27 results about "Embryo rescue" patented technology

The invention discloses a breeding method of a drought-resistant machine-harvested cotton variety. The breeding method comprises the following steps: step A1, hybridizing multiple parents; the method comprises the following steps: constructing a basic hybrid combination by taking a cotton line with strong drought resistance as a female parent and a cotton line with outstanding mechanical-harvest character as a male parent, introducing a wild cotton germplasm and a high-quality fiber cotton germplasm as innovative parents, and polymerizing drought-resistant, mechanical-harvest-suitable, high-quality fiber and stress-resistant related genes through composite hybridization by combining distant hybridization with an embryo rescue technology to obtain F1-generation seeds; the invention relates to the technical field of cotton breeding. According to the breeding method for the drought-resistant machine-harvested cotton variety, parent combinations are scientifically configured, a stable drought-resistant line and a machine-harvested line are used for constructing a basic hybrid combination, meanwhile, a wild cotton stress-resistant germplasm and a high-quality fiber cotton germplasm are introduced, and a quaternary composite hybridization and special embryo rescue technology is innovatively adopted; the seedling rate of the distant hybridization embryo of the wild cotton is increased to 60% or above, and the problem of abortion of the distant hybridization embryo is effectively solved.

This application relates to a method and primer pairs for identifying allotetraploids from intergeneric hybridization of Chinese cabbage and arugula based on KASP technology. The method includes obtaining a sample to be identified, wherein the sample is obtained by intergeneric hybridization of Chinese cabbage '72049' as the female parent and arugula 'ASTRO' as the male parent, followed by embryo rescue and chromosome doubling; using the genomic DNA of the material as a template, amplification is performed using KASP primer pairs; fluorescence signal detection is performed on the amplified product; the authenticity of the allotetraploid is determined; and distant hybrids can be predicted based on the genotyping results, significantly improving identification efficiency and saving costs.

This invention discloses a method for producing a new variety, "Zheliang," through distant hybridization of sugarcane with sorghum and maize. It relates to the field of agricultural biotechnology. The method mainly includes: selecting specific wild sugarcane species as the female parent and wild sorghum and maize species as the male parents; cultivating the sugarcane under controlled environmental conditions using a specially formulated potting mix; artificially emasculating the female parent and spraying it with a specific treatment solution before pollination to improve hybrid compatibility; obtaining tri-haploid hybrids through artificial pollination, embryo rescue, and chromosome doubling techniques; and performing multiple generations of backcrossing and screening on the hybrids to ultimately obtain a genetically homozygous, trait-stable new variety, "Zheliang." This method solves the technical problems of mismatched flowering periods and hybridization incompatibility between sugarcane and sorghum / maize, successfully cultivating a "Zheliang" variety that combines the strong ratooning ability of sugarcane with the excellent resistance and biomass of sorghum and maize. It allows for two harvests per year, with double yields and double harvests, and boasts advantages such as high yield, high quality, strong resistance, and a short growth cycle, resulting in significant economic benefits.

The invention discloses a wheat haploid accurate identification and breeding method based on molecular markers, and belongs to the technical field of agricultural biology. The method comprises the following steps: hybridizing a created multifunctional haploid induction line HIP-Plus serving as a male parent with a target female parent, and saving immature embryos to obtain seedlings; a three-stage identification system comprising visual preliminary screening, molecular confirmation and early selection is adopted to carry out accurate screening on seedlings: firstly, preliminary screening is carried out based on embryo color, then haploid confirmation and transgenic component detection are carried out by utilizing TaMTL allele specific markers, and finally genetic typing is carried out through important character function markers to screen out elite haploids; and carrying out chromosome doubling to obtain a double haploid homozygous line. According to the method, the haploid identification accuracy is improved, the breeding period is shortened, the early directional selection of the target agronomic traits in the haploid stage is realized, and the breeding efficiency and accuracy are greatly improved.

