50 results about "Screening cultures" patented technology

The invention relates to the technical field of plant tissue culture and plant transgenosis, in particular to a method for co-transformation of an LcSUCLA alpha-LcSUCLA beta-pEGFC binary expression vector and Leymus chinensis double genes. The method comprises the following steps: obtaining leymus chinensis tissue culture sterile calluses, transforming agrobacterium competent by a freeze thawing method, activating agrobacterium tumefaciens, infecting the leymus chinensis calluses and co-culturing, screening and culturing plant transformation materials, differentiating and screening a culture medium and culture conditions, and differentiating and screening a culture medium and culture conditions as well as a rooting culture medium and culture conditions. According to the invention, double-gene co-transformation of leymus chinensis is realized, and a leymus chinensis double-transgenic plant is obtained. The cotransformation of the LcSUCLA alpha and the LcSUCLA beta in the leymus chinensis can synthesize more succinic acid than the single transformation, so that the tolerance of the leymus chinensis under the stress of basic salt is further improved.

The application discloses a method for improving genetic transformation rate of high-altitude meadow grass, comprising the following steps: (1) taking mature seeds of high-altitude meadow grass as explants to induce callus, and obtaining the callus; (2) adding a surfactant Silwet L-77 with a mass fraction of 3‰ of the bacterial liquid to the agrobacterium bacterial liquid carrying a hygromycin resistance gene, and obtaining a culture bacterial liquid; (3) placing the regenerated plant in a screening culture medium containing 40 mg / L of hygromycin resistance for culture, screening the survived plants, and obtaining the genetically transformed plants. The method can improve the transformation efficiency of the high-altitude meadow grass by about 4.54 times through adding the surfactant in the genetic transformation process and cooperating with the screening of the obtained differentiation culture medium.

The application provides a genetic transformation method of sugarcane variety Liucheng 1541 and belongs to the technical field of plant genetic engineering. The genetic transformation method is that leaf tissues of the sugarcane variety Liucheng 1541 are induced and cultured on a callus induction culture medium containing 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine; the obtained embryogenic callus is infected by agrobacterium; after co-cultivation and recovery culture, the obtained recovered callus is transferred to a screening culture medium containing glufosinate ammonium for screening culture; then the screened resistant embryogenic callus is inoculated on a differentiation culture medium containing glufosinate ammonium for differentiation culture; and after rooting culture, the transgenic plant of Liucheng 1541 is obtained. The method improves the embryogenic callus induction rate, solves the problem of callus browning in the tissue culture process of the variety Liucheng 1541, determines the glufosinate ammonium screening concentration of the embryogenic callus, and establishes the genetic transformation system of Liucheng 1541.

The invention discloses a genetic transformation method suitable for a traditional Chinese medicine divaricate saposhnikovia root, and belongs to the technical field of biology. According to the genetic transformation method, the leaf is taken as an infected object, the infected leaf is differentiated to obtain the callus, and then the regenerated seedling is differentiated, so that the test period of genetic transformation is shortened, and the differentiation rate of the regenerated seedling is improved. According to the method, the bacteriostasis rate, the differentiation rate and the positive conversion rate are improved through callus induction, bacteriostasis and resistance screening culture media of specific mass concentration components and limitation of the infection time of agrobacterium tumefaciens, an efficient and stable genetic transformation system of the radix saposhnikoviae is established, and the method has a good application prospect in precise improvement of variety characters of the radix saposhnikoviae. And a support is provided for cultivating a high-quality, high-yield and multi-resistance new divaricate saposhnikovia root variety.

The invention belongs to the technical field of disease control in aquaculture, and particularly relates to a hippocampus enteritis pathogen separation and identification and medication screening method based on a high-throughput sequencing technology, which comprises the following steps: collecting intestinal tissues of a diseased hippocampus and a healthy hippocampus, respectively preparing tissue homogenate, and freezing for preservation; the method comprises the following steps: extracting DNA (Deoxyribose Nucleic Acid) of microorganisms of an intestinal tissue sample, performing 16SrDNA high-throughput sequencing, analyzing intestinal flora composition, and determining dominant bacteria; selecting and screening a culture medium according to dominant bacteria, and performing pathogenic bacterium separation and purification on intestinal tissues of the diseased hippocampus; dNA of the purified strain is extracted, 16S rDNA amplification and sequencing are carried out, and the species of pathogenic bacteria are identified through sequence comparison; according to the method, the diversity and abundance differences of microorganisms in the sick hippocampus and the healthy hippocampus are compared through a high-throughput sequencing technology, and targeted separation, identification and drug sensitivity screening are further carried out by taking statistically significant difference bacteria as the guidance of potential pathogenic bacteria.

