1666 results about "Callus" patented technology

The present invention relates to a method for visualizing GFP expression in callus, various tissue and organ of the transgenic plants as image and system using the same. The said method needs no other additional genetic product, substrate or cofactor and can detect very simply and quickly GFP expression by using the said system of the present invention consisting of a CCD camera, a light source, band-pass filter and data processing computer, so it provides many advantages for selection of transgenic seeds, for studying of gene expression in the tissue or organ of plants, or for studying of specificity of each development step.

The invention discloses application for using an LbCpf1 - RR mutant in a CRISPR / Cpf1 system in plant gene editing. The application for using the LbCpf1 - RR mutant in the CRISPR / Cpf1 system in plant gene editing uses an OsPDS gene and an OsSBEIIb gene as target genes, constructs a target double loci of one gene and a series of vectors of two genes, and utilizes an agrobacterium-mediated transformation method to import the vectors into rice callus, and rice plants with the target gene knockout are successfully obtained by using the LbCpf1-RR mutant. The only difference between the LbCpf1-RR mutant and the protein LbCpf1 is that the 532nd amino acid changes from G to R, and the 595th amino acid changes from K to R. The LbCpf1-RR mutant provided by the application for using the LbCpf1 - RR mutant in the CRISPR / Cpf1 system in plant gene editing expands the PAM site sequence identified by the LbCpf1-RR mutant, so that the editing range of the CRISPR / Cpf1 system in rice genome is expanded, great significance for promoting the application of the system in the field of plant genome editing is achieved. The application for using the LbCpf1 - RR mutant in the CRISPR / Cpf1 system in plant geneediting has great application value.

The invention belongs to the field of manually planted Aquilaria senensis (Lour.) Gilg and relates to a microbial flora preparation for producing Chinese eaglewood wood and a matched bottle interpolation method. Eight endophytic strains of the Chinese eaglewood wood such as colored Botryodiplodia theobromae Pat are extracted and separated from natural Chinese eaglewood wood tissues, and are respectively disinfected and purified to obtain liquid flora preparations with Chinese eaglewood wood production capacity, and the obtained liquid flora preparations jointly act on the Aquilaria senensis (Lour.) Gilg. Meanwhile, the conventional matched infusion method is improvement on the bottle interpolation method; and by the ingenious bottle interpolation method, the disadvantages that a complete set of infusion apparatus is depended and the dripping speed is required to be constantly observed and adjusted in the conventional method are overcome. By the method, a microbial inoculum can be manually immersed into calluses of healthy Aquilaria senensis (Lour.) Gilg in batches and penetrate and infect protoxylem to promote a micro-circulation system in the protoxylem to secrete resin and produce the Chinese eaglewood wood in an abnormal metabolism mechanism.

Compositions and methods include a new strain of Bacillus amyloliquefaciens having activity against plant pathogens. The compositions are useful for benefiting plant growth and / or conferring protection against a pathogenic infection when applied to plant foliage, flowers, fruits, bark, roots, seeds, callus tissue, grafts, cuttings, surrounding soil or growth medium, and soil or growth medium concomitant with sowing seed and planting callus tissue, grafts, and cuttings. The compositions containing the Bacillus amyloliquefaciens RTI472 strain ca n be applied alone or in combination with other microbial, biological, or chemical insecticides, fungicides, nematicides, bacteriocides, herbicides, plant extracts, plant growth regulators, or fertilizers. In one example, the Bacillus amyloliquefaciens RTI472 strain can be delivered to the plant as part of an integrated pest management program, with other microbial or chemical insecticides, fungicides, nematicides, bacteriocides, herbicides, plant extracts, and plant growth regulators.

The present invention is directed to the introduction of molecules, including nucleic acids, carbohydrates, plant growth regulators and peptides into cells and tissues. The present invention is also directed to media and methods for enhancing embryogenic callus production of elite lines of soybean.

