CN110305810A - A kind of Bacillus thuringiensis subsp. israelensis culture medium and its application - Google Patents
A kind of Bacillus thuringiensis subsp. israelensis culture medium and its application Download PDFInfo
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- CN110305810A CN110305810A CN201910553544.3A CN201910553544A CN110305810A CN 110305810 A CN110305810 A CN 110305810A CN 201910553544 A CN201910553544 A CN 201910553544A CN 110305810 A CN110305810 A CN 110305810A
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- 241000193365 Bacillus thuringiensis serovar israelensis Species 0.000 title claims abstract description 27
- 239000001963 growth medium Substances 0.000 title claims abstract description 25
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 24
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 24
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 24
- 235000005822 corn Nutrition 0.000 claims abstract description 24
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims abstract description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 22
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000001888 Peptone Substances 0.000 claims abstract description 12
- 108010080698 Peptones Proteins 0.000 claims abstract description 12
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 12
- 229930006000 Sucrose Natural products 0.000 claims abstract description 12
- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 12
- 235000013312 flour Nutrition 0.000 claims abstract description 12
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 12
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 12
- 235000019319 peptone Nutrition 0.000 claims abstract description 12
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 12
- 235000010333 potassium nitrate Nutrition 0.000 claims abstract description 12
- 239000004323 potassium nitrate Substances 0.000 claims abstract description 12
- 239000000843 powder Substances 0.000 claims abstract description 12
- 239000005720 sucrose Substances 0.000 claims abstract description 12
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims abstract description 12
- 229960001763 zinc sulfate Drugs 0.000 claims abstract description 12
- 229910000368 zinc sulfate Inorganic materials 0.000 claims abstract description 12
- 239000011780 sodium chloride Substances 0.000 claims abstract description 11
- 239000008367 deionised water Substances 0.000 claims abstract description 10
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 10
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 10
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 10
- 240000008042 Zea mays Species 0.000 claims description 22
- 239000002994 raw material Substances 0.000 claims 4
- 244000068988 Glycine max Species 0.000 claims 2
- 235000010469 Glycine max Nutrition 0.000 claims 2
- ZGBSOTLWHZQNLH-UHFFFAOYSA-N [Mg].S(O)(O)(=O)=O Chemical compound [Mg].S(O)(O)(=O)=O ZGBSOTLWHZQNLH-UHFFFAOYSA-N 0.000 claims 2
- 241000256118 Aedes aegypti Species 0.000 abstract description 5
- 241000144210 Culex pipiens pallens Species 0.000 abstract description 5
- 230000001018 virulence Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 239000007788 liquid Substances 0.000 abstract description 2
- 238000009629 microbiological culture Methods 0.000 abstract description 2
- 238000011020 pilot scale process Methods 0.000 abstract description 2
- 241000209149 Zea Species 0.000 abstract 2
- 230000003698 anagen phase Effects 0.000 abstract 1
- 241000726221 Gemma Species 0.000 description 7
- 239000013078 crystal Substances 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 241000255925 Diptera Species 0.000 description 4
- 230000028070 sporulation Effects 0.000 description 4
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 3
- 102000002322 Egg Proteins Human genes 0.000 description 3
- 108010000912 Egg Proteins Proteins 0.000 description 3
- 235000014103 egg white Nutrition 0.000 description 3
- 210000000969 egg white Anatomy 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 241000256059 Culex pipiens Species 0.000 description 2
- 241000256113 Culicidae Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000000749 insecticidal effect Effects 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 241000256054 Culex <genus> Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000193386 Lysinibacillus sphaericus Species 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 244000144987 brood Species 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000009655 industrial fermentation Methods 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000035806 respiratory chain Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to microbiological culture media technical fields, specifically disclose a kind of Bacillus thuringiensis subsp. israelensis culture medium, the weight percent of each component is respectively as follows: sucrose 0.5~1%, corn flour 0.6~1.2%, beancake powder 1~5%, peptone 0.1~0.5%, corn pulp 0.4~1.2%, calcium carbonate 0.1~0.5%, potassium dihydrogen phosphate 0.05~0.1%, magnesium sulfate 0.05~0.1%, zinc sulfate 0.05~0.1%, sodium chloride 0.1~0.3%, potassium nitrate 0.01~0.05%, deionized water 89.95~97.04%.The invention also discloses application of the above-mentioned culture medium in culture Bacillus thuringiensis subsp. israelensis.Culture medium of the present invention can promote Bacillus thuringiensis subsp. israelensis to rapidly enter logarithmic growth phase; zymocyte liquid living spores ratio is greater than 90%, high to Aedes aegypti and culex pipiens pallens larvae virulence, to greatly improve product quality; production cost is reduced, is suitable for high-volume pilot scale culture and produces.
