30results about How to "Improve embryo emergence rate" patented technology

The invention relates to a method for obtaining collard regenerated plant and special cultivation base. Wherein, said method comprises (1), planting the collard into induced cultivation base, until the density is 5*10<4-5*10<5n/mL, at 30-35Deg.C and in dark, treating it at high temperature for 24-72hours, and under 24-26Deg. C and in dark, inducing idiosome; (2), when the idiosome has been formed, under 24-26Deg. C, 1500-2500Lux, cultivating for 5-10days while each day lights 14-18hours; then grafting the seed into differentiation cultivation base, to be cultivated in same conditions; (3) when grows out branch and leaf, transplanting it into rooting cultivation base, under 24-26Deg. C, 1500-2500Lux, lighting 14-18hours each day, to root, to obtain the collard regenerated plant.

The invention discloses a method for culturing isolated microspore of a common head cabbage to obtain a regeneration plant, which belongs to the field of biotechnology. The method comprises the following steps: choosing a flower bud of appropriate length, pre-treating the flower bud under the low temperature of 4 DEG C for 24 hours, using the squeezing and pressing method to make the microspore isolated after sterilization, filtering and centrifuging the microspore, suspending the purified microspore in a liquid culture medium, and thermally pre-treating the microspore under the high temperature of 32.5 DEG C for 24 hours and transferring to dark cultivation under the temperature of 25 DEG C for 15-20 days; when a macroscopic embryoid emerges, placing in the sunshine for cultivation for 5-7 days until the emergence of a leaf type embryoid, after the greening of the embryoid, switching to a secondary culture medium for germination and transferring to a root culture medium to root, and obtaining the integrated regeneration plant; and then opening the bottle for hardening off, and hardening and transplanting into a culture pan with sterilization matrix. In case of using the method for cultivation, the embryoid with high frequency can be obtained, the obtained haploid regeneration plant is not only a good breeding material, but also an excellent material for research on molecular markers, genetic mapping, gene clone, trans gene and the like.

The invention provides a culture medium formulation for regenerated plantlets of Brassica isolated microspores, which relates to improvement of a culture medium formulation for regenerated plantlets of Brassica microspores as horticultural crops, and belongs to the field of bioengineering. The invention provides the culture medium formulation for regenerated plantlets of Brassica isolated microspores, which is high in induced embryo formation rate and differentiation rate. A culture medium of the invention comprises an embryoid induction culture medium, a differentiation culture medium and a rooting culture medium, wherein the embryoid induction culture medium, the differentiation culture medium and the rooting culture medium all comprise macroelements, trace elements and organic elements. The formulation has the advantages of providing a large number of new genotype pure lines for breeding and greatly enriching breeding resources of Brassica.

The invention discloses a method for improving the embryogenesis rate of Raphanus sativus L. sinoruber makino. The method comprises the following steps: sterilizing Raphanus sativus L. sinoruber makino flower buds, then adding B5 wash medium to prepare a suspension liquid, filtering to obtain a filtrate, and centrifuging to obtain precipitate; sequentially adding NLN-13 induction medium and an activated carbon mixed solution to obtain microspore suspension liquid; subpackaging into a sterile culture dish, then adding sterile broccoli anther, performing Parafilm sealing and then conducting heat-shock treatment; performing routine culture to obtain cotyledonous embryoid; inoculating to an embryoid differentiation medium until differentiation is performed, and regenerating buds; cutting and inoculating regenerated buds to a rooting medium for rooting culture, and hardening seedling and transplanting, to obtain regenerated plants. According to the method, the broccoli anther easy to form embryos and the Raphanus sativus L. sinoruber makino isolated microspore difficult to form embryos are mixed for culture according to certain proportion, so that the embryonic development of the Raphanus sativus L. sinoruber makino can be promoted to obtain a great amount of regenerated plants, and the culture efficiency can be improved.

The invention discloses a rape microspore culture method utilizing mannitol as an osmotic regulator, belonging to the technical field of plant tissue culture. The method comprises the following steps: fetching cabbage type rape as a donor plant for microspore culture; fetching the flower bud on the donor plant; sterilizing and adding an NLN-13 liquid medium to obtain a suspension; filtering to obtain filtrate, and centrifuging to obtain precipitate; adding a mannitol-containing NLN liquid medium, a colchicine solution and active carbon mixed liquid to obtain a microspore mixed suspension; performing heat shock treatment or direct culture to obtain a cotyledon-stage embryoid; inoculating into a regeneration medium till bud regeneration; cutting the regenerated bud and inoculating to a rooting medium for rooting culture; performing seedling hardening and transplanting to obtain a regenerated plant; and fetching the tender leaves of the regenerated plant and detecting the ploidy of each regenerated plant. Through the method disclosed by the invention, the microspore quality, regeneration rate and doubling rate are remarkably improved.