The invention provides a rapid rhizoma polygonati propagation technology method. The method comprises the following steps: breaking seed dormancy by changing temperature treatment and hormone treatment, performing induction in vitro to culture callus by utilizing seeds subjected to dormancy breakage as materials, performing differentiation culture and root induction, and performing vegetative propagation on rhizoma polygonati seedlings. By adoption of the technology, the rhizoma polygonati propagation factor can be greatly improved, the production period is shortened, the sowing quantity is reduced, and the production cost is reduced.

The invention belongs to the technical field of plant vegetative propagation, and in particular relates to a cuttage seedling raising method for twigs of prunus maritime. The method solves the problems of cuttage seedlings of prunus maritime of rotten calluses of roots, low rooting rate, low rate of survival, week seedlings and susceptibility to water loss and withering of prunus maritime and strengthens the growth and rejuvenation of the roots, stems and leaves of the seedlings during cuttage through hardening seedlings, improves the rate of survival in wild field planting and lays firm good function fur special planting seedlings of pruns maritime.

The invention relates to a method for cultivating saussurea involucrate cell, wherein it comprises that using MS basic culture medium wit some NNA (fruitone), 6-BA (6-aminopurine) as the culture medium of callus; inducing the seed of saussurea involucrate into plant; using said plant as expalnt to be differented at special condition, to induce callus; subculturing the callus periodically, to increase the callus; using part of callus to function the seed and collecting the left to be dried as the saussurea involucrate culture; collecting said culture to be dried as the cell culture. The invention can improve the content of effective components.

The invention discloses application of panax japonicus transcription factor gene PjWRKY1, namely, application in improving the expression quantity of key enzyme genes in the biological synthesis of panax japonicus saponins, and increasing the content of saponins in panax japonicus callus. The nucleotide sequence of the PjWRKY1 gene is shown as SEQ ID NO: 1, and WRKY transcription factors are coded. Through the adoption of functional genomics and techniques related to metabolic engineering, a panax japonicus PjWRKY1 transcription factor is proved to have the effect of positively regulating the biological synthesis of panax japonicus saponins; when the panax japonicus PjWRKY1 transcription factor gene disclosed by the invention is established on a plant expression vector and transferred into the panax japonicus callus for over expression, the expression quantity of the key enzyme genes in the synthetic route of panax japonicus saponins is increased, and the yield of panax japonicus saponins is increased.

The invention discloses a method for improving the culturing efficiency of japonica rice anther. The method comprises the steps that firstly, culture medium mother solutions of the japonica rice anther are prepared according to formulas of the culture medium mother solutions of the japonica rice anther, then the various mother solutions are utilized to prepare an induction culture medium, a differentiation culture medium and a rooting culture medium all of which are used in different periods, and the culture media in the different periods are correspondingly applied to induction, green plantlet differentiation and rooting of calluses of the japonica rice anther respectively. By the adoption of the method, the percentage of seeding emergency of culturing of the japonica rice anther can be improved, the rice breeding progress is quickened, and the seed selection efficiency of good variety of rice is improved.

The invention discloses a cutting seedling-culturing method for France Grasse roses. The purpose is to solve the problem that rooting of ex-vivo branches of the France Grasse roses is difficult. The method includes the following steps that step1, according to preparation at the early stage, a cutting bed is laid, and cutting media are loaded into non-woven cloth seedling-culturing net bags and comprise, by weight, 30%-35% of vermiculite, 30%-35% of perlite and 35%-40% of grass carbon or rice hull carbon; step2, cutting preparation is performed; step3, according to growth promoting, low cut portions of sheared cutting mother branches are immerged into a growth promoting mixed solution, and soaking is performed; step4, cutting is performed; step5, management after cutting is performed; step6, seedling hardening and transplanting are performed. By the adoption of the method, callus tissue of the roses grows rapidly, and the survival rate is high. The callus tissue starts to be generated at the roots of cuttings after one week from cutting, callus tissue is overgrown at the roots after 15 days, and callus tissue is generated on 95% of branches after 20 days. Meanwhile, rooting and sprouting are rapid at a large quantity; rooting starts on bud eyes on the lowest portion after 15 days from cutting, and the rooting rate reaches above 85% after 30 days.

