CN101440362A - Use of UV induction mutation for reinforcing cracking performance of Bdellovibrio - Google Patents

Use of UV induction mutation for reinforcing cracking performance of Bdellovibrio Download PDF

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CN101440362A
CN101440362A CNA2008102204883A CN200810220488A CN101440362A CN 101440362 A CN101440362 A CN 101440362A CN A2008102204883 A CNA2008102204883 A CN A2008102204883A CN 200810220488 A CN200810220488 A CN 200810220488A CN 101440362 A CN101440362 A CN 101440362A
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bdellovibrio
ultraviolet
application
cracking
bacteria
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CNA2008102204883A
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CN101440362B (en
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蔡俊鹏
韩晓宁
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华南理工大学
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Abstract

The invention belongs to the technical field of mutation breeding of microorganisms, and particularly relates to application of ultraviolet-induced mutation in the strengthening pyrolysis performance of bdellovibrio bacteria. The application takes the ultraviolet-induced mutation as a main way and is assisted with an infrared light source and LiCl enhanced mutagenesis to perform induced mutation under the slow stirring, the time is between 2 and 8 minutes, and the radiation distance is between 5 and 20 centimeters; and the bdellovibrio bacteria is preserved under dark light and ice bath conditions and is accumulatively mutagenized until the requirement is achieved. The application has a simple method and convenient operation, has remarkable effect on strengthening the pyrolysis performance of the bdellovibrio bacteria, can pyrolyze two salmonella strains which fail to be pyrolyzed originally and a plurality of Gram-positive bacteria (including bacillus aerogenes capsulatus, clostridium difficile, streptococcus suis and enterococcus) proven by the experiment, solve the problems that usually the bdellovibrio bacteria can not pyrolyze the Gram-positive bacteria and the pyrolysis ability is extremely weak even if the bdellovibrio bacteria can pyrolyze the Gram-positive bacteria, and widen the application range of the bdellovibrio bacteria.

