69results about How to "Rich germplasm resources" patented technology

The invention belongs to a plant breeding method and in particular relates to a breeding method for a high-yield and high-resistance green Chinese onion variety applicable to multi-season cultivation. The breeding method comprises the following processing steps of: breeding crossing parents, carrying out a matching breeding test for cross combination, propagating the parents and propagating a generation F1. According to the breeding method provided by the invention, the problems that varieties are complex, characters of a strain degenerate, individual uniformity is poor, resistance is low and all-year production can not be realized, and the like in the prior art can be solved; the bred green Chinese onion hybrid is high in yielding capability, high in lodging resistance and flooding tolerance and applicable to multi-season cultivation, and disease resistance is obviously enhanced; and application amount of pesticide is reduced, production cost is reduced, land cropping index is increased, environmental protection is facilitated, pollution is reduced, vegetable basket safety is guaranteed, and income is increased.

The invention discloses a method for preparing a cabbage type rape BnPABP3 promoter (PBnPABP3) and the application thereof. The method comprises the following steps of: cloning PBnPABP3 sequence by utilizing the genome walking method, extracting genome DNA by utilizing the SDS cracking method, carrying out enzyme digestion on DNA by respectively using endonuclease DraI, EcoRV, PvuII and StuI in different systems, carrying out the first round of PCR amplification by adopting the product as a template after joint segments are connected with the product, and carrying out the second round of PCR amplification by adopting the 50 times diluent of PCR products obtained in the first round as a template so as to obtain the PBnPABP3. The PBnPABP3 is applied to anther and pollen grains of plant. The promoter has the function of driving exogenous gene expression, the expression parts of the promoter are in the anther and pollen grains of plant, and the promoter has the application potentiality in the aspects of improving the safety of rape edible oil as well as the quality of crops, artificially creating sterile line and restorer line, enriching germ plasm resources, and the like.

The invention relates to a successful cutting propagation method of desert caragana, which comprises the following steps of: selecting a strongly growing bush as a maternal plant, and respectively scissoring a twig, a hard branch and a taproot of the desert caragana for cutting; putting the twig of the desert caragana into 100ppm of growth regulator solution, wherein the soaking depth is 2-3 centimeters, and the soaking time is 6.5-10 hours; putting the hard branch into 500ppm of growth regulator solution, wherein the soaking depth is 2-3 centimeters, and the soaking time is 1-3 hours; soaking the cutting of the taproot in 50ppm of growth regulator solution for 18-20 hours; and selecting a position with flat land and good drainage for arranging a cutting bed, selecting sandy soil as a cutting ground mass, cutting at the depth of 4-5 centimeters, watering after cutting, setting up a shading net in time, watering once every five days, tearing off the shading net after one month, and counting the survival rate after three months. An experiment result indicates that the processing method can ensure the success of the cutting propagation of the desert caragana, and provide abundant germ plasm resources for ecological restoration and stockbreeding production in desert regions.

The invention discloses a brassica napus BnPABP8 promoter and a preparation method and use thereof. The preparation method comprises the following: A, a step of primer design, which is to design specific walking primers according to known sequences; B, a step of brassica napus promoter preparation, which is to extract brassica napus DNA by using a sodium dodecyl sulfonate (SDS) lysis method, perform genome walking according to genome walking kits, digest 5 micro liters of DNA by using DraI enzyme, detect the purity of the DNA, perform enzyme digestion of the DNA by using the incision enzymes provided by four kits, and purify the product of the enzyme digestion; C, a step of purification, which is to add ethanol into enzyme digestion solution, precipitate, centrifuge, dry in the air, dissolve in sterile water and add specific connectors after purification is accomplished, wherein GSP1 and AP1 are used as primers in primary polymerase chain reaction (PCR) reaction; and D, a step, which is to perform amplification by using the diluted solution of the product of the primary PCR reaction as a template and GSP2 and AP2 as primers in a secondary PCR reaction, recover purified product of the secondary PCR product, connect with a T vector, and obtain the PBnPABP8. The invention also discloses the use of the PBnPABP8 in roots, stems, leaves, buds and anthers of plants. When the promoter is used, the safety of the edible brassica napus oil can be improved, and the disease resistance, stress resistance and lodging resistance in crops can be improved by transgenosis.