The present invention discloses the process of culturing adventitious root tissue of licorice, and belongs to the tissue culture of licorice as one kind of Chinese medicinal materials. The process of the present invention includes soaking licorice seed in solution with ethanol in 70-75 vol% and sodium hypochlorite in 2 vol% to sterilize and planting in MS culture medium to induce bacteria-free bud; inoculating the cut cotyledon and bud in MS culture medium containing different plant growth regulator to induce callus; transferring the callus growing for 20 days in MS culture medium with indolebutyric acid and kinin to induce adventitious root; and finally transferring the adventitious root to liquid culture medium containing the same plant growth regulator for fast growth. The present invention has simple process, high inducing rate, fast adventitious root proliferating speed and other features.

The invention discloses a method for cuttage reproduction of a horseradish tree. Through preparation of matrix, preparation of rooting agent, section of cuttings, cuttage and disposal, after-cuttage management and protection, transplanting, and other stps, the cuttage reproduction of the horseradish tree are completed. By adopting the method provided by the invention, the cutting slips start to form a callus for about 10 days generally, and successively grow fibrous roots; after 4 weeks, about 80% of cutting slips take root. Compared with a routine method, when the cuttage reproduction of the horseradish tree is performed, the callus forming time of the root part of the cutting slips is short, the side roots of the cutting slips are multiple, and the root length is relatively long, an the rooting rate of cutting reaches 86% above. The method for cuttage reproduction solves the problems that it is difficult to take root in the cuttage process of the horseradish tree, and the root part is rotted for too long rooting time; besides, the cuttage survival rate is effectively improved.

The invention discloses a method for improving yield of salvianolic acid B in savia miltiorrhiza suspension culture cells by inducing, specifically comprising the following steps: 1) cultivating the aseptic seedlings; 2) inducing the callus; 3) sub-cultivating the callus; 4) cultivating the callus by suspending; 5) inducing to produce the salvianolic acid B; and 6) harvesting the culture cells. The method solves the problem that the salvianolic acid B is not produced after the inducing factors, such as yeast extract, galacturonic acid, mycelia extract and the like are added. The content of the salvianolic acid B in the savia miltiorrhiza suspension culture cells is increased by about 5 times by using the method. The technology condition provided by the invention is suitable for industrial production of the salvianolic acid B on a large scale by a cell culture method after being expanded.

A cuttage raising method for olive seedlings on a greenhouse hotbed includes: preparing a cuttage bed, acquiring scions, treating cuttings, performing cuttage, selecting cuttage time and performing post-cuttage management; setting the cuttage bed in a solar greenhouse; selecting an annual young tree as a female tree to acquire the scions, and selecting highly mature shoots without disease and insect damage as the cuttings; cutting each cutting into a cutting 10-12cm long with 4-6 segments, reserving 2-4 leaves at the top end of the cutting, smoothing the position, 0.5-1cm away from a bud, of the top end by cutting, and cutting the lower end of the cutting into an inclined horse-ear-shaped end; soaking the root of each cut cutting in 1000mg / kg-2000mg / kg indolebutyric acid solution at a 5cmdepth for one night; on the next day, dipping each cutting with mixture of indolebutyric acid and talcum powder before cuttage; reserving row spacing of the cuttings to be 5-6cm and plant spacing to be 3-4cm; keeping interior temperature of the solar greenhouse to be 18-20 DEG C, humidity to be 90%-95%, cuttage bed soil temperature to be 23-25 DEG C and water content to be 60%-70%; moving the seedlings, which root for 45-55 days, off the bed. By the method, the rooting rate of cuttage seedlings reaches more than 90%, calluses emerge 35-40 days after cuttage, and the seedlings can root in 45-55 days.