The invention relates to an improved embryo rescue technology for cucumber haploid breeding, and aims to solve the problem of low embryo rescue operation efficiency in a technology of inducing haploid by radiation pollen. By using a male parent material carrying a green fluorescent protein (GFP) marker, the haploid screening process in the seedling stage is simplified, and efficient and simple large-scale haploid cultivation is realized in combination with an optimized rapid embryo rescue method. The method comprises the specific steps of radiation pollen preparation, pollination and subsequent embryo rescue processes, particularly emphasizes the technical key points of disinfection, seed taking, cleaning and inoculated culture, and ensures that the operation speed and the success rate are improved while the pollution risk is strictly controlled. Compared with a traditional method, the method has the advantages that the obtaining efficiency of haploid cucumber plants is remarkably improved, the method is suitable for large-scale haploid breeding of cucurbitaceae crops, breeding of new cucumber varieties is accelerated, and agricultural yield increase and income increase of farmers are promoted.

The invention relates to a method for distant sexual hybridization of plants, which comprises the following steps: selecting a good variety of a sexual propagation plant type expected to be obtained as a hybridization female parent, castrating before male flowers are mature, treating female flowers before insemination and carrying out reproduction isolation, selecting sperms of any interspecific and farther organisms with characters of the variety expected to be obtained, and carrying out distant sexual hybridization on the selected sperms. And adding a distant hybridization agent, uniformly stirring to form a sperm, inseminating the female parent, taking back a fertilized embryo, carrying out embryo rescue, culturing into a seedling, carrying out chromosome doubling on the obtained seedling, and transplanting into soil. In the mature period, harvesting true hybrids according to individual plants, and cultivating F2 and later generations by using a pedigree method until homozygosis. The bottleneck of distant sexual hybridization is broken through, high-yield and high-quality varieties can be cultivated, and the method has wide application prospects.

The present invention discloses a method for rescuing peony distant hybrid embryos and forming seedlings, belonging to the technical field of plant breeding. The present invention provides a culture medium specifically for rescuing peony distant hybrid embryos and forming seedlings, the culture medium comprising a germination medium, an embryo callus induction medium, a differentiation medium, and a rooting medium; the present invention also provides a method for rescuing peony distant hybrid embryos and forming seedlings using the dedicated culture medium. The present invention overcomes the common problem of embryo abortion in the process of distant hybridization between tree peonies and peony by performing an embryo rescue operation in a timely manner after pollination. By optimizing seed disinfection, embryo separation, culture medium formulation, and culture conditions, the normal development of embryos can be effectively promoted. The staged induction and differentiation culture strategy ensures the formation of embryo callus and its effective transformation into adult plants, significantly improving the success rate of embryo rescue in the process of distant hybridization between tree peonies and peony, and providing technical support for the cultivation of new varieties of tree peonies for both oil and ornamental purposes.

The present invention discloses a method for identifying Chinese persimmon genotypes and its application, belonging to the technical field of genotype identification. The method for identifying Chinese persimmon genotypes provided by the present invention generates a fluorescent signal through a specific hydrolysis reaction between a probe and a target molecule, resulting in smaller errors, more accurate identification results, and higher result stability. Only two reference genomes are required, and identification is easy to perform, which can significantly reduce the workload of Chinese persimmon genotype identification, reduce the consumption of reagents and consumables, and lower the identification cost. The method can significantly shorten the time required for identification and improve identification efficiency. The identification of Chinese persimmon genotypes can provide a reference for the selection of hybrid parents and can also be used for embryo rescue of young plants, greenwood grafting, and marker-assisted selection of hybrid combinations, significantly improving PCNA breeding efficiency and having important significance for the breeding of new persimmon varieties with intellectual property rights and for the improvement of persimmon varieties.