The invention discloses a breeding method of sterile and maintainer lines of north glutinous sorghum. The breeding method comprises the following specific steps: 1, hybridizing by taking non-glutinous sorghum LgB/R5M874B of a maintainer line as a female parent and a glutinous sorghum maintainer line 45B as a male parent, and inbreeding generations by selecting single plants with three good characters of glutinousness, large spikes and early mature; 2, hybridizing by taking a non-glutinous sorghum sterile line A2V4A as a female parent and an F2 generation in the step one as a male parent, carrying out four-generation backcross transformation by adopting a method of carrying out stable hybridization together with backcross transformation and taking good single plants of a filial generation as a male parent, and breeding a glutinous sorghum sterile line by taking good sterility, glutinous grains and heat smut resistance as breeding standards, wherein in glutinousness identification in the step one or step two, only endosperm is cut for carrying out iodine solution reaction, so that normal sprouting of the glutinous grains is ensured, and embryo is prevented from being damaged due to length cutting of the grains. According to the breeding method disclosed by the invention, good characters of yield, quality, resistance, combining ability, appearance and the like of south and north sorghum are organically integrated, so that the sterile and maintainer lines can become basic parent lines of the north glutinous sorghum, and the germplasm resource of the glutinous sorghum is enriched.

The invention discloses a preparation method and an application of a brassica napus BnPABP5-3 promoter (PBnPABP5-3). A genome walking method is utilized to clone a PBnPABP5-3 sequence: an SDS (sodium dodecyl sulphate) cracking process is applied to extract genome DNA; in different systems, endonucleases DraI, EcoRV, PvuII and StuI are used to digest DNA, and the digested DNA is respectively connected with joint segments, then primary PCR (Polymerase Chain Reaction) amplification is carried out by taking the DNA connected with the joint segments as templates, and secondary PCR amplification is carried out by taking 50-time diluted solution of products of the primary PCR as a template, thus the PBnPABP5-3 is obtained. The PBnPABP5-3 can be applied to the anther, pollen grain, root tip and ovule of a plant. The promoter has the function of driving expression of a foreign gene, the expression part is in the anther, pollen grain, root tip and ovule of the plant, and the promoter has application potential in the improvement of the safety of rape edible oil, the improvement of the stress resistance and lodging resistance, improvement of the quality of crops, artificial sterile line creation, germ plasma resource enrichment and the like.

The invention discloses a method for preparing a promoter (PBnPABP2) of Brassica napus BnPABP2 and the application thereof. The operation of cloning a PBnPABP2 sequence by using a genome walking method comprises the following steps: extracting genome DNA by using a SDS cracking process; in different systems, carrying out restriction endonuclease reaction on the extracted DNA respectively by using endonucleases DraI, EcoRV, PvuII and StuI; after the obtained DNA is connected with a linker fragment, carrying out first-round PCR amplification by taking the linker fragment as a template; and carrying out second-round PCR amplification by taking a50-time diluent of the obtained first-round PCR product as a template, thereby obtaining the PBnPABP2. The PBnPABP2 is applied in the roots, stems, leaves, flower buds, anthers and peels of plants. The promoter has the function of driving the expression of exogenous genes, and the expression sites thereof are respectively in the roots, stems, leaves, flower buds, anthers and peels of plants; and the promoter has application potentials in the aspect of safety of edible oil of Brassica napus and the aspects of improving the disease resistance, stress resistance and lodging resistance of crops, improving the quality of crops, artificially creating a male sterile line and a restoring line, enriching germ plasm resources, and the like through transgenosis.

The invention discloses a sanqingshan cold-resistant dendrobium officinale seedling culturing method. The culturing method sequentially comprises the following steps: hybridizing and breeding dendrobium officinale, improving a dendrobium officinale population, breeding high-quality dendrobium officinale variety, and carrying out expanding propagation on high-quality dendrobium officinale seedlings. The group improvement is combined with breeding of high-quality individual plant, the breeding of a high-quality new variety strain and fast propagation of high-quality seedlings by applying plant tissue cell culture technology through adopting the progeny obtained from hybridization of sanqingshan wild dendrobium officinale and dendrobium huoshanense, varieties can be screened out according to cold resistance and content of active ingredients by utilizing low-temperature induction, and the breeding speed of the new strain can be greatly accelerated; and the breeding method has importance values on promoting economic development by forming high-quality dendrobium officinale variety and expanding the resources of the dendrobium officinale seeds.

