180 results about "Gametophyte" patented technology

The invention discloses a paraffin section method for fern gametophytes, which comprises the steps of material selection and fixation, washing, coloration and bluing, dehydration and hyalinizing, waxing and embedding, sectioning, patching, dewaxing and mounting, as well as microscopic examination and photographing, wherein improved FAA (formalin, acetic acid and alcohol) stationary liquid is adopted in the fixation, and the stationary liquid consists of formalin, acetic acid and 30% alcohol (formalin:acetic acid:30% alcohol = 1:1:18). The method is adopted, so that the disadvantage of much youngness of the fern gametophytes can be effectively overcome; the fixation can be well performed for cell divisions at all stages; from a paraffin section in a later stage, a plurality of mitotic sections can be observed, so that a whole development process can be reflected detailedly; therefore, the method has high development and application values.

The present invention provides an introduction method of the laminaria japonica, which can resolve the problems of the prior art that the selected frond section having the mature sporangia is easy to decay and the spore is easy to release, thereby leading to the failure of the introduction. The introduction method adopted by the present invention is to firstly dry and incite the mature sporangia leaf of the laminaria japonica in a temperature-controllable laboratory, then put the mature sporangia leaf of the laminaria japonica into a water tank that is provided with the fresh sterilized sea water and the temperature of the water is controlled between eight cent degrees and twelve cent degrees to release the spores, then cultivate the spore to form the gametophyte, to put the seedling curtain attached with the gametophyte into a low-temperature enclosed bag and finally transport in low-temperature storage to the destination. The shape of all gametophytes on the laminaria japonica seedling curtain is normal, which has high successful rate and the survival rate is nearly one hundred percentages.

An artificial reproduction method for drynaria includes asexual production and spore reproduction. Said asexual reproduction includes such steps as taking its rhizome segment with 3-5 bud points and 3-5 sporophyllary leaves, fixing it to tree or mountain rock, covering it by bryophyte, spraying water and management. Said spore reproduction includes such steps as sterilizing the burnt earth, immersing in cold boiled water, loading the sporanges in bag, drying in air, preparing the suspension of sporanges, spraying it onto said burnt earth, optical culture to obtain gametophytes, transplanting, culturing to obtain sporophytes, and transplanting them on tree or mountain rock.

The present invention relates to a method for propagation and cultivation of platycerium. It is characterized by that said invention adopts sterile spore germination technique and makes it be combined with test-tube seedling-cultivating technique to implement propagation and cultivation of platycerium. Said method includes the following several steps: collecting storing, health and mature spore, making sterile spore germination, gametophyte growth culture, insemination to form zygoite, sporophyte enrichment culture, rooting culture and transplantation, etc.

The invention relates to a method for screening real-time fluorescence quantification PCR internal reference molecules of syntrichia caninervis in a desert. The method comprises the following steps: selecting 15 internal reference candidate genes by utilizing a syntrichia caninervis transcriptome database, designing an internal reference gene specific primer of real-time fluorescence quantification PCR by taking 15 internal reference genes as templates; carrying out a fluorescence quantification PCR experiment by selecting syntrichia caninervis gametophytes stressed by 10 non-living things and free of stress (in contrast) as experimental materials; carrying out fluorescence quantification data analysis by using geNorm, NormFinder and Refinder software, so as to screen out the best fit and the most stable internal reference molecules CDPK and alpha-TUB2 of the syntrichia caninervis for developing fluorescence quantification research under various abiotic stresses. By adopting the method disclosed by the invention, the errors of the syntrichia caninervis transcriptome material under different abiotic stresses in ribonucleic acid (RNA) quality, yield, reverse transcription efficiency and the like can be avoided, the real-time fluorescence quantification detection data can be better corrected and standardized, and the accuracy and the reliability of the gene quantification research are improved.

This invention discloses a clonic seeding method of sea-tangle gametocyte. Comparing with former mature sea-tangle seed source, using gametocyte clone as seed source can solve these problems: seeding time is limited by sea areas and water temperature; seeding time is long; sea-tangle seeding causes its varieties complex; the growth degree and dote condition of sea-tangle are different, which causes various diseas occur during seeding period. The seeding-pick are proceed at twice and two curtains are employed, these heightens sea-tangle seeding output.