The invention relates to the field of biotechnology breeding, in particular to a method for obtaining an interspecific filial generation of elephant trunk orchid and calanthe calyx orchid by using embryo rescue. According to the method, the calanthe ciliata is used as a male parent, the elephant trunk is used as a female parent, the embryo rescue technology is combined, in the embryo rescue technology, every 1 L of embryo rescue culture medium is composed of 2 g / L of Huabao No.1, 100 g / L of potato homogenate, 30 g / L of cane sugar, 0.5 g / L of activated carbon, 7.0 g / L of agar and the balance of water, and the pH is 5.6-5.8. According to the method, germination of the hybrid embryo of the calanthe ciliata and the elephant trunk can be remarkably promoted, new genetic characteristics are introduced potentially, and the hybrid embryo is obtained. The genetic diversity of the phalaenopsis variety is enhanced, and a new germplasm resource is provided for breeding of the phalaenopsis.

The invention provides an embryo rescue method for hybrid generations of chestnut plants and application of the embryo rescue method, and belongs to the technical field of plant breeding. According to the method, hybrid embryos obtained after hybridization pollination of the female parent Japanese chestnuts and the male parent Chinese chestnuts are subjected to in-vitro culture and rescue, the rescue success rate can be increased through the embryo rescue technology, and the abortion rate of interspecific filial generation embryos is effectively reduced. The invention further provides application of the embryo rescue method for the hybrid generation of the chestnut plants in hybrid breeding of the chestnut plants, a certain platform is provided for increasing germplasm resources of the chestnut plants, creating new varieties and transforming old varieties, and a certain theoretical basis is provided for screening the hybrid generation with excellent quality.

The invention discloses a method for obtaining Leymus chinensis and New Wheat Straw distant hybridization offspring using young embryo rescue technology, comprising the following steps: 1) hybridization: Leymus chinensis is as female parent, and New Wheat Straw is as male parent; Collect male parent pollen, pollen is pointed on female parent pistil stigma with tweezers for pollination, and immediately bag; 2) disinfection: take hybrid spike after pollination, strip out non-aborted ovary, use volume fraction 75% alcohol to soak 1min, sterile water rinse three times, sodium hypochlorite soak 10min, rear sterile water rinse 5 times; 3) use one-step seedling method or induced callus to expand and multiply, differentiate into seedling method seedling; 4) practice seedling transplanting: when tissue culture seedling grows to 10cm 15cm, ultrapure water is added in culture bottle, after indoor seedling hardening 7d, plant root culture medium is rinsed clean, transplanted into sterilized nutrient soil and cultivated. The present invention has higher young embryo germination rate, induced callus rate and differentiation rate, obtains the hybrid offspring of Leymus chinensis and New Wheat Straw in a relatively short time.

The invention relates to the technical field of plant sexual reproduction, in particular to a plant embryo in-vitro rescue method which comprises the following steps: selecting a plant female parent needing embryo rescue; preparing a treating fluid containing a plant hormone regulator; fixing a continuous release device on a branch below the silique or legume of the maternal plant; the treatment liquid is applied to the continuous release device, so that active ingredients in the treatment liquid are absorbed by the branches and transported to the siliques or pods. The externally applied plant hormone regulator can be upwards transported to ovules through the phloem and the xylem and directly acts on the hybrid embryo with development stagnation. Hormonal balance in the embryo is remodeled through cooperative regulation and control of hormones, and a stagnation signal is reversed into a development signal, so that the normal development process of the embryo is rescued and completed in situ without in-vitro operation, and the method is simple to operate, high in success rate, time-saving and labor-saving, and convenient to popularize and apply.

The invention relates to the field of plant conservation and breeding optimization, in particular to a hybrid embryo rescue method and a method for creating a large number of interspecific hybrid seedlings of Populus nigra by using the hybrid embryo rescue method. The method comprises the following steps: establishing an embryo rescue system, namely observing a separation layer between infructescence and branches after pollination for about 2 months, cutting part of infructescence to develop the embryo rescue system, washing capsules with running water for 120 minutes, lightly wiping peels with 75% alcohol for about 2 minutes, washing the capsules with sterile water for 5-6 times in a super clean bench, lightly absorbing moisture on the surfaces of the capsules with sterile filter paper, and taking out the capsules; the method comprises the following steps of: cutting a capsule by using a sterile scalpel, thoroughly removing floc, carefully clamping seeds by using sterile tweezers, putting the seeds on a 1 / 2MS culture medium containing auxin analogues for embryo rescue, and screening out an optimal culture medium which is 1 / 2MS + 0.06 mg / L naphthylacetic acid (NAA) + 0.08 mg / L indolebutyric acid (IBA) and has the pH value of 5.8-6.0 by adopting a culture dish germination method and taking 1 / 2 MS as the culture medium.