The invention discloses a neutralizing antibody with an AQP3 channel blocking function as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The neutralizing antibody can be specifically combined with an extracellular domain of AQP3, after combination, a water channel and a small molecule transport function of the AQP3 are blocked through steric hindrance and conformation change, and meanwhile, antigen combination specificity is reserved. The preparation method is realized through'recombinant antigen-immunization-B cell screening culture-recombinant antibody expression and affinity purification-ELISA detection ', and the functions of the antibody are ensured. The antibody can be efficiently combined with AQP3 positive cells, is strong in targeting property and high in specificity (the serum titer reaches 1: 1024 * 10, and the purified antibody titer is greater than or equal to 1: 128 * 10), and can be independently used or combined with a nano-carrier for treating related diseases, so that the technical bottleneck that a common AQP3 antibody can only be combined and cannot efficiently play a blocking role is solved.

The invention discloses a method for constructing a genetic transformation system of cymbidium kanran, which belongs to the technical field of plant genetic engineering, and comprises the following steps of: carrying out monoclonal propagation on rhizomes after germination of cymbidium kanran seeds to obtain vegetative propagules with stable characters, taking the vegetative propagules as transformation receptors, taking GFP (Green Fluorescent Protein) as marker genes, and constructing the genetic transformation system of cymbidium kanran. The method comprises the following steps: adding agrobacterium EHA105 into a YEB liquid culture medium containing 50 mg / L of kanamycin and 25 mg / L of rifampicin as a dip dyeing solution; placing the injured transformation receptor in dip dyeing for transformation, and then carrying out regeneration culture. The invention relates to a method for carrying out genetic transformation on cymbidium kanran by utilizing the innovative receptor and the screening culture medium, and the method can realize efficient and stable genetic transformation without genotype dependence and can solve the problem that the genetic transformation of cymbidium kanran varieties is difficult to carry out. According to the invention, an efficient genetic transformation and regeneration system of cymbidium kanran is established for the first time, and a foundation is laid for verification of related functional genes of cymbidium kanran and further development of biological research of cymbidium transgenic breeding and the like.

This invention belongs to the field of microbial technology, and particularly relates to a screening culture medium for precise intestinal flora transplantation, its preparation method, and a method for enriching Enterobacteriaceae bacteria. The application of the composition provided by this invention in the preparation of intestinal flora screening culture media, through the synergistic effect of the basic nutritional conditions provided by the oligofructose, chlorogenic acid, and CDC anaerobic agar basal medium, enables effective screening and enrichment of Enterobacteriaceae bacteria in fecal samples under in vitro culture conditions, showing excellent application prospects in the isolation, purification, and functional studies of Enterobacteriaceae bacteria.

The invention discloses a stable genetic transformation method of floating moss. The genetic transformation method comprises the following steps: (1) crushing thallus tissues of the floating moss, (2) pre-culturing explants, (3) infecting and co-culturing, (4) screening and culturing for the first time, (5) screening and culturing for the second time, (6) enlarging and culturing, and (7) verifying transformation. According to the genetic transformation method, a thallus of floating moss is taken as a starting explant, an agrobacterium-mediated hygromycin and kanamycin screening transformation system of the floating moss is established for the first time, the blank of a floating moss genetic transformation system is filled, and experimental results show that after multi-generation culture, a transformed plant can still stably express a target gene. The method has the advantages of high transformation efficiency and simplicity in operation, and can be used for subsequent genetic transformation research of unknown floating moss genes. The method has a wide application prospect.

