159 results about "Agar plate" patented technology

The invention discloses a gene engineering bacterial strain used for producing phenazine-1-methanamide and its purpose; which is characterized in that knockout of psrA or rpeA gene in Pseudomonas chlororaphis HT66(Pseudomonas chlororaphis HT66) CCTCC NO: M 2013467 genome is carried out. According to the invention, a strain of Pseudomonas chlororaphis is performed with seed culture, fermentation, extraction, concentration and crystallization to obtain phenazine-1-methanamide with purity of more than 96%. After the deletion of psrA and rpeA gene in the bacterium genome, the fermentation output of phenazine-1-methanamide can reach as high as 1800ppm. The agar plate bacteriostasis tests found that phenazine-1-methanamide has obvious inhibition effect on common plant pathogenic fungi such as Rhizoctonia solani, fusarium oxysporum, pythium wltmum and Stevia Septoria, has good stability and activity, and has good application prospect.

The invention discloses an Enterococcus faecalis strain SBD, of which the collection number is CGMCC No.4848. The strain is a Gram positive strain and is spherical according to observation with microscope, and the strain does not form spores; the strain can well grow on a Mann, Rogosa and Sharpe (MRS) agar plate and can grow into a round, smooth and raised bacterial colony like a grey white dew and with a diameter of 0.5 to 1 millimeter within 48 hours; and the strain can grow in a facultatively anaerobic environment, the growth temperature range is from 10 to 50 DEG C, the optimal growth temperature is 30 to 40 DEG C, and the growth pH value range is from 4 to 10 and the optimal pH value is 6.5. The bacterial preparation made by the strain is nontoxic and safe, can resist gastric juice and cholate, has a strong inhibition effect on various harmful bacteria and can be widely used in livestock breeding industry (by directly adding into daily ration or drinking water of animals) to increase the disease resistance in animals; and the preparation is expected to replace antibiotic for feed purpose and can obviously improve average weight of weaned piglets, reduce a feed-to-meat ratio and improve living environment of livestock and therefore has a bright prospect.

The invention provides a method for separating and screening a saline-alkaline-resistant pure bacterial strain degrading petroleum hydrocarbon from oily soil. The method comprises the steps: firstly collecting soil polluted by salinized petroleum hydrocarbon from an oil field, adding the collecting soil according to an amount of 10% into an inorganic salt culture medium which takes crude oil as a unique carbon source, and carrying out enrichment culture in a constant temperature shaker; then inoculating a 10% enriching liquid into a fresh crude oil inorganic culture medium and continuously carrying out habituated culture under a same condition for 4-5 times, wherein the pH, salinity and crude oil content of the culture medium in the domestication process are gradually raised; after diluting the culture liquid after domestication in a gradient manner, coating the culture liquid to a LB-agar panel, after bacterial colonies grow, picking up single colonies in different shapes, and lineating and purifying and separating single bacteria; and finally, spreading the separated single bacteria to a crude oil inorganic salt-agar panel and detecting whether the bacterial colonies grow or not, if so, the bacterial strain is the bacteria degrading petroleum hydrocarbon; determining the petroleum hydrocarbon degradation rate of the single bacterial strain and carrying out 16Sr DNA molecular identification on the bacterial strain so as to determine the species relationship of the bacterial strain.

Systems for preparing cell culture dish media are provided which are well adapted for preparing cell culture media, supplements, additives and/or other ingredients in an efficient and versatile manner to form sterilized cell culture dish media having a variety of ingredients well adapted with respect to uniformity and nutritional content to culture microorganisms in a particularly effective manner. The systems include an array of filtration and steam sterilization devices to sterilize various types of media for selective combination of one or more of the types of media in a mixing chamber to form cell culture dishes (e.g., agar plates). Methods of preparing cell culture dish media and cell culture dishes are also provided.