Description

The application of uv induction sudden change in strengthening the Bdellovibrio cracking performance
Technical field
The invention belongs to the microorganism mutation breeding technical field, be specifically related to the application of uv induction sudden change in strengthening the Bdellovibrio cracking performance.
Background technology
To the microorganism of occurring in nature, if just separate simply and purifying, choose bacterial classification according to qualifications, be difficult to obtain excellent species, and generally be difficult to meet the needs of production.And Microbial Breeding is meant under artificial condition, and the hereditary property of utilizing factors such as physics, chemistry to bring out microorganism morphs, and therefrom selects satisfactory excellent species with specialized character to cultivate, and makes microorganism effectively be applied to industrial production.
Bdellovibrio can be infected, the characteristic of cracking host bacteria makes it to be suitable as the biopurification factor of restraining or removing pathogenic bacterium in food and the environment.But, studies show that, generally speaking, Bdellovibrio can the most of sections of cracking, the Gram-negative bacteria of genus, but its cracking performance is different because of different strains, and they can not the cracking gram-positive microorganism, even if can cracking only a few gram-positive microorganism, cracking ability also extremely weak (Cao Haipeng, Yang Xianle etc. separation of hybridized prussian carp enteron aisle Bdellovibrio and biological Characteristics Study [J] thereof. microorganism journal, 2007 (34): 52~56; Thank to heroes, Fang Wenhong etc. Bdellovibrio seawater bacterial strain Bdh5221 crack characteristic and growth effect factor research [J]. marine fishery, 2007 (29): 97~102).Therefore, in the dominant environment of Gram-positive pathogenic bacterium, Bdellovibrio can't be brought into play the ability of its environment purification, and its application will be restricted.
Strengthen the cracking performance of Bdellovibrio, acquisition is the Bdellovibrio of cracking gram-positive microorganism effectively, to ensureing that the human physical and mental health is significant.This so, at present Chinese scholars adopts the nature method for screening more, not only the time long, and be difficult to screen and get.Ultraviolet mutagenesis is prominent to be still one of the most frequently used and effective means in the Microbial Breeding so far, it is higher and be difficult for regressive advantage that it has mutation frequency, but to reach the research that strengthens the Bdellovibrio cracking performance, still there is not any report at present by uv induction sudden change method.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, the method for suddenling change with uv induction is means, is purpose to strengthen the Bdellovibrio cracking performance, and the application of a kind of uv induction sudden change in strengthening the Bdellovibrio cracking performance is provided.With the cracking ability of effective raising Bdellovibrio, thereby widen the range of application of Bdellovibrio, satisfy need of production practice better to Gram-negative and positive pathogenic bacterium.
For achieving the above object, main technical schemes of the present invention is as follows:
Should be with being to sport the master with ultraviolet induction, auxiliary simultaneously with infrared source and LiCl reinforcement mutagenesis, it comprises following concrete steps and condition thereof:
(1) stablizes light wave
In the darkroom, ultraviolet source is placed the operating table surface top, auxiliary infrared source places the operating table surface both sides, operates preceding 30~60min and opens ultraviolet source, stablizes light wave, and the power of its ultraviolet source is 10~25W, and the power of infrared source is 15~50W;
(2) aseptic induced mutation
Filtering under the visible light state, earlier with concentration greater than 10 5The Bdellovibrio concentrated solution of pfu/mL adds in the sterile petri dish, add LiCl solution and its final concentration is maintained 0.1~0.5% to culture dish again, add aseptic magnetic force rotor and open magnetic agitation, under slowly stirring, culture dish is placed under the ultraviolet source, open auxiliary infrared source simultaneously, the beginning induced mutation, mutation time is 2~8min, irradiation distance is 5~20cm; Described Bdellovibrio concentrated solution is to be obtained through centrifugal concentrating by Bdellovibrio wild strain Bdellovibrio sp.BDJ01, Bdellovibrio wild strain Bdellovibrio sp.BDJ01 is preserved in Chinese typical culture collection center in the Chinese Wuhan City Wuhan University on January 13rd, 2008, and deposit number is CCTCC NO:M 208011;
(3) ice bath is preserved
Close ultraviolet, infrared source, take out bacterium liquid lucifuge immediately, and place ice bath to preserve at least 1 hour;
(4) mutagenesis that adds up
With the existing enhanced Bdellovibrio mutagenic fungi of cracking ability in the step (3), (2), 1~10 mutagenesis that adds up of (3) repetitive operation set by step is until the Bdellovibrio mutagenic fungi that obtains at least two kinds of gram-positive microorganisms of energy cracking.
Gram-positive microorganism described in the step (4) is meant Clostridium perfringens (Clostridium perfringens), clostridium difficile (Clostridium difficile), swine streptococcus (Streptococcus suis) and faecalis (Enterococcus sp.);
Bdellovibrio concentrated solution concentration is 10 described in the step (2) 5~10 7Pfu/mL.
Ultraviolet source is the ultraviolet lamp of power 15~20W described in the step (1), and auxiliary infrared source is the infrared(ray)lamp of power 20~25W.
The final concentration of LiCl solution is 0.2~0.3% described in the step (2).
Mutation time is 4~6min in the step (2), and irradiation distance is 8~15cm.
The present invention compared with prior art has following advantage:
1, the present invention adopts uv induction sudden change method to strengthen the cracking performance of Bdellovibrio, for the cracking performance that strengthens Bdellovibrio provides a kind of novel method.Improved the cracking ability of Bdellovibrio effectively to Gram-negative and positive pathogenic bacterium, thus widened Bdellovibrio range of application, can satisfy need of production practice better.