The invention provides a genetic transformation method of an excellent sugarcane variety Liucheng 1541, and belongs to the technical field of plant genetic engineering.The genetic transformation method comprises the steps that leaf tissues of the sugarcane variety Liucheng 1541 are taken and subjected to induction culture on a callus induction culture medium containing 2, 4-dichlorphenoxyacetic acid and 6-benzylaminopurine; the method comprises the following steps: infecting embryogenic calluses with agrobacterium, co-culturing, carrying out recovery culture, transferring the recovered calluses to a screening culture medium containing glufosinate-ammonium, carrying out screening culture, inoculating the screened resistant embryogenic calluses to a differential culture medium containing glufosinate-ammonium, carrying out differentiation culture, and carrying out rooting culture to obtain the glufosinate-ammonium resistant embryogenic callus. The transgenic plant of the Liucheng 1541 is obtained. According to the method, the embryogenic callus induction rate is increased, the problem of callus browning in the variety Liucheng 1541 tissue culture process is solved, the glufosinate-ammonium screening concentration of the embryogenic callus is determined, and a Liucheng 1541 genetic transformation system is determined.

The invention discloses an agrobacterium tumefaciens-mediated soybean sprout tip transformation method. The method comprises the following steps of explant infection and co-culture, bud induction and screening culture, elongation recovery and rooting culture and transgenic plant domestication culture. According to the method, the soybean sprout tips are selected as the explants, the soybean genetic transformation process is simplified, direct budding of apical meristem and synchronous rooting of underground parts can be achieved, only one month is needed from explant infection to obtaining of a complete regenerated transgenic plant with a developed root system, the transformation efficiency is high, the soybean transformation period is greatly shortened, and the method is suitable for large-scale popularization and application. The cost is effectively reduced. Meanwhile, the problem of soybean genotype limitation and the problem that northeast varieties bloom in advance in the tissue culture stage are solved, the method is applied to more than twenty main cultivated varieties difficult to transform, the transformation efficiency is high and stable, the highest efficiency can reach 81%, and reliable technical support is provided for soybean basic research and gene editing breeding work.

The invention discloses a pseudo-sorghum genetic transformation method and application. The pseudo-sorghum genetic transformation method comprises the steps of pseudo-sorghum mature seed explant pretreatment, induction culture, gene gun transformation, screening culture, differentiation culture, budding culture, rooting culture and the like. According to the method disclosed by the invention, the callus is successfully induced from the pseudo-sorghum seed immature embryo, the inductivity is high, a simple and efficient genetic transformation method is established, target editing of a pseudo-sorghum gene editing plant can be realized, and the transformation efficiency and the regeneration capacity are high.

The invention discloses a screening culture medium and a preparation method and application thereof, the screening culture medium comprises a basic culture medium and additives, and the additives comprise amphotericin B, vancomycin, ox bile salt, sodium deoxycholate and No.3 bile salt. According to the screening culture medium provided by the invention, a special infectious microbe inhibitor is added on the basis of a trisaccharide iron culture medium, so that most gram-negative enterobacter, staphylococcus aureus and part of fungi except salmonella enteritis subspecies and shigella cannot grow; the interference of the infectious microbes on the detection of the salmonella enteritis subspecies and the shigella is avoided, so that the fluorescent probe can be applied to the detection of the salmonella enteritis subspecies and the shigella in a clinical anus swab, and the sensitivity of color development is improved.

A yeast dual-expression vector and a construction method therefor, a protein-protein interaction identification method and the use. The yeast dual-expression vector allows two exogenous genes to be cloned into the same vector and expressed simultaneously in yeast, thereby enabling interaction identification. The present invention greatly reduces the procedures in original yeast two-hybrid systems, such as plasmid construction and extraction, library preparation, yeast mating and screening culture, and significantly improves the identification efficiency of protein-protein interaction.

The application discloses a jujube genetic transformation method using anther-induced callus as a receptor, comprising the following steps: culturing sterile anthers to obtain callus; constructing a plant recombinant overexpression vector containing a target gene and transferring the plant recombinant overexpression vector into agrobacterium to obtain an agrobacterium immersion liquid; immersing the callus in the agrobacterium immersion liquid, and then co-culturing the immersed callus under dark conditions; and then placing the callus on a culture medium containing hygromycin for screening, so that a homozygous resistant callus can be obtained after 20-30 days of screening culture. The application firstly uses the anther-induced callus of Jing 39 jujube as a receptor to establish a method of the jujube genetic transformation system, the method has a high transformation frequency, and meanwhile, the method can be assisted by 35S::EGFP fluorescent reporter gene marking for screening, and is convenient and fast. The method provides convenience for jujube gene function analysis and jujube genetic improvement.