Methods are provided for producing apomictic plants from sexual plants divergent with respect to responses to different photoperiods and schedules of megaspore and gametophyte development. A preferred system is to identify divergent lines from within a species or closely related group of species, accentuate the divergence by breeding where necessary, and produce artificial amphiploids that contain genomes from the apposing divergent lines. Apomixis results from the asynchronous expression of female developmental programs induced by combining the reproductively divergent lines. The procedures for manipulating the expression of apomixis described herein permit the development of true-breeding hybrids of various cultivated crops.

A tissue culturing method includes following steps: providing a chopped gametophyte, generating calluses by culturing the chopped gametophyte, and performing apogamic regeneration of sporophytes, by culturing the calluses in a culture fluid to develop the sporophytes from the calluses. The present invention also provides a tissue culturing method of ferns and an explant.

The present invention discloses a seedling cultivation method of sea tangle, wherein the method comprises the following steps: a. cultivating the collected zoospores under the white radiation irradiation with a certain illumination intensity for forming gametophyte; b. cultivating the gametophyte with the blue light illumination with a certain illumination intensity for converting the gametophyte to micro sporinite; c. growing the micro sporinite under the red light illumination with a certain illumination intensity for a certain period of time; and d. cultivating the sporinite under blue light illumination with a certain illumination intensity so that the sporinite obtains the dimension required for stored cargo delivery. The healthy growing of sporinite can be guaranteed under a higher temperature through adopting the combination irradiation with various light qualities for executing seedling cultivation of sea tangle. The method is simple for applying and the seedling cultivation cost is low.

It relates to a method of quickly selecting and breeding new species of radish. In the breeding method, the haploid embryo bodies developed from male gametophyte cells are acquired through culturing anthers of radish in glass. Then haploid embryo bodies are cultured, doubled chromosomes, and pollinated independently, thereby the different genotypic clones are obtained. The field characters of these clones are compared, and the super clone could be screened out as routine new strains or mating parents. The breeding method avoids the insemination process for radish, simplify the isozygoty process of heterozygotes, shorten the breeding process, thereby improve the breeding efficiency.

The invention relates to a method for rock green coverage by using moss. The method comprises the following steps: collecting moss in the field, selecting moss which has good growth vigor and relatively high rock drought resistance as a moss material, and selecting an adhesive with no toxicity, environment friendliness, water and air permeability, and good stickiness, so that the moss cannot be scoured out by rainwater before survived on rocks; selecting peat soil suitable for the growth of moss as a nutrient agent, or mixing peat soil with soil where the moss is collected to obtain a nutrientagent; stirring moss gametophytes, the adhesive and the nutrient agent according to a certain proportion to obtain a jelly, and smearing the jelly on rock walls; and carrying out watering every day to guarantee the humidity required by the growth of the mos. The provided method for rock green coverage is simple and easy to operate, is low in cost, is beneficial to ecological greening and can be used to improve the surrounding environment.

The invention discloses a cultivation method of a moss and desert algae composite crust. The cultivation method comprises the first step of collecting in summer a superior kind of moss in a natural habitat of an area to be restored; the second step of returning activities of the moss; the third step of using distilled water to wash moss crust the activities of which are returned and obtaining moss gametophyte; the fourth step of putting the gametophyte into a solid culture medium to be subjected to cultivation after the gametophyte is subjected to sterilization and obtaining moss protonema; the fifth step of picking the moss protonema and conducting static liquid cultivation to obtain differentiated gametophyte, and putting the gametophyte into a fluid culture medium to conduct aerobic cultivation; the sixth step of conducting aerobic cultivation on the desert algae through the fluid culture medium till a logarithmic growth phase, and obtaining algae thick liquid after removing supernatant; the seventh step of mixing the moss gametophyte with the desert algae thick liquid and afterwards inoculating the moss gametophyte and the desert algae thick liquid to the surface of desertification soil, and cultivating the moss and desert algae composite crust. The cultivation method of the moss and desert algae composite crust is easy to carry out and convenient to operate, evolvement process of biological soil crust in a desert area is accelerated, and fast treatment of desertification soil in arid and semi-arid lands is realized.