The invention relates to a sugarcane callus protoplast separation and purification method, which comprises sugarcane callus culture, sugarcane suspended cell culture, sugarcane protoplast separation, and sugarcane protoplast purification. By the method, suspended cell lines can be obtained by sugarcane embryogenic callus culture and an adequate amount of complete protoplasts with high quality and uniformity can be obtained by enzyme digestion, separation and purification; therefore, ideal experimental materials can be provided for study transgenetic transient expression such as sugarcane exogenous gene expression, promoter analysis and gene silencing.

The present invention belongs to the field of plant tissue culture, and specifically relates to a tissue culture and rapid propagation method of white crane lily. Sterilized with 0.1% mercuric chloride, ②cut the stems into 1-2mm thick slices, and put them together with the terminal buds in the induction and differentiation medium supplemented with 6‑BA and VC for culture, ③callus or adventitious buds were inoculated Add 6‑BA and IAA to the medium, wherein the adventitious bud mass used to induce rooting seedlings is directly inserted into the medium added 6‑BA, IAA and TIBA, ④Single buds with plant height ≥30mm or Three clumps with a main bud plant height ≥ 25mm were inserted into the rooting medium supplemented with activated carbon. The invention provides a method for tissue culture and rapid propagation of white crane lily, which can obtain a large amount of asexually propagated white crane lily tissue culture seedlings in a short time.

Disclosed is a culture composition containing micro and macro-ingredients, vitamins and other supplements such as phytagel, sucrose, picloram. The medium is useful in the formation and development of callus lines of Taxus sp., and resulting in production of taxol in high amounts.

The invention belongs to the field of crop growing and particularly relates to a method for cultivating seedlings by means of green branches under the full exposure and light mist condition. According to the method for cultivating seedlings by means of green branches under the full exposure and light mist condition, seedling cultivation facilities are simple, investment is low, the influence on blooming, in the next year, of a female parent is small, plants which can grow under natural condition can be cultivated directly at a time, and the survival rate can reach 90%. The method for cultivating the seedlings by means of green branches under the full exposure and light mist condition comprises the following steps of (1) seedbed manufacturing, (2) branch selection and ear trimming, (3) hole forming and cuttage, (4) mist spraying control, (5) management after cuttage, and (6) seedling hardening, wherein according to mist spraying control, after cuttage, secondary sterilization is conducted, a high-pressure light mist control system is started at the same time, five billion mist particles are generated by a nozzle per second, the diameter of each mist particle ranges from 0.5 micron to 5 microns, and a water film exist on the surface of each leaf all the time under the full exposure condition; according to management after cuttage, sterilization is conducted in the day when cuttage is conducted, mist spraying is conducted every seven days since then, mist spraying is conducted every ten to fifteen days after the seedlings strike roots, mist spraying is stopped in the later stage, foliage dressing is conducted once every ten days, embryogenic calluses appear after seven to eight days generally, and a large amount of roots are generated after fourteen days.

The invention discloses a tissue culture and rapid propagation method for kadsura coccinea. The method includes the step A of selection and processing of explants, the step B of induction of explants, the step C of browning prevention processing of calluses, the step D of differentiation of calluses, the step E of subculture multiplication culture, the step F of strong seedling culture, the step G of rooting culture and the step H of seedling hardening and transplanting, wherein in the step A, young and tender leaves close to tips of kadsura coccinea stems are taken and disinfected to obtain sterile explants. By means of the method, the browning rate of explants of kadsura coccinea is controlled at 7% or below, the rooting rate reaches 95%, and the seedling formation rate reaches 97% or above.

The invention provides a method for effectively reducing environmental risk caused by pollen propagation of transgene plant. In the method, two tightly linked gene expression cassettes of 1) a pollen lethal gene ZM-AA1 driven by a pollen growth anaphase singular promoter PG47 and 2) a fluorescence screening mark gene FP driven by a callus / seed coat singular promoter END 2 are transferred to a Zhonghua 11 wild type material. During selfing fructification of a single plant carrying a single copy transgene, a pollen grain without carrying the transgene can be fertilized with a female gamete normally, but a pollen grain carrying the transgene is abortive in pollen growth anaphase and can not be fertilized with a female gamete, so as to reduce risk of transgene drift in the environment.

