1064 results about "Callus" patented technology

The invention discloses a pennisetum purpureum gene PpbHLH148L and application thereof in improving the cold resistance of plants, and relates to the technical field of gene engineering. The nucleotide sequence of the gene is as shown in SEQ ID NO.5. After the gene is subjected to subcellular localization and expression condition analysis after cold stress, the gene is related to low-temperature tolerance. An overexpression vector of the gene is constructed, Agrobacterium tumefaciens is transferred, then Arabidopsis thaliana infection is carried out, cold tolerance determination is carried out on overexpressed Arabidopsis thaliana, phenotypic difference and physiological index difference between an overexpressed strain after cold treatment and a wild type are observed, and it is found that the gene can significantly improve the low-temperature stress tolerance of Arabidopsis thaliana. And in the low-temperature treatment of the pennisetum alopecuroides transgenic callus, the transgenic callus also shows higher tolerance than the wild callus at low temperature. The pennisetum purpureum gene PpbHLH148L can be used for low-temperature tolerance modification of plants, and has an application prospect in creation of low-temperature-resistant plant materials.

The invention relates to a same-variety grafting method for Tibetan pinus bungeana, and belongs to the technical field of forest vegetative propagation. In order to solve the technical problems that in the high-altitude environment, the grafting survival rate of the white bark pine is low, a connector is prone to being infected by microorganisms, and callus is formed slowly, efficient grafting is achieved through collaborative optimization of scion pretreatment and stock incision treatment. The method comprises the steps that a one-year-old branch with the diameter being 4-6 mm is selected and subjected to temperature and humidity control and composite antibacterial liquid dipping treatment; a sterile healing interface is formed in cooperation with rootstock incision slope cutting and four times of composite antibacterial liquid brushing; a one-year-old branch is cut, at least two or more bud points exist on the branch, two plump bud points are reserved, the branch is cut open from the position between the two plump bud points, and the scion is obtained. According to the method, the grafting survival rate and the interface stress resistance of the pinus bungeana in the alpine region are remarkably improved, the method is suitable for rapid breeding and hereditary characteristic retention of excellent germplasm of the pinus bungeana in Tibet plateau ecological restoration engineering, and more scion resources of the pinus bungeana can be utilized and protected.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a method for establishing an elymus nutans seed callus induction and regeneration system. The invention provides a tissue culture medium and a tissue culture method for culturing elymus nutans, and in the tissue culture medium provided by the invention, 2, 4-D can induce explants to form calluses and promote cell division and growth. The CH has a good effect on differentiation of embryoid and adventitious buds. ABA can promote the development of seed embryos. The KT can promote the generation of calluses and the growth of adventitious buds. NAA can promote plant growth and promote generation of adventitious roots. Under the combined action of NAA, 2, 4-D, ABA, KT and CH, the callus induction rate of the mature embryo is improved. The invention also provides a regeneration tissue culture method of elymus nutans, the mature embryo is used as an explant for tissue culture, and the callus induction rate of the mature embryo is high.

The invention relates to the technical field of plant tissue culture, in particular to a Rhododendron pulchrum tissue culture method based on callus regeneration. According to the method, the tender cotyledons of aseptic seedlings are taken as explants, and an efficient Rhododendron pulchrum tissue culture and rapid propagation technical system is established through induction of calluses, subculture multiplication of the calluses, differentiation of adventitious buds, bud strengthening and rooting culture, so that complete tissue culture progeny plants are formed. The induction rate of the callus tissue of the system is 100.0%, the proliferation multiple of the callus tissue in a subculture cycle reaches 5.2 times, 28.5 adventitious buds can be differentiated from a callus tissue block with the size of about 0.5 mm * 0.5 mm * 0.5 mm, the rooting rate reaches 95.8%, the system is not limited by factors such as seasons, climate and regions, and efficient and rapid propagation can be achieved all the year round. Meanwhile, the establishment of the system also lays a technical foundation for genetic transformation improvement, mutant screening breeding and the like of the Rhododendron pulchrum based on the callus.