The invention relates to a method for soilless planting of mosses on a net shed sand bed in a solar greenhouse, and belongs to the technical field of moss planting. According to the technical scheme, a disease and insect pest control system is arranged in the solar greenhouse; a suspended sandy planting bed and a built-in sun-shading net tent are built in the solar greenhouse, wild mosses attached to the surface of a shady moist stony rock mass are collected, and moss seedlings are cultivated on a seedbed of a three-layer composite structure; the moss seedlings are taken out and smashed into moss pulp, and the moss pulp is applied on gauze flat laid on the surface of the suspended sandy planting bed to cultivate a lawn-shaped moss gametophyte colony. The innovative moss soilless planting technology has the advantages that industrialized production of the mosses is achieved by utilizing the solar greenhouse with a regulable environment in the North, a produced lawn-shaped moss gametophyte rhizoid intertwining layer is thick, tight and not easy to loosen, the leaves are bright green, the market selling price is high, and an ecological green environment-friendly moss decoration material is utilized sustainably and widely.

The invention discloses a method for generating macrogametocytes by somatic cells of inducing undaria pinnatifida parthenogenesis juvenile sporophytes, which has easy operation, low cost and high maturation rate of the generated macrogametocytes and can provide a large quantity of the macrogametocytes for the large-scale seedling production of undaria pinnatifida. The method comprises the following steps of: cutting undaria pinnatifida macrogametocytes stored and cultured in a room into pieces and then culturing under the conditions of temperature suitable for the undaria pinnatifida macrogametocytes to grow to be mature, illumination and nutritive salt, wherein when the undaria pinnatifida macrogametocytes grow to be mature, form oocysts in large quantities and ovulate with ovums, a small quantity of the fertilized ovums are germinated to seedlings through parthenogenesis because unfertilized ovums are died by being unfertilized by spermatozoa; and when the somatic cells of the parthenogenesis seedlings are grown to 0.4-0.6 cm, adopting a rhizoid cutting method to enable fronds to lose polarity and constraint on the somatic cells, and inducing the somatic cells of the seedlings to generate the macrogametocytes.

The invention discloses a mass pteridophyta reproduction method. The mass pteridophyta reproduction method includes the steps of (1), harvesting pteridophyta spores; (2), sowing, namely, mixing collected spores with a hormone solution, spraying the mixture into a sowing medium, and reproducing by keeping relative air humidity above 85% and temperature ranging from 25 DEG C to 27 DEG C, wherein the hormone solution consists of 1 / 2 MS (Murashige and Skoog) culture medium, NAA (naphthalene acetic acid) and GA3; (3), performing 'two-time transplant', namely, transplanting gametocytes on the culture medium after the gametophytes mature and before sporophytes arise, and transplanting seedlings into yellow soil when the sporophytes have 2-3 leaves. By the aid of the mass pteridophyta reproduction method, spore germination rate can be increased, and prothallus development can be promoted; production sowing quantity is guided effectively, and sporophyte growth and transplant management are benefitted; gametophyte fertilization is further benefited by 'two-time transplant', and seedling growth uniformity can be improved while seedling survival rate can be increased.

The invention discloses a method for controlling turf degradation by using bryophytes and herb-bush plants combined with high-order granule technology, and belongs to the technical field of vegetationecological restoration. The method uses a combination of bryophyta gametophyte and herb-bush plants as a novel grass seed spraying agent, which conforms to a natural ecological succession process. The bryophytes can rapidly multiply under suitable conditions by utilizing the superiority of the bryophytes, and the germination rate and coverage of herbaceous plants are improved to form a more stable ecosystem so as to prevent current excessive turf degradation and large-scale open-air rock and soil and accelerate the formation of an organic whole of slope rock mass and vegetation matrix. A soilculture medium that is more suitable for plant growth is produced by using the high-order granule technology, and spray seeding is carried out by using professional equipment so as to provide betterprotection for the plant growth. Plant species are determined according to local climate, soil and water and site conditions, and the spray seeding is carried out according to the mass ratio of bush:herb: bryophytes=7:2:1. Artificially forced greening changes into self-reproduction of natural vegetation. The method of the invention is applicable to places where turf is seriously degraded or unableto grow, such as various slopes, barren mountains and acid-alkaline soil.

The invention discloses a multi-control sterility expression vector which is constructed through an Ms7 gene and used for recovering, keeping and reproducing corn recessive genic male sterile lines, and an application method of the multi-control sterility expression vector, and belongs to the field of plant genetic engineering in the biotechnology. The multi-control sterility expression vector mainly comprises four functional elements, namely, the fertility reproducing gene functional element for corn male sterility, the functional element used for restraining corn male gametophyte formation, the cone peel color screening and marking functional element, and the herbicide resistance gene functional element. The constructed vector is introduced into corn receptors such as cells, callus or organs, can be used for recovering, keeping and reproducing the male sterility lines of corn, is expected to be applied to corn sterile cross breeding and cross hybrid seed production, and generates great economical value in the corn production.

The invention discloses a pteridophyte seedling raising device. The pteridophyte seedling raising device comprises a seedling raising basin, a sowing clapboard, and a culture medium pushout constructional element. The seedling raising basin comprises a basin body (2) and a cover body (3) which is matched with the basin body. The basin body is composed of a bottom wall and side walls. Through-holes are formed in the bottom wall. The cover body is composed of a top wall and side walls. Convection holes are formed in the side wall of the cover body. The sowing clapboard (4) is arranged in the basin body and is composed of clapboard side walls and side walls of griddings arranged inside the clapboard side walls. The culture medium pushout constructional element (5) comprises a base seat and a convex circular cylinder which is arranged on the base seat and matched with the shape of gridding cavities. The pteridophyte seedling raising device can improve efficiency of seedling and transplanting, reduce cross infection probability between sporophytes, reduce damage of gametophytes and sporophytes in transplanting process, and has the advantages that cultivating room is made full use of, stability of development environment of spores and seeding transplanting process standardization are guaranteed, management cost is reduced and the pteridophyte seedling raising device is suitable for pteridophyte seedling standardization and large-scale production.

The invention relates to a kelp germplasm separation method. According to the kelp germplasm separation method, when kelp zoospores are diffused, kelp is inside and outside wrapped by medicine, gauze and bolting-silk, so that kelp mucus and impurities can be effectively prevented from influencing water quality; a pipettor separation gametophyte can be used for effectively improving the separation efficiency and avoiding pollution; and a culture medium is improved, so that the rapid growth requirement of the kelp zoospores is satisfied, and the pollution can be effectively avoided. Therefore, the kelp germplasm separation method is safe and high-efficiency.

The invention discloses a multi-control sterile carrier constructed on basis of an Ms30 gene and used for restoring, maintaining and breeding a maze male sterile line and its application method and belongs to the field of biotech genetic engineering. The multi-control sterile carrier comprises four functional gene elements: a maze male sterile fertility restoration functional gene (1), a maze male gametophyte formation preventive functional gene (2), a corn peel color screening mark functional gene (3), and an herbicide resistance functional gene element (4). Such a construction can be introduced into a maze acceptor (such as a cell, callus or organ) to restore, maintain and breed the maze male sterile line corresponding to the Ms30 gene. The method is expected to be applicable to novel sterile hybridization and hybrid seed production of maize and has great application value in maize production.

The invention relates to a novel method for screening microRNA (Ribose Nucleic Acid) target genes at a high flux, in particular to a novel method for screening microRNA target genes at a high flux by using a biotin-modified microRNA mimic. The method is mainly characterized in that: the biotin-modified microRNA mimic is designed and synthesized, and the target gene mRNA which is directly combined with the biotin-modified microRNA mimic is enriched and selected by using a magnetic bead which contains a gametophyte with high affinity to biotins. By adopting the method, the mRNA target genes which directly interact with microRNA are enriched, so that problem of false positive existing in other methods is solved; moreover, the method is easy and convenient to operate, has relatively low cost and is suitable for most laboratories.

The invention discloses a method for quickly breeding pteris vittata by a spore semi-tissue culture method, comprising the following steps of: sterilely treating the spore of the pteris vittata, culturing on a culture medium to form into gametophyte, and seeding the gametophyte in soil to be cultured into the young seedling of the pteris vittata; the process for forming the gametophyte comprises the following steps of inoculating the sterile spore of the pteris vittata onto a 1 / 2 MS culture medium for cultivation at the temperature of 22-28 DEG C, periodically irradiating in the process of cultivation under the condition that the light intensity is 2000-2500lx, irradiating for 12-14 hours every day, and cultivating 5-8 days, i.e. germinating the spore to form into the gametophyte. The process for cultivating the gametophyte into the young seedling of the pteris vittata comprises the following steps of soaking the gametophyte in a 1 / 10 MS culture medium for 0.5-1.5 hours, seeding on the surface of soil, carrying out surface coating, and cultivating under the irradiation condition of natural light for 3-5 days. According to the breeding method, the pteris vittata breeding period can be shortened, the operation steps can be simplified, the pteris vittata is high in survival rate, and the breeding coefficient can be greatly improved.

The invention discloses a multivalence recombinant protein vaccine for chicken coccidiosis, and a preparation method and application of the multivalence recombinant protein vaccine for chicken coccidian. The multivalence recombinant protein vaccine for chicken coccidiosis comprises 4 chicken coccidian proteins: an eimeria tenella rhoptry protein 41, an eimeria tenella gametophyte protein 22, an eimeria maxima gametophyte protein 56 and an eimeria acervulina merozoite protein Ea3-1E. The four chicken coccidian proteins are respectively connected with escherichia coli expression carriers to obtain recombinant expression carriers for expressing recombinant proteins. Different recombinant proteins are mixed with an adjuvant in proportion, when descendant chicks of immunized hens are inoculatedwith coccidian are compared with descendant chicks of hens which are not immunized, the survival rate and the relative weight increment of chicks inoculated with the coccidian can be effectively increased, the discharge quantity of oocysts is reduced, and favorable maternal immunity effects are shown.

The invention discloses a method for establishing a racomitrium japonicum gametophyte regeneration system. The method provided by the invention comprises the following steps of shearing racomitrium japonicum subjected to appropriate culture into small racomitrium japonicum segments from top to bottom, carrying out surface disinfection, inoculating a conventional MS culture medium with the sterilized small racomitrium japonicum segments, carrying out culture, carrying out screening to obtain aseptic explants, transferring the aseptic explants to a medium comprising a conventional MS medium and30g / L of cane sugar, inducing gametophytes to produce protonemata, transferring the protonemata to a propagation medium, carrying out culture for 50 days, transferring the protonemata obtained by theprevious step to a medium for producing gametophytes by induction, and carrying out culture for 70 days. The method for establishing a racomitrium japonicum gametophyte regeneration system is a novelmethod, has simple processes and a low cost, can realize propagation of a mass of racomitrium japonicum gametophytes in a short time, and guarantees the success of follow-up experiments.

The invention discloses a pinus massoniana embryogenic callus proliferation and maintenance culture method. The method includes: placing a filamentous embryogenic callus cell cluster obtained by induction of 3-4 weeks of a conventional somatic embryo and an embryogenic callus obtained by separation of a female gamete into a petri dish with a culture medium I; removing dead cells and unnecessary tissues in embryogenic calli in 4-6 weeks; transferring into a centrifugal tube, adding a culture medium II, mixing and well shaking; adding an embryogenic culture obtained after well shaking into a petri dish with a culture medium III; after 6-8 weeks, directly transferring an appropriate size of the embryogenic calli into a culture medium IV for normal subculture in a subculture period of 10-14d to finally obtain a pinus massoniana embryogenic cell line which is fast in growth, vigorous and available for long-term subculture. Evident browning and water soaking of the embryogenic calli in long-term subculture are avoided, the embryogenic calli grow fast, proliferation effects are stably high, the proliferation coefficient reaches 25 or above, and high economic benefits and social benefits are achieved.