The invention provides an excised mutagenesis tetraploid method of water melon and ploidy early stage certification technique, wherein dinitro toluene herbicide (DNH) is employed to substitute the conventional colchicines as inducer, whose function is to suppress the Mitosis in the metaphase of cell division through the mechanism of interfering spindle, so as to double the tissue cell chromosome. The method has the advantages of increased inducement success rate and substantially shortened time required for inducement.

The invention discloses a Bose factor polypeptide anti-wrinkle eye cream and a preparation method thereof. The eye cream provides a conditioner with main active ingredients, and the conditioner comprises tocopheryl acetate, hexapeptide-9, palmitoyl pentapeptide-4, tripeptide-1, palmitoyl palmitoyl tripeptide-5, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, hydroxypropyl tetrahydropyrantriol (Bose factor), carnosine, glycosphingolipid, illicium verum callus culture filtrate and rosa damascena flower oil. The eye cream can effectively promote the generation of eye skin collagen, increase theelasticity and compactness of the skin, prevent wrinkles from deepening and repair the damaged barrier of the eye skin, and has the effects of wrinkle resistance, whitening and oxidation resistance.

The invention discloses a tissue culture method of haworthia retusa. The tissue culture method comprises the following steps: taking a haworthia retusa fleshy leaf as an explant; washing, sterilizing and disinfecting; then cutting the explant to the length of about 1.5cm-2cm and upwards and obliquely inserting a notch into a starting culture medium, wherein the culture temperature is 25+ / -2 DEG C, the illumination time is 24h / d, and the illumination intensity is 2000lx; after the haworthia retusa explant is cultured for 7 days and the flesh of the haworthia retusa explant is gradually expanded, transferring the explant into an induction callus culture medium so as to be cultured; after 20 days, forming calluses by the explant; transferring the explant into a differential culture medium to be cultured; after about 35 days, gradually germinating and differentiating buds by the calluses; after 50 days, transferring the buds into a bud proliferation culture medium to be proliferated into budlets; gradually forming cluster seedlings by cluster buds and generating small haworthia retusa; and dividing the cluster seedlings by using a dissecting knife, continually proliferating and culturing, and putting the small haworthia retusa into pots after 70 days. According to the method, leaf flesh tissues of succulent plants are taken as the explants and a lot of plant materials can be obtained in short time, so that the succulent plants including the haworthia retusa can be rapidly proliferated and popularized.

The invention provides a winkle removal composition with a symbiotic bacterium combined ferment substance, a wrinkle removal eye cream and a preparation method of the wrinkle removal eye cream. The wrinkle removal composition comprises the following components in parts by weight: 0.01-5 parts of acetyl hexapeptide-8, 0.1-10 parts of oligopeptide-4, 0.1-10 parts of a crithmum maritimum callus culture filtrate, 0.1-10 parts of dipeptide diamido-butyryl benzyl amide diacetate, 0.01-5 parts of a camellia sinensis leaf extract, 0.1-10 parts of a coffea arabica extract, 0.1-10 parts of a retinol type compound and 1-10 parts of a symbiotic bacterium combined ferment substance. The winkle removal composition is capable of not only promoting c cell proliferation and regeneration and improving skinelasticity, but also effectively preventing oxidation and preventing collagen decomposition. The wrinkle removal eye cream is not only capable of removing dynamic wrinkles, static wrinkles and dry wrinkles which are already formed around eyes, and preventing formation of wrinkles, but also providing long-lasting moisture retention, and making skin tough and healthy.

