129 results about "Rhizogene" patented technology

The invention belongs to the technical field of genetic engineering, and particularly relates to an agrobacterium rhizogenes-mediated visualized genetic transformation method for cucurbitaceae plants, which comprises the step of transfecting the cucurbitaceae plants by using agrobacterium rhizogenes containing recombinant plasmids, and the recombinant plasmids are expression cassettes containing Ruby reporter genes. A visual agrobacterium rhizogenes-mediated transformation system of the cucurbitaceae plants is established by utilizing the RUBY gene, professional equipment or chemical treatment is not needed, red hairy roots can be visually screened to identify and induce the positive plants, and the method is simple, rapid and efficient and can be used for identifying and inducing the positive plants of the cucurbitaceae plants such as bottle gourds and the like. And the gene is transferred into an exogenous gene or is subjected to gene editing on the cucurbitaceae plant, so that the gene has important significance.

The invention provides a SmbHLH130 gene and an application of the SmbHLH130 gene in increasing the content of tanshinone and salvianolic acid in salvia miltiorrhiza. According to the invention, a transcription factor SmbHLH130 is obtained through multi-omics data analysis in the early stage. A CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) technology is utilized, a SmbHLH130 gene editing carrier is constructed, mutant hairy roots of gene knockout SmbHLH130 are obtained through agrobacterium tumefaciens transformation mediated by agrobacterium rhizogenes ATCC15834, and whether knockout of the mutant hairy roots is successful or not is identified through sequencing. And determining the content of tanshinone and salvianolic acid in the hairy roots of the transgenic salvia miltiorrhiza by using a high performance liquid chromatography (HPLC). The tanshinone content and the salvianolic acid content of the obtained gene knockout hairy roots are remarkably improved. Real-time fluorescent quantitative PCR (RT-qPCR) analysis shows that the expression quantity of a key enzyme gene participating in the biosynthesis of the active component of the salvia miltiorrhiza is also remarkably increased. The invention provides a method for increasing tanshinone and salvianolic acid contents of hairy roots of salvia miltiorrhiza, which can provide a new thought and target gene for production of active components of salvia miltiorrhiza and molecular design and breeding of salvia miltiorrhiza, and has higher application value and research value.

The invention provides a genetic transformation method suitable for shiny-leaved yellowhorn hairy root induction. The genetic transformation method comprises the following steps: culturing shiny-leaved yellowhorn seedlings infected by agrobacterium rhizogenes, preparing agrobacterium rhizogenes infection liquid, infecting the agrobacterium rhizogenes, inducing transgenic hairy roots by the agrobacterium rhizogenes and culturing. According to the method, a receptor material, a red tag gene RUBY and a green fluorescent gene GFP which are rapidly cultured and are suitable for agrobacterium rhizogenes infection are used as reporter genes, induction and transformation of hairy roots are carried out through agrobacterium rhizogenes mediation by utilizing a non-tissue culture infection and induction culture method, and the hairy roots are obtained. The cultivation period and cost of traditional plant hairy root induction are reduced, and a hairy root genetic transformation method suitable for shiny-leaved yellowhorn is established.

The invention relates to the field of asexual propagation of avocados, in particular to a method for inducing rooting of avocado cutting slips by utilizing agrobacterium rhizogenes. The method comprises the following steps: under open experimental conditions, taking current-year-old semi-lignified shoots of grafting plant scions of persea americana L. of 2-3 years old as experimental materials, taking high-temperature sterilized coco coir as a cutting medium, carrying out vacuum infiltration infection on the shoots in an agrobacterium rhizogenes K599 infection solution for 20 minutes, infecting wounds of seedlings, inducing rooting, and carrying out rooting. A system for inducing avocado cutting rooting by using agrobacterium rhizogenes is established. According to the method, the technical problem that the avocado cutting slips are difficult to root is solved, and the technical effect of rapidly breeding the avocado clonal rootstock seedlings is achieved. According to the method, clonal breeding, popularization and application of the excellent stock variety can be realized, a more convenient and efficient way is provided for breeding, popularization and application of the excellent persea americana stock variety, and the method has important significance for breeding, popularization and application of the excellent persea americana stock.