The invention relates to the technical field of biology, and provides a novel technology for rapidly screening and identifying high-yield poly-gamma-glutamic acid bacillus. The method comprises the following steps: preparing a solid screening culture medium, sterilizing, subpackaging into a square sterile culture dish with scales, diluting a to-be-separated substance, coating the solidified culture medium with the diluted to-be-separated substance, culturing for 24-96 hours, and observing whether a liquid product exists on a cover plate or not; if so, determining that the solid screening culture medium region corresponding to the region contains the high-yield poly-gamma-glutamic acid strain, and determining the single colony of the high-yield poly-gamma-glutamic acid through a sterilization gun head wire drawing experiment. According to the method, the traditional process of screening the high-yield gamma-polyglutamic acid bacillus is simplified, and the workload in the screening process is greatly reduced.

The invention discloses a suspension culture cell monoclonal screening culture method. The method comprises the following steps: selecting cells under suspension culture conditions and inducing the cells to recover adherence; separating the single cells developed into obvious clones, and carrying out radioactive ray irradiation on the remaining cells, so that the remaining cells lose the multiplication capacity and become feeder layer cells; culturing the separated monoclonal cells, recording the culture time and culture environment, establishing a high-density cell bank, and establishing a growth time prediction formula for the multiple monoclonal cells according to the culture time; and calculating the specific time for culturing the required number of cells according to the growth time prediction formula established in the step S3. According to the method, the remaining cells are subjected to radioactive ray irradiation and are converted into feeder layer cells with important effects, so that the waste of cell resources is avoided, the flexible utilization of the cell resources is realized, and the treatment cost of the remaining cells is also remarkably reduced.

The utility model provides a high-yield ergothioneine strain screening culture bin relates to strain screening technical field, including base, culture bin and cover plate, the upper end of base is provided with the jacket seat, in the jacket seat is provided with the thermal insulation layer, the upper part of the thermal insulation layer is provided with the neck, and the culture bin is embedded in the neck, the cover plate is provided with the cover plate. The upper end of the cultivation bin is covered with a cover plate, the cover plate is detachably connected with the cultivation bin, the cultivation bin is detachably connected with the base, multiple sets of anti-slip strips are arranged on the outer ring face of the base, the anti-slip strips are of a semi-cylindrical anti-slip strip structure, and an anti-slip pad is arranged on the bottom face of the base and made of silica gel. The heat preservation layer is made of heat preservation cotton materials, and the outer sleeve base is made of explosion-proof glass materials. The strain screening device is safe and stable in hand taking and placing, tight isolation is achieved, heat preservation and heat insulation are better kept in the screening process, the influence of external environment temperature factors is reduced, therefore, the strain screening quality is guaranteed, and the strain screening device is more efficient and practical.

The utility model discloses a melon planting resource salt tolerance gradient screening culture device, which belongs to the technical field of plant culture screening equipment, and comprises a culture box, a culture drawer arranged in the culture box, a culture basin arranged on the culture drawer, a fixing mechanism and an adjusting mechanism, the fixing mechanism comprises a sliding groove block fixed to the inner wall of the incubator and a sliding block fixed to the outer wall of the culture drawer, the sliding block is in sliding fit with the sliding groove block, and the fixing mechanism further comprises a buckle installed in the sliding groove block in an up-down sliding mode and a limiting ring used for preventing the buckle from being disengaged. The problems that the culture drawer is not firmly fixed and inconvenient to store and take, and the humidity in the box is difficult to control are solved, fixation is stable, operation is convenient and fast, and the disease risk can be reduced.

The invention belongs to the technical field of biology, and particularly relates to a preparation method for synthesizing feeding single-cell protein independent of photosynthesis, a strain and application, and the feeding single-cell protein is synthesized through a renewable energy electrolysis water hydrogen production step, a CO2 hydrogenation methanol synthesis step and a methanol fermentation step; in the microbial fermentation step, screening culture conditions of high temperature and high-concentration methanol are creatively adopted, and only the growth requirement of the bacillus methylophilus is met, so that the cost for maintaining the sterile culture condition is reduced, the singleness of the fermentation flora is ensured, and the product with stable quality is obtained. According to the invention, instability of renewable energy sources is decoupled through multi-stage energy conversion and metabolic engineering by using unstable electric energy produced by the renewable energy sources, so that high-safety feeding single-cell protein is produced, and the device also has the characteristics of high energy transfer efficiency and low water consumption per unit product.