The invention discloses an in-vitro rapid propagation method of macromitrium cavaleriei card and ther, which comprises the following steps: (1) carrying out surface sterilization of mature capsula of the macromitrium cavaleriei card and ther which grows in the natural environment; inoculating the mature capsula on Knop's culture medium without any hormone to culture and obtain protonemata; and then culturing for 30 days to obtain gametophyte; (2) inoculating single gametophyte on a culture medium for subculture multiplication of the gametophyte to obtain new gametophyte. The invention builds the in-vitro rapid propagation method of macromitrium cavaleriei card and ther for the first time; by using the in-vitro rapid propagation method, large-scale artificial propagation of the gametophyte of the macromitrium cavaleriei card and ther can be realized in a short time. The method has obvious scientific significance and practical value, and can provide a large number of uniformly-growing ideal test materials for indicating and monitoring atmospheric pollution by utilizing bryophytes. Simultaneously, the method continuously provides rich abundant seedlings for garden landscapes which use the bryophytes, such as establishing of a moss garden.

A new method for culturing kelp seedlings in scale without substratum includes steps of (1) obtaining kelp embryoid sporophore by means of gametophyte clone cross breeding, (2) moving mixed liquid containing the kelp embryoid sporophore into sterile PES culture media to prepare embryoid sporophore culture solution and breeding the kelp embryoid sporophore into kelp young sporophore of 200+ / -20 micrometers, and (3) expanding a kelp young sporophore culture system gradually by a three-step culture method, increasing culture temperature, breeding the young sporophore into seedlings of 1-2cm gradually in culture containers by a ventilation suspension method. By the ventilation suspension method, the survival rate can be increased greatly and loss caused by rotten seedlings can be reduced. Due to the ventilation suspension method for culture of kelp, operation in kelp culture is simple, laborsaving and timesaving, limitation in mature main weed is avoided, the kelp seedlings can be cultured anytime according to actual requirements, and the survival rate and the culture density of unit water body are higher than those of conventional methods.

The invention discloses a novel method for quickly screening high-yield kelp strains, and belongs to the technical field of kelp seedling breeding. Photosynthetic system parameters of photosynthetic systems II of gametophytes of different strains of kelps are detected and compared by a chlorophyll fluorescence technology, and the strains with large parameter values are preferably selected to be retained. The chlorophyll fluorescence technology adopted by the novel method disclosed by the invention is a high-efficiency and lossless detection technology; according to the fluorescence detection, the number of required gametophyte samples is small (fresh weight: 0.05g), and the gametophytes cannot be badly affected; the detected samples can be continued to be used for gametophyte cloning breeding; the new strains of the high-yield kelps are screened by detecting the photosynthetic parameters of the gametophytes; compared with the conventional seedling breeding method, the novel method has the advantages that the breeding time is greatly saved, and a breeding result is high in pertinence.

The invention discloses a method for simultaneously extracting high quality DNA / RNA of kelp gametophyte, and belongs to the field of molecular biology. The method comprises steps of material selection, material grinding and cracking for several times, extraction, precipitation, dissolution, and DNA / RNA purification. The provided method can solve the problems that in the prior art the material consumption is huge, the materials should be grinded by liquid nitrogen, the obtained DNA / RNA amount is little, the DNA section is small, and the polysaccharide pollution rate is high. The invention aims to provide a simple and practical preparation method, which is suitable for being carried out in a lab, of kelp gametophyte DNA / RNA.

The invention discloses a rapid propagation method of a vesicularia protonema and gametophyte. The method includes disinfecting vesicularia sporule; preparing a spore suspension; inoculating the sporesuspension to an LQ culture medium for culture; germinating the spores to generate the protonema; subculturing and expanding the protonema to obtain a large number of new gametophytes; grinding and crushing the vesicularia protonema or / and gametophytes by using a homogenizer; and inoculating the crushed moss suspension to the LQ culture medium for culture. The method establishes a vesicularia tissue culture system for the first time, can obtain a large number of the vesicularia protonemata and gametophytes in a short time, provides a large amount of experimental materials and technical support for genetic improvement of vesicularia, and further provides rich provenances for gardening, landscaping and the like by applying the vesicularia.