The invention discloses a high-survival-rate method for grafting pinus sylvestris on pinus sylvestris in an alpine region. The grafting adaptability and the survival rate are improved through multi-dimensional innovation. A rising slope is arranged on the sunny side of a pinus sylvestris rootstock, double connectors of a falling slope are arranged on the shady side of the pinus sylvestris rootstock and matched with double wedge-shaped parts of Tibetan white bark pine scion buds, and the rootstock-scion contact face is increased; a photo-thermal response shape memory polymer bandage is adopted for fixation, germs are inhibited, and callus differentiation is promoted; a double-layer protection structure is constructed, a humidity control group is arranged in an inner-layer moisturizing breathable film to maintain the humidity of a connector to be larger than or equal to 60%, and an outer-layer anti-UV windproof net is intelligently adjusted to cope with sand wind and strong light. Furthermore, the callus differentiation is accurately regulated and controlled by pre-embedding pH responsive cellulose microspheres, or an intelligent temperature control film and an aerogel layer are additionally arranged to stabilize the temperature difference, and the freezing resistance is enhanced in combination with scion bud low-temperature domestication and abscisic acid treatment. The method remarkably improves the grafting survival rate, is suitable for efficient introduction of the Tibetan white bark pine in the alpine region, and is high in process synergy and high in environmental adaptability.

The invention discloses a large-scale preparation method of ginseng exosomes, which comprises the following steps: culturing calluses: inoculating explants into an induction culture medium until ginseng calluses are generated; proliferation of the calluses: inoculating the newborn calluses into a proliferation culture medium for proliferation culture; inducing germination and rooting: after the ginseng callus is amplified, transplanting the ginseng callus into a germination induction culture medium until the ginseng callus is germinated, and then transferring the ginseng callus into a rooting culture medium for culturing and rooting; and preparing the exosome, namely grinding root hairs grown from the ginseng callus regeneration plant, uniformly mixing the ground root hairs with PBS (Phosphate Buffer Solution) or performing liquid culture on ginseng suspension cells, and finally performing centrifugal treatment to obtain the ginseng exosome. According to the large-scale preparation method of the ginseng exosome, the endotoxin pollution in the growth and development process is reduced through ginseng callus culture rooting, and then the ginseng exosome is prepared, so that the large-scale preparation requirement in modern production and processing is met.

The invention relates to a method for producing notoginsenoside and ginsenoside by utilizing panax notoginseng cotyledon embryos, and belongs to the technical field of plant adventitious root tissue culture. The method comprises the following steps: inducing embryogenic calluses by utilizing immature zygotic embryos of panax notoginseng; proliferating the panax notoginseng embryogenic callus on a plate culture medium; s2, putting the pseudo-ginseng embryogenic callus obtained in the step S2 into a liquid culture medium, carrying out cell dispersion, and sieving by using a sieve with the aperture of 24-30 meshes; a mechanical stirring type bioreactor is filled with a liquid culture medium, and the globular embryo and the heart-shaped embryo are placed in the mechanical stirring type bioreactor to be cultured; s3, recovering the filtrate in the step S3, filtering by using a screen with the aperture of 30-80 meshes and 80-150 meshes to obtain uniform panax notoginseng embryogenic cells, and differentiating the panax notoginseng embryogenic cells in a triangular flask for the step S4; and drying the cotyledon embryo obtained in the step S4 at 45 DEG C, and grinding into powder. According to the method, large-scale generation of cotyledon embryos can be completed only in 4-6 weeks, the production process is remarkably accelerated, the method is suitable for industrial batch production requirements, and the production efficiency is high.

