55 results about "Rhizogene" patented technology

The invention specifically discloses a salix cheilophila in-situ genetic transformation method independent of a tissue culture system, and belongs to the field of plant genetic engineering. According to the method, semi-lignified branches of salix cheilophila are taken as receptors, vacuumizing treatment is performed by utilizing agrobacterium rhizogenes infection liquid, hairy roots are directly induced under a water culture condition after short-term dark culture, and positive identification is performed through reporter genes. According to the method, tedious sterile tissue culture operation is completely avoided, the transformation period is shortened to about 22 days, and the technical threshold and the time cost are remarkably reduced. By adopting the system, the positive rate can reach 54.33% when the pBI121 vector is used for transformation, and the efficiency is remarkably improved compared with that of an existing method. The method provides an efficient and reliable technical platform for root system gene function research, expression analysis and genetic improvement of salix cheilophila and related species thereof, and has high application potential.

The invention relates to the technical field of plant genetic engineering, in particular to application of an algae NySAT2 gene in regulation and control of alfalfa crude protein and detergent fiber. According to the invention, a sequence after codon optimization of an NySAT2 gene coding region is obtained by using gene synthesis and codon optimization technologies, an algae NySAT2 gene overexpression vector is constructed, a target gene is introduced into an alfalfa genome by using an agrobacterium rhizogenes mediated genetic transformation method, and a transgenic plant is obtained by using a genetic transformation system. Molecular identification and phenotypic analysis show that on the premise of keeping normal growth vigor (plant height and biomass have no significant difference from those of a wild type), the crude protein content of a medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased compared with that of the wild type medicago sativa plant, and meanwhile, the contents of acid detergent fibers and neutral detergent fibers are remarkably reduced, so that the content of the crude protein in the medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased. The forage quality and the feeding value of the medicago sativa are effectively improved.

The invention belongs to the technical field of genetic engineering, and particularly relates to an agrobacterium rhizogenes-mediated coastal chrysanthemum indicum tissue culture-free genetic transformation method and application thereof in enhancing the salt tolerance of a root system. The agrobacterium rhizogenes-mediated coastal chrysanthemum indicum tissue culture-free genetic transformation method comprises the following steps: transfecting agrobacterium rhizogenes with plasmids containing to-be-transformed gene expression cassettes to obtain recombinant agrobacterium rhizogenes, transfecting stems with terminal buds of coastal chrysanthemum indicum with the recombinant agrobacterium rhizogenes, and then cutting. The method provided by the invention is short in required period, and the transgenic plant can be obtained only in 30-60 days. The problems of long chrysanthemum conversion rate period, complex operation and tedious process are solved. A tissue culture mode is not needed, so that the tedious process of tissue culture is reduced; local materials can be used according to the characteristic of cutting rooting of chrysanthemum, chrysanthemum seedlings do not need to be cultivated from seeds, and time cost is reduced.

This invention discloses a banana-specific alkaline biocontrol organic fertilizer and its preparation method, belonging to the field of bio-organic fertilizer technology. The banana-specific alkaline biocontrol organic fertilizer of this invention comprises the following raw materials: 25-30 parts modified tobacco straw carbonized powder, 20-25 parts moringa seed fermentation product, 5-10 parts cactus fermentation product, 10-14 parts seaweed residue, 8-12 parts humic acid powder, and 3-5 parts shell powder. The moringa seeds are fermented using enzymatic hydrolysis and microbial agents, effectively breaking down large molecules into smaller molecules that can be directly absorbed by banana plants, promoting plant growth. Simultaneously, *Achromobacterium solani* and *Acinetobacter rhizogenes* secrete various active substances, significantly enhancing banana disease resistance and growth promotion. This invention's organic fertilizer, with its synergistic effect of multiple substances, reduces the use of chemical fungicides, effectively inhibits banana wilt disease, neutralizes soil acidity, increases banana yield, and improves economic and social benefits.

The application discloses a soybean alkali-resistant gene GmFPS1 and application thereof. The soybean farnesyl pyrophosphate synthase gene GmFPS1, and a nucleotide sequence of the GmFPS1 gene is shown as SEQ ID NO. 1. The application also discloses application of the soybean farnesyl pyrophosphate synthase gene GmFPS1 in construction of alkali-resistant transgenic crops. The application carries out genetic transformation of a plant alkali-resistance related protein coding gene GmFPS1 into transgenic Arabidopsis and Agrobacterium rhizogenes-mediated soybean chimera, and studies alkali stress resistance of the transgenic Arabidopsis and Agrobacterium rhizogenes-mediated soybean chimera. The transgenic Arabidopsis and Agrobacterium rhizogenes-mediated soybean chimera obtained have improved alkali stress resistance. The result of qRT-PCR also shows that the soybean GmFPS1 is induced in response to alkali stress, and the expression level is significantly up-regulated. Therefore, the plant alkali-resistance related protein coding gene GmFPS1 is introduced into crops by a genetic engineering method, and a new alkali stress-resistant transgenic plant variety can be obtained.