The invention discloses an agrobacterium rhizogenes-mediated wild jujube seedling hypocotyl in-vivo transgenosis method, and belongs to the technical field of woody plant genetic engineering. The method mainly comprises the following four steps: cultivating wild jujube seedlings, preparing agrobacterium rhizogenes infection liquid containing a GUS label and a ZjMAPKK4 gene, scratching hypocotyl, smearing the agrobacterium rhizogenes infection liquid on a wound to induce hairy roots, and identifying and screening transgenic positive hairy roots. According to the method, wild jujube seedlings serve as materials, agrobacterium rhizogenes serve as mediation, the hypocotyl position is selected to be scratched and infected, and a transgenic material specifically expressed at the root can be rapidly obtained. The method is easy to operate, short in period and low in cost, the process of wild jujube gene function verification can be greatly accelerated, a new method is provided for wild jujube gene function verification, and the method has important significance on wild jujube gene functions and genetic engineering breeding.

The invention relates to the technical field of plant genetic engineering, and provides an agrobacterium rhizogenes-mediated sunflower transient transformation system and a construction method thereof, sterile conditions and tissue culture are not needed, a vector to be transformed is transferred into agrobacterium rhizogenes, positive strains are cultured and screened, and a dip dyeing solution is prepared; cutting a stem section of the sunflower, and pricking holes near a cut; immersing the stem segments in the dip dyeing liquid for a period of time; taking out and inserting into sterilized vermiculite soil mixed with orobanche coerulescens for rooting to obtain an instantly transformed sunflower plant; the method is easy and convenient to operate and short in period, the expression level of the HaCERK1 gene can be effectively improved, the resistance of sunflowers to orobanche coerulescens infection is enhanced, and the gene function research period is shortened.

The invention relates to the technical field of biology, and particularly discloses an agrobacterium rhizogenes mediated broad bean hairy root induction method without tissue culture, which comprises the following steps: germination of broad bean seeds, transformation of agrobacterium, infection of explants, co-culture of the explants and identification of transgenic broad bean hairy roots. According to the broad bean hairy root induction method provided by the invention, transgenic hairy roots can be obtained only in 2-4 weeks, the method is simple to operate, and tedious sterile operation is avoided; besides, the recombinant agrobacterium rhizogenes of a betaine biosynthesis system RUBY are utilized to infect the broad bean hypocotyl and then culture, so that the broad bean hairy root induction method is successfully established, and the transgenic hairy root can be clearly observed by naked eyes without special equipment or chemical treatment.

The invention provides an agrobacterium rhizogenes-mediated papaya hairy root genetic transformation method and a papaya transgenic positive plant, and belongs to the technical field of plant genetic engineering. The invention provides a papaya genetic transformation method independent of tissue culture, which can directly utilize agrobacterium rhizogenes to treat explants for culture to obtain papaya transgenic positive plants without a series of tedious and complicated sterile processes such as callus induction and induced differentiation rooting. The problems of long tissue culture transformation period, large manpower and material resource consumption, low positive rate and the like are avoided. The constructed agrobacterium rhizogenes mediated genetic transformation system has the characteristics of high efficiency, stability and rapidness, the agrobacterium rhizogenes K599 cultured by using the solid TY culture medium can directly infect wounds, the operation is simple, the hair root incidence rate and the positive rate are relatively high, additional experimental equipment is not needed, and the method is suitable for most laboratories.

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 an induction method for obtaining a transgenic persea americana composite plant without tissue culture, and belongs to the technical field of persea americana biology. The induction method provided by the invention comprises the following steps: germinating avocado seeds by using quartz sand to obtain avocado seedlings, infecting rootless seedlings from the avocado seedlings by adopting a vacuum infiltration method, and inserting the rootless seedlings subjected to dip dyeing into a sterilized coco coir matrix for culturing to obtain the avocado composite plant. According to the method, an efficient avocado genetic transformation system mediated by agrobacterium rhizogenes is established, tissue culture is not needed, rootless seedlings are adopted as materials, the transformation efficiency is improved, meanwhile, consumption of experimental equipment, consumables and manpower is reduced, the problem that the avocado genetic transformation system is not established yet is solved, and the method is suitable for popularization and application. The technical effect of quickly, efficiently, simply and easily obtaining the transgenic material with low consumption is achieved, and a more convenient and efficient way is provided for genetic transformation of avocados.

