191 results about "Cotyledon" patented technology

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 provides a crop population three-dimensional reconstruction and organ phenotypic character analysis method, and relates to the field of three-dimensional reconstruction, and the method comprises the steps: collecting cotton envelope image data, carrying out the data preprocessing, and carrying out the motion recovery processing and Gaussian sputtering of the cotton envelope image data, the method comprises the following steps of: preprocessing a group cotton point cloud, removing a ground point cloud, obtaining a complete cotton group point cloud model, segmenting a cotton individual point cloud model and a corresponding organ point cloud model from the complete cotton group point cloud model, automatically analyzing cotton key phenotypic characters, and performing correlation analysis. According to the method, a cotton original habitat three-dimensional point cloud model is constructed based on mobile visual equipment, and key characters such as cotyledon node height, plant height and leaf area can be automatically analyzed, so that the problems that phenotypic character extraction in the prior art is high in cost, low in flux, easy to make mistakes, inconsistent in standard, generally destructive, labor-consuming, time-consuming and the like are solved.

A method for applying at least one spray product to agricultural land having crop plants and weeds. The crop plants and weeds can be assigned to a cotyledon class of monocotyledons or to a cotyledon class of dicotyledons. The at least one spray product is applied to the acquired field section of the agricultural land by means of at least one spray nozzle unit of an agricultural spraying device depending on the cotyledon class of the crop plants and a first ascertained weed identification number and / or a second ascertained weed identification number.

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 present disclosure provides methods of gene transformation of sunflower based on Agrobacterium-mediated gene delivery to meristem cells of fresh imbibed / germinated sunflower seeds. One embodiment of the present disclosure is a method of directly transforming a sunflower from a mature seed with a heterologous polynucleotide, comprising: generating an explant from a germinated sunflower seed by removing cotyledons and primary leaves having intact meristem and tender shoot radicles; contacting the sunflower explant comprising the cotyledon with Agrobacterium comprising a heterologous polynucleotide in an infection medium supplemented with a plant hormone using vacuum and ultrasound treatment; co-culturing the explant from a germinating sunflower plant in a culture medium lacking the plant hormone in the presence of the Agrobacterium genus; culturing the mature seed explant in a regeneration medium comprising a selective agent; and culturing the seed with the regenerated tender shoots and with the secondary roots in a culture medium lacking the selective agent.

The invention discloses an application of a gene GmAGO5 of a soybean ARGONAUTE family. The gene GmAGO5 of the soybean ARGONAUTE family is shown The nucleotide sequence of the soybean GmAGO5 protein coding gene GmAGO5 is as shown in SEQ ID NO. 1. The invention also discloses a preparation method of the soybean GmAGO5 protein coding gene The constructed plant gene editing knockout vector pSCM-GmAGO5 is transferred into an acceptor material Tianlong No.1 by using a cotyledonary node transformation method. The GmAGO5 gene is knocked out, so that the plant height of the gene edited soybean is reduced, the branch included angle is reduced, and the protein content of the soybean seeds is remarkably increased. Therefore, the soybean GmAGO5 protein coding gene GmAGO5 disclosed by the invention can be applied to reducing soybean plant height, reducing branch included angle and improving the protein content of soybean seeds through a genetic engineering means.

The invention belongs to the technical field of biology, particularly relates to the technical field of plant genetic engineering, and more particularly relates to application of an MYC gene in regulation and control of chloroplast development and composite stress resistance. Specifically, the invention innovatively discovers that the combined action of warm and JA can significantly induce seedling cotyledon chloroplast dysplasia, and deletion / reduction of MYC gene expression can significantly inhibit chloroplast dysplasia of plants under the combined action of warm and JA. The discovery provides guidance for remarkably improving plant seedling chloroplast development in a complex dynamic environment and improving the seedling survival rate and photosynthetic productivity, can promote optimization of a chloroplast artificial regulation and control technology, and realizes directional design of efficient photosynthetic organs in facility agriculture and an artificial photosynthetic system.