This invention relates to a genetic transformation method for Ningxia wolfberry using germinating seeds as explants, belonging to the field of plant genetic transformation technology. The invention mainly includes the following steps: (1) After disinfecting Ningxia wolfberry seeds, inoculate them onto a germination medium, and obtain germinating seeds through vernalization and dark culture treatment; (2) Activate and culture Agrobacterium rhizogenes and Agrobacterium tumefaciens strains carrying target gene expression frames to obtain a bacterial solution for infection; (3) Take the germinating seeds obtained in step (1) and mix them with the bacterial solution obtained in step (2) for infection, then transfer the infected germinating seeds to a co-culture medium or substrate for co-culture; (4) Cultivate the material after co-culture in step (3) to induce root growth, and detect the expression level of the target gene or reporter gene on the roots or plants to obtain transgenic positive plants. This invention has the advantages of simple operation, significantly shortened transformation cycle, and high transformation efficiency.

The invention discloses a method for efficiently inducing adventitious buds and rooting through transgenic hairy roots of Populus albus Nanlin 895, and relates to the technical field of biological breeding. The plant material used in the invention is a transgenic Nanlin 895 poplar hairy root generated by mediation of agrobacterium rhizogenes; dividing the hairy roots into root sections of 1-2 cm by using a sterile blade, and putting the hairy roots on a culture medium added with 0.01 mg / L TDZ and 0.2 mg / L 6-BA to induce adventitious buds; placing the adventitious buds in a culture medium added with 0.01 mg / L of TDZ for bud elongation; and cutting off the elongated adventitious buds, placing the adventitious buds in a culture medium added with 0.1 mg / L IBA, and carrying out rooting culture so as to obtain a positive Nanlin 895 poplar transgenic plant. Experiments prove that the adventitious bud induction rate of the plant material is 70%, the adventitious bud elongation rate is 40%, the rooting rate is 90%, and the method has good practicability.

The application provides application of a GmERF5 gene in drought resistance and salt stress resistance, and belongs to the technical field of genetic engineering.The GmERF5 gene is cloned into a pCAMBIA3301 vector to obtain a recombinant vector; the obtained recombinant vector is transformed into Agrobacterium rhizogenes, and a transgenic complex plant is obtained by using the obtained Agrobacterium rhizogenes to infect seeds, so that overexpression of the GmERF5 gene is realized.The overexpression of the GmERF5 gene can improve the antioxidant enzyme activity of the transgenic soybean plant, and the drought and salt stress resistance of the soybean is enhanced.The soybean GmERF5 of the application provides a new regulation gene resource for improving the drought resistance and salt tolerance of soybean, can be used for cultivation and improvement of stress-resistant soybean materials, lays a theoretical foundation for drought resistance and salt molecular mechanism, and simultaneously provides a theoretical basis and gene resource for soybean drought resistance and salt molecular breeding.

The invention provides a flora and an application of the flora in biodegradation of polyurethane plastics. The flora is prepared from raoultella ornithinolytica, klebsiella pneumoniae, coxakella rhizogenes and achromobacter xylosoxidans. According to the method, the polyurethane degrading flora is screened from the collected soil sample, the flora can efficiently degrade waterborne polyurethane and thermoplastic polyurethane elastomers with large chemical structure difference at the same time to degrade polyurethane plastic, the process is greatly simplified, the operation cost is reduced, traditional landfill incineration is replaced, green circulation of polyurethane is assisted, and the method has the advantages of being environmentally friendly and environmentally friendly. The method has important practical significance for solving the problem of environmental pollution caused by polyurethane waste and promoting green cyclic utilization of polyurethane materials.