The invention discloses a culture device for a white fly and tetranychid mite feeding experiment and a method for conducting experiment using the same. The culture device comprises a culture dish and a sealing device covering the culture dish, wherein a water agar plate is arranged in the culture dish; the sealing device covering the culture dish is provided with multiple air holes; and the water agar comprises agar and water in a mass ratio of 1:50. When the culture device for the white fly and tetranychid mite feeding experiment is used to conduct experiment, the leave is separated and can be sheared into any shape without using the entire leave as the experimental material, thereby avoiding influence on the experiment caused by the limitation under space, temperature and humidity when the living plant or cut plant is used as the experimental material; and the experimental conditions are easy to control, and the operation is simple. Moreover, the problem of relatively large error between the repeated experiments for the same treatment caused by differences in height, size and strongness of the living plants or cut plants can be overcome.

The invention relates to an analytical method of components of fatty acids contained in listeria cells, in particular to a method for carrying out bacteria classification by adopting a cell chemical analysis method, belonging to the technical field of biological engineering. The method mainly comprises the following steps of: rejuvenating listeria; respectively separating and purifying the listeria by using an OXA agar plate and a trypticase soy yeast extract agar plate; culturing the listeria of a single typical colony, and preparing into a bacterial suspension; carrying out inactivation processing by using formaldehyde; averagely filling the bacterial suspension processed by the formaldehyde into centrifuge tubes for washing; carrying out methyl esterification by using a mixed solution of hydrochloric acid and the formaldehyde; and carrying out GC-MS (Gas Chromatograph-Mass Spectrometer) analysis on the prepared fatty acids. The method can not only break through the limit of the traditional bacterial taxonomy and reduce the errors brought about by anthropic factors on the traditional morphological taxonomy, but also provide a strong and powerful tool for the classification and the identification of new strains and virus types; and in addition, the method can be used for identifying the listeria from other bacteria and foods.

The invention relates to a single spore isolation and purification method of verticillium dahliae. The method mainly comprises the following steps: an agar medium cylinder is taken from a water agar plate by the use of a glass capillary with diameter being 1mm and is picked onto a sterilized glass slide with a sterilization needle; a verticillium dahliae strain is activated in a PDA solid medium and the activated strain is cultured in a Czapek-Dox liquid medium; spore concentration is counted under a microscope; and the spore concentration is adjusted to 102-106 cfu to prepare a spore suspension; 1 microlitre of the spore suspension is sucked up and dripped onto the surface of the water agar cylinder on the glass slide; the glass slide is moved into a culture dish for constant temperature incubation at 25 DEG C; the glass slide is detected under the microscope, and the cylinder containing a single conidiospore which has germinated germ tubes is transferred onto a PDA plate to be cultured at 25 DEG C so as to finish single spore isolation. The method is simple to operate, can be implemented by general operation staff and is suitable for mass separation. By the method, separation success rate is high. The method is also suitable for production of other fungi of microconidia.

The invention relates to a Xanthobacter and its snail killer. A snail bacterium, namely Xanthobacter autotrophicus is a Xanthobacter autotrophicus II whose collection unit. It uses beef peptone agar plate or slope culture medium to separate bacterial strains, obtains snail-killing Xanthobacter autotrophicus solution (a hundred million grades/ml) by bottle-shaking fermentation process; and the application of Xanthobacter autotrophicus as a snail killer is that: making ultrasonic processing on the Xanthobacter autotrophicus solution and plant material forming agent, then mixing them to prepare a novel biological snail killer, which has the characters: (1) quickly killing the snail, and effectively preventing the snails from climbing up to flee; (2) harmless to non-target living things and fish, fast to naturally degrade and no pollution; (3) no toxic and side effect, and time of maintaining snail-killing pesticide effect 15 days long; (4) convenient to store, transport and use; etc.

