53 results about "Protonema" patented technology

The invention discloses a semi-leaf laver seedling cultivating method. The method includes the following steps that semi-leaf laver nutritious trichome is obtained; the semi-leaf laver nutritious trichome is cultivated in nutritious liquid until a protonema of a sporangium branch with the set proportion is formed and the cultivating conditions include that the temperature ranges from 20 DEG C and 30 DEG C, the light intensity ranges from 1000 lx and 2600 lx, the lighting period is 10 hours to 16 hours and the nutritious liquid sea water ratio ranges from 1.026 to 1.031. The protonema of the sporangium branch with the set proportion is placed on an attachment basal body and stimulated by water currents until conchospores of which the density reaches the seedling collecting requirement are formed and seedling cultivating of the semi-leaf laver is finished. Thus, the problem that the seedlings of the semi-leaf laver can not be rapidly and efficiently cultivated with the prior art is solved.

The invention provides nostoc capable of degrading polychlorinated biphenyls. A frond of the nostoc is multicellular protonema, has algal filaments and a colloid sheath, and can fix inorganic nitrogen. The nostoc can degrade polychlorinated biphenyl (PCB) 1254 efficiently; after the nostoc is exposed for a week, the degradation efficiency of the nostoc on the PCB1254 is over 60 percent, and the content of residual chlorine ions tested in a culture medium after the degradation and dechloridation of the PCB1254 is 2.26mg/L and 3.01mg/L; the one-month degradation efficiency of the nostoc on six typical indicative polychlorinated biphenyl (PCB52, PCB101, PCB118, PCB138, PCB137 and PCB180) in environment is more than 70 percent, wherein the degradation rate of the PCB52 is 98.7 percent; and after the PCB1254 is degraded for one month, the residual amount of the PCB1254 in the culture medium is only 15.65 percent. Therefore, the nostoc can be widely applied to the degradation of the PCBs insoil environment.

The invention relates to a stereoscopic culturing system and method of porphyra yezoensis shell protonema. The stereoscopic culturing system comprises a plurality of culturing tanks which are distributed stereoscopically, a water storage tank is arranged under the culturing tanks and leads water into the culturing tanks through a water circulation system, and the culturing tanks are each further provided with an artificial light source used for culturing shell protonema. The culturing method includes the steps that light is regulated, wherein 400-500 nm of blue light and 600-700 nm of red light are the main, and light sources at other spectral lines are also taken into account; the intensity ratio of the red light to the blue right is regulated between 4:1 and 12:1; light intensity is controlled between 500 Lux and 5000 Lux; the daily illumination time of the artificial light sources is regulated between 8 hours and 18 hours. The stereoscopic culturing system and method of porphyra yezoensis shell protonema can improve the reliability of seedling culturing production, shorten the culturing period of shell protonema, flexibly arrange the harvesting time of conchospore, and largely reduce the production cost.

The invention provides a method for obtaining and cultivating sphagnum protonema. The method comprises the following steps: carrying out tissue smashing on sphagnum cormus used as a raw material; cultivating in a liquid culture medium; growing new protonema from fragments of sphagnum tissue cormus, and carrying out tissue smashing on the protonema; carrying out liquid cultivation, so as to obtain the sphagnum protonema. By adopting the method, a lot of protonemata of the sphagnum can be obtained; the cormus with uniform growth can be largely obtained by using the protonema within a short period of time; application of the sphagnum in a tissue culture physiological experiment and sphagnum protoplast preparation, transformation or regeneration is facilitated.

The invention discloses a rapid breeding method of high-quality peat moss. According to the rapid breeding method, a large amount of the young and tender cormus of peat moss can be cultured by using peat moss protonema as a material, and the young and tender cormus are sowed in a field environment, and can grow in a large-quantity and high-quality manner under properly-controlled environmental conditions, wherein the regeneration survival rate reaches 100%. According to the present invention, with the artificial propagation method of peat moss, the harvesting amount can be regulated by controlling the planting material, the swing material has strong regeneration ability, the yield is high, the quality consistency is high, and the method can be used for large-scale production planting of peat moss, wetland repair with peat moss, and other fields.