The invention belongs to the technical field of plant genetic engineering and biotechnology, and discloses a construction method of a Hainan camellia oleifera seedling transgenic hairy root system, which comprises the following steps: selecting a 3-month-old camellia oleifera seedling explant Haidao camellia oleifera No.4, pretreating, transferring a target plasmid into agrobacterium rhizogenes K599 by an agrobacterium-mediated method, and culturing to obtain the Hainan camellia oleifera seedling transgenic hairy root system. Preparing an infection solution, optimizing infection conditions, inducing hairy roots to generate, and finally verifying the transgenosis effect through fluorescence observation and molecular identification. According to the method, an infection liquid formula, infection conditions and culture conditions are optimized, the induction efficiency of the transgenic hairy roots is remarkably improved, technical support is provided for functional gene research and genetic improvement of Hainan camellia oleifera, and the method has important application value.

The invention belongs to the technical field of genetic transformation, and particularly relates to a poplar genetic transformation method suitable for an outdoor environment. The method comprises the following steps: obtaining a poplar scion, cutting the poplar scion into 18cm cutting slips, soaking the cutting slips in an activated agrobacterium rhizogenes solution for 30 minutes to obtain an infected scion, performing soil culture or water culture, and transplanting the scion into a large container for continuous culture after transgenic positive roots grow out. The method provided by the invention does not need to depend on sterile conditions, genetic transformation can be realized through cuttage and water culture in an outdoor environment, and the method has the advantages of high throughput and large scale.

An induction method for an Antirrhinum majus L. hairy root producing an anthocyanin and a glycoside derivative thereof, a method for screening an inducing gene, and production and use, specifically belonging to the technical field of plant tissue culturing. The induction method for an Antirrhinum majus L. hairy root producing an anthocyanin and a glycoside derivative thereof comprises the following steps: using Agrobacterium rhizogenes containing an inducing gene to infect an Antirrhinum majus L. explant, co-culturing the Agrobacterium rhizogenes and the Antirrhinum majus L. explant, and performing sterilization culture to obtain the Antirrhinum majus L. hairy root producing the anthocyanin and the glycoside derivative thereof. The inducing gene comprises a gene encoding an MYB transcription factor, or a mutant thereof. The induction method can quickly and effectively obtain the hairy root synthesizing the anthocyanin and the glycoside derivative thereof while realizing the acquisition of the anthocyanin and the glycoside derivative thereof in a short time without being affected by seasons in the case of limited germplasm resources.

The invention provides a method for verifying gene functions based on combination of agrobacterium rhizogenes and a movable element. The provided method comprises the following steps: (1) constructing an expression vector fused by a target gene and a movable element, wherein the target gene is a functional gene to be verified of the plant; (2) transferring the expression vector into agrobacterium rhizogenes to obtain transformed agrobacterium rhizogenes; (3) infecting a plant by utilizing the transformed agrobacterium rhizogenes to obtain a positive plant with hairy roots; (4) determining the function of the target gene in the plant through the expression of the target gene in the upper end tissue of the positive plant and the phenotypic difference between the target gene and the control plant; wherein the movable element is a genetic material capable of transferring the target gene from the hairy root of the positive plant to the upper tissue. The method provided by the invention is simple to operate, high in transformation efficiency and stable and reliable in phenotype, and can be widely applied to plant gene function research.

The application provides a method for inducing transgenic hairy root formation of Lagenaria vulgaris by non-tissue culture, comprising the following steps: (1) Lagenaria vulgaris culture: sowing Lagenaria vulgaris seeds in a seedling substrate, and culturing in a light incubator until the Lagenaria vulgaris plant grows to a stage between one heart and one leaf and one heart; (2) infection by a hole-punching and smearing method: taking the cotyledon node to 1 cm below the cotyledon node of the Lagenaria vulgaris plant as an infection site, punching a hole around the infection site, and smearing the hole with Agrobacterium rhizogenes; (3) nodule induction: continuing to culture the Lagenaria vulgaris plant in the light incubator, and observing that the infection site is swollen; and (4) inducing hairy root formation by cutting and culturing: obliquely cutting the swollen position of the infection site of the Lagenaria vulgaris plant, and inserting the lower end of the obtained upper plant, i.e. the obliquely cut end, into a growth carrier for culture to induce hairy root formation. The application can induce transgenic hairy root formation of Lagenaria vulgaris, has a high induction rate, and can be applied to promoter function research, functional gene analysis, secondary metabolite research and the like.