This invention discloses a method for introducing exogenous genes into Gynostemma pentaphyllum, comprising: S1: Selecting Gynostemma pentaphyllum seeds with seed coats within 3 months of harvest, disinfecting, and culturing to obtain aseptic seedlings from seedlings, or selecting stem segments with axillary buds after leaf removal, disinfecting, and culturing using a subculture proliferation medium containing cephalosporin to obtain aseptic rooted seedlings; S2: Selecting the aseptic seedlings or aseptic cuttings obtained in S1, culturing in a rooting medium containing cephalosporin to obtain aseptic rooted seedlings; S3: Transforming plasmid Pcambia1301-PMI into Agrobacterium and culturing in a medium containing kanamycin and rifampin to obtain an Agrobacterium resuspension; S4: Infecting the aseptic rooted seedlings obtained in S2 with the Agrobacterium resuspension, and performing co-culture, antibacterial culture, and screening culture to obtain transgenic adventitious shoots. This invention can successfully introduce exogenous genes into Gynostemma pentaphyllum, obtaining Gynostemma pentaphyllum positive plants.

The application discloses a high-efficiency genetic transformation method of pinellia ternate, and belongs to the technical field of plant tissue culture and genetic transformation, and comprises the following steps: (1) preparing a genetic transformation receptor; (2) preparing an infection solution; (3) infection; (4) co-culture; (5) screening culture; and (6) seedling culture. The method is simple in process and convenient in operation, and can obtain pinellia ternate transgenic plants with high genetic transformation rate.

The invention provides a method for efficient and stable genetic transformation of an algae strain by agrobacterium tumefaciens-mediated porphyra yezoensis, and belongs to the technical field of plant transgenosis. According to the method, a mixed suspension of porphyra yezoensis thallus fragments, monospores and thallus plantlets is used as a transformation receptor, and an efficient and stable genetic transformation technology is established through the steps of infection solution optimization, gradient screening culture and the like. The technical problems that existing porphyra yezoensis genetic transformation is low in efficiency, poor in repeatability, difficult in transformant screening and the like are effectively solved, the transformation efficiency can reach 30%-40%, the genetic stability of positive plants is close to 100%, a reliable technical platform is provided for gene function verification, character improvement and molecular breeding of porphyra yezoensis, and important scientific research value and application prospects are achieved.

The invention discloses an agrobacterium tumefaciens-mediated sweet osmanthus callus genetic transformation method which comprises the following steps: (1) collecting sweet osmanthus seeds, disinfecting and sterilizing, inducing zygotic embryos to form calluses, and carrying out multiplication culture; (2) activating and propagating agrobacterium tumefaciens, resuspending the agrobacterium tumefaciens until OD600 is 0.4-0.6 by using a buffer solution, infecting healthy calluses, performing vacuum treatment and co-culture, and performing screening culture; and (3) verifying the integration and expression of the target gene through visual identification, DNA identification, real-time fluorescent quantitative PCR or western blot analysis. According to the method, a stable genetic transformation system of the osmanthus fragrans callus is established for the first time, the hormone proportion, infection conditions and screening parameters are optimized, overexpression of OfPSI1 and RUBY genes is successfully realized, the transformation efficiency is high, and the operation is simple and convenient. The invention provides a reliable technical platform for sweet osmanthus gene function research and molecular breeding, and has important scientific research and application values.

The application provides a method for efficiently and stably transforming algae strains of Porphyra yezoensis by Agrobacterium mediation, and belongs to the technical field of plant transgenesis. The application uses a mixed suspension of leaf thallus fragments, single spores and leaf thallus seedlings of Porphyra yezoensis as a transformation receptor, and establishes an efficient and stable genetic transformation technology through steps such as optimization of an infection solution and gradient screening culture. The application effectively solves the technical problems of low genetic transformation efficiency, poor repeatability and difficulty in screening of transformants of the existing Porphyra yezoensis, and the transformation efficiency can reach 30% to 40%, and the genetic stability of positive plants is close to 100%, thereby providing a reliable technical platform for gene function verification, trait improvement and molecular breeding of Porphyra yezoensis, and having important scientific research value and application prospect.