The invention provides a distant hybridization method of common head cabbage and cabbage type rape and belongs to the field of biotechnological breeding. The method comprises the following steps: (1)selecting cytoplasmic male sterile line cultivated species of common head cabbage as female parents, taking restorer line cultivated species of cabbage type rape as male parents, carrying out artificial pollination and then isolating the female parents and the male parents; (2) cutting ovaries which are pollinated before 15-18 days, and disinfecting the ovaries; (3) peeling ovules of the disinfected ovaries and carrying out induction culture until the ovules are germinated; (4) carrying out differentiation culture of the germinated ovules to obtain an F1 generation of seedlings; and (5) separating lateral buds of the F1 generation of seedlings and inoculating a rooting culture medium with the lateral buds to obtain complete hybridized seedlings. According to the distant hybridization method, an embryo rescue technique is used, the problem that the hybrid plants cannot be obtained because of high possibility of abortion of young embryos when the cultivated species of common head cabbageand cabbage type rape are distantly hybridized can be overcome; excellent restoration genes of the rape are transferred into cytoplasmic male sterile line cultivated species of common head cabbage, so that the fertility of the cabbage can be restored; the technical basis is provided for innovation and application of genetic resources of the cabbage.

The invention provides a culture method for regenerated plantlets of Brassica isolated microspores, which relates to improvement of a culture technique for regenerated plantlets of Brassica isolated microspores as horticultural crops, and belongs to the field of biological breeding techniques. The invention provides the culture method applicable to regenerated microspore plantlets of Brassica crops. The culture method comprises the following steps: (1) culturing microspores; (2) performing differentiation culture; and (3) performing rooting culture. The culture method has the advantages of providing a large number of new genotype pure lines for breeding, greatly enriching breeding resources of Brassica, accelerating breeding pace, greatly shortening breeding time and having broad application prospects.

The invention discloses a method for increasing non-heading Chinese cabbage microspore embryo induction rate. The method includes the steps: (1) flower bud collection; (2) microspore embryo culture: sterilizing, cleaning and rinsing the flower buds collected in the step (1), grinding the rinsed flower buds, filtering and centrifuging the grinded flower buds, diluting yellow-green precipitation bya first culture medium, performing heat shock treatment, and performing dark culture for 14-16 days at the temperature of 24-26 DEG C to form an embryoid; (3) enabling the embryoid to sprout to form aplant. The first culture medium is an NLN-13 culture medium added 0.04-0.08mg/L 6-BA, 10-20mg/L ascorbic acid and 1mg/L activated carbon. According to the method, operation steps are simplified whengerm extraction rate can be ensured, operation time is greatly shortened, and the production efficiency of plants is improved.

The invention discloses a method for cultivating non-heading Chinese cabbage microspore plants. The method comprises the following steps: (1) selecting flower buds; (2) cultivating microspore derivedembryos, wherein the flower buds are sterilized and washed and rinsed with a 1/2NLN-13 culture medium; the rinsed flower buds are added into the 1/2NLN-13 culture medium and fully ground, filtered andcentrifuged, a yellow-green precipitate is diluted with a NLN-13 culture medium and split-charged into a culture dish after treatment with ethylmethane sulfonate, activated carbon is added into the culture dish to enable the concentration of the activated carbon to be 1 mg/mL, heat shock treatment is carried out, and then dark culture is carried out for 14 days to form embryoid bodies; (3) germinating mature embryos into plants, wherein the embryoid bodies undergo shaking culture, the mature embryos becoming green are transferred to an improved solid 1/2MS culture medium for 40 days, and thenacclimatization and transplanting are directly carried out. According to the method, under the condition that the embryo yield can be guaranteed, the operation steps are simplified, the operation time is greatly shortened, and the plant production efficiency is improved.

The invention discloses a method for inducing a pumpkin haplobiont. The method comprises the following steps that surface disinfection is carried out on an unfertilized ovary of a pumpkin, the parts,without ovule, at two longitudinal ends of the ovary and skin of the ovary are cut off and then the remaining ovary is transversely cut into ovary slices with the thickness of 1-2 mm, and the ovary slices are inoculated into an induction culture medium for induction culture after being sterilized; when an obtained embryo has an obvious morphological upper end and a morphological lower end, the embryo is transplanted into a seedling culture medium for seedling culture until a regenerated plant is obtained; the pumpkin haplobiont is screened out through ploidy identification. The method is easyto operate, the haplobiont can be obtained within 60 days, the embryo yield and the plant regeneration rate are relatively high, the breeding period is shortened, the breeding efficiency is greatly improved, and the haploid rate is high. The invention also provides the induction culture medium. Only one hormone is added on the basis of an MS culture medium, preparation is convenient, the price islow, and the induction culture medium can be effectively applied to unfertilized ovary isolated culture of the pumpkin.