The invention discloses application and a preparation method of an alfalfa tissue culture hairy root tip chromosome slice, and the preparation method comprises the following steps: (1) preparing an explant by using mature alfalfa leaves, and then mediating hairy roots to transform the explant by using agrobacterium rhizogenes to obtain hairy roots; and (2) carrying out chromosome slide preparation by using the hairy roots obtained in the step (1). According to the method disclosed by the invention, the hairy roots cultured in vitro are used as chromosome slide preparation materials, and the hairy roots cultured in vitro are generated by induction of agrobacterium rhizogenes, have the characteristics of hormone autotrophy and infinite growth, are rapid in growth, stable in heredity and easy to operate, can be subcultured and preserved for a long time under an aseptic condition, realize material taking as required, are not limited by seasons and growth cycles, and are suitable for large-scale production. And the flexibility and repeatability of the experiment are greatly improved.

This invention relates to a compound antibacterial agent and its application in open plant tissue culture. The active ingredients of the compound antibacterial agent include rifampin, kasugamycin, ticarcillin sodium clavulanic acid, and tetrachloroisophthalonitrile. The compound antibacterial agent can synergistically inhibit Agrobacterium rhizogenes or Rhodotorula glutinis, contaminating bacteria that are easily generated during plant tissue culture.

The invention discloses an agrobacterium rhizogenes-mediated arrowhead genetic transformation method, and belongs to the technical field of plant genetic engineering. An agrobacterium rhizogenes-mediated arrowhead genetic transformation system is established through stem tip tissue culture without inducing calluses, and the method comprises the following steps: sterilizing arrowhead stem tips and preparing sterile stem tips; culturing and activating agrobacterium rhizogenes; infecting the stem tips with agrobacterium rhizogenes; performing stem tip tissue culture to induce rooting; performing positive identification on new roots; acclimatizing and transplanting to obtain positive seedballs, and accelerating germination and regenerating to obtain positive plants. The arrowhead genetic transformation method based on agrobacterium rhizogenes, established by the invention, has important significance on gene function research and molecular breeding of arrowhead.

The invention relates to a construction method of an Agrobacterium rhizogenes mediated acacia melanoxylon genetic transformation system, which comprises the following steps: by taking an acacia melanoxylon SR17 sterile single plant as an explant, selecting an Agrobacterium rhizogenes strain Ar1193, infecting for 45 minutes in a heavy suspension of which the bacterial liquid concentration OD600 is equal to 0.9 and the acetosyringone concentration is 100 [mu] mol.L <-1 >, co-culturing for 3 days, and then transferring into a bacteriostatic culture medium to induce hairy roots; and taking the transgenic hairy roots obtained by induction as explants, and inoculating the explants into a WPM culture medium containing 0.05 mg.L <-1 > TDZ and 200 mg.L <-1 > casein hydrolysate to induce regeneration of adventitious buds so as to obtain complete transgenic plants. The screening concentrations of the four antibiotics, namely kanamycin, hygromycin, glyphosate and glufosinate-ammonium, are determined; a stable and efficient acacia melanoxylon genetic transformation system is established, the hairy root induction rate reaches 63.17%, the transgenic positive rate reaches 15.42%, and the adventitious bud regeneration rate reaches 52%.

The invention discloses a construction method and application of a chassis cell line for directionally synthesizing anthocyanin, and belongs to the technical field of cell line construction. The invention provides a construction method of a chassis cell line for oriented synthesis of anthocyanin, the chassis cell line obtained by the method has all genes for oriented synthesis of anthocyanin, and particularly, the purpose of oriented synthesis of a large amount of anthocyanin can be achieved by overexpressing anthocyanin in plants to synthesize transcriptional regulatory factors. According to the invention, an overexpression hairy root strain is obtained by constructing a plant overexpression vector of a transcription factor and utilizing an agrobacterium rhizogenes mediated method based on a chassis cell hairy root genetic transformation system, namely the chassis cell line is obtained. By utilizing the chassis cell line, the effect of efficiently and directionally synthesizing cyanidin through gene overexpression can be achieved, the content of cyanidin is high, and the purity of extracted anthocyanin is remarkably improved. The chassis cell line provided by the invention has the advantages of short growth cycle and simple culture mode, and can be subjected to industrial mass production.

A genetic transformation method for transferring exogenous genes into Prunella vulgaris and its application include the following steps: using Prunella vulgaris seedlings growing in substrate soil before flowering as infection material; introducing an expression vector containing the exogenous gene into Agrobacterium rhizogenes to prepare Agrobacterium rhizogenes infection solution; before infection, obliquely cutting and injuring the base of the Prunella vulgaris stem, removing the original root system, placing the Prunella vulgaris in the Agrobacterium rhizogenes infection solution, and then infecting using ultrasound; after infection, planting the Prunella vulgaris in the substrate, culturing in the dark, and continuing to grow under long-day conditions until hairy roots are produced. This invention achieves efficient, simplified, tissue culture-free, and visualized hairy root genetic transformation in Prunella vulgaris, with a genetic transformation rate of over 70% after 8 weeks of culture, providing a novel technical approach for the genetic improvement and functional gene research of Prunella vulgaris.