The present invention relates to the technical field of molecular markers, in particular to a molecular marker associated with cold resistance of corn and the use thereof. The molecular marker associated with the cold resistance of corn is ZmTIP4;3, and is obtained by amplification using primers shown as SEQ ID NO. 1 and SEQ ID NO. 2. Since the cold resistance of corn is a quantitative trait, phenotype analysis is time-consuming and labor-consuming. The molecular marker can be applied to cold-resistant corn breeding, and identification can be achieved in the corn seed stage or the early stage of cotyledon emergence, which saves time and is accurate, thus accelerating the process of breeding cold-resistant corn varieties.

The invention relates to an agrobacterium tumefaciens-mediated cork tree genetic transformation method. The method comprises the following steps: preparing an infection solution from an agrobacterium solution; the method comprises the following steps: taking in-vitro cotyledons or stems as explants after cork tree seeds germinate into seedlings, infecting the cotyledons or stems of the cork tree by adopting an infection solution, and carrying out co-culture, recovery culture and screening culture after infection is completed, so as to obtain transgenic hairy roots or transgenic calluses. According to the scheme provided by the invention, stable transformation of balsa roots can be realized, verification of functional genes is realized, the functional genes exist in a living body form for a long time, and the genetic transformation efficiency of balsa is improved.

The present invention provides a special cold-tolerant regulator for cotton seedling stage, its preparation method and application method, belonging to the technical field of crop cultivation. The special cold-tolerant regulator for cotton seedling stage includes chlorophyllin iron and chitosan; the mass ratio of chlorophyllin iron to chitosan is 28-38:500-700. The preparation method of the present invention includes the following steps: (1) Mix chlorophyllin iron with water to obtain solution A; (2) Mix chitosan with glacial acetic acid solution and dissolve to obtain solution B; (3) Add solution A to solution B, and make up the volume with water to obtain the special cold-tolerant regulator for cotton seedling stage. The application method of the present invention is to perform foliar spraying before the occurrence of chilling injury from the cotyledon stage to the five-leaf stage of cotton seedlings. Applying the cold-tolerant regulator prepared by the present invention before the occurrence of chilling injury in the cotton seedling stage can significantly reduce the chilling injury index of cotton seedling leaves and improve the survival rate of cotton seedlings.

The present disclosure provides an application method in rice callus differentiation based on an Oryza sativa Leafy Cotyledon 1 (OsLEC1) gene, including the steps of: selecting guide ribonucleic acid (gRNA) target sites; cloning tandem fragments including gRNA; ligating each gRNA fragment; performing polymerase chain reaction (PCR) amplification on a ligation product; performing enzyme digestion on the purified product and a target vector; transforming the ligated vector; performing Agrobacterium-mediated genetic transformation of rice; and screening and identifying transgenetic plants. In the present disclosure, the differentiation of callus directly affects the emergence efficiency of transgenic plants. The knockout of OsLEC1 can promote the differentiation of rice callus, suggesting that OsLEC1 can serve as an important target gene for improving the transformation efficiency of rice and even gramineous crops. OsLEC1 can be used as a starting point to construct various molecular tools to enhance transformation efficiency.

The invention discloses a method for synergistically improving the lycopene content and the sugar-acid ratio in tomato fruits, belongs to the technical field of biology, and particularly relates to a method for knocking out or silencing an INVINH1 gene to synergistically improve the lycopene content and the sugar-acid ratio in the tomato fruits, and a nucleotide sequence of a protein coding region of the INVINH1 gene is shown as SEQ ID NO.1. The invention further discloses a method for synergistically improving the lycopene content and the sugar-acid ratio in the tomato fruits. A carrier for knocking out or silencing the INVINH1 gene is constructed by utilizing a CRISPR / Cas9 gene editing technology, the carrier is transferred into tomato cotyledons through agrobacterium tumefaciens mediation, INVINH1 gene knockout homozygous strains invinh1-1 and invinh1-2 are obtained through screening by utilizing a tissue culture technology, the lycopene content and saccharic acid in tomato fruits of a mutant plant are remarkably improved compared with those of wild plants, and the gene knockout or silencing method is suitable for industrial production. The invention provides an important gene resource for cultivating tomato varieties for synergistically improving lycopene and sugar-acid ratio.