The invention discloses a gene for improving wheat immature embryo transformation efficiency and application thereof, and belongs to the technical field of plant genetic engineering. According to the invention, a new wheat regeneration gene TaDJA gene is found, and the TaDJA gene is over-expressed in a wheat variety 'Kefeng No.3' which is extremely difficult to transform, so that the callus induction rate and the transformation efficiency of wheat immature embryos can be improved; the method is beneficial for integrating exogenous nucleic acid molecules into the genome of the wheat variety difficult to transform, and provides technical support for genetic improvement of the wheat variety difficult to transform.

The invention discloses a dendrobium officinale tissue culture and rapid propagation seedling raising method, which belongs to the field of biotechnology, and comprises the following steps: S1, explant treatment: selecting a dendrobium officinale plant with good growth vigor and no obvious pest and disease damage, carrying out high temperature stress treatment at 40-45 DEG C for 6-8 days, then taking the tender stem of the dendrobium officinale plant subjected to high temperature stress treatment as the explant, and carrying out tissue culture and rapid propagation on the explant; directly transferring the explant to a low temperature of 2-6 DEG C for stress treatment for 8-12 hours; s2, callus induction: taking the explant treated in the step S1, disinfecting the explant, taking the explant without axillary buds, cutting the explant into small blocks, inoculating the small blocks to a callus induction culture medium, and carrying out dark culture for 12-15 days; the callus induction culture medium comprises 1.5 to 3 mg / L of MS + 2, 4-D, 2 to 3 mg / L of KT and 0.1 to 0.3 mg / L of NAA; s3, bud differentiation induction and rooting induction. According to the tissue culture and rapid propagation method for the dendrobium officinale, the browning rate of the callus during callus induction can be effectively reduced, and the tissue culture seedling raising efficiency is improved.

The invention relates to the technical field of plant extraction, in particular to a preparation method of a plant cell exosome, the plant cell exosome and application of the plant cell exosome. The preparation method comprises the following steps: after the surface of a plant leaf is scratched, inoculating the plant leaf into a first culture medium for induction culture, then transferring the plant leaf into a second culture medium for subculture to obtain a callus, and then adding the callus into a buffer solution for grinding to obtain a filtrate; centrifuging the filtrate again to obtain supernate; and adding an exosome complexing agent into the supernate, incubating overnight, centrifuging, and freeze-drying to obtain the exosome. The first culture medium comprises an MS culture medium and the following components added into the MS culture medium: pullulan, 2, 4-D, agar and NAA; the second culture medium comprises an MS culture medium and the following components added into the MS culture medium: pullulan, 2, 4-D, agar, NAA and sodium phytate. The preparation method can increase the protein content in the exosome.

The invention belongs to the technical field of plant biology, and particularly relates to a genetic transformation method based on oat immature embryos, which comprises the following steps: preparing an infection solution containing a target gene; taking immature caryopses of the oats 21-23 days after pollination; infecting the immature embryos with infection liquid, and co-culturing the immature embryos after infection is completed; after co-culture is completed, immature embryos are sequentially transferred into an L3-T culture medium and an L3-B5 culture medium to induce formation of calluses, and after the calluses are formed, the calluses are sequentially induced to be differentiated, rooted and hardened. According to the method, efficient genetic transformation of exogenous genes is realized by selecting proper oat immature embryos, proper culture conditions and proper transformation conditions. By means of the genetic transformation method, the genetic transformation efficiency of the oats is improved to 25% from 2%-3%. And meanwhile, the immature embryo is used as a transformation acceptor material, and the complete plant can be efficiently and stably regenerated by the method disclosed by the invention.

The invention provides an AbAIL5 gene and application of the AbAIL5 gene in improvement of genetic transformation efficiency of amorphophallus bulbifer, and belongs to the technical field of biology. The invention provides an AbAIL5 gene. The nucleotide sequence of the AbAIL5 gene is shown as SEQ ID NO: 1. Genetic transformation is performed on the amorphophallus bulbifer somatic embryo by constructing an overexpression vector of the AbAIL5 gene, so that the genetic transformation efficiency of the amorphophallus bulbifer callus can be improved, and candidate genes are provided for application of a large-scale breeding technology and a genetic transformation technology of the amorphophallus bulbifer callus in good varieties.