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 belongs to the technical field of genetic engineering, and discloses a method for increasing the content of camptothecin in hairy roots of ophiorrhiza pumila and transgenic plants of the ophiorrhiza pumila by using CaORA1. Comprising the following steps: acquiring a CaORA1 coding sequence, expressing, regulating and forming an overexpression vector, transforming agrobacterium rhizogenes, transforming ophiorrhiza pumila, inducing hairy roots, inducing a ophiorrhiza pumila transgenic plant, and measuring the content of camptothecin. A method for inducing a transgenic ophiorrhiza pumila plant through the hairy roots of the ophiorrhiza pumila is established in the invention, the overexpressed CaORA1 can promote the expression of key enzyme genes OpSTR and OpSLS in a camptothecin biosynthetic pathway, and HPLC (High Performance Liquid Chromatography) results show that the camptothecin content of the transgenic hairy roots and the transgenic plant is obviously improved.

The invention discloses an agrobacterium rhizogenes mediated sunflower hairy root transformation system, and belongs to the technical field of genetic transformation. The transformation system comprises the following steps: accelerating germination of sunflowers, transferring germinated sunflower seeds into a seedling tray, wrapping with a preservative film for moisturizing, and culturing in an illumination incubator; the method comprises the following steps: culturing an agrobacterium rhizogenes Ar. Qual strain containing a pBI121-GUS (glucuronidase) plasmid; preparing a bacterial liquid; the method comprises the following steps of: beveling sunflower seedlings along a position 3cm below cotyledons, and immersing cuts into a bacterium solution containing pBI121-GUS plasmid agrobacterium rhizogenes Ar.qual; the sunflower seedlings are transferred to a seedling raising plate to be cultured; culturing for 5-6 days, and transferring the seedlings into a filter paper system of a culture dish to promote root growth. According to the agrobacterium rhizogenes-mediated sunflower hairy root transformation system, the genetic transformation efficiency of sunflowers is improved, the transformation period is greatly shortened, and obtained hairy root plants are good in growth consistency, robust in seedlings and capable of being used in molecular breeding research.

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 discloses a method for inducing apocarya stems to generate transgenic hairy roots, and belongs to the field of forest tree genetic engineering. According to the method, the semi-lignified or lignified stems of the apocarya are infected by utilizing the agrobacterium rhizogenes to obtain stable transgenic hairy roots, rooting can be induced within 1-2 months, the rooting rate is as high as 94.43%, and the transgenic positive rate can reach 98.38%. The invention provides an economical and efficient non-tissue culture dependent transgenic method, it is clear that transgenic roots can be used for gene function verification, the method breaks through the technical obstacle that stable genetic transformation of the carya illinoensis is difficult, and technical support is provided for research of functional genes.

The invention provides an induction method of sesbania hairy roots, which comprises the following steps: beveling hypocotyl of a sesbania seedling, dipping recombinant agrobacterium rhizogenes in the oblique incision of the seedling on the ground, inserting the seedling into vermiculite, and culturing until a new root system is generated, namely the new root system is a sesbania transgenic root system. The method is simple to operate, low in cost and short in time consumption, avoids tedious sterile operation, provides important technical support for molecular biology research of sesbania, and has a relatively great application prospect.

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 provides a method for carrying out genetic transformation on safflower carthamus in a tissue-free culture mode, which comprises the following steps: cutting off radicles of newly germinated safflower carthamus seedlings, taking the radicles as explants, infecting the explants in a mode of smearing agrobacterium rhizogenes containing a target gene on wounds, burying the infected explants in soil for culturing, and carrying out tissue-free culture on the explants to obtain the safflower carthamus. The transgenic safflower hairy roots are obtained. The method fills the blank of a safflower non-tissue culture genetic transformation system, the whole transformation period is 20-30 days, and the method can be applied to rapid and efficient genetic transformation, gene editing and variety improvement of safflower.

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.

The invention discloses application of an MrDRRP gene in regulating and controlling drought resistance of hairy roots of medicago ruthenica, and belongs to the technical field of biology. According to the invention, a disease-resistant related gene, named as MrDRRP, derived from medicago ruthenica is utilized, the medicago ruthenica is transformed through an agrobacterium rhizogenes mediated transgenic technology, the MrDRRP transgenic hairy roots of the medicago ruthenica are obtained, and drought stress experiments prove that the drought tolerance of the obtained hairy roots of the medicago ruthenica is remarkably enhanced, and the length and fresh weight of the hairy roots are remarkably improved. Therefore, the method is provided for cultivating the drought-resistant medicago ruthenica, the drought resistance of the medicago ruthenica is enhanced, the planting range of the medicago ruthenica is widened, and the method has good application value.

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.