The invention discloses a cold water coagulable culture medium coagulator and a preparation method thereof. The culture medium coagulator is mainly prepared from the following raw materials by weight portion: 1 portion of xanthan gum and 0.5 to 1.5 portions of guar gum which are mixed together; 1 portion of polyacrylate sodium and 7 to 10 portions of locust bean gum which are mixed together, wherein the mass ratio of the mixture of the xanthan gum and the guar gum to the mixture of the polyacrylate sodium and the locust bean gum is 7-9:1. The cold water coagulable culture medium coagulator can be dissolved and coagulated in cold water, has good water absorption, short reaction time and high gel strength and viscosity, is suitable for the growth of most microbes, and can form gel. The gel can be used as the coagulator for a microbe culture medium, thus the cold water coagulable culture medium coagulator avoids the defects that the prior agar culture medium needs heating and consumes more time for cooling, and an agar plate is easy to dehydrate and check.

The invention provides a Korean pseudomonas and application thereof. The Korean pseudomonas is identified as Korean pseudomonas CJ-361, and the preservation number is CGMCC NO.12122. The Korean pseudomonas CJ-361 is obtained through separation from radix pseudostellariae rhizosphere soil by adopting a pseudomonas selectivity culture medium, an antagonistic experiment is performed on fusarium oxysporum and fusarium moniliforme separated to the radix pseudostellariae root lesion portion through an agar plate opposite culture method, and it is found that the Korean pseudomonas has the strong antagonism on the fusarium oxysporum and the fusarium moniliforme. Meanwhile, the invention provides the optimum culture medium for antagonistic bacterium growth, the optimum formula is 1/4LB+1/20MS (containing no ferric salt)+0.1 wt% brown sugar, and the fermented liquor has the obvious antagonistic effect. The antagonistic strain and the microbial agent thereof can effectively prevent and treat radix pseudostellariae root rot diseases, and the environment-friendly, ecological and safe biological prevention and control potential strain is provided for overcoming or relieving replantation diseases of radix pseudostellariae.

Belonging to the technical field of biological pesticides, the invention provides a Gentiana nubigena endophyte applicable to biological control of plant diseases and its fermentation process. The strain involved in the invention is Bacillus pumilus LD-b1, which is preserved in the General microbiological center of China Committee for Culture Collection of Microorganisms on May 14, 2012, and has a preservation number of CGMCC No.6112. Mixed strain agar plate punching method experiments show that the strain LD-b1 can inhibit the growth of a plurality of plant disease fungi. LD-b1 can substantially inhibit Valsa mali from eroding apple fruits. In greenhouse pot experiments, the LD-b1 can achieve a cucumber Mycosphaerella arachidicola control effect of 81.2%. The fermentation process consists of: culturing LD-b1 in a seed tank for 24h to a logarithmic growth phase, inoculating 5% of the LD-b1 into a production tank according to an inoculation amount, controlling the initial pH at 7.5 and the fermentation temperature at 30DEG C, maintaining dissolved oxygen over 5%, and performing fed-batch fermentation for 68h, thus obtaining a bacterial amount over 10<10>cfu/mL. With the advantages of fast growth and wide antibacterial spectrum, the strain LD-b1 has good application prospects in biological control of plant diseases.

The invention discloses a method for screening and preparing culture seed for dirhamnolipids, which comprises the following steps: choosing the soil sample polluted by grease chronically as the strain source in strain screening; adopting vegetable oil as the liquid culture medium of single carbon source, and coating with the vegetable oil on the hexadecyl trimethyl ammonium bromide agar plate with stronger selectivity after enrichment; then selecting suspect strain depending on the morphology and growth vigour of the bacterial colony; testing the surfactivity of the strain under primary screening after fermentation culture; analyzing the fermentation broth of the strain with high activity directly with TLC method; then selecting the strain with high dirhamnolipids productivity; extracting the rhamnolipids from the strain fermentation broth under screening with extraction method; obtaining purer dirhamnolipids with column chromatographic separation method. The invention overcomes the disadvantage that the strain is screened only based on the total output of rhamnolipids, and the strain with high dirhamnolipids output is obtained difficultly in the prior screening method, and has the advantages of high accuracy, high rhamnolipids output of the screened strain, more particularly higher dirhamnolipids output, and simple preparation technology.