The application belongs to the field of biology and modern agricultural technology, and discloses a rooting aid invasion agent, and a formula thereof is as follows: B5 culture medium 3.0-3.5 g / L, sucrose 28-32 g / L, MES 3.8-4.0 g / L, acetyl-syringone 38-42 mg / L, 6-BA 1.5-1.8 mg / L, gibberellin 0.02-0.03 mg / L, and surfactant Silwet L-77 100 ml / L, and pH is 5.4. The application also discloses a soybean hairy root induction and transformation method mediated by Agrobacterium rhizogenes by using the rooting aid invasion agent, and the method comprises the following steps: disinfection and germination of soybean seeds; resuspension of Agrobacterium rhizogenes with a target expression plasmid in the rooting aid invasion agent to obtain an Agrobacterium rhizogenes aid invasion liquid; infection; co-culture; and screening of positive roots.

The invention discloses a tissue-culture-free agrobacterium rhizogenes-mediated genetic transformation system method for crape myrtle hairy roots, and relates to the technical field of molecular biology.The method comprises the steps that an overexpression vector carrying an RUBY visual tag is constructed, agrobacterium rhizogenes K599 is introduced, and a positive engineering strain is obtained through resistance screening and molecular verification; propagating the positive engineering strain, centrifugally collecting thalli, resuspending by using a buffer solution containing acetosyringone, adjusting the concentration of a bacterial solution, and standing and activating to form a bacterial suspension for infection; the method comprises the following steps: selecting current-year semi-lignified branches of crape myrtle to prepare cutting slips, retaining two pairs of bud points, and performing incision treatment; the cutting slips are completely immersed in the bacterial suspension for infection, and agrobacterium tumefaciens-mediated genetic transformation is completed in combination with vacuum-assisted treatment; taking out the cutting slips, temporarily dipping the cutting slips in a rooting aqueous solution, and then cutting the cutting slips in a sterile substrate formed by mixing vermiculite and perlite; and culturing for 24 hours under a dark condition, and continuously culturing for 30 days in a high-temperature and high-humidity environment.

This invention discloses a method for transforming cinnamon into transgenic hairy roots mediated by Agrobacterium rhizogenes, belonging to the field of plant genetic engineering technology. The method includes the following steps: (1) using sterile cinnamon seedlings as explants, preparing an Agrobacterium rhizogenes inoculum for infection, and immersing the explants in the inoculum for infection treatment; (2) co-culturing the infected explants, then transferring them to an antibacterial culture medium for further culture to induce hairy roots; (3) screening and identifying the induced hairy roots to obtain transgenic hairy roots. This invention utilizes rootless sterile cinnamon seedlings as explants, which is convenient to obtain. Using these explants as hairy root induction materials can shorten the induction period and increase the induction rate. The transgenic hairy root system established by this invention lays the foundation for future analysis of cinnamon gene function; at the same time, it also provides stable materials for subsequent induction of complete transgenic regenerated plants using hairy roots.

The invention discloses a sweet potato root system K + transport channel gene IbSKOR as well as a recombinant overexpression vector and application thereof. The nucleotide sequence of the gene IbSKOR is as shown in SEQ ID NO. 10. The method comprises the following steps: constructing a sweet potato root system K < + > transport channel key gene IbSKOR to an overexpression vector, and then transforming the recombinant overexpression vector into agrobacterium rhizogenes K599; then, transforming the expression vector containing the sweet potato IbSKOR gene into a sweet potato block through an agrobacterium rhizogenes mediated in-situ genetic transformation system; and growing seedlings from the positive transgenic potato blocks to obtain stable transgenic plants. It is further found that the drought tolerance of IbSKOR overexpressed sweet potato plants is remarkably improved. Therefore, a new drought-enduring sweet potato strain can be obtained through overexpression of the IbSKOR gene, the research blank of a sweet potato K + transport mechanism is filled, a new sweet potato genetic transformation technology method is provided, and reference is provided for cultivation and screening of new drought-enduring plant varieties.

