 24th, 2014 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: Beijing
North Star West Road, Chaoyang District 1 No. 3 Institute of Microorganism, Academia Sinica of institute, deposit number is CGMCC No.9375.
A kind of microbial insecticide, containing above-mentioned bacillus thuringiensis (Bacillus thuringiensis) bacterial strain
JQ23。
Mentioned microorganism insecticide, its dosage form is liquid dosage form.
Mentioned microorganism insecticide, containing above-mentioned bacillus thuringiensis (Bacillus in its described liquid dosage form
Thuringiensis) spore concentration of bacterial strain JQ23 is 107～108cfu/ml。
Mentioned microorganism method for producing insecticide, comprises the steps:
(1), actication of culture: bacillus thuringiensis (Bacillus thuringiensis) the bacterial strain JQ23 preserved is existed
12～16h are activated on LB plating medium, at 29～31 DEG C；Picking list bacterium colony is on LB slant medium, at 29～31 DEG C
Lower cultivation 3～5 days, obtain the bacterial strain of activation；
(2), prepared by seed liquor: scrape the inoculation activated in a little ring step (1) to LB liquid with aseptic inoculation ring
In culture medium 50ml, 29～31 DEG C, rotating speed be to cultivate 10～14h under the conditions of 200～220rpm/min, obtain seed liquor；
(3), fermentation culture: according to volume ratio be 1～2% ratio the seed liquor of step (2) gained is linked into Semen Glycines
In cake starch powder culture medium, 29～31 DEG C, rotating speed be to cultivate 48～60h under the conditions of 200～220rpm/min, fermented
Liquid；
(4), the fermentation liquid of dilute with water step (3) gained, regulation pH value to 6.6～6.8, adds 0.02～0.1%
Sodium benzoate, obtains microbial insecticide；Or by the fermentation liquid of step (3) gained 4 DEG C, rotating speed be 5000rpm/min bar
Under part, centrifugal 30min, takes precipitation, obtains precipitate, then dilute with water precipitate, and regulation pH value, to 6.6～6.8, adds 0.02%
～the sodium benzoate of 0.1%, obtain microbial insecticide.
LB plating medium described in above-mentioned preparation method step (1) or LB slant medium are those skilled in the art
Known conventional culture medium, its constituent and part by weight thereof be: tryptone 10g, yeast extract 5g, sodium chloride
10g, agar powder 15g, water 1L, pH7.2.
The conventional cultivation that LB fluid medium described in above-mentioned preparation method step (2) is well known to those skilled in the art
Base, its constituent and part by weight thereof be: tryptone 10g, yeast extract 5g, sodium chloride 10g, water 1L, pH7.2.
Constituent and the percentage by weight thereof of the soybean cake powder culture medium described in above-mentioned preparation method step (3) be:
Soybean cake powder 2.5%, cottonseed meal 1.2%, corn starch 1.5%, yeast powder 0.5%, KH2PO40.03%,
MgSO40.05%, water 1L.
Above-mentioned bacillus thuringiensis JQ23 application on preventing and treating vegetable-crop pest-insect.
Described vegetable-crop pest-insect refers to Diptera eye Sciaridae insect.
Described eye Sciaridae insect refers to Bradysia odoriphaga (Bradysia odoriphaga).
The process of bacterial strain JQ23 of the present invention collection screening and taxonomic identification thereof:
(1) process and the typoiogical classification that gather screening are identified:
Bacterial strain JQ23 of the present invention is that in April, 2013 is gathered by Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie in Henan Province
Soil sample in the inoculation that is separated at LB solid medium (tryptone 10g, yeast extract 5g, sodium chloride 10g, fine jade
Cosmetics 15g, water 1L, pH7.2) on cultivate under the conditions of 30 DEG C, it is special that different time sampling microscopy observes bacterium colony (see Fig. 1) form
Levy, crystal characteristic etc..LB culture medium is cultivated different phase observed result as follows, trophosome: in shaft-like, the blunt circle in two ends, greatly
Little about 1.1～1.5 × 0.5 μm；Single or two or more is that chain exists.Spore: oval, size about 1.0～1.2 × 0.5 μ
M, for hypopus；To high temperature or be dried etc. poor environment have stronger resistance.Parasporal crystal: spherical, diameter about 0.4 μm,
There is insecticidal activity.(the elegant pearl in east etc. are write, Science Press for these morphological characteristics and " common bacteria system identification handbook "
.2001 year) described in bacillus cereus morphological characteristic basically identical, tentatively judge that this bacterial strain belongs to bacillus thuringiensis
(Bacillus thuringiensis), and by its named JQ23.