The invention discloses a dihaploid induction method of head cabbage with high efficiency, which belongs to the filed of biological breeding. The method relates to a regeneration of microspore plant of the horticultural plant head cabbage and an improvement of dihaploid induction technology thereof. The method comprises the following steps: (1) selecting bud and separating microspore; (2) doubling inducing and culturing formed embryo; (3) differencing seedlings and subculturing; (4) cultivating strong seedling rooting. According to the invention, the dihaploid of head cabbage can be applied in a mass production with a large scale, a plurality of novel genetype pure lines can be provided for breeding. The method is affected slightly of the growth environment and genotype, and has the advantages of high embryo generation and embryo formation, good doubling effect, strong growth of seedlings plant, less subculture frequency, strong root and seedings after rooting and high survival rate of the field transplanting.

The invention relates to the technical field of Chinese cabbage breeding, and particularly relates to a method for creating diploid germplasm resources by using tetraploid Chinese cabbages. The tetraploid Chinese cabbages are subjected to isolated microspore culture, the obtained tetraploid isolated microspores are subjected to heat shock treatment and dark culture temperature regulation culture to obtain embryoids, and the embryoids are subjected to tissue culture and acclimatization and transplantation to obtain regenerated plants and germplasm resources. According to the method, the advantage that the number of chromosomes of the tetraploid Chinese cabbages is multiplied is utilized, the frequency of interchromosomal gene interchanges is significantly higher than the frequency of diploid intergenic interchanges, and the formed microspore genotypes are richer, so that a new diploid material with more variant characters can be obtained.

The invention discloses a method for improving the embryonic induction rate of free microspores of medicago sativa l., wherein the method includes the steps: selecting robust disease-free and insect-free flower buds from a donor plant at a full-blossom period of medicago sativa l., selecting flower buds of the free microspores in a mononuclear metaphase, and cultivating. The method has the advantages that the microspore embryos of the medicago sativa l. can be obtained quickly and efficiently, the embryonic induction rate reaches 44.22 per bud, and obtained regenerated plants can be used for haploid breeding of medicago sativa l.

The invention discloses a method for improving germplasm creation efficiency of radish, which comprises the following steps: S1, preparation of rootstocks and scions: selecting a hybrid variety easy to generate embryos as a rootstock material, selecting a hybrid variety with brittle green skin color and flesh color as a scion material, and sowing and vernalizing rootstock and scion seeds; S2, field planting; S3, grafting environment management: controlling the environment temperature to enable plants to grow, and irrigating roots with a chlormequat chloride diluent; S4, grafting time selection: taking the second and later primary branches of the rootstocks as grafting parts, and selecting primary branches with the length of 6-10 cm as the scions; S5, grafting: grafting the scion buds on rootstock plants; and S6, management after grafting: after grafting is completed, managing grafted plants by controlling the growth environment temperature. The method can effectively improve the embryogenesis rate of free microspores of the material difficult to embryogenesis, accelerate the purification speed of the radish material and improve the germplasm creation efficiency.

The invention discloses a method for rapidly obtaining haplobiont of Brassica oleracea var. botrytis. The method comprises the following steps: sterilizing Brassica oleracea var. botrytis flower buds, then adding B5 wash medium to prepare a suspension liquid, filtering to obtain a filtrate, and centrifuging to obtain precipitate; sequentially adding an NLN-13 induction medium and activated carbon mixed solution to obtain microspore suspension liquid; subpackaging into a sterile culture dish, then adding sterile brassica oleracea anthers, performing Parafilm sealing and then conducting heat-shock treatment; performing routine culture to obtain cotyledonous embryoid; inoculating to an embryoid differentiation medium until differentiation is performed, and regenerating buds; cutting and inoculating regenerated buds to a rooting medium for rooting culture, hardening seedlings and transplanting, to obtain regenerated plants. According to the method, the brassica oleracea anthers easy to form embryos and the Brassica oleracea var. botrytis isolated microspores difficult to form embryos are mixed for culture according to certain proportion, so that the embryonic development of the Brassica oleracea var. botrytis can be promoted to obtain a great amount of regenerated plants, and the culture efficiency can be improved.