The invention belongs to the technical field of epsilon-polylysine production, and particularly relates to a screening culture method and application of tolerant streptomyces hygroscopicus. The screening culture method of the tolerant streptomyces hygroscopicus comprises the following steps: subculturing streptomyces hygroscopicus engineering bacteria in a culture medium taking straw cellulose hydrolysate as a carbon source for at least 30 generations to obtain primary streptomyces hygroscopicus. The primary streptomyces hygroscopicus is subcultured for 10-25 generations in a fermentation culture medium with the epsilon-polylysine concentration being 0.5 g / L-5g / L and straw cellulose hydrolysate as a carbon source, and the tolerant streptomyces hygroscopicus is obtained. Wherein the tolerance means that epsilon-polylysine is produced while an inhibitor is resisted. The straw cellulose hydrolysate contains an inhibitor. According to the screening culture method of the tolerant streptomyces hygroscopicus, disclosed by the invention, the yield of epsilon-polylysine in virus-free cellulose hydrolysate can be up to 1.35 g / L at most, and the yield of epsilon-polylysine can be up to 30.57 g / L at most under the condition of fed-batch fermentation in a 5L fermentation tank.

The invention provides a method for constructing a stable genetic transformation system of Chinese pears, and belongs to the technical field of genetic transformation system construction. A construction method of a Chinese pear stable genetic transformation system comprises the following steps: infecting in-vitro petioles of Chinese pears by using an agrobacterium solution containing a target gene segment, co-culturing the infected Chinese pear in-vitro petioles, removing bacteria, screening, culturing, subculturing and identifying to obtain positive and stable genetic Chinese pear tissue culture seedlings. The technical scheme provided by the invention accelerates the research of the Chinese pear gene function, and has important significance for accelerating the genetic improvement work of Chinese pears.

The invention discloses a rapid screening method of cholesterol-lowering lactic acid bacteria, and belongs to the technical field of biology. Comprising the following steps: (1) coating a fermented food sample solution into an MRS-maize germ oil solid culture medium for preliminary screening culture, and screening single colonies with acid producing circles; (2) carrying out catalase inspection and gram staining microscopy on the strains obtained by primary screening, and screening out bacterial colonies with positive gram staining results and negative catalase experimental results; and (3) inoculating the primarily screened strain into a liquid culture medium containing cholesterol micelles for secondary screening culture, measuring the cholesterol concentration before and after culture, calculating the degradation rate, and selecting the strain with the strongest cholesterol degradation capability as a target strain. According to the method, in the screening process of the cholesterol-reducing lactic acid bacteria, the lactic acid bacteria can be effectively screened, experimental steps are simplified, and the cholesterol-reducing lactic acid bacteria can be primarily screened, so that the overall screening efficiency is improved.

The invention discloses an agrobacterium tumefaciens-mediated bottle gourd stable genetic transformation method. The invention provides a bottle gourd stable genetic transformation system mediated by agrobacterium tumefaciens. The system provides a key technical platform for gene function research and molecular breeding of bottle gourds. An agrobacterium tumefaciens-mediated genetic transformation method is adopted, and the principle is that a target gene is inserted into a modified T-DNA region, an agrobacterium tumefaciens infection mechanism is utilized to realize transfer and integration of an exogenous gene to a plant cell, and a transgenic plant is obtained by combining a tissue culture technology. According to the method, screening culture is not carried out, after co-culture is carried out for two days, a fluorescence microscope is used for observing the fluorescence area of a groove part of a seed notch, the agrobacterium infection efficiency is judged, screening is carried out, and detection is carried out again after seedlings are obtained, so that the time is saved by 30-40 days compared with genetic transformation of plants needing screening culture; the method has the advantages of good hereditary stability and simple operation.

The application belongs to the technical field of plant genetic engineering, and particularly relates to a genetic transformation method of 'Hanfu' apple seed callus and application. The transformed callus is obtained through induction culture, proliferation culture, immersion culture, co-culture and screening culture of the explant, wherein the explant is 'Hanfu' apple seed; the uninfected 'Hanfu' apple seed callus induced by the method has weak fluorescence signal under ultraviolet excitation light. When the callus is subjected to genetic transformation mediated by Agrobacterium and the pRI101 GFP vector is introduced, the fluorescence signal intensity of the callus is significantly increased, indicating that the 'Hanfu' apple callus can be subjected to genetic transformation containing a GFP label.