2, the present invention is by sporting the master with ultraviolet induction, auxiliaryly simultaneously strengthens the mutagenesis effect with infrared source and LiCl, and method is simple, easy to operate, applicable to industrial production requirement, to the research of Bdellovibrio and use realistic meaning is all arranged.
3, to strengthen Bdellovibrio cracking performance effect remarkable in the present invention, in embodiment 1,2, adopt the present invention to strengthen verifying of Bdellovibrio cracking performance: can cracking originally can not cracked two strain Salmonellass, and several gram-positive microorganisms, comprise Clostridium perfringens, clostridium difficile, swine streptococcus and faecalis.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiment is not limited in this.
Embodiment 1
Sport the master with ultraviolet induction, the auxiliary simultaneously application with infrared source and LiCl reinforcement mutagenesis comprises following concrete steps and condition thereof:
1. stablize light wave
In the darkroom, the ultraviolet lamp of 15W is placed the operating table surface top, be that the infrared lamp of 20W places the operating table surface both sides as the power of auxiliary infrared source, 30min opens ultraviolet source in advance, stablizes light wave;
2. aseptic induced mutation
With the Chinese typical culture collection center that is preserved in the Chinese Wuhan City Wuhan University, deposit number is Bdellovibrio wild strain Bdellovibrio sp.BDJ01 nutrient solution 100mL centrifugal 20min under 4 ℃, the condition of speed 5000 * g of CCTCC NO:M 208011, get supernatant liquor after centrifugal under 4 ℃ of conditions again through the centrifugal 20min of the speed of 12000 * g, precipitation suspends with 10mL DNB liquid nutrient medium.With the suspension via hole diameter is the cellulose microporosity filter membrane filtration of 1.2 μ m, and obtaining concentration is 3 * 10 5Pfu/mL Bdellovibrio wild strain concentrated solution; Wherein nutrient solution is to cultivate by the cultural method of the little ecological Studies progress of animal (p190,, China Agricultyre University Press in 2000), DNB liquid nutrient medium: nutrient broth 0.8g, tyrosine acid hydrolysis thing 0.5g, yeast extract 0.1g, NaCl 30g, distilled water 1000mL, pH7.2~7.6.
Filtering under the visible light state, above-mentioned 3 * 10 5Pfu/mL Bdellovibrio wild strain concentrated solution 5mL adds in the 9cm sterile petri dish, in culture dish, add concentration again and be 1.0% 1.25mL LiCl solution, make its final concentration reach 0.2%, add aseptic magnetic force rotor and open magnetic agitation, under slowly stirring, culture dish is placed under the ultraviolet lamp, open auxiliary infrared source simultaneously, its irradiation distance is 15cm; The beginning induced mutation, mutation time is 4min.
3. ice bath is preserved
Close ultraviolet lamp and infrared lamp, take out bacterium liquid, wrap up lucifuge with tinfoil, and place ice bath to preserve at least 1 hour at once to suppress reparation.
4. mutagenesis adds up
With the existing institute of cracking host ability in the step 3 enhanced Bdellovibrio mutant strain, six mutagenesis that adds up of 2,3 repetitive operations set by step.
The method of inspection of Bdellovibrio mutant strain cracking host ability is as follows:
With Gram-negative bacteria-schwann's bacterial of alga, Salmonellas bacterial strain S029, S394, gram-positive microorganism-Clostridium perfringens, clostridium difficile, swine streptococcus and faecalis are inoculated in respectively in the 100mL2216E assistant Bel marine bacteria liquid nutrient medium, placing rotating speed is 200rpm, temperature is to cultivate 14h in 28 ℃ the constant temperature shaking table, and concentration is about 10 8~10 11Cfu/mL.Nutrient solution is under 4 ℃ of temperature condition, and with the centrifugal 20min of 5000 * g speed, the thalline that precipitates suspends again with the 5mLDNB liquid nutrient medium, and it is standby to place 4 ℃ of refrigerators to preserve.Wherein 2216E helps Bel marine bacteria liquid nutrient medium component: peptone 5g, yeast extract 1g, high ferric phosphate 0.01g, NaCl 30g, distilled water 1000mL, pH7.2~7.6.
With the Bdellovibrio mutagenic fungi bacterium liquid after the present embodiment uv induction sudden change with mix with the concentrated solution 0.5mL of above-mentioned three strain Gram-negative bacterias and four strain gram-positive microorganisms respectively without each 0.5mL of wild strain bacterium liquid of mutagenesis, leave standstill 30min, mix with 50 ℃ DNB upper strata substratum 3.5mL, be poured on the DNB solid medium and pave and make double-layer plate, put into 28 ℃ of cultivations of constant incubator, observe the situation that occurs bacterial plaque on the flat board.DNB upper strata substratum wherein: NaCl 15g, distilled water 500mL, pH7.2~7.6, agar powder 3g; DNB solid medium: add the 15g agar powder in the 1000mL DNB liquid nutrient medium.
Be published in research report (Jurkevitch on " use and environmental microbiology " in 2000 according to Israel scholar Jurkevitch, E., Minz, D.and Ramati, B.2000.Prey range characterization, ribotyping, anddiversity of soil and rhizosphere Bdellovibrio spp.isolated on phytopathogenic bacteria.Appl.Environ.Microbiol.66,2365-2371.), the quantity that occurs plaque on the double-layer plate can be represented the quantity of Bdellovibrio indirectly, the diameter of plaque can be represented the cracking ability of Bdellovibrio indirectly, the quantity of plaque is many more, diameter is big more, can illustrate that the cracking ability of Bdellovibrio is strong more.
Resulting Bdellovibrio mutagenic fungi of the 3rd step in the present embodiment is tested by above method, and the quantity and the size of the plaque on the double-layer plate of being made by Bdellovibrio bacterium liquid before and after the mutagenesis are judged its cracking performance, see Table 1 by comparing result.