The invention discloses application of an OsWRKY28 gene in regulating and controlling the number of regenerated buds of rice callus, and belongs to the technical field of plant genetic engineering. According to the invention, a key regulation factor OsWRKY28 for regulating and controlling the regeneration of the rice callus is found. By constructing the rice OsWRKY28 knockout mutant and the overexpression strain, compared with a wild type, the number of regenerated buds of the callus of the OsWRKY28 knockout mutant is remarkably increased, and the number of regenerated buds of the callus of the OsWRKY28 overexpression strain is remarkably reduced. Therefore, it is determined that the OsWRKY28 gene plays an important role in regulating and controlling the regeneration process of the callus. Therefore, the regeneration capacity of the rice callus can be improved by knocking out the OsWRKY28 gene, so that the transgenosis efficiency is improved.

The invention relates to the technical field of plant breeding, and particularly discloses a metabonomics-based alfalfa stress resistance gene analysis function and a breeding method. The core of the method is that screening is carried out by combining a tissue culture technology with metabolic intervention; an alfalfa explant is inoculated into an induction culture medium containing a specific metabolic pathway regulator for metabolic intervention culture; then transferring the obtained callus to a verification culture medium containing the metabolism regulator and an abiotic adversity stress factor at the same time, and carrying out metabolism mark verification and screening; and finally regenerating the screened cell line into a complete plant. According to the method, metabolic pressure is actively applied to the cellular level, and dual verification screening is implemented, so that early-stage, efficient and accurate breeding of the stress-resistant alfalfa material is realized, and the technical problems of long period, low efficiency and insufficient utilization of metabolic information in traditional breeding are solved.

The invention discloses an efficient genetic transformation method of stem tumor sesbania, and belongs to the technical field of plant genetic transformation. According to the efficient genetic transformation method of the stem tumor sesbania, the cotyledonary node is used as the initial explant, callus induction is not needed, an optimized culture medium component is combined, a genetic transformation system of the stem tumor sesbania can be rapidly and effectively established, the genetic transformation efficiency can be improved and can reach 25% or above, the transformation time is shortened, and the cost is reduced. And a foundation is laid for domestication and accurate improvement work of the stem tumor sesbania.

The invention discloses a method for obtaining leymus chinensis mutants by using heavy ion beams to irradiate seed calluses, relates to the technical field of leymus chinensis breeding biology, and solves the three-low problems of low heading rate, low maturing rate and low germination rate of leymus chinensis seeds. After the leymus chinensis callus is subjected to induction culture, selecting faint yellow granular callus obtained by induction culture, and transferring the faint yellow granular callus into a culture dish filled with an induction culture medium for subculture; carrying out irradiation treatment by using a 5Gy carbon ion beam, transferring the callus subjected to irradiation treatment into a sterile culture bottle filled with a differential culture medium, and culturing to obtain adventitious buds; cutting off callus adhered to the base under a sterile condition, and inserting into a rooting culture medium for rooting culture after a rooting point is exposed; and after obvious root systems grow out, carrying out potting culture to obtain the leymus chinensis mutant. According to the invention, new genetic variation can be created, early in-vitro selection is realized, more effective mutants are obtained as new germplasm resources of leymus chinensis, and the breeding process of leymus chinensis is accelerated.

The invention relates to the technical field of biology, in particular to a vitis davidii VdbHLH51 gene as well as an encoding protein and application of the vitis davidii VdbHLH51 gene. The vitis davidii VdbHLH51 gene is located on a chromosome 13 of vitis davidii and is distributed in a 772522-773497 region, and the nucleotide sequence of the vitis davidii VdbHLH51 gene is as shown in SEQ ID NO. 1. The amino acid sequence of the protein coded by the gene is as shown in SEQ ID No.2. The invention also discloses an application of the vitis davidii VdbHLH51 gene in improvement of resistance of grapes to pathogenic bacteria after overexpression of grape calluses. The VdbHLH51 gene provided by the invention can improve the resistance of grapes to grape anthracnose, and provides a theoretical basis for grape anthracnose resistance breeding.