This invention belongs to the field of plant genetic engineering technology and discloses a method for constructing transgenic hairy roots with multi-gene co-expression of the tanshinone biosynthesis pathway. The method first constructs a multi-gene expression vector containing the SmCPS1, SmKSL1, and SmCYP76AH1 genes, and then introduces the multi-gene expression vector into Agrobacterium rhizogenes. Subsequently, recombinant Agrobacterium rhizogenes is used to infect tanshinone explants, and transgenic hairy roots are induced after co-culture, sterilization culture, and resistance screening. Positive hairy roots are then identified and their expression analyzed by PCR and qRT-PCR, achieving stable co-expression of key genes in the tanshinone biosynthesis pathway. This invention achieves a single co-transformation of multiple key genes, simplifies the multi-gene transformation process, reduces the risk of gene segregation and expression instability caused by multiple transformations, and can provide stable transgenic materials for research on the tanshinone biosynthesis pathway and the metabolic regulation of active components in tanshinone.

The application provides application of a soybean GmERE1 gene in drought resistance and salt stress resistance, and belongs to the technical field of plant genetic engineering.The nucleotide sequence of the GmERE1 gene is shown as SEQ ID NO.1, and the protein sequence coded by the GmERE1 gene is shown as SEQ ID NO.2.The application also provides a recombinant vector for overexpressing and knocking down GmERE1 gene expression, and the recombinant vector is transformed into Agrobacterium rhizogenes;transgenic complex plants are obtained by using the obtained Agrobacterium rhizogenes to infect seeds, so that overexpression and knockdown expression of the GmERE1 gene are realized.Combination of GmERF5 and GmERE1 can improve the activity of peroxidase in soybean plants by promoting the expression of GmPOD1, and then improve the drought resistance and salt tolerance of the plants.

The invention provides application of agrobacterium rhizogenes in promoting rooting of fructus forsythiae, and belongs to the technical field of rapid vegetative propagation of plants. The method for promoting rooting of fructus forsythiae comprises the following steps: (1) inoculating agrobacterium rhizogenes into a culture medium for activation culture; 2) performing amplification culture on the activated and cultured bacterial liquid, collecting the bacterial liquid, and re-suspending the bacterial liquid to obtain a re-suspended bacterial liquid; according to the method, complex equipment is not needed, operation is easy and convenient, the cost is low, the rooting rate of fructus forsythiae is remarkably increased, the rooting time is shortened, and the method is suitable for large-scale seedling production.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a construction method and application of a hairy root system of dracocephalum heterophyllum benth with high rosmarinic acid content. According to the method, dracocephalum heterophyllum benth aseptic seedling stem segments are used as explants, agrobacterium rhizogenes LBA9402 carrying a rosmarinic acid synthase (RAS) gene expression vector is used for infection, co-culture and induction, and then positive transgenic hairy roots integrated with the RAS gene are obtained through multi-stage screening and identification. The dracocephalum heterophyllum hairy root system established by the invention has the characteristics of genetic stability, rapid growth and strong rosmarinic acid synthesis capability, the rosmarinic acid content is obviously increased, and an efficient biotechnology platform is provided for realizing large-scale and sustainable production of rosmarinic acid.

The application discloses a method for quickly obtaining a transgenic raspberry plant. The application selects 3-6 mature leaves at the top of a raspberry plant as a receptor material, contacts the receptor material with Agrobacterium rhizogenes ATCC15834 for infection, inserts the infected receptor material into a soil matrix for co-culture, applies an antibiotic every week after the co-culture, forms a callus at a petiole, differentiates the callus into a root, grows a bud at the root, and finally obtains the transgenic raspberry plant. The method has the advantages of simplicity, short culture period and no need for tissue culture.

The invention relates to the technical field of plant breeding, in particular to an agrobacterium rhizogenes-mediated grape plant genetic transformation method under a non-sterile environment and application of the agrobacterium rhizogenes-mediated grape plant genetic transformation method. The genetic transformation method comprises the following steps: infecting grape seeds with agrobacterium rhizogenes carrying a target gene under the vacuum condition of 0.6-1 kg / cm < 2 > for more than 20 minutes, and ensuring that the OD600 value of the agrobacterium rhizogenes is 0.4-1.2; the grape seeds are grape immature zygotic embryos in a germination state; and the agrobacterium rhizogenes are K599 agrobacterium rhizogenes. The invention provides a genetic transformation method based on a specific seed growth period and agrobacterium tumefaciens, which can obviously improve the transformation efficiency and shorten the period from seed germination to seedling, and has the characteristics of high repeatability and wide applicability. And an efficient and stable genetic transformation technical basis is provided for gene function research and variety improvement of grapes and other grape plants.