The Bdellovibrio mutagenic fungi of table 1: embodiment 1 and the cracking performance of wild strain be (mutagenesis) relatively
Annotate: the unit of plaque diameter is cm in the table 1; The unit of plaque quantity is individual/flat board, and is as follows.
The result that plaque occurs on the double-layer plate is observed in table 1 explanation, and the plaque quantity that the Bdellovibrio mutagenic fungi cracking schwann's bacterial of alga after the mutagenesis is occurred is increased to 375 with respect to wild strain BDJ01 by 306; And plaque also occurred on the flat board of cracking Salmonellas S029, promptly be increased to 83 by no spot.Performance of this Bdellovibrio mutagenic fungi cracking schwann's bacterial of alga of screening behind a ultraviolet mutagenesis of explanation increases to some extent, and beginning can cracking wild strain institute can not cracked Salmonellas S029, but also do not show the ability of cracking positive bacteria this moment.Bacterial strain after the utilization mutagenesis for the first time continues the mutagenesis that adds up, and its cracking performance maybe will continue to improve.For this reason, resulting Bdellovibrio mutagenic fungi of the 3rd step in the present embodiment is gone on foot the mutagenesis six times that adds up again through the 4th, then by testing with quadrat method, plaque situation on the double-layer plate of being made by Bdellovibrio bacterium liquid before and after the mutagenesis by contrast is judged its cracking performance, Bdellovibrio mutagenic fungi that screens and wild strain comparison the results are shown in Table 2.
The Bdellovibrio mutagenic fungi of table 2 embodiment 1 and the cracking performance of wild strain be (six mutagenesis that adds up) relatively
The result that plaque occurs on the double-layer plate is observed in table 2 explanation, and with respect to wild strain, the plaque quantity that the Bdellovibrio mutagenic fungi cracking schwann's bacterial of alga after six mutagenesis that adds up is occurred is increased to 450 by 328; And can the cracking wild strain institute can not cracked Gram-negative bacteria-Salmonellas S029, S394 and gram-positive microorganism-faecalis and clostridium difficile.The Bdellovibrio mutagenic fungi that this explanation screens behind six ultraviolet mutagenesises improves a lot than the cracking performance of wild strain.
Particularly the range of application of Bdellovibrio has been expanded in the appearance of mutagenic fungi cracking gram-positive microorganism-faecalis and clostridium difficile ability, and being applied to for it that industry such as food provides may.
Embodiment 2
Sport the master with ultraviolet induction, the auxiliary simultaneously application with infrared source and LiCl reinforcement mutagenesis comprises following concrete steps and condition thereof:
1. stablize light wave
In the darkroom, the ultraviolet lamp of 20W is placed the operating table surface top, be that the infrared lamp of 25W places the operating table surface both sides as the power of auxiliary infrared source, 60min opens ultraviolet source in advance, stablizes light wave.
2. aseptic induced mutation
The concentration method of Bdellovibrio wild strain BDJ01 is with embodiment 1, and obtaining concentration is 2 * 10 7The Bdellovibrio wild strain concentrated solution of pfu/mL; Filtering under the visible light state, above-mentioned 2 * 10 7The concentrated solution 5mL of pfu/mL adds in the 9cm sterile petri dish, in culture dish, add concentration again and be 1.0% 1.67mL LiCl solution, make its final concentration reach 0.25%, add aseptic magnetic force rotor and open magnetic agitation, under slowly stirring culture dish is placed under the ultraviolet lamp, open auxiliary infrared source simultaneously, irradiation distance is 8cm, the beginning induced mutation, mutation time is 6min.
3. ice bath is preserved
Close ultraviolet lamp and infrared lamp, take out bacterium liquid, wrap up lucifuge with tinfoil, and place ice bath to preserve at least 1 hour at once to suppress reparation.
4. mutagenesis adds up
With the existing enhanced Bdellovibrio mutagenic fungi of cracking ability in the step 3, ten mutagenesis that adds up of 2,3 repetitive operations set by step.
The method of inspection of Bdellovibrio mutagenic fungi cracking ability is with embodiment 1, and the plaque situation on the double-layer plate of being made by Bdellovibrio bacterium liquid before and after the mutagenesis is judged its cracking performance, sees Table 3 by comparing result.
The Bdellovibrio mutagenic fungi of table 3 embodiment 2 and the cracking performance of wild strain be (mutagenesis) relatively
In the table 3, the plaque diameter that the Bdellovibrio mutagenic fungi cracking schwann's bacterial of alga after the mutagenesis is occurred increases to 1.8cm by 1.5, and quantity increases to 410 by 306.The cracking ability of expression ultraviolet mutagenesis strain slightly strengthens.
Utilize bacterial strain after the mutagenesis for the first time continue the to add up spot situation that of the Bdellovibrio mutagenic fungi after the mutagenesis ten times to see Table 4:
The Bdellovibrio mutagenic fungi of table 4 embodiment 2 and the cracking performance of wild strain be (ten mutagenesis that adds up) relatively
The result of table 4 shows, Bdellovibrio mutagenic fungi cracking performance after ten mutagenesis that adds up continues to improve, compare with wild strain, not only the ability of cracking Gram-negative bacteria-schwann's bacterial of alga has enhancing, and can cracking original institute can not cracked Gram-negative bacteria-Salmonellas S029 and S394 and gram-positive microorganism-Clostridium perfringens, clostridium difficile, swine streptococcus and faecalis.
So far, the ability of mutagenic strain cracking Gram-negative bacteria has not only obtained enhancing, and has also shown the ability of stronger cracking gram-positive microorganism.
Among the embodiment 2, the Bdellovibrio mutagenic fungi called after Bdellovibrio sp.BDM01 that obtains behind ten ultraviolet mutagenesises is preserved in Chinese typical culture collection center (CCTCC) in the Chinese Wuhan City Wuhan University on April 28th, 2008, and deposit number is CCTCC NO:M 208066.