The present invention relates to a method for producing a composition for promoting hair growth, and more particularly, to a raw material mixing step for mixing ground pine needles, ground pine cones, heartleaf houttuynia herbs, green tea leaves, cotyledons and an ethanol aqueous solution, and to a method for producing a composition for promoting hair growth. A fermentation step: inoculating the mixture cured in the curing step with lactic acid bacteria and fermenting; and mixing the additives, namely mixing the additives containing the hair root cell growth factors. A fermentation product produced by the fermentation step. The composition for promoting hair growth prepared by the method provides nutrition for hair, improves the health of scalp and hair roots, and shows an excellent hair growth effect.

The application relates to the technical field of bean kernel processing, and particularly discloses a bean kernel skin and cotyledon separation device, which comprises a microwave heating device and a mounting frame, the microwave heating device is mounted at the top of the mounting frame, both ends of the microwave heating device are provided with blocking plates, a conveying belt is mounted in the mounting frame, a plurality of placing boxes are mounted on the surface of the conveying belt, each of the plurality of placing boxes is provided with a turnover mechanism, two fixing frames are mounted at the top of the mounting frame, a driving mechanism is arranged at the bottom of each of the two fixing frames, a separation box is arranged at one end of the mounting frame, and an automatic separation mechanism is mounted in the separation box. In the application, the microwave heating device and the separation box are arranged, the skin and the cotyledon of the bean kernel can be automatically separated by using the microwave peeling method, and the efficiency is extremely high; and the turnover mechanism is arranged, the bean kernel can be conveniently turned over, and the separation efficiency of the skin and the cotyledon of the bean kernel is improved.

The invention discloses application of apple MdMYB41L in regulation and control of seed size, and belongs to the technical field of plant genetic engineering. According to the invention, an MdMYB41L gene is cloned from apple GL-3, the CDS nucleotide sequence of the MdMYB41L gene is shown as SEQ ID NO: 1, and a plant overexpression vector is constructed. A transgenic plant is obtained by heterologous transformation into arabidopsis thaliana through an agrobacterium tumefaciens-mediated method. Compared with wild arabidopsis thaliana, the weight, the length, the width and the surface area of the transgenic plant seeds are obviously improved. In addition, the cotyledon area of a transgenic plant is obviously higher than that of a wild type. Therefore, the MYB transcription factor MdMYB41L provides a new thought in the aspects of improving the plant seed size and molecular breeding.

The invention belongs to the technical field of biology, and particularly relates to a soybean hairy root genetic transformation method which comprises the following steps: taking soybean cotyledons as explants, infecting wounds of the explants by utilizing agrobacterium rhizogenes carrying reporter genes, transferring the infected explants into vermiculite, culturing, rooting, and screening positive roots. According to the method disclosed by the invention, a simple and efficient soybean hairy root genetic transformation system is established, tissue culture is not needed, and the problems of long transformation period, high cost and low transformation rate in the prior art are solved.

The invention discloses a method for realizing efficient in-vitro regeneration of tea tree cotyledons through a somatic embryo indirect generation way, and belongs to the technical field of plant tissue culture. The tea tree somatic embryo indirect generation system is successfully constructed by screening culture conditions, a culture medium formula and other methods, and a foundation is laid for construction of a tea tree genetic transformation system and industrialized seedling production; according to the method, immature cotyledons of tea trees are used as explants, the cotyledons are induced to generate calluses, then the calluses are proliferated and differentiated to generate somatic embryos, and the somatic embryos are further differentiated to form complete plants, so that efficient and synchronous propagation of tea tree seedlings is realized, and technical support is provided for high-quality variety propagation and new variety breeding of tea trees; the problems of low propagation coefficient, long growth cycle, serious descendant degeneration and the like in traditional seed propagation of tea trees are solved, and the technical bottlenecks of serious explant browning, high bacterial contamination rate, poor plant growth synchronism and the like in other tissue culture technologies are solved.