The application relates to a method for efficiently creating a far-cross hybrid between Chinese cabbage and the genus Eruca, which comprises the following steps: taking the selfing line 'ASTRO' of the genus Eruca as a male parent, wherein the selfing line 'ASTRO' has excellent properties such as drought and flood resistance, disease resistance, salt and alkali tolerance, and the like; taking an excellent Chinese cabbage material '72049' as a female parent; spraying magnesium chloride hexahydrate; and then performing intergeneric hybridization. After embryo rescue and colchicine doubling, a doubled material is obtained, and the doubled material is transplanted into a field. Through cytological identification, morphological observation and pollen fertility identification, the doubled material is finally determined as a true hybrid plant.

The invention discloses a distant hybridization cultivation method of a hexaploid common wheat disease-resistant and stress-resistant variety. The method comprises the following steps: parent screening and determination, cross pollination and fruiting promotion, hybrid immature embryo rescue and test-tube plantlet cultivation, molecular marker-assisted screening, and homozygous stability and comprehensive character identification. The invention relates to the technical field of agricultural crop breeding. The distant hybridization cultivation method of the hexaploid common wheat disease-resistant and stress-resistant variety reaches the parent screening standard, hybridization and embryo rescue parameters, a molecular marker screening system and the like are clear and controllable, and the distant hybridization cultivation method can be popularized to other wheat distant hybridization breeding scenes; a standardized technical normal form is provided for improving wheat characters by using excellent genes of related species such as rye, and the genetic basis and the resource utilization range of wheat breeding are widened.

This invention discloses a method for aseptic rapid propagation of *Cephalotaxus fortunei* based on embryo rescue and its application, belonging to the field of plant tissue culture technology. The method includes the following steps: sterilizing the embryos of *Cephalotaxus fortunei* to obtain aseptic embryos; inoculating the aseptic embryos into a germination medium for germination culture to obtain aseptic seedlings; inoculating the cotyledons and hypocotyl regions of the aseptic seedlings into a callus induction medium to induce callus formation; transferring the callus to an adventitious bud induction medium to induce the differentiation of adventitious buds; inoculating the adventitious buds or stem segments obtained from them into an adventitious bud proliferation medium for subculture; inoculating the proliferated adventitious bud stem segments into a rooting medium to induce rooting, obtaining fully rooted seedlings; and finally, acclimatizing and transplanting the fully rooted seedlings. This invention uses embryos as starting material, which can improve the initial success rate of aseptic culture and broaden the genetic background of regenerated plants.

The application discloses a method for improving the transformation efficiency of grape embryo rescue breeding abnormal seedlings, and comprises the following steps: inoculating the embryo rescue abnormal seedlings in a transformation culture medium to culture, transferring the small seedlings with grown true leaves to a seedling culture medium to culture, and obtaining normal seedlings; for the abnormal seedlings which are not transformed into the normal seedlings, performing the rescue through an adventitious bud regeneration approach again; cutting the abnormal seedlings to obtain hypocotyls, inoculating the hypocotyls in an induction culture medium to perform the adventitious bud regeneration culture, and transferring the explants differentiated with the adventitious buds to the seedling culture medium to obtain the normal seedlings. The application provides a technical scheme for effectively transforming the abnormal seedlings generated in the process of the seed rescue breeding of the seedless grape into the normal seedlings, the abnormal seedlings generated in the process of the seed rescue of the seedless grape are transformed and reused by using two rescue methods, and then the seed rescue breeding efficiency of the seedless grape is improved, the seed rescue efficiency of the seedless grape can be improved from about 4-10% to about 25%.