Claims (6)

1, the application of uv induction sudden change in strengthening the Bdellovibrio cracking performance is characterized in that, should be with being to sport the master with ultraviolet induction, and auxiliary simultaneously with infrared source and LiCl reinforcement mutagenesis, it comprises following concrete steps and condition thereof:
(1) stablizes light wave
In the darkroom, ultraviolet source is placed the operating table surface top, auxiliary infrared source places the operating table surface both sides, operates preceding 30~60min and opens ultraviolet source, stablizes light wave, and the power of its ultraviolet source is 10~25W, and the power of infrared source is 15~50W;
(2) aseptic induced mutation
Filtering under the visible light state, earlier with concentration greater than 10 5The Bdellovibrio concentrated solution of pfu/mL adds in the sterile petri dish, add LiCl solution and its final concentration is maintained 0.1~0.5% to culture dish again, add aseptic magnetic force rotor and open magnetic agitation, under slowly stirring, culture dish is placed under the ultraviolet source, open auxiliary infrared source simultaneously, the beginning induced mutation, mutation time is 2~8min, irradiation distance is 5~20cm; Described Bdellovibrio concentrated solution is to be obtained through centrifugal concentrating by Bdellovibrio wild strain Bdellovibrio sp.BDJ01, Bdellovibrio wild strain Bdellovibrio sp.BDJ01 is preserved in the Chinese typical culture collection center in the Chinese Wuhan City Wuhan University, and deposit number is CCTCC NO:M208011;
(3) ice bath is preserved
Close ultraviolet, infrared source, take out bacterium liquid lucifuge immediately, and place ice bath to preserve at least 1 hour;
(4) mutagenesis that adds up
With the existing enhanced Bdellovibrio mutagenic fungi of cracking ability in the step (3), (2), 1~10 mutagenesis that adds up of (3) repetitive operation set by step is until the Bdellovibrio mutagenic fungi that obtains at least two kinds of gram-positive microorganisms of energy cracking.
2, the application of uv induction sudden change according to claim 1 in strengthening the Bdellovibrio cracking performance is characterized in that gram-positive microorganism is meant Clostridium perfringens, clostridium difficile, swine streptococcus and faecalis described in the step (4).
3, the application of uv induction sudden change according to claim 1 in strengthening the Bdellovibrio cracking performance is characterized in that Bdellovibrio concentrated solution concentration is 10 described in the step (2) 5~10 7Pfu/mL.
4, the application of uv induction sudden change according to claim 1 in strengthening the Bdellovibrio cracking performance is characterized in that ultraviolet source is the ultraviolet lamp of power 15~20W described in the step (1), and auxiliary infrared source is the infrared(ray)lamp of power 20~25W.
5, the application of uv induction sudden change according to claim 1 in strengthening the Bdellovibrio cracking performance, the final concentration that it is characterized in that LiCl solution described in the step (2) is 0.2~0.3%.
6, the application of uv induction sudden change according to claim 1 in strengthening the Bdellovibrio cracking performance is characterized in that mutation time is 4~6min in the step (2), and irradiation distance is 8~15cm.
CN2008102204883A 2008-12-26 2008-12-26 Use of UV induction mutation for reinforcing cracking performance of Bdellovibrio CN101440362B (en)