The application discloses a method for high-frequency regeneration of Rhododendron chrysanthum based on cotyledon node indirect organogenesis, and belongs to the field of plant biotechnology. The method takes the cotyledon node of Rhododendron chrysanthum sterile seedling as an explant, and obtains a complete regenerated plant through the induction of callus, differentiation of adventitious buds, elongation of adventitious buds, strong seedling and rooting culture in sequence. The application realizes the super-high-frequency differentiation of adventitious buds by combining the cotyledon node explant with a specific hormone combination for the first time, the differentiation coefficient is as high as 96, and the rooting rate is 92.7%, thereby effectively solving the technical bottleneck of low propagation efficiency of Rhododendron chrysanthum, and providing reliable technical support for the conservation and industrialization development of Rhododendron chrysanthum germplasm resources.

The present invention discloses a method for regulating soybean rapid multiplication and generation addition by LED light, which includes: after soybean seeds germinate and enter the cotyledon stage, regulating the light quality, light intensity and photoperiod based on the growth stage of soybean until the soybean is completely mature; the growth stages of soybean include the cotyledon stage, the stage from the cotyledon stage to the second branching stage, the stage from the second branching stage to the flower and pod stage, the stage from the flower and pod stage to the maturity stage, and the maturity stage; the pods harvested after the soybean matures are dried and then stored at low temperature. The present invention provides an LED light regulation method and a cultivation method for significantly shortening the breeding generation propagation time of dryland soybeans based on the dynamic LED signal spectrum. Compared with the traditional simple photoperiod and light intensity regulation methods, while significantly shortening the breeding generation propagation time, it can ensure the germination rate of seeds during the rapid propagation process, providing an effective technical approach and method for ensuring sufficient high-quality offspring samples required for the purification process of rapid breeding generations of crops in a controllable environment.

The invention belongs to the field of gene engineering, and discloses a method for rapidly evaluating the plant growth capacity of transgenic herbicide-resistant cotton in the seedling stage, and the method comprises the following steps: S1, sowing transgenic cotton seeds to be detected; s2, when cotton cotyledons are completely flattened, glyphosate with the working concentration of 0 time or 2 times or 4 times is sprayed; s3, 7 days after spraying, respectively taking the overground part and rhizosphere soil of the cotton, and detecting the dry weight of the overground part and the water content and microbiome of the rhizosphere soil; and S4, judging the plant growth capability of the transgenic herbicide-resistant cotton in the seedling stage. The method provided by the invention is a rapid, standardized and repeatable seedling stage plant growth ability evaluation method, is suitable for multi-material and large-scale determination, is easy to popularize and apply in the agricultural field, and has urgent and important practical significance for accelerating glyphosate-resistant cotton variety breeding and industrial application.

The application belongs to the field of in vitro rapid propagation of Chinese medicine seedling, and particularly relates to a high-efficiency liquid suspension culture method of pinellia ternate, which comprises obtaining of pinellia ternate explants under sterile conditions, induction of embryogenic callus, liquid suspension culture, somatic embryo development and maturation, somatic embryo germination and transplanting of regenerated seedlings. By using the method, the induction rate of pinellia ternate embryogenic callus can reach 81.6%, stable mature cotyledon embryos can be obtained, and the embryos can be regenerated into complete plants, a high-efficiency and stable regeneration system is successfully established, favorable conditions are created for industrial production of pinellia ternate fine seedlings, and a good foundation is laid for genetic transformation of pinellia ternate.