The invention discloses a method and application of knocking out a BnMAX1 gene in Brassica napus by a CRISPR-Cas9 system, two sgRNAs of specifically targeting Brassica napus BnMAX1 gene are designed and synthesized into oligo dimer, are connected with Cas9 vector, and introduced into hypocotyl callus of Brassica napus by Agrobacterium tumefaciens-mediated transformation to regerated into shoots. Cas9 nuclease was guided by sgRNA to cleave target sequence. Each sgRNA could pass through CRISPR-Cas9 system mediates the cleavage of BnMAX1 gene in A and C genomes, and achieves the goal of gene knockout. Phenotypic identification showed that homozygous mutant lines increased the number of branches and pods per plant, decreased plant height and increased yield.

The invention discloses a method for breeding locusts, which uses zygotic embryo of different embryonic ages as explants, and uses MS+2, 4-D+BA+cane sugar+agar as an induction medium of an embryonic callus. The method comprises: using MS basal medium + MES + glutamine + casein hydrolysate + naphthylacetic acid + 6-Benzylaminopurine+cane sugar+agar as a somatic cell embryo medium, by inducting thecallus, transferring the formed globular embryo onto MS+casein hydrolysate medium, carrying out maturation culture of somatic embryo, and bourgeoning the cultured somatic embryo on a basal medium to from a complete small plant. The method of the invention has the advantages of high callus inductivity, high somatic embryo inductivity and high germination rate, and can cultivate a large numbers of locust test-tube plantlets in a short period for large scale and factory production.

The invention priovidese a new Broussonetia papyrifera mulberry tree hybrid distant hybridization and polyploidization breeding method. The method is characterized in that an inductive hybrid fruit callus doubling distant hybridization polyploid hybrid line is established by an embryo engineering and tissue culture technique of distant hybrids through a reproduction engineering kind improvement technology. The method comprises following steps: 1, obtaining distant hybrid fruits; 2, carrying out callus induction of the hybrid fruits; 3, carrying out colchicine treatment of the callus of the hybrid fruits; 4, carrying out recovery culture of treated calluses; 5, differentiating the calluses into bud seedlings; 6, differentiating the bud seedlings into roots; 7, transplanting tissue culture plants; 8, carrying out comparative observation and selection of survival plants; and 9, carrying out asexual propagation of the selected plants to form a stable kind (line). The method solves the problems comprising slow growth,weak adaptability, unclear trunk and bad material quality of present Broussonetia papyrifera, and allows the new polyploid Broussonetia papyrifera mulberry tree hybrid having the advantages of obvious trunk, strong adaptability and good material quality to be cultivated.

The invention provides a kiwi fruit tissue culture method and an application thereof. According to the method, axillary buds are germinated under an aseptic condition to form a leaf stalk serving as an explant, formation of callus tissues is induced, and then the callus tissues are cultured into kiwi fruit seedlings. According to the culture method, the contamination rate of a tissue culture process can be reduced, and the induction rate, the germination rate and the rooting rate of the callus tissues are increased. In addition, the kiwi fruit seedlings cultured by the method have the advantages of strong growth, developed root systems and high transplantation survival rate.

The invention relates to a culture medium suitable for a large-scale sympodial bamboo including sinocalamus affinis and Mianzhu and a culture method, in particular to a large-scale sympodial bamboo callus induction and plant regeneration high efficiency culture medium and a culture method thereof. The culture medium comprises an MS (Murashige and Skoog) culture medium and a callus induction and plant regeneration high efficiency culture medium and is characterized in that the MS culture medium is prepared from the following raw materials: (a) macroelements: potassium nitrate, ammonium nitrate, potassium dihydrogen phosphate, magnesium sulfate and calcium chloride; (b) microelements: potassium iodide, boric acid, manganese sulfate, zinc sulfate, sodium molybdate, copper sulfate and cobalt chloride; (c) organic components: inosite, glycine, aneurin hydrochloride, puridoxinehydrochloride and niacin; and (d), ferric salts: disodiumedetate and ferrous sulfate.