The invention provides an exosome from rice as well as a preparation method and application thereof. The exosome has a double-layer membrane spherical structure, and the particle size range is 20-1000nm. The exosome is obtained from rice calluses, and has the characteristics of high biological safety, high stability and sustainable mass production. And tests prove that the exosome has the functions of resisting aging, promoting cell regeneration and tissue repair, protecting and the like, and also can remarkably improve the cell absorption efficiency and the delivery efficiency of active ingredients such as peptide cells.

The invention relates to the technical field of plant genetic engineering, in particular to a ginkgo GbGRF2 gene and application of a GbGRF2-GbGIF2 fusion gene of the ginkgo GbGRF2 gene in plant tissue culture. According to the invention, the GbGRF2 gene coding region full length of the GRF gene family of gingko is successfully cloned from gingko, and the gene presents a specific expression mode in the regeneration process of gingko callus; the overexpression of the fusion gene of GbGRF2 and GbGRF2-GbGIF2 can significantly promote the regeneration efficiency of adventitious buds in the in-vitro culture process of plants; the gene provided by the invention has important theoretical significance and application potential in a plant tissue regeneration regulation network, and plays a key role in improving in-vitro regeneration efficiency of gymnosperm.

The invention discloses a construction method and application of an in-vitro callus-like organ, the survival rate of the organ is improved by improving freeze-formed bio-ink, and a stable directional channel structure is further prepared, so that the internal mass transfer problem of the callus-like organ is improved, and the material exchange efficiency is improved. The forming mode of the callus-like organ is simple and effective, the stent material is easy to prepare, and the callus-like organ has mature cross-linking and printing characteristics, is expected to be applied to drug screening research or bone defect repair, is clear in process and simple in composition, and has relatively high operability.

The invention relates to the technical field of functional food and biological medicine, in particular to a modified pseudo-ginseng exosome and an intestinal conditioning gel composition and application thereof. According to the gel beverage, an exosome derived from panax notoginseng callus cultured in a laboratory is used as a core active component, is modified through phosphatidylserine-aminated pectin-genipin and is embedded into a hydrogel matrix composed of carboxymethyl chitosan and sodium alginate, so that the exosome is protected from being degraded in the stomach, and the activity of the exosome in the stomach is improved. The exosome can be effectively released under the intestinal pH and flora environment, so that the targeted delivery of the intestinal tract is realized, and the preparation method is suitable for improving the intestinal inflammation and maintaining the intestinal health. The invention can effectively solve the technical problems of uncontrollable raw material quality and safety, low oral delivery stability and active ingredient availability and poor medication compliance of the existing plant exosome.

The invention discloses an application of an ALKBH9B gene in promotion of plant callus formation, belongs to the technical field of gene engineering, and provides an application of the ALKBH9B gene in promotion of plant callus formation. The invention also provides an application of an expression cassette, a vector or a recombinant bacterium containing the ALKBH9B gene in promoting formation of poplar leaf calluses. The ALKBH9B gene is knocked out, the expression quantity of the ALKBH9B gene is reduced, it is found that when the gene editing strain with the knocked-out gene is cultured on a callus induction culture medium for 5 days, callus tissue starts to be formed, the callus forming speed is high, and the net weight of the callus and the callus generation rate are remarkably higher than those of a wild strain. The result shows that ALKBH9B has an inhibition effect in the process of inducing the callus by the leaf, and the deletion of ALKBH9B can promote the formation of the callus.