The invention relates to the technical field of cucumber grafting, and particularly discloses a cucumber single-shoulder oblique splitting grafting method which comprises the following steps: step 1, a preparation stage: S1, selecting pumpkin seedlings which grow robustly, have developed root systems and are high in disease resistance as rootstocks; s2, cucumber seedlings which grow well and are free of plant diseases and insect pests are selected as scions; according to the method, one cotyledon is cut off through an oblique cleaving method with growing points in stock treatment, the grafting speed is greatly increased, the later problem that the growing points are not thoroughly removed is avoided, tight and stable combination of scions and stocks is achieved by accurately controlling the oblique cleaving angle and the attaching connection mode, and the grafting efficiency is improved. Water loss and germ invasion in the grafting process are effectively reduced, so that the grafting survival rate is remarkably increased; compared with a traditional grafting method, the method has the advantages that the operation steps are simplified, the strict requirements for the sizes, shapes and the like of scions and stocks are reduced, the difficulty of the grafting technology is lowered, and more farmers and gardening workers can easily master and apply the method.

This invention discloses a method, system, and device for branching the gene sequence of influenza A virus, relating to the field of bioinformatics. The method includes the following steps: reading a user-input phylogenetic tree structure file and a Fasta file; dividing the phylogenetic tree into multiple branches based on a set first custom parameter and the gene sequence data of each cotyledon node; verifying the relationship between branches based on a set second custom parameter, and merging branches whose genetic distance is within a set threshold; verifying the merged branches; and if there are unassigned cotyledon nodes, assigning them to the branch with the closest genetic distance that satisfies the custom parameter, thereby obtaining the final branching result of the influenza A virus gene sequence. The method employed in this invention enables targeted phylogenetic branching based on the characteristics of influenza A virus, improving the accuracy of the branching results.

The application belongs to the field of plant biotechnology, and particularly relates to a cucumber CsODO1 gene and application thereof, and is a MYB transcription factor capable of being combined with a CsCSE1 promoter, which is screened through a yeast single hybridization. It is proved through EMSA and tobacco transient expression analysis that the CsODO1 gene directly and specifically combines with a "CAACCA" sequence in the CsCSE1 promoter, and inhibits the expression of the CsCSE1 gene. The CsODO1 gene can be induced to express by a powdery mildew fungus, and is differentially expressed in the cucumber infected by the powdery mildew fungus. It is found through a cucumber cotyledon transient transformation system mediated by an agrobacterium that the CsODO1 gene negatively regulates the biosynthesis of lignin and the resistance of the cucumber to the powdery mildew fungus. The application provides a new reference gene resource for the cultivation of a cucumber disease-resistant variety, and provides a new thought for the research on the molecular mechanism of the plant immunity mediated by the lignin pathway.

The invention relates to a construction method of an efficient regeneration system of idesia polycarpa, which comprises the following steps: soaking idesia polycarpa seeds in 3% gibberellic acid diluted by 1000 times for 3 days, sterilizing, inoculating to a germination culture medium of 1 / 2MS, 1.0 mg / L 6-benzylamino adenine (6-BA) and 0.5 mg / L 3-indolebutyric acid (IBA), and culturing for 25 days to obtain seedlings; a hypocotyledonary axis at the position 2 cm below the cotyledon of the seedling is obliquely cut and inoculated to a culture medium containing MS, 1.0 mg / L of thidiazuron (TDZ) and 0.5 mg / L of IBA, and calluses are induced after 30 days; inoculating the calluses to a culture medium containing MS, 2.0 mg / L TDZ and 0.05 mg / L IBA, and differentiating regenerated buds after 30 days; after seedling strengthening and rooting culture, the seedlings are transplanted to a sterilization matrix with the ratio of nutrient soil to vermiculite to perlite being 4: 1: 1. The differentiation rate of regenerated seedlings reaches 64.82% and is remarkably higher than that in the prior art, the seed germination rate and the transplanting survival rate are high, and an idesia polycarpa regeneration system can be efficiently constructed.