The invention discloses a brassica napus BnCP51 promoter as well as a preparation method and an application of the promoter. The preparation method comprises the following steps of: performing polymerase chain reaction (PCR) on the basis of a primer designed according to a BnCP51 gene upstream 2kb sequence obtained from the all genomic sequencing of brassica napus, thus obtaining the sequence of a 5' upstream promoter of the brassica napus BnCP51; and designing a primer according to the 5' upstream 2kb sequence of the BnCP51 obtained from the all genomic sequencing of the brassica napus, extracting genomic DNA of brassica napus leaves, performing PCR amplification on the genomic DNA, connecting the genomic DNA with a pMD18-T carrier after performing purification and recovery, transformingcompetent cells, picking positive clone, and sequencing after PCR positive detection and digestion verification, thus obtaining the upstream 2kb flanking sequence of target genes, namely PBnCP51. Dueto the efficient expression of the PBnCP51 in rape anther and pollen grains, the expression efficiency of foreign genes in transgenic plants is improved, thus the PBnCP51 can be used in the gene engineering study of the plants and in the transgenic safety study of seed rape.

The invention discloses a method for performing sterilization by using a mixed antibacterial agent in the cultivation process of chlamydomonas reinhardtii and belongs to a method for removing bacteria and fungi from chlamydomonas reinhardtii. The method comprises the following steps of: A. inoculating mixed infectious microbes polluted chlamydomonas on a TAP solid medium plate and cultivating for 3 to 5 days under the conditions of 23 +/- 0.5 DEG C, 14/10 hour light/dark period and light intensity of 8000 Lx; B. preparing a mixed antibacterial agent of azoxystrobin of 10 mu g/ml and nalidixic acid of 15 mu g/ml for later use on the TAP solid medium plate; C. transferring and lining the cultivated infectious microbe-containing chlamydomonas to the TAP plate containing the mixed antibacterial agent and cultivating for 5 to 8 days under the conditions of 23 +/- 0.5 DEG C, 14/10 hour light/dark period and light intensity of 8000 Lx; and D. transferring and lining the microbe-removed chlamydomonas to the common TAP solid medium plate again, cultivating for 3 to 5 days under the conditions of 23 +/- 0.5 DEG C, 14/10 hour light/dark period and light intensity of 8000 Lx, and performing subsequent experiments or performing conservation for standby application. The method has the advantage that fungi are removed by azoxystrobin and bacteria are removed by nalidixic acid, so that the problem about infectious microbe pollution in scientific experiments and application process of the chlamydomonas reinhardtii is solved.

The invention discloses an inducing method of gracilaria verrucosa callus. The method is characterized by comprising the steps of inoculating gracilaria verrucosa thallus which is adopted as an explant and sterilized by a detergent and mercury bichloride in an inducing culture medium, and adding 2mg of 2,4-D, 1mg of IAA, 1mg of KT and 30g of cane sugar in a 1L PESI culture medium; inducing the explant for 7d in darkness, and continuously culturing the explant under the illumination condition to form a great amount of callus. By adopting the method, the callus can be successfully induced by adopting the gracilaria verrucosa as the explant, the induction rate of the callus can reach 82 percent, the callus can further develop to form bud or thallus of the gracilaria verrucosa, and not only can a great amount of gracilaria verrucosa thallus be formed, but also rich seed resource can be provided for the gracilaria verrucosa.

The invention discloses a method for breeding high-yield high-quality stress-resistant good new common andrographis herb varieties. The method includes the steps: taking common andrographis herb seedsas raw materials, taking 60 Co-gamma rays as radiation sources, radiating the common andrographis herb seeds, dividing the radiated seeds into three groups, soaking the seeds by distilled water, germinating the seeds in the salt stress environments containing NaCl solution and the drought stress environments containing PEG 6000 solution, and calculating germination rate, germination potential, germination indexes, vigor indexes, root length and bud length or fresh weight; selecting new salt-tolerant and drought-resistant high-yield high-quality stress-resistant varieties according to germination rate, germination potential, germination indexes, vigor indexes, root length and bud length or fresh weight of each sample. According to the method, the common andrographis herb seeds are radiatedby the aid of suitable dosages of 60 Co-gamma rays, good tolerance improvement methods of common andrographis herbs under salt stress and drought stress are expected to be achieved in practice, and application potential is high. According to the method, injury of adverse situations to plