91 results about "Rhizogene" patented technology

The invention provides an agrobacterium rhizogenes-mediated forsythia suspensa hairy root genetic transformation method and application thereof, and belongs to the technical field of plant genetic engineering. The method comprises the following steps: introducing an expression vector into agrobacterium rhizogenes, and mixing with an infection solution to prepare an agrobacterium rhizogenes infection solution; the forsythia suspensa stem is put into the agrobacterium rhizogenes infection liquid to be infected for 0.5-1 h through a vacuum infiltration method, and the infected forsythia suspensa stem is cut into a matrix to be subjected to dark culture and then continues to grow till hairy roots are generated; the OD600 value of the agrobacterium rhizogenes infection liquid is 0.5 to 1.0. The method is suitable for an agrobacterium rhizogenes mediated genetic transformation system of the fructus forsythiae, and the system provides technical support for functional gene research, medicinal component analysis and molecular breeding of the fructus forsythiae, and has important significance for promoting genetic improvement and resource utilization of the fructus forsythiae.

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 discloses a method for high-yield production of rare saponin Rg3 through gene editing of a panax notoginseng adventitious root saponin synthesis pathway, which comprises the following steps: 1) inputting a gene CYP716A53V2 into an online tool CRISPR to find a series of potential Cas9 target sites, and carrying out random selection; (2) respectively designing downstream primers according to different selected Cas9 target sites, and carrying out PCR (Polymerase Chain Reaction) amplification by using a U6p.4-F primer as an upstream primer to respectively obtain different sgRNA sequences; (3) respectively connecting the sgRNA-n with a linearized pP1C.4 carrier, so as to obtain a recombinant plasmid; 4) respectively transforming the recombinant plasmids into agrobacterium rhizogenes, and culturing to obtain different recombinant bacterial liquids; and (5) respectively putting the pseudo-ginseng adventitious roots into different positive recombinant bacteria solutions, carrying out vacuum infection, and screening out the transgenic pseudo-ginseng adventitious roots with high Rg3 content. The method has the advantages of controllable steps, accurate regulation and control, greenness, environmental protection and high yield of saponin Rg3.

The invention relates to the technical field of genetic engineering, in particular to an agrobacterium tumefaciens-mediated radix pseudostellariae hairy root genetic transformation method and application thereof. The method comprises the following steps: preparing a radix pseudostellariae explant; activating and culturing agrobacterium rhizogenes; constructing an engineering bacterium containing a target gene recombinant vector; infecting explants and inducing hairy roots; carrying out resistance screening and sterilization on transgenic hairy roots; carrying out amplification culture on hairy roots; and carrying out RT-PCR detection. The hairy root genetic transformation method provided by the invention is simple and rapid to operate and high in gene expression level, and can provide technical support for gene function research. Meanwhile, by using the genetic transformation method provided by the invention, through overexpression of the radix pseudostellariae cyclic peptide coding gene, a transgenic material with significantly improved content of the pharmacodynamic substance cyclic peptide of the radix pseudostellariae can be obtained, and a research material is provided for researching a biosynthetic pathway of the cyclic peptide.

The invention discloses a rubus cochinchinensis efficient genetic transformation method based on RUBY reporter gene and agrobacterium rhizogenes mediation, which realizes that the transformation period is shortened to be within 15 days and the transformation efficiency is improved to be more than 60% through direct explant treatment under a non-tissue culture condition. The method comprises the following specific steps: 1) preparing an explant material; (2) constructing an RUBY carrier and transforming agrobacterium rhizogenes; (3) infecting the explant by utilizing agrobacterium rhizogenes carrying a target gene; (4) co-culturing after infection and carrying out RUBY visual screening; 5) positive root molecular biology verification; (6) inducing positive roots into buds; according to the method, induction of the positive callus, the positive root and the positive bud of the rubus coreanus stem segment under the non-tissue culture condition is successfully achieved, a stable and efficient genetic transformation system is constructed, the method has the remarkable advantages of being easy and convenient to operate, low in cost, wide in applicability, high in reproducibility and the like, and a key technical support is provided for molecular breeding of the rubus coreanus.

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 relates to the technical field of agricultural product production, in particular to a pepper cuttage method and application. The invention provides a pepper cuttage method which comprises the following steps: taking 8-leaf-age lateral branches of pepper as cutting slips, punching the base parts of the cutting slips, then infecting the base parts of the cutting slips by using agrobacterium rhizogenes, and culturing for 36-60 hours under a dark condition. According to the method, agrobacterium rhizogenes induction and punching technologies are combined, the rooting speed of pepper cuttage can be greatly increased, the cuttage survival rate is increased, the seedling raising time is remarkably shortened, and the purpose of rapid and effective seedling raising is achieved. According to the pepper cutting method, limitation of traditional cutting seedling raising is broken through, additional nutrient solution does not need to be added, pepper root systems can rapidly grow in vermiculite, the cutting seedling raising time is shortened, and the effect of rapid seedling emergence and transplanting is achieved.

The invention belongs to the technical field of genetic transformation, and particularly relates to an agrobacterium rhizogenes-mediated genetic transformation method for catalpa bungei. The genetic transformation method of catalpa bungei comprises the following steps: obtaining stem segments of catalpa bungei after co-culture; after co-culture is finished, the stem segments of the catalpa bungei after co-culture are soaked with an antibiotic solution and then inserted into a degerming culture medium until transgenic hairy roots or positive roots grow out, and catalpa bungei plants with the transgenic hairy roots or positive roots grow out are obtained; transplanting the catalpa bungei plants with the transgenic hairy roots or positive roots, taking down the positive roots of the catalpa bungei plants after the catalpa bungei plants grow up, and performing root segment cuttage to obtain the whole transgenic catalpa bungei plants. The catalpa bungei genetic transformation method provided by the invention has relatively high infection efficiency on different genotypes of catalpa bungei, and the positive rate of genetic transformation is relatively high; and the catalpa bungei genetic transformation method can be suitable for different genotypes of catalpa bungei, and has genotype universality.

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 present invention discloses a method for inducing transgenic hairy roots in the stems of Carya serrata, belonging to the field of forest genetic engineering. The method utilizes Agrobacterium rhizogenes to infect semi-lignified or lignified stems of Carya serrata to obtain stable transgenic hairy roots. These roots can be induced to develop within one to two months, with a rooting rate as high as 94.43% and a transgene-positive rate of 98.38%. This method provides a cost-effective, tissue culture-independent transgenic method, and clearly demonstrates that transgenic roots can be used to verify gene function. This method overcomes the technical barriers to stable genetic transformation in Carya serrata, providing technical support for the research of functional genes.

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.

An Agrobacterium rhizogenes mediated method for high-efficiency genetic transformation and gene editing in Brassica crops. A genetic transformation and gene editing system for Brassica crops is established by means of Agrobacterium-rhizogenes-mediated delivery of three growth and development regulatory factors, i.e. ZmWUS2, AtIPT and AtPLT5. By means of the Agrobacterium-rhizogenes-mediated delivery of the three growth and development regulatory factors, i.e. ZmWUS2, AtIPT and AtPLT5, explants are induced to form calli, from which shoots are directly formed, so that the problem of difficult regeneration of roots of Brassica crops is solved, thereby achieving high-efficiency transformation in species with poor root regeneration capacity.

The invention discloses a Ningxia wolfberry genetic transformation method based on adventitious bud induction. The agrobacterium rhizogenes are adopted for genetic transformation, and the problem that buds of traditional Ningxia wolfberry leaves are difficult to regenerate is effectively solved. The agrobacterium rhizogenes can effectively induce formation of hairy roots of the Ningxia wolfberry, so that the technical bottleneck in the regeneration process of the Ningxia wolfberry is broken through, and the transformation efficiency is remarkably improved. In the transformation process, although the Ningxia wolfberry hairy roots induced by agrobacterium rhizogenes successfully obtain transgenic markers, the original adventitious bud regeneration capacity is inhibited. By means of the dedifferentiation and repeated induction culture technology, the redifferentiation capacity of the hairy root induced callus is effectively recovered, the regeneration bud forming efficiency of the hairy root induced callus is remarkably improved, and a transgenic Ningxia wolfberry plant is obtained.

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 provides a genetic transformation method for obtaining a bamboo transgenic plant and a construction method of a bamboo transgenic plant regeneration system, and particularly belongs to the technical field of plant transgenic regeneration. The invention provides a genetic transformation method for obtaining a bamboo transgenic plant. The genetic transformation method comprises the following steps: cutting off a bamboo seedling, infecting a stubble wound position by adopting agrobacterium rhizogenes carrying a target gene, moisturizing and culturing the infected wound, and growing a transgenic bud at the wound position. According to the genetic transformation method, the bamboo can be helped to directly generate transgenic buds in the juvenile period through agrobacterium, the problems that a traditional transgenic bamboo process is complex, time consumption is long and efficiency is low can be solved, and thoughts are provided for research of a transgenic technology of the bamboo.

The present invention discloses a trifoliate orange cold-resistant gene CtrBBX32 and its application in genetic improvement of plant cold resistance. The CtrBBX32 gene is a gene of a subfamily of the zinc finger structural protein family isolated and cloned from the extremely cold-resistant citrus material trifoliate orange, and its sequence is shown in SEQ ID No. 1. Interference vectors and overexpression vectors are constructed with this gene, and it is introduced into trifoliate orange through genetic transformation mediated by Agrobacterium tumefaciens and Agrobacterium rhizogenes to obtain transgenic plants. Biological function verification shows that the cloned CtrBBX32 gene of the present invention has the function of regulating plant cold resistance. The discovery of this gene provides a new gene resource for plant stress resistance molecular design and breeding, and provides a new genetic resource for the implementation of green agriculture and water-saving agriculture. The development and utilization of this genetic resource is conducive to reducing agricultural production costs and achieving environmental friendliness.

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 present invention discloses an induction method for obtaining a transgenic avocado composite plant without tissue culture, and belongs to the field of avocado biological technology. The induction method provided by the present invention comprises the following steps: germinating avocado seeds with quartz sand to obtain avocado seedlings, infecting rootless seedlings from the avocado seedlings with a vacuum infiltration method, inserting the rootless seedlings after the infection into a sterilized coconut husk matrix for cultivation, and obtaining the avocado composite plant. The present invention establishes an efficient genetic transformation system for avocado mediated by Agrobacterium rhizogenes, which does not require tissue culture and uses rootless seedlings as materials, thereby improving the transformation efficiency and reducing the consumption of experimental equipment, consumables and manpower, solving the problem that the avocado genetic transformation system has not yet been established, achieving the technical effect of obtaining transgenic materials quickly, efficiently, simply and with low consumption, and providing a more convenient and efficient approach for avocado genetic transformation.

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%.