Description
Technical field
The present invention relates to a kind of microbiological culture media, in particular to a kind of Bacillus thuringiensis subsp. israelensis culture medium.
Background technique
Bacillus thuringiensis subsp. israelensis is gram-positive bacteria, and thallus both ends blunt circle has flagellum, and movement is active, spore
Capsule does not expand, the short ellipse of gemma or oval, parasporal crystal subcircular.The main mosquito eradication of Bacillus thuringiensis subsp. israelensis is living
Property substance be crystalline protein, generated in the sporulation II phase, there are three types of different type crystal form, be located at exitine outside,
It falls off in growth course with the release of gemma.Crystalline protein by after larval feeding under insect midgut basic protein enzyme effect
Toxic peptide is dissociateed, respiratory chain normal function is destroyed, causes mosquito larvae dead.Bacillus thuringiensis subsp. israelensis is to a variety of mosquito childrens
Worm poisoning power with higher, is the microorganism for being widely used in mosquito control in world wide.It the advantage is that controllable 1~
4 instar larvaes, quick after application (4~for 24 hours), holding effect is long (7~30d), is suitable in addition to drinking water various water bodys and (including pours
Water) mosquito class prevention and treatment, in water body can natural decomposition, it is pollution-free, do not influence nontarget organism, to operator, wild animal and
Domestic animal is without side-effects.
The fermentation medium of Bacillus thuringiensis subsp. israelensis at present, sporulation relatively low in the prevalence of fermentation level
It is inconsistent, the defects of unstable product quality.It is general in actual production to take the temperature adjusted in fermentation process more, pressure, lead to
The process conditions such as gas agitating are to improve living bacteria count amount, or use high-temperature process to the brood cell of strain, are allowed in industrial fermentation
It can synchronize to form gemma in the process, to improve the quantity and ratio of sporulation, to shorten fermentation time, but these methods are real
Border is complex for operation step, and increases production cost.In addition, Bacillus thuringiensis subsp. israelensis is to albopictus larvae toxic effect
Preferably, Bacillus sphaericus is worse than to Ability of Culex Pipiens Larvae toxic effect, improves it at present and the research of culex virulence is mostly sieved from bacterial strain
Choosing, transgenic engineered bacteria building etc. are implemented, and increase research and development cost, and operating difficulties should not succeed.
Summary of the invention
The purpose of the present invention is to provide a kind of culture medium of suitable Bacillus thuringiensis subsp. israelensis growth and its application,
The quantity and ratio of sporulation are improved, fermentation time is shortened, the insecticidal effect to Ability of Culex Pipiens Larvae is improved, enhances its application.
In order to achieve the above objectives, the present invention provides a kind of Bacillus thuringiensis subsp. israelensis culture mediums, wherein each component
Weight percent be respectively as follows: sucrose 0.5~1%, corn flour 0.6~1.2%, beancake powder 1~5%, peptone 0.1~
0.5%, corn pulp 0.4~1.2%, calcium carbonate 0.1~0.5%, potassium dihydrogen phosphate 0.05~0.1%, magnesium sulfate 0.05~
0.1%, zinc sulfate 0.05~0.1%, sodium chloride 0.1~0.3%, potassium nitrate 0.01~0.05%, deionized water 89.95~
97.04%.