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Cited By (8)

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CN101955888A (en) * 2010-06-25 2011-01-26 朱笃 Mutant strain of trichosporon cutaneum B3 for producing grease at high yield, EMS thereof and ultraviolet ray compound mutagenesis breeding method
CN101961083A (en) * 2010-08-31 2011-02-02 华南理工大学 Use of bdellovibrio bacteriovorus bdelloplast as bacteria remover in fruit cleaning
CN101978860A (en) * 2010-08-31 2011-02-23 华南理工大学 Application of bdellovibrio mixture as bactericide to cleaning of fruits
CN101978861A (en) * 2010-08-31 2011-02-23 华南理工大学 Application of bdellovibro nectophore serving as bacteria remover to cleaning of fruits
CN102007941A (en) * 2010-08-31 2011-04-13 华南理工大学 Application of Bdellovibrio nectophore in preparing bactericide for controlling red mouth disease of scophthalmus maximus
CN102027883A (en) * 2010-08-31 2011-04-27 华南理工大学 Energy-saving type sea cucumber culturing method
CN103725625A (en) * 2010-08-31 2014-04-16 华南理工大学 Bdellovibrio preparation, and fermentation method and application thereof
CN107988136A (en) * 2017-12-14 2018-05-04 华南理工大学 One plant of ocean Bdellovibrio and the promotion plastidogenetic application of leech under ampicillin

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CN1273584C (en) * 2003-12-03 2006-09-06 北京大学 Bdellovibrio sp. and its application

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101955888A (en) * 2010-06-25 2011-01-26 朱笃 Mutant strain of trichosporon cutaneum B3 for producing grease at high yield, EMS thereof and ultraviolet ray compound mutagenesis breeding method
CN101961083A (en) * 2010-08-31 2011-02-02 华南理工大学 Use of bdellovibrio bacteriovorus bdelloplast as bacteria remover in fruit cleaning
CN101978860A (en) * 2010-08-31 2011-02-23 华南理工大学 Application of bdellovibrio mixture as bactericide to cleaning of fruits
CN101978861A (en) * 2010-08-31 2011-02-23 华南理工大学 Application of bdellovibro nectophore serving as bacteria remover to cleaning of fruits
CN102007941A (en) * 2010-08-31 2011-04-13 华南理工大学 Application of Bdellovibrio nectophore in preparing bactericide for controlling red mouth disease of scophthalmus maximus
CN102027883A (en) * 2010-08-31 2011-04-27 华南理工大学 Energy-saving type sea cucumber culturing method
CN102027883B (en) * 2010-08-31 2013-03-20 华南理工大学 Energy-saving type sea cucumber culturing method
CN102007941B (en) * 2010-08-31 2013-04-10 华南理工大学 Application of Bdellovibrio nectophore in preparing bactericide for controlling red mouth disease of scophthalmus maximus
CN103725625A (en) * 2010-08-31 2014-04-16 华南理工大学 Bdellovibrio preparation, and fermentation method and application thereof
CN103725625B (en) * 2010-08-31 2015-09-16 华南理工大学 A kind of bdellovibrio bacteriovorus preparation and fermentation process thereof and application
CN107988136A (en) * 2017-12-14 2018-05-04 华南理工大学 One plant of ocean Bdellovibrio and the promotion plastidogenetic application of leech under ampicillin

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