The invention relates to the technical field of plant breeding, and discloses a culture medium and method for promoting masson pine somatic embryo maturation, masson pine embryogenic callus is inoculated into a somatic embryo maturation culture medium containing betaine, polyvinylpyrrolidone or dimethyl thiourea for somatic embryo induction culture, and the masson pine somatic embryo can be obviously promoted to mature in advance. Test results show that morphological observation is carried out on the embryogenic callus at different somatic embryo induction time points, when the embryogenic callus is cultured for 60 days, most cotyledon embryos are mature and a large number of mature embryos appear, the number of embryos is counted, the number of mature somatic embryos is 250-3500 / g embryogenic callus, and the number of total embryos is 3333-9000 / g embryogenic callus; the method can efficiently promote maturing of the masson pine somatic embryo, the somatic embryo induction efficiency is high, and the method has good practicability.

The invention discloses a method for establishing a gleditsia sinensis tissue culture regeneration system, and belongs to the technical field of plant tissue culture. Comprising the following steps: (1) inoculating explants from wild gleditsia sinensis seeds into an adventitious bud induction culture medium, and performing induction culture to generate adventitious buds; and (2) cutting off the adventitious bud from the base, inserting the cut end into a root induction culture medium, and inducing rooting. According to the method, hypocotyls and cotyledons of gleditsia sinensis aseptic seedlings are used as explants, and the influence of different plant growth regulator combinations, browning inhibitors and the like on the regeneration efficiency is discussed. According to the method, an efficient gleditsia sinensis regeneration system is successfully established, and a theoretical basis and a technical support are provided for high-quality seedling breeding and genetic engineering genetic improvement.

The invention provides a tissue culture method based on big-leaf tea cotyledons, which at least comprises the following steps: collecting mature fruits of a big-leaf tea tree variety in Yunnan province, peeling off peels, taking out seeds, and peeling off testa of the seeds to take out complete cotyledons; the cotyledons are sequentially subjected to callus culture, adventitious bud induction culture, adventitious bud multiplication culture and rooting culture. According to the tissue culture method based on the big-leaf tea cotyledons, the cotyledons of the mature fruits of the Yunnan big-leaf tea trees are used for tissue culture, so that the propagation time of the tea seedlings of the Yunnan big-leaf tea trees is shortened, the labor cost is reduced, and the tea seedlings can be cultured all the year round. Compared with a traditional sowing and seedling raising method, the method has the advantages that the cotyledons are used for tissue culture induction, the proliferation rate is high, 3-5 tea seedlings can be bred through each cotyledon, the breeding speed is high, the period is short, and seedlings can be transplanted after 100-120 days.

The invention discloses a culture method for rapid rooting of gingko aseptic seedlings, and relates to the technical field of gingko tissue culture. The culture method for rapid rooting of the gingko aseptic seedlings comprises the following steps: inoculating the gingko aseptic seedlings into a first rooting culture medium for culture, and then transferring the gingko aseptic seedlings into a second rooting culture medium for culture, the second rooting culture medium is an MS culture medium containing 0.1 to 0.2 mg / L of 6-benayl aminopurine, 0.5 to 1.5 mg / L of indolebutyric acid, 15 to 25 mL / L of gingko sterile main root juice, 7 to 9 mL / L of gingko sterile cotyledon juice, 25 to 35 g / L of sucrose and 6 to 8 g / L of agar. The invention effectively solves the problems of lack of staged regulation and control, single culture medium component, poor functionality, unreasonable hormone proportion and the like in the prior art.

The invention discloses a pea cotyledon protoplast separation and instantaneous conversion method and application. The method discloses steps of pea cotyledon protoplast separation and instantaneous conversion and used reagents. The method comprises the steps of material acquisition, protoplast preparation, purification, activity detection, instantaneous conversion and the like. The method is easy to operate and short in digestion time, and a large amount of early-stage preparation work is saved. According to the method disclosed by the invention, the high-quality pea cotyledon protoplast suspension can be quickly and efficiently obtained, the activity of the protoplast can reach 90% or above, and a powerful technical support is provided for scientific researches such as subsequent gene function verification, gene editing and single cell transcriptome sequencing.