(2) utilize gyrB gene order that JQ23 bacterial strain carries out molecular identification and classification:
By JQ23 inoculation in LB culture fluid, 30 DEG C of shaken overnight, take 1mL bacterium solution in Eppenddorf pipe, centrifugal,
Abandoning supernatant, 200 μ L ultra-pure waters suspend, and 100 DEG C process 10min, 12000rpm and are centrifuged 5min, take supernatant and be DNA profiling.Utilize
Bacillus cereus gyrB-F and gyrB-R is that primer carries out PCR amplification, obtains pcr amplification product；Wherein said primer sequence is:
GyrB-F:5 ' TTGRCGGHRGYGGHTATAAAGT3 ' (SEQ ID No.1),
GyrB-R:5 ' TCCDCCSTCAGARTCWCCCTC3 ' (SEQ ID No.2)；
The pcr amplification reaction system (50 μ L) of gyrB: 10 × PCR Buffer (Mg2+)5μL；dNTP Mixture
(2.5mM) 5 μ L, each 1 μ L of primer (10 μm ol/L), template 1 μ L, Taq polymerase (5U/ μ L) 1 μ L, ultra-pure water is mended to 50 μ L.Instead
The program is answered to be: 95 DEG C of 5min；95 DEG C of 30s, 55 DEG C of 45s, 72 DEG C of 1min, 30 circulations；72℃10min.Amplified production is delivered
Shanghai Sheng Gong biological engineering company limited checks order, and obtains the gyrB gene order (see SEQ ID No.3) of JQ23 bacterial strain.Will
The gyrB gene order of JQ23 bacterial strain carries out tetraploid rice in Genbank, found that JQ23 and bacillus thuringiensis
GyrB gene homology the highest, reach 99%；Utilize MEGA software building phylogenetic tree, result (see Fig. 3) simultaneously
JQ23 bacterial strain condenses together with bacillus thuringiensis, illustrates that JQ23 bacterial strain is bacillus thuringiensis (Bacillus
Thuringiensis), and different from any other bacterial strain, it is a new strains.
(3) utilize phoR gene order that JQ23 bacterial strain carries out molecular identification and classification:
Extract the DNA of JQ23 bacterial strain, then with the DNA of JQ23 bacterial strain as template, enter for primer with phoR-F and phoR-R
Performing PCR expands, and obtains pcr amplification product；Wherein said primer sequence is:
PhoR-F:5 ' TTYARYTCATGRGAVACATT3 ' (SEQ ID No.4)；
PhoR-R:5 ' GGNTAYAAANARGAGGAGCC3 ' (SEQ ID No.5)；
The PCR reaction system (50 μ L) of phoR: 10 × PCR Buffer (Mg2+) 5 μ L, dNTP Mixture (2.5mM) 5 μ
L, each 2 μ L of primer (10 μm ol/L), template 1 μ L, Taq polymerase (5U/ μ L) 1 μ L, ultra-pure water is mended to 50 μ L.Response procedures is:
95℃5min；95 DEG C of 30s, 50 DEG C of 45s, 72 DEG C of 1min, 30 circulations；72℃10min.Gained amplified production is delivered Shanghai raw
Work biological engineering company limited checks order, and obtains the phoR gene order (see SEQ ID No.6) of JQ23 bacterial strain.By JQ23's
PhoR gene order (see SEQ ID No.6) carries out tetraploid rice in Genbank, found that JQ23 bacterial strain and Su Yunjin
The phoR gene homology of bacillus cereus is the highest, reaches 99%；Utilize MEGA software building phylogenetic tree, result simultaneously
(see Fig. 4) JQ23 bacterial strain condenses together with bacillus thuringiensis, illustrates that JQ23 bacterial strain belongs to Su Yun gold spore in classification
Bacillus (Bacillus thuringiensis), and different from any other bacterial strain, it is a new strains.
The using method of JQ23 microbial insecticide of the present invention: JQ23 microbial insecticide is directly watered and is diluted to 1 ×
107Cfu～1 × 108Cfu/ml, prevents and treats in fragrant-flowered garlic maggot early period of origination pouring root, continuous pouring root 3 times, every minor tick 5-7d.
The present invention has the advantage that or beneficial effect: bacillus thuringiensis bacterial strain JQ23 of the present invention is to Bradysia odoriphaga
Larva has the highest virulence, and its bacterial concentration is 1 × 107During more than cfu/mL, to the olaiphage larva in Chinese Chive correction of 7 days
Mortality rate reaches more than 83.84%；JQ23 microbial insecticide 1 × 107Under cfu/mL concentration, field Revision insect recluced rate reaches
66.03%, prevention effect reaches 85.09%；The present invention is that the preventing and treating of Bradysia odoriphaga provides a kind of effective, harmless
, the method for environmental protection；Secondly, bacillus thuringiensis of the present invention belongs to natural product, nontoxic to people and animals, environmentally safe,
Prevent and treat for vegetable-crop pest-insect as microbial insecticide, belong to environmentally friendly insecticide, opening of this microbial insecticide
Send out and utilize and again occur effectively stopping " poison Folium Allii tuberosi event ", ensure food safety；Additionally, the microorganism of JQ23 of the present invention is killed
Worm agent preparation method is simple, easy to use.