The formula of the Bacillus thuringiensis subsp. israelensis culture medium includes following weight hundred as a preferred technical solution,
Divide the component of ratio: sucrose 0.5~1%, corn flour 0.8~1%, beancake powder 2~4%, peptone 0.1~0.5%, corn pulp
0.6%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.05~0.1%, magnesium sulfate 0.05~0.1%, zinc sulfate 0.05~0.1%,
Sodium chloride 0.2%, potassium nitrate 0.01~0.05%, deionized water 92.25~95.54%.
The formula of the Bacillus thuringiensis subsp. israelensis culture medium includes following weight hundred as a preferred technical solution,
Divide the component of ratio: sucrose 0.6%, corn flour 0.8%, beancake powder 3%, peptone 0.3%, corn pulp 0.6%, calcium carbonate
0.1%, potassium dihydrogen phosphate 0.08%, magnesium sulfate 0.1%, zinc sulfate 0.1%, sodium chloride 0.2%, potassium nitrate 0.01%, go from
Sub- water 94.11%.
The formula of the Bacillus thuringiensis subsp. israelensis culture medium includes following weight hundred as a preferred technical solution,
Divide the component of ratio: sucrose 0.9%, corn flour 0.8%, beancake powder 3%, peptone 0.5%, corn pulp 0.6%, calcium carbonate
0.1%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.08%, zinc sulfate 0.08%, sodium chloride 0.2%, potassium nitrate 0.03%, go from
Sub- water 93.61%.
The present invention also provides application of the above-mentioned culture medium in culture Bacillus thuringiensis subsp. israelensis.
The beneficial effects of the present invention are: culture medium of the present invention can promote Bacillus thuringiensis subsp. israelensis to rapidly enter logarithm
Growth period, zymocyte liquid living spores ratio are greater than 90%, high to Aedes aegypti and culex pipiens pallens larvae virulence, to greatly improve
Product quality reduces production cost, is suitable for high-volume pilot scale culture and produces.
Specific embodiment
In order to solve above-mentioned purpose, the present invention enumerates following embodiment and is illustrated.
Embodiment 1
With the following culture media shaking vase culture of weight percentages of components: sucrose 0.5%, corn flour 0.6%, beancake powder 1%, egg
White peptone 0.1%, corn pulp 0.4%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.05%, magnesium sulfate 0.05%, zinc sulfate 0.05%,
Sodium chloride 0.1%, potassium nitrate 0.01%, deionized water 97.04%.
It is sub-packed in 500mL triangular flask, every bottle of 50mL, 0.1MPa sterilizing 30min.1 ring lawn of picking is inoculated in shaking flask, rotation
Rotatable shaking table shake culture, 30 DEG C of temperature, revolving speed 220r/min, culture to gemma and crystal 40% separate shaking table at present, microscopy
Spore development situation.Microscopy shows that thalli morphology is preferable, and sporangiocyst formation rate 98%, crystal size is consistent, and development sync rates reach
93%.
Embodiment 2
With the following culture media shaking vase culture of weight percentages of components: sucrose 0.6%, corn flour 0.8%, beancake powder 3%, egg
White peptone 0.3%, corn pulp 0.6%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.08%, magnesium sulfate 0.1%, zinc sulfate 0.1%, chlorine
Change sodium 0.2%, potassium nitrate 0.01%, deionized water 94.11%.
It is sub-packed in 500mL triangular flask, every bottle of 50mL, 0.1MPa sterilizing 30min.1 ring lawn of picking is inoculated in shaking flask, rotation
Rotatable shaking table shake culture, 30 DEG C of temperature, revolving speed 220r/min, culture to gemma and crystal 40% separate shaking table at present.