The present invention relates to a Streptomyces violaceoruber and its molluscicides, wherein said Streptomyces violaceoruber is Streptomyces violaceoruber I which is preserved at China typical culture preservation center (CCTCC) with a preservation identification serial number of M203102 and a preservation date at 20030803. The bacterial strain is separated and activated by adopting Gao synthesized number 1 vegetable gelatine flat plate or inclined surface culture medium, and the Streptomyces violaceoruber bacterium solution with molluscidal functions is then cultured through fermentation by using a swing bottle; the Streptomyces violaceoruber bacterium solution and the plant pelleting additive are mixed to constitute a molluscicide. The Streptomyces violaceoruber in accordance with the present invention possesses stronger molluscidal effect, and the molluscicide formulated by the Streptomyces violaceoruber bacterium solution and plant pelleting additive possesses the following advantages: 1, without toxic and side effects and pollutions for the environment, it can be naturally degraded rapidly; 2, without toxic and side effects for persons and animals, it can maintain a long molluscidal time (longer than 15 days); 3, it is convenient to store, convey and use; 4, it has more improved molluscidal effect than pure bacterium solution and pure bacterium powder, with reduced drug amount; 5, relative pure plant powder, it has improved medical effects and prolonged time for maintaining the medical effects.

The invention provides ribosome resistance mutagenesis breeding vancomycin production bacterial strain and an application thereof, specifically the mutagenesis breeding method of mutagenesis bacterial strain of Amycolatopsis orientalis and the application thereof in preparing vancomycin, relating to breeding of microorganism. The method comprises the following steps: inoculating the Amycolatopsis orientalis to Gause-1 agar culture medium panel to be cultured; taking out spore and using sterile water to prepare spore suspension and coating the spore suspension in Gause-1 agar culture medium panel featuring mixing resistance of vancomycin and streptomycin to be cultured; randomly selecting produced single bacterial colony and streak-inoculating the single bacterial colony to the Gause-1 agar culture medium panel to be cultured, carrying out biological activity determination on produced mutagenesis bacterial strain; selecting mutagenesis bacterial strain with inhibition zone diameter thereof being 2mm larger than the inhibition zone diameter of original strain and inoculating the mutagenesis bacterial strain to liquid fermentation medium to be cultured; determining the content of vancomycin in fermentation liquor by HPLC method to verify the obtained vancomycin mutagenesis bacterial strain; subjecting the obtained vancomycin mutagenesis bacterial strain to 5 times of continuous passage stability inspection to obtain the vancomycin production bacterial strain.

The invention discloses a bacilluscereus (Bacilluscereus1205) and an application thereof. The morphological characteristics are as follows: the 1205 bacterial strain is cultivated on an ordinary agar plate culture medium at 37 DEG C for 36 hours, the bacterial colony is nearly round, grey white, non-transparent and as large as 3-7mm, the edge is irregular and expanded, the surface is rough, granulated and is like ground glass or wax, no pigment is secreted; the bacilluscereus is a gram positive bacteria, rod-shaped, 1.0-1.3*3.0-5.0 micrometer, chaining and moveable; the spore is ellipse, terminal or secondary terminal and unclearly expanded; and the bacilluscereus can endure the cultivation of 7% NaCl. The bacilluscereus has the characteristics of strong stress resistance, simple nutritional requirement, stabilization, easy colonization, stability to light and ultraviolet, low cost, long storage period, is easy to apply and realize industrialization, and has a certain growth promoting effect on tobaccos.