Accompanying drawing explanation
Fig. 1. for JQ23 bacterial strain bacterium colony photo.
Fig. 2. the olaiphage larva in Chinese Chive for JQ23 strain infection contrasts photo, and wherein left side is slow for healthy Folium Allii tuberosi
Eye fungus gnat larva, right side for the olaiphage larva in Chinese Chive by JQ23 strain infection.
Fig. 3. according to gyrB gene order build JQ23 phylogenetic tree.
Fig. 4. according to phoR gene order build JQ23 phylogenetic tree.
Detailed description of the invention
The preparation of embodiment 1:JQ23 strain microorganism insecticide
Carry out as follows:
(1) actication of culture: bacillus thuringiensis (Bacillus thuringiensis) the bacterial strain JQ23 that will preserve
((it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 24th, 2014, preservation address:
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, deposit number is CGMCC No.9375))
At LB plating medium, (its constituent and part by weight thereof be: tryptone 10g, yeast extract 5g, sodium chloride 10g, fine jade
Cosmetics 15g, water 1L, pH7.2) upper 29 DEG C of activation 13h, picking list bacterium colony is in LB slant medium ((its constituent and weight thereof
Ratio is: tryptone 10g, yeast extract 5g, sodium chloride 10g, agar powder 15g, water 1L, pH7.2) upper 30 DEG C cultivate 3 days,
The bacterial strain that must activate.
(2) preparation of seed liquor: scrape, with aseptic inoculation ring, the inoculation that a little ring step (1) activates and train to LB liquid
Support base (its constituent and part by weight thereof are: tryptone 10g, yeast extract 5g, sodium chloride 10g, water 1L, pH7.2)
In 50ml, 30 DEG C, shaking speed be 200rpm/min under conditions of cultivate 12h, obtain seed liquor.
(3) fermentation culture: the seed liquor of step (2) is linked into soyabean cake starch powder according to the ratio that volume ratio is 1%
Culture medium (its constituent and percentage by weight thereof be: soybean cake powder 2.5%, cottonseed meal 1.2%, corn starch 1.5%,
Yeast powder 0.5%, KH2PO40.03%, MgSO40.05%, add water mixed dissolution, adjusts pH7.2,121 DEG C of sterilizing 30min, then drops
Standby to 30 DEG C) in, at 30 DEG C, rotating speed is that 48h cultivated by 200rpm/min shaking table, detects thalline and Number of spores in fermentation liquid,
In fermentation liquid, born of the same parents' crystalline substance separates and stops fermentation culture when reaching 95%, obtains fermentation liquid (hereinafter referred to as " bacterium solution ")；Through dilution plate
Viable count in count method fermentation liquid is more than 5.46 × 108cfu/ml。
(4) by step (3) gained fermentation liquid regulation pH value to 6.6～6.8, the sodium benzoate of 0.02%～0.1% is added,
Obtain microbial insecticide；Or by fermentation liquid 4 DEG C, rotating speed be 5000rpm/min under the conditions of centrifugal 30min, discard supernatant
Liquid, gained is precipitated as JQ23 spore crystalline substance mixture, and dilute with water thalline JQ23 spore crystalline substance mixture is to 1 × 107Cfu/ml concentration, regulation
PH value, to 6.6～6.8, adds the sodium benzoate of 0.02%～0.1%, is microbial insecticide.
Embodiment 2: JQ23 bacterial strain of the present invention preventing and treating olaiphage larva in Chinese Chive indoor biometrics test
(1) EXPERIMENTAL DESIGN:
JQ23 bacterium solution water embodiment 1 prepared is diluted to 5 × 10 respectively8、1×108、1×107、1×106、1×
105Five Concentraton gradient of cfu/ml.
Comparison: sterilized water
(2) test method:
(1) pouring in disposable plastic culture dish by 1.5% water agar of sterilizing, thickness is 3mm, upper berth two layers of filter paper；
(2) Folium Allii tuberosi is cut to the Folium Allii tuberosi section that size is 10mm, is immersed in 20min in JQ23 bacterium solution prepared by embodiment (1),
Taking-up was soaked the Folium Allii tuberosi section of JQ23 bacterium solution and was put in plate.
(3) picking olaiphage larva in Chinese Chive in the same size, bouncing is seeded in step (2) the Folium Allii tuberosi section containing bacterium
On, with sterilized water for comparison.It is placed in 25 DEG C, raises 7d under the conditions of photoperiod 14h:10h (L:D).Often process 30 examination worms, if 3
Secondary repetition, regular inquiry death borer population after dispenser.Data statistic analysis application SPSS18.0 software carries out data process.