The invention relates to a method for applying and popularizing porphyra yezoensis fine seeds. The method includes the steps of (1) pure fine seed protonema germplasm, wherein shell protonema cultivation is conducted in a nursery pond isolated from a seed source, and conchospore seedling picking is conducted when the protonema is mature, (2) pure fine seed seedling net sea area cultivation, wherein breeding trait recovery is conducted, (3) picking mature fine seed F1 generation algal bodies which serve as the seed algae, (4) cleaning the seed algae with clean sea water, conducting dehydration on the seed algae, drying the seed algae in shade for later use or storing the seed algae at -20 DEG C for later use, wherein the obtained seed algae can be used for fine seed productive carpospores collection, and seedling picking and seedling cultivation can be conducted. The method for applying and popularizing the porphyra yezoensis fine seeds solves the production problems that when the fine seed free protonema is inoculated to shell seedling, later-period maturity promotion measures are not easy to control, conchospore seedling picking efficiency is low, the fine seeds are not easy to cultivate and popularize, and large-scale fine seed seedling and seed supply are achieved. Through the technology, fine seed application has more operability and higher popularization, and the mass production progress of the porphyra yezoensis fine seed is promoted.

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 present invention discloses a trans-regional relay seedling method for porphyra haitanensis. The method comprises: inoculating protonema or carposporophyte of porphyra haitanensis on shell strings in southern part of Zhejiang and sea area of Fujian; after the protonema of shells is cultivated to conchosporangium by adopting a three-dimensional seedling method, promoting maturity by way of shortening the illuminating time, reducing the illuminating intensity and adjusting the adding proportion of a nutritive salt; and transporting mature shells with sporangia to the north of Jiangsu, releasing conchospore by virtue of the thermal expansion principle, and culturing the conchospores attached to a net cord into a sea area. The seedling method disclosed by the present invention has the advantages that shell protonema is cultured rapidly in the south, and is transported to the north if being mature to perform spore dispersion and seedling collection for the first time. The seedling method disclosed by the present invention is suitable for large-scaled porphyra haitanensis cultivation in the north in advance, or performing efficient cultivation by transporting the seedling nets back to the south. Furthermore, the collected seedling shells can be transported back to the south for continuous secondary cultivation, secondary seedling collection is performed in shells in the same batch and are cultured in the south. The method is suitable for being widely applied in porphyra haitanensis seedling fields.

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.

The invention discloses a hermaphrodite laver dihaploid colony building method. The method comprises taking hermaphrodite laver thalluses (gametophytes) with obvious color difference as cross parents, carrying out mixed cultivation on father and mother thalluses with antheridiums, collecting carpospores, selecting the carpospores with germ tubes, carrying out single culture to obtain protonema (sporophore), carrying out induction to obtain conchospore of the protonema, carrying out culture to obtain macroscopic F1 thalluses, wherein if 90% or more of the thalluses contain amphiphilic color chimeras, the thalluses are used for building of a dihaploid colony, separating all color blocks of the four-color block chimera, acquiring monospores or single isolated cells of each one of the color blocks, carrying out culture to obtain macroscopic thalluses, carrying out single thallus culture and autofertilization to obtain pure thallus of each one of color blocks of the four-color block chimera and building a dihaploid colony. The method solves problems of hermaphrodite laver dihaploid colony building and lays the foundation of genetic linkage map construction.

The invention relates to a method for cultivating and breeding pangenesis tortula protonema by using cell engineering, which belongs to the technical field of biology and comprises following concrete steps: a. mature spores are sterilized, and capsulea surface is sterilized for 0.5 to 1 minutes by a solution that contains 1.0% of mercury bichloride and 1.0% of a washing agent; b. germination is implemented on a solid culture medium, the spores are inoculated to an MS culture medium that has the pH value of 5.8 and is added with 2 to 5 mg/L of 2,4-D, 2.0g/L of sodium silicate and 2.0% of sucrose, and cultivated at the temperature of 25 plus or minus 3 DEG C and illumination intensity of 4000lx for 10 to 20 days; c. mass propagation of the protonema is implemented in the MS fluid culture medium for spore germination that has pH value of 5.8 and is added with 2 to 5 mg/L of 2,4-D, 2.0g/L of sodium silicate and 2.0% of sucrose, shaking cultivation is implemented at the temperature of 25 plus or minus 3 DEG C and the speed of 120rpm for 7 to 10 days and the power source is diffused lights; or augmentation cultivation is implemented in a fermentation flask charged with sterile air at the speed of 200L per hour; a small amount of the protonema is collected by 800rpm centrifugation, and a large amount of the protonema is collected through the filtration by 4 to 8 gauze layers. The invention has the advantages of simple and convenient preparation method, low cost and consistent maturity of the obtained protonema.