The invention provides a method and a kit for identifying Vibrio harveyi quickly by means of strong vibrio harveyi phage and belongs to the technical field of identification of cultures of Vibrio harveyi. The method comprises the following steps: 1) coating 10<7>-10<9>cfu/mL to-be-detected culture to a LB agar plate, leaving the LB agar plate to stand for 15-20min to obtain a to-be-detected plate;2) dropwise adding a strong vibrio harveyi phage mixed liquid to the to-be-detected plate to obtain an identification system; and 3) leaving the identification system obtained in the step 2) to standfor 8-16h at 28-35 DEG C, observing whether bacteriophage plaques appear or not in the identification system, and identifying the to-be-detected culture as the Vibrio harveyi. The method is low in cost and time- and labor-saving, the time needed is only 16-18h, and the method is particularly suitable for initially screening a lot of Vibrio harveyi.

The invention relates to a method for detecting the total number of bacterial colonies in an activated lactobacillus drink, relating to a microorganism detecting method. The method comprises the following steps of: filtering diluent of the activated lactobacillus drink with a filter membrane, flushing the filter membrane with physiological saline, pasting the flushed filter membrane on a TTC nutrient agaragar tablet with the filtering face facing upwards, converting the nutrient agaragar tablet and culturing at 37 DEG C, and counting after culturing for 48+/-2h. By adopting the method, the total number of pollution bacterial colonies in the activated lactobacillus drink can be accurately, simply and quickly detected so as to provide an effective method for detecting the sanitation quality of the of the activated lactobacillus drink.

The invention provides a method for counting the effective viable bacterium content of Bdellovibrio bacteriovorus powder, which comprises the following steps: appropriately diluting a sample to be detected, so that Bdellovibrio bacteriovorus in the sample to be detected is sufficiently dispersed into individual cells; then, carrying out incubation culture in an aseptic nutrient broth diluted by 10 times to promote growth and propagation, thus forming a megascopic turbid liquid; further determining that a large amount of Bdellovibrio bacteriovorus is contained in the turbid liquid having the maximum dilution through plaque detection; and finally, determining the total amount of effective viable bacteria in the sample to be detected. Compared with a microscopic direct observation method anda dilution-by-tap-water double-layer agar plate method, the method provided by the invention is more accurate, and overcomes the defects that detection is inaccurate due to dilution in the dilution-by-tap-water double-layer agar plate method and killed bacteria and viable bacteria can not be distinguished in the microscopic direct observation method.

The invention discloses a puncher for a countercurrent immunoelectrophoresis agar plate. The puncher comprises a punching tube, a fixing plate, a fixing plate frame, a handle and a negative-pressure barrel, wherein the punching tube is communicated with the negative-pressure barrel through a hose; the inner side of the opposite side of the fixing plate frame is provided with a slide path; the two ends of the fixing plate can be arranged in the slide path in a sliding mode; at least one end of the fixing plate is provided with a stirring block; the stirring block is positioned at the outer side of the fixing plate frame; a slide slot for allowing the stirring block to slide is formed in the fixing plate frame; a displacement sensor is arranged in the slide path; and a displacement display screen is arranged on the fixing plate frame or the handle. The puncher can be used for punching a plurality of holes in the agar agarose gel plate once and can automatically suck out agar in the holes, so that the whole operation can be completed by only one hand, and therefore, the puncher is efficient and convenient.

The invention provides an evaluation method for a disinfection effect of a disinfectant with a bacteria test sheet. The method comprises the following steps of: (1) preparing and grouping samples to be tested; (2) disinfecting: uniformly mixing the samples to be tested and diluted disinfectant and disinfecting; (3) inoculating: inoculating the samples to be tested and the disinfected samples on a total bacterial colony number test sheet; (4) culturing: packaging the bacterial colony test sheet on which the samples are inoculated into a sealed bag and placing in a thermostat of 37 DEG C to culture for 24-48 hours; and (5) observing a result: counting the bacteria number on each test sheet and calculating sterilization rate of each disinfectant. The evaluation method for the disinfection effect of the disinfectant with the bacteria test sheet is simple and convenient, has high accuracy and is an effective method which can substitute the conventional agar plate sterilization rate evaluation method.