The invention provides a dendrobium devonianum epiphytic tea tree cultivation method, which belongs to an epiphytic plant planting technology, and is characterized in that a dendrobium devonianum root system can be closely fused with a tea tree trunk through treatment of'minimal trauma + callus inducer ', and a structural foundation is laid for material exchange. The jasmonic acid methyl ester stress with a specific concentration is applied to the dendrobium nobile in the key phenological period, the dendrobium nobile is accurately induced to synthesize and secrete beneficial secondary metabolites, the theanine content in the tea can be remarkably increased, the proportion of ester type catechin can be reduced, and therefore the fresh degree of the tea is remarkably increased, and the astringent taste of the tea is remarkably reduced. The stress resistance of the dendrobium devonianum and the tea trees treated by the method is also enhanced, and the disease occurrence rate is reduced. While high-quality dendrobium nobile is harvested, the quality and economic benefits of the produced tea leaves are synchronously improved.

The invention discloses an artemisia apiacea tissue culture and one-step seedling forming method, which comprises the following steps: cutting stems with axillary buds of artemisia apiacea as explants, and carrying out sterilization and inoculation pretreatment; inoculating the explants on an induction culture medium for culturing, and inducing the explants to generate cluster buds; transferring the germinated cluster buds to a proliferation culture medium for proliferation culture of the cluster buds; and cutting the proliferated non-rooted seedlings, and transferring to a rooting culture medium for rooting culture. According to the method, single high-concentration excitation 6-BA is used as a hormone for inducing the culture medium, axillary bud stem explants can be directly induced to generate cluster buds, and the generation of calluses is remarkably reduced. Meanwhile, in the subculture multiplication culture, after the subculture stage and the concentration of the used 6-BA are flexibly adjusted, the generation of calluses and the generation of vitrification phenomena can be further reduced. According to the method, the cluster buds can be rapidly rooted, so that a large number of seedlings are obtained, and the method has the remarkable advantages of saving time and manpower and material resource cost in production.

The invention relates to the technical field of plant genetic engineering. The invention provides a beta-ketoester acyl-CoA reductase KCR gene, wherein the nucleotide sequence of the KCR gene is as shown in SEQ ID NO. 1. The invention also provides a recombinant vector. The invention also provides a recombinant bacterium. The invention also provides an application of the KCR gene, the recombinant vector or the recombinant bacterium in improving the salt tolerance of plants. The invention also provides application of the KCR gene, the recombinant vector or the recombinant bacterium in plant salt tolerance genetic improvement. The invention further provides a method for obtaining the salt-tolerant plant. After the KCR gene is over-expressed in the grape callus, the salt tolerance of the obtained transgenic grape callus is obviously improved. And after the gene is over-expressed in arabidopsis thaliana, the obtained transgenic arabidopsis thaliana epidermis wax is accumulated and the salt tolerance is obviously increased, so that a new gene resource can be provided for salt tolerance improvement and germplasm resource breeding of a grape cultivation variety.

The invention discloses a podophyllum hexandrum callus rapid proliferation method, and belongs to the technical field of plant tissue culture, the podophyllum hexandrum seed dormancy is broken through a sand storage method, the seeds are soaked in distilled water and a gibberellin solution and then are fully and uniformly mixed with river sand, after storage and germination acceleration, the seeds are sown in soil added with an equal amount of matrix, and podophyllum hexandrum seedlings are obtained through culture; fresh leaves of podophyllum hexandrum seedlings are taken as explants, the explants are disinfected and cut into small pieces, the small pieces are cultured on an MS culture medium added with hormone for 30 d to induce callus, the induced callus is cultured and proliferated at the optimal temperature of 15 DEG C, and the callus is inoculated to the same fresh culture medium after being cultured for 20 d to prevent browning, so that the high-quality callus is obtained. And then replacing a fresh culture medium every 20 days to realize rapid and mass proliferation of the podophyllum hexandrum callus. Through the method, a large amount of high-quality podophyllum hexandrum callus can be rapidly obtained, and sufficient materials are provided for podophyllotoxin extraction and medicinal value development.