The application relates to a method for identifying a heterologous tetraploid of Brassica rapa and Eruca sativa intergeneric hybridization based on a molecular marker technology and a primer pair. The method comprises the following steps: obtaining a sample to be identified, wherein the sample is obtained from embryo rescue and chromosome doubling after intergeneric hybridization with Brassica rapa '72049' as a female parent and Eruca sativa 'ASTRO' as a male parent; taking genomic DNA of the material as a template, and performing amplification by using specific primers; performing gel electrophoresis on the amplified product; and observing an electrophoresis strip to determine the authenticity of the heterologous tetraploid.

The application relates to a method for identifying Brassica rapa interspecific hybrid allopolyploids based on InDel technology and a primer pair. The method comprises the following steps: obtaining a sample to be identified, wherein the sample is obtained by performing interspecific hybridization with Brassica rapa '72049' as a female parent and Eruca vesicaria 'ASTRO' as a male parent, and then performing embryo rescue and chromosome doubling; taking genomic DNA of the material as a template, and performing amplification by using an InDel primer pair; performing gel electrophoresis on the amplified product; and observing an electrophoresis strip to determine the authenticity of the allopolyploid.

The invention belongs to the technical field of plant breeding, and particularly discloses a method for preparing watermelon hybrid seeds by bee pollination, which is characterized in that a watermelon dominant sterile female parent line is created based on a watermelon dominant male sterile material, and haploids can be quickly, accurately and efficiently obtained by using a gene editing or immature embryo rescue culture technology. According to the watermelon hybrid seed production method, dominant male sterile double haploids are obtained through doubling, the dominant male sterile double haploids are hybridized with fertile female parents to efficiently obtain batches of completely sterile female parent seeds, and then bee pollination replaces artificial pollination to produce watermelon hybrid seeds, so that the labor cost of watermelon seed production can be reduced by 44%, and the yield of watermelon hybrid seeds is improved. And the purity of the hybrid seeds is improved to 100%. The problem of insufficient labor force can be effectively solved, the production cost of hybrid seeds is reduced, and the method has high application value in large-scale seed production of watermelons.

The invention discloses a method for creating a novel chrysanthemum germplasm with high linarin content based on distant hybridization between opisthopapus and chrysanthemum, which comprises the following steps: inducing ovules after pollination for 13-18 days to generate calluses and obtain distant filial generation plants by combining artificial cross breeding with an immature embryo rescue technology, and carrying out morphological and cytological identification to obtain the novel chrysanthemum germplasm with high linarin content. And obtaining a true hybrid filial generation. The content of linarin in inflorescences and leaves of true hybrid filial generations is obviously increased and is obviously higher than that of the female parent Jinlingzirobe. The invention provides a new way for efficient utilization of opisthopapus and chrysanthemum germplasm resources, and has significant breeding and application values.

The invention discloses a lisinaceae fruit tree distant hybridization immature embryo rescue method. The lisinaceae fruit tree distant hybridization immature embryo rescue method comprises the following steps: step (1), determining a key period of lisinaceae fruit tree distant hybridization immature embryo rescue; step (2), collecting hybrid young fruits according to the key period of rescuing distant hybrid young embryos of the Limiaceae fruit trees, disinfecting the hybrid young fruits, and taking out hybrid seeds with endopleura from the hybrid young fruits; cutting out a wound on the endopleura of the hybrid seed but not completely cutting the seed, and taking the treated hybrid seed as an explant; (3) inoculating an embryo rescue culture medium with the explant, and carrying out dark culture until buds grow out; step (4), transferring the buds to a proliferation culture medium for light culture until young branches grow; and (5) transferring the young branches to a rooting culture medium for light culture until rooting, and transplanting rooted seedlings for seedling hardening. The method effectively solves the problem of low crossbreeding efficiency caused by abortion of the hybrid immature embryos in distant hybridization of the Liapaceae fruit trees.