Embodiment 3
With the following culture media shaking vase culture of weight percentages of components: sucrose 0.9%, corn flour 0.8%, beancake powder 3%, egg
White peptone 0.5%, corn pulp 0.6%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.081%, zinc sulfate 0.08%,
Sodium chloride 0.2%, potassium nitrate 0.03%, deionized water 93.61%.
It is sub-packed in 500mL triangular flask, every bottle of 50mL, 0.1MPa sterilizing 30min.1 ring lawn of picking is inoculated in shaking flask, rotation
Rotatable shaking table shake culture, 30 DEG C of temperature, revolving speed 220r/min, culture to gemma and crystal 40% separate shaking table at present.
Experimental subjects
The Bacillus thuringiensis subsp. israelensis shake flask fermentation culture solution of Example 1-3, bioassay is to 3 age of Aedes aegypti
Larva, 4 instar larvae of Culex pipiens pallens virulence effect.
Experimental result
Table 1 is toxicity test result of each group to Aedes aegypti and culex pipiens pallens larvae.
As can be seen from Table 1, the Bacillus thuringiensis subsp. israelensis of three embodiments is equal to two kinds of mosquito larvae virulence effects
Preferably, wherein in embodiment 2 to Aedes aegypti larva LC50 up to 0.47ppm, be to culex pipiens pallens larvae LC50 in embodiment 3
0.52ppm。
To sum up, culture medium of the invention is used for the culture of Bacillus thuringiensis subsp. israelensis, and gemma yield is high, at low cost,
And obtained culture solution insecticidal activity with higher, stability is good, application value with higher.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification made, equivalent replacement, improvement etc. be should all be included in the protection scope of the present invention.
Claims (5)
1. a kind of Bacillus thuringiensis subsp. israelensis culture medium, which is characterized in that described to be trained by Bacillus thuringiensis subsp. israelensis
Support base be made by each raw material of following weight percent: sucrose 0.5~1%, corn flour 0.6~1.2%, beancake powder 1~5%,
Peptone 0.1~0.5%, corn pulp 0.4~1.2%, calcium carbonate 0.1~0.5%, potassium dihydrogen phosphate 0.05~0.1%, sulfuric acid
Magnesium 0.05~0.1%, zinc sulfate 0.05~0.1%, sodium chloride 0.1~0.3%, potassium nitrate 0.01~0.05%, deionized water
89.95~97.04%.
2. Bacillus thuringiensis subsp. israelensis culture medium according to claim 1, which is characterized in that the Dipel
Israel subclass culture medium is made by each raw material of following weight percent: sucrose 0.5~1%, corn flour 0.8~1%, soya-bean cake
Powder 2~4%, peptone 0.1~0.5%, corn pulp 0.6%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.05~0.1%, sulfuric acid
Magnesium 0.05~0.1%, zinc sulfate 0.05~0.1%, sodium chloride 0.2%, potassium nitrate 0.01~0.05%, deionized water 92.25
~95.54%.
3. Bacillus thuringiensis subsp. israelensis culture medium according to claim 1 or 2, which is characterized in that the Su Yunjin
Thuringiensis israelensis kind culture medium is made by each raw material of following weight percent: sucrose 0.6%, corn flour 0.8%, beancake powder
3%, peptone 0.3%, corn pulp 0.6%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.08%, magnesium sulfate 0.1%, zinc sulfate
0.1%, sodium chloride 0.2%, potassium nitrate 0.01%, deionized water 94.11%.
4. Bacillus thuringiensis subsp. israelensis culture medium according to claim 1 or 2, which is characterized in that the Su Yunjin
Thuringiensis israelensis kind culture medium by each raw material of following weight percent by being made: sucrose 0.9%, corn flour 0.8%, soya-bean cake
Powder 3%, peptone 0.5%, corn pulp 0.6%, calcium carbonate 0.1%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.08%, zinc sulfate
0.08%, sodium chloride 0.2%, potassium nitrate 0.03%, deionized water 93.61%.
5. application of the described in any item culture mediums of claim 1-4 in culture Bacillus thuringiensis subsp. israelensis.
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Cited By (1)
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