The result of the test of table 1 bacterial strain of the present invention JQ23 preventing and treating Bradysia odoriphaga 2 instar larvae
Note: with string difference letter representation significant difference.
Result (being shown in Table 1) JQ23 bacterial concentration is more than 1 × 107During spore/mL, JQ23 was to olaiphage larva in Chinese Chive 3 days
Corrected mortality be more than 78.9%.Illustrate that JQ23 bacterial strain has the highest insecticidal activity to olaiphage larva in Chinese Chive.
Embodiment 3, bacillus thuringiensis JQ23 of the present invention prevent and treat Bradysia odoriphaga field comparison test
(1) test medicine
(1), processing JQ23 bacterium solution prepared by I: embodiment 1, concentration is 1 × 107cfu/ml。
(2), II: chlorpyrifos ec (480g/L) (pesticide company limited of Qingdao Nintaus), 1000 times of liquid of dilute with water are processed.
(3), III: clear water (blank) is processed
(2) test period and place: in April, 2014 is big in the greenhouse of Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie
Carry out in canopy.
(3) test method: by the Chinese chive field of chamber planting, be divided into 20m2/ community, often process be repeated 3 times, take with
Machine group arranging method is placed.At fragrant-flowered garlic maggot early period of origination, using pouring root method to process by each process in (), every community is used
Water yield 1.5L, interval 7d is repeated once, and dispenser 3 times, uses random 5 sampling methods, every some 0.2m altogether2, investigation is used for the first time
7d larva insect population number after the medicine prelarva last dispenser of insect population cardinal sum, calculates average population decline rate and averagely prevents and treats effect
Really.Computing formula is:
Insect density × 100% before Revision insect recluced rate %=(insect density after insect density-dispenser before dispenser)/dispenser.
(Revision insect recluced rate of process-blank Revision insect recluced rate)/(1-blank insect population subtracts prevention effect %=
Move back rate) × 100%.
Result (being shown in Table 2) bacillus thuringiensis JQ23 bacterial concentration is 1 × 107During cfu/ml, right after continuous dispenser 3 times
The Revision insect recluced rate of fragrant-flowered garlic maggot reaches 66.03%, and prevention effect reaches 85.09%, and field control effect is notable；Meanwhile, Su Yunjin
Bacillus cereus belongs to biological pesticide, to person poultry safety's nonhazardous, ensures food safety, environmentally safe, meets China and can hold
Continuous demand for development, therefore has preferable application prospect.
Table 2 JQ23 of the present invention bacterial strain preventing and treating Bradysia odoriphaga comparative test result
Claims (5)
1. bacillus thuringiensis (Bacillus thuringiensis) bacterial strain JQ23, this bacterial strain is in June, 2014
Within 24th, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.9375.
2. a microbial insecticide, it is characterised in that containing the bacillus thuringiensis (Bacillus described in claim 1
Thuringiensis) bacterial strain JQ23.
3. according to the microbial insecticide described in claim 2, it is characterised in that its dosage form is liquid dosage form.
4. according to the microbial insecticide described in claim 3, it is characterised in that its described liquid dosage form is wanted containing having the right
The spore concentration seeking the bacillus thuringiensis described in 1 (Bacillus thuringiensis) bacterial strain JQ23 is 107~
108cfu/ml。
5. the bacillus thuringiensis bacterial strain JQ23 described in claim 1 is at preventing and treating Bradysia odoriphaga (Bradysia
Odoriphaga) application on.
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| CN105145488B (en) * | 2015-07-08 | 2019-09-06 | 山东省农业科学院植物保护研究所 | A method of it traps and monitors bradysia odoriphaga larva |
| RU2586720C1 (en) * | 2015-08-10 | 2016-06-10 | Олег Иванович Квасенков | Method for producing kvass |
| CN105936644B (en) * | 2016-07-09 | 2021-11-16 | 河北省农林科学院植物保护研究所 | Insecticidal protein and nucleotide sequence and application thereof |
| CN108061782A (en) * | 2018-02-09 | 2018-05-22 | 滨州学院 | A kind of Dipel is to the indoor biometrics method of Bradysia odoriphaga |
| CN110004098B (en) * | 2019-05-22 | 2022-07-29 | 江苏省农业科学院 | Bacillus thuringiensis and application thereof |
| CN112175051B (en) * | 2020-09-03 | 2022-05-20 | 武汉科诺生物科技股份有限公司 | Rapid purification method of low-content Bt crystal protein |
| CN112501083B (en) * | 2020-12-22 | 2023-01-03 | 武汉科诺生物科技股份有限公司 | Bacillus thuringiensis liquid fermentation culture medium and fermentation culture method |
| CN116064321A (en) * | 2022-11-25 | 2023-05-05 | 江苏师范大学 | Chinese chive endophytic Paenibacillus polymyxa and application thereof |
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