The invention discloses a method for realizing plant regeneration through hybrid embryo rescue of cymbidium faberi, and belongs to the field of reproductive biology. The cymbidium faberi with novel floral leaf characters is high in price due to extremely high ornamental value, but the varieties and the quantity of the cymbidium faberi with the novel floral leaf characters are rare due to dependence on natural variation and adoption of a division propagation mode, and the collection pressure of the wild cymbidium faberi is caused. According to a hybrid in-vitro embryo rescue technology, parents with novel characters can be combined, regenerated plants of hybrid embryos are obtained through the embryo rescue technology, selectable hybrid progenies with novel phenotypes are provided, and the breeding period is shortened in a nutritional line breeding mode.

The invention relates to the technical field of plant biology, in particular to a dianthus chinensis crossbreeding method which comprises the following steps: S1, parent plant screening and flowering phase adjustment; s2, artificial castration and moisturizing directional pollination; s3, performing ovary low-temperature pretreatment and immature embryo in-vitro stripping sterile inoculation; s4, carrying out induction culture and grading screening on calluses; s5, subculture and adventitious bud differentiation; s6, rooting culture and test-tube plantlet molecular marker preliminary screening; and S7, carrying out gradient acclimatization and transplantation and final screening of field characters. Through the synergistic effect of ovary low-temperature pretreatment, immature embryo oxidation resistance and dynamic embryo rescue opportunity, the success rate of interspecific distant hybridization and intraspecific different ploidy hybridization is increased, reproduction isolation of dianthus dianthus distant and ploidy hybridization is successfully broken through, filial generations which cannot be obtained through traditional breeding are obtained, and the breeding efficiency is improved. The polymerization of excellent characters of different species and different ploidy dianthus chinensis is realized, and the breeding gene resources of the dianthus chinensis are enriched.

The present application relates to the field of plant biotechnology, in particular to a method for rescuing seedlings from Malus robusta and Malus baccata far-off hybridization zygotes and application, and the method is summarized as follows: a method for rescuing seedlings from Malus robusta and Malus baccata far-off hybridization zygotes is provided.The present application carries out interspecific far-off hybridization by taking Malus robusta as female parent and Malus baccata as male parent.According to the degeneration of far-off hybridization zygotes, the zygotes are rescued.The present application determines the suitable sampling period for rescuing zygotes of hybrid combinations, the concentration and ratio of exogenous hormones required for zygote germination, proliferation and expansion, rooting culture, and the suitable hardening and transplanting method.The zygote rescue system for interspecific hybridization of Malus robusta and Malus baccata established by the present application has a zygote germination rate of 74.3%, a proliferation coefficient of 5.1, a rooting rate of 95.5% in rooting culture, and a hardening and transplanting survival rate of 96.4%.

This invention discloses a method for inducing haploids in Camellia oleifera through radiation pollen pollination, belonging to the field of plant tissue culture. The method includes the following steps: irradiating Camellia oleifera pollen; determining the viability and germination rate of the irradiated pollen; performing hybridization pollination using the irradiated pollen; selecting the pollinated embryos and inoculating them into a culture medium for embryo rescue; and screening the plants obtained from the embryo rescue for ploidy identification to identify haploids. This invention creates a new approach for cultivating haploids in Camellia oleifera, applicable to both the Camellia oleifera and Camellia brevicornuate groups, and can be applied to the breeding of Camellia oleifera varieties, accelerating the Camellia oleifera breeding process.

The present application relates to the field of biotechnology breeding, and particularly relates to a method for obtaining interspecific hybrid offspring of Rhynchostylis retusa and Vanda teres by embryo rescue. The present application takes Vanda teres as the father and Rhynchostylis retusa as the mother, and combines with the embryo rescue technology. In the embryo rescue technology, the composition of the embryo rescue medium is as follows: 2g / L of Hua Bao No.1, 100g / L of potato homogenate, 30g / L of sucrose, 0.5g / L of activated carbon, 7.0g / L of agar, and the rest is water, and pH is 5.6-5.8. The method of the present application can significantly promote the germination of the hybrid embryo of Rhynchostylis retusa and Vanda teres, potentially introduce new genetic traits, enhance the genetic diversity of Phalaenopsis varieties, and provide a new germplasm resource for Phalaenopsis breeding.