The invention relates to the technical field of plant cell engineering, and particularly discloses a plant protoplast semi-fusion breeding method which comprises the following three stages of seedling material selection, preparation, fusion, induction and cultivation. A large number of fused plant seedlings are obtained through a series of operations and used for large-scale planting. The method comprises the following steps: selecting excellent varieties of a plant A and a plant B, respectively preparing the plant A and the plant B into protoplasts in a preparation process, culturing the protoplasts into corresponding calluses, and performing half-cell fusion on a fusion part to obtain mosaic calluses. Then, the callus of the plant A to be used as the root is induced to locally induce adventitious roots, the callus of the plant B to locally induce adventitious buds, and finally, breeding is carried out to obtain the super seedlings. According to the method, the grafting principle is combined, the degree and conditions of cell fusion are optimized, the breeding cost is reduced, meanwhile, it is ensured that the fusion body can stably express the excellent characters of parents at different parts, and agricultural large-scale production is facilitated.

The invention discloses a method for enhancing squalene synthesis through oil tea HMGR gene editing. According to the method, phosphorylation regulation and control sites Ser47 and / or Thr208 recognized by MAPK are targeted, and enzymatic activity inhibition is relieved and metabolic flux is enhanced by constructing phosphorylation resistance mutants (such as Ser-> Ala and Thr-> Val). The constructed editing vector can be introduced into camellia oleifera tissues in manners of PEG-mediated protoplast transient transfer, agrobacterium infection of calluses and the like, so that stable expression is realized. Experiments prove that the enzyme activity of HMGR in the editor is remarkably improved, the squalene accumulation amount is increased by 50% or above, the editing efficiency is high, expression is stable, and the method is suitable for high-value development of plant triterpenoid functional components and excellent strain breeding. The method also has the potential of popularization and application in other triterpenoid synthetic crops.

The invention discloses a method for inducing multiple types of somatic cells to change fate and directly develop into somatic embryos and application, and belongs to the technical field of plant genetic engineering. According to the invention, functional genes SPCH (AT5G53210) and ARF5 (AT1G19850) capable of promoting conversion of various types of somatic cells into the totipotent stem cells are found. Overexpression of the totipotent regulatory factor LEC2 can induce formation of somatic embryos on arabidopsis sinapis leaves, under the background, the functional genes are subjected to ectopic expression, more arabidopsis sinapis leaf cells can be converted into totipotent stem cells through SPCH overexpression without calluses, and therefore the totipotent stem cells can be used for inducing formation of the somatic embryos on the arabidopsis sinapis leaves. According to the method, more arabidopsis sinapis leaf cells can be transformed into totipotent stem cells, stem tip cells, hypocotyl cells and root cells can be transformed into totipotent stem cells, more arabidopsis sinapis leaf cells can be transformed into totipotent stem cells through ARF5 overexpression, the stem tip cells and the root cells can be transformed into totipotent stem cells, and finally somatic embryos are formed through induction.

According to the method, a plant explant callus culture mode is adopted, a carbon source containing a fluorophore is added into a culture medium, the callus which is provided with the fluorophore, thin in cell wall, easy to dissociate by a single enzyme and uniform in cell shape and size is generated through induction, and the cell dissociated by the single enzyme is detected by a flow cytometry, so that the cell quality is improved. The single cell purity and activity of the prepared leaf are 98.5% and 99.95% respectively, the clotting rate is only 1.5%, the cell purity and activity are improved by 32.6% and 21.05% respectively compared with those of a traditional technical scheme, and the clotting rate is reduced by 31.0%; according to the method, rice seeds are used as explants, seed single cells with the cell purity and activity being up to 99.5% and 100% respectively are obtained, and the limitation that the seed single cells cannot be prepared by a traditional enzymolysis method is broken through; besides, through co-induction of the safflower leaves and the rice seeds, the purity and activity of the prepared cells are as high as 99.98% and 100.0 respectively, and the limitation of single cell preparation caused by heterogeneity among species is overcome.