The invention discloses a preparation method of moss-physcomitrella patens protoplast, comprising the following steps: mechanically polishing and grinding the raw material-protonema of the protoplast and aseptic ultra-pure water, transferring to a 10-dish BCD culture medium, controlling the pH value at 4.5, the temperature at 23 DEG C and the light intensity at 80mu mol photons m<-2>s<-1>, culturing for 5 days under long irradiation of 16 hours, then repeating the processes once, 5 days later, examining the cell growth condition by a microscope, and collecting 20-dish protonema; freshly preparing 20ml of driselase with the concentration of 0.7% and dissolved in 8% mannitol, dissolving the collected protonema for 20 minutes at room temperature under a dark condition, centrifuging for 5 minutes at1000rpm, collecting cells, washing with 8% mannitol three times, dissolving in 5ml of mannitol, and microscopically counting to obtain about 9.6*10<6>; and transferring to a regeneration culture medium for culturing for 3 days, and then microscopically observing so as to discover that the regeneration efficiency is about 99%. Compared with the prior art, the obtained regeneration protoplast is doubled.

The invention discloses a porphyra yezoensis mutation breeding method based on pigment mutant selection. The porphyra yezoensis mutation breeding method comprises the steps of: carrying out induced mutation, carrying out enzymolysis and separation on differentiated cells, selecting good regenerated thallus according to an improved variety selection target for carrying out single-strain culture, qualitatively and quantitatively determining the selected good pigment mutant regenerated thallus; carrying out enzymolysis, culturing into regenerated thallus to obtain diploid pure-line protonema through a parthenogenesis technology, and culturing into F1-generation thallus; and crushing the good pure-line protonema and then carrying out a shell seedling culturing test, picking seedlings from shell protonema on a culture web curtain, and carrying out a thallus pigment mutant good strain culturing test in a sea area. According to the invention, the breeding of the large cultivation algae porphyra yezoensis is switched into early-stage prediction and judgment in a laboratory from the later-stage sea cultivation test detection, stable inheritance of the porphyra yezoensis pigment mutant is obtained by using ray radiation treatment, and mutants relevant to the breeding target trait are selected to be used as breeding targets for carrying out breeding.

The invention discloses a disinfection method of bryophyte explants. By use of the method, the sterilization effect is good, and disinfection waste liquor does not pollute environment, is harmless tothe human body, has low toxicity and cannot damage bryophytes easily. According to the disinfection method, the bryophyte explants are cleaned and then subjected to soaking disinfection treatment by alcohol, sodium hypochlorite, alcohol and sodium hypochlorite in sequence, next, the explants are cleaned up by sterile water and placed in a culture dish to be cultured for 14-21 days until protonemaand cormus are formed on the surface of a culture medium, the protonema and the cormus formed on the surface of the culture medium are sheared and subjected to soaking disinfection treatment by alcohol, sodium hypochlorite, alcohol and sodium hypochlorite in sequence, finally, the explants are cleaned up by sterile water, and the sterile bryophyte explants are obtained. The disinfection method issimple to operate, low in operation threshold, harmless to environment and good in test effect, requires a small quantity of explants, does not use high-pollution disinfectants and is suitable for promotion and application in the technical field of plant tissue culture.

The invention discloses a method for promoting diffusion of altar laver shell protonema conchospore. The method is characterized by comprising the following steps that 1, brassinolide or gibberellin is dissolved in pure water, and mother liquor with the concentration of 1 mol/L is prepared; 2, the brassinolide mother liquor or the gibberellin mother liquor is added in a common culture solution to be prepared into a brassinolide culture solution with the concentration of 1-12 umol/L or a gibberellin culture solution with the concentration of 6-10 umol/L; 3, mature altar laver shell protonemata are suspended and completely immersed in the brassinolide culture solution or the gibberellin culture solution, then shake cultivation is carried out for 12 h to 14 h, cultivation is finished at 7:00 of the next day, then the mature altar laver shell protonemata are placed in seawater, standing is carried out for 2-10 hours, and altar laver conchospore is collected. The method has the advantages that the diffusion speed of the altar laver conchospore can be effectively increased, and meanwhile the diffusion quantity is remarkably increased.

The invention relates to a double haploid breeding method of porphyra haitanensis. The method comprises the following steps of: selecting cross parents; promoting pure protonema with mature parents for diffuse conchospore culture to obtain parent thalli; selecting male and female thalli to hybridize; after culturing the male and female thalli, removing the male thalli, keeping the female thalli to culture singlely till the carpospores is released; culturing the single carpospores to be germinated to a synthesized protonema, and crushing the protonema for continuous culture; taking out part of the protonema to promote manurity, releasing the conchospore and filtering the same to culture to obtain F1 generation thalli, picking out the thalli, putting algae blocks in different colors in a glucose solution to be soaked for enzymolysis, and when single cells or multicellular aggregates are dissociated from the algae fragments, picking out single cells to culture; performing further cultivation till the culture liquid is restored to normal disinfecting seawater, wherein the double haploid is successfully induced; sucking out the pure protonema for amplified cultivation and storing the same as germplasm; and then taking out part of the protonema to promote maturity, promoting matured pure protonema to diffuse the conchospore, and collecting the conchospore to culture to obtain F2 generation pure thalli.

The invention provides a rapid propagation method for physcomitrella patens protonemata. Physcomitrella patens protonema materials growing 5 days are subjected to mechanical grinding crushing and subculturing, test plates which take a BCD medium as a basic medium and have different calcium ion concentrations are arranged, the BCD medium plates with different CaCl2 concentrations are inoculated with the protonema materials after the protonema materials are subjected to grinding subculturing, growing status observation is conducted, and finally the calcium ion concentration at which the optimal protonema growing and differentiating states are achieved is determined. A culture and formula improvement experiment for physcomitrella patens protonema culturing shows that on the conditions that the temperature of an incubator is controlled at 23 DEG C, the light intensity is 80 micromoles photons m<-2>s<-1>, and the sun illumination time is 16 hours, the plant protonemata which grow most vigorously and generate the most materials are obtained at the calcium ion concentration of 5-mM, and a technical basis is provided for a later protoplast preparation and conversion recombination experiment.

The invention discloses a treatment method for crushing microalgae with ultrasonic waves. The method comprises the following steps of mixing a concentrated algae solution enriched with microalgae witha PBS buffered solution to obtain a mixed solution, placing the mixed solution in an ultrasonic generation instrument, and performing treatment for 20-40min, so as to microalgae of which the length is 2-30 [mu]m, wherein the shape of the microalgae is any one of an aggregation group, protonema and rectangular strips. The method disclosed by the invention overcomes the technical defect that a conventional physical mechanical crushing method generates uneven and too big cutting force, so that the integrity of the cell body of the microalgae is damaged. According to the method, the cell wall ofthe microalgae is not destroyed, microalgae cenobium in the shape of the aggregation group, protonema and rectangular strips is subjected to intercellular breaking and separation, so that cell massesin rows, groups or clusters are broken to obtain cell rows or cell groups being shorter in length, even the cell masses are broken into single algae cells, and algae intra-cellular nutrient componentsare effectively reserved. The microalgae breaking solution obtained by the method can meet nutrient requirements of aquatic animals for ingestion cabibre at the seedling raising early stage.

The invention relates to an agrobacterium tumefaciens-mediated moss stable genetic transformation method. According to the method, the moss is the protonemata or gametophyte of the Physcomitrella patens and the Bryum argenteum. The method comprises the steps of culture of moss protonema and gametophyte, genetic transformation and co-culture, screening culture and identification. By taking moss protonema and gametophyte as infection materials, the agrobacterium engineering bacterium liquid carrying a target gene is filled into a culture dish for genetic transformation; the moss is directly infected with agrobacterium, the transformation period is short, and a large number of transgenic materials can be obtained in a short time. Compared with a protoplast transformation method, the method is simple and easy to operate and control, short in period, high in transformation efficiency, good in expression intensity and low in cost, and lays a foundation for gene function research.

The invention discloses a liquid culture method for rapidly increasing the biomass of didymodon tectorum protonemata. The liquid culture method comprises the following steps: taking didymodon tectorum spores, carrying out plant tissue rapid propagation culture on a solid Knop culture medium, carrying outsterile culture ofdidymodon tectorum on a solid culture medium forsubculture at a temperature of 24 DEG C, under a condition of 24 hours of illumination and 24 hours of dark, culturing in an incubator with the light intensity of 3000 lx for 1 week, taking the material, mechanically grinding and crushing, inoculating the material into an improved liquid Knop culture bottle by a pipette, culturing in the incubator with the temperature of 24 DEG C, the illumination of 12 hours, the darkness of 12 hours and the light intensity of 3000 lx for one month, observing the growth state of protonemata by using a microscope during the period, and measuring and counting the biomass of protonemata. According to the invention, a large amount of didymodon tectorum protonemata can be obtained in a short time, and the application potential of didymodon tectorum protonemata as a transgenic material subsequently is greatly improved.

The invention relates to protonema engineering blue algae with high fat content and high essential fatty acid content and constructed by adopting a reverse vector method. According to the invention, the engineering anabaena is constructed, the expression of a pepc gene contained in the blue algae is regulated and controlled by adopting gene manipulation which includes the fishing of a target gene, the construction of a vector, the transformation of the blue algae, the screening of transformants, the culture and detection of the engineering blue algae and the content measurement of total fat and fatty acid. A result proves that the fat content of the anabaena with the pepc gene reversely expressed is enhanced by 93% compared with that of a wild type, the total fat content is increased from 11.5% of the wild type to 22.2%, and the fat yield is double increased and achieves the mean fat level of eukaryotic algae. Thus, the constructed anabaena with the pepc gene reversely expressed can be taken as a more ideal raw material used for preparing biodiesel and essential fatty acid.

The invention relates to a cultivation method of polyploidy of triptonium nigrum, and belongs to the technical field of polyploidy induction. The method disclosed by the invention comprises the following steps of soaking the leucotinum leucotinoides protonemata by using a colchicine solution to obtain mutagenized leucotinum leucotinoides protonemata; carrying out continuous subculture on the mutagenized hypnum macranthum protonemata for three times to obtain subcultured protonemata; carrying out ploidy identification on gametophytes differentiated from the subcultured protonemata to obtain genomic doubled mutants; carrying out subculture on the genome doubled mutant for five times, and screening out a genome doubled and stably inherited mutant; and observing the phenotype of the mutant with doubled genome and stable inheritance, and screening the triptonia melanoides mutant with a brand new phenotype as a new triptonia melanoides strain. The cultivation method is simple to operate, the occurrence rate of chimeras can be reduced, and the obtained polyploidy is stable in heredity.

The invention provides an artificial large-scale cultivation device for porphyra haitanensis shell protonemata. The top of an ultrasonic cleaner is connected to a cover plate, wherein the cover plate is provided with a sprayer on the bottom surface. A storage battery and a power interface are connected to the bottom of a side of the ultrasonic cleaner. The ultrasonic cleaner is connected through a return pipe with a water tank which is connected with a pump through a suction pipe. The pump is connected with the ultrasonic cleaner through a flow pipe. A controlling panel with a display screen and a control switch is disposed on the outside surface of the ultrasonic cleaner. A batten is disposed on an upper end of the ultrasonic cleaner internal wall. The ultrasonic cleaner is connected through the batten with a wooden board provided with rectangle grooves evenly distributed on the surface. Ropes provided with evenly distributed shells are evenly distributed on the bottom surface of the wooden board. The bottom surface of the cover plate is provided with the evenly distributed sprayers whose positions are matched with the rectangle grooves on the wooden board surface. The device can clean shells with an excellent cleaning effect, has the storage battery with distinct performance, and possesses good aging and corrosion resistances.

The invention discloses a tissue culture propagation method of bryum caespiticium. According to the tissue culture propagation method of the bryum caespiticium, the sterilization condition of gametophytes is that 75% ethanol, 1% NaClO and 0.01% HgCl2 are used for treating the bryum caespiticium for 30 seconds. The optimal culture conditions of a regeneration system for the bryum caespiticium are that the photoperiod is 12h/d, the pH value is 6, and 1/2MS, 3.0% sucrose and 0.1 mg/L 6-BA are adopted. The optimal culture conditions of the regeneration system of the gametophytes of the bryum caespiticium are that the photoperiod is 16h/d, the pH is 6.0, and 1/2MS, 3.0% cane sugar, 1.0 mg/L 6-BA and 0.5 mg/L IBA are adopted. According to the tissue culture propagation method of the bryum caespiticium, the disinfection mode and the growth condition which are most suitable for the tissue culture propagation method of bryum caespiticium are screened out, the breeding quantity of the bryum caespiticium is increased, so that a foundation is laid for fully exerting multiple ecological functions of the bryum caespiticium, and an ideal material is provided for vegetation restoration of an ecological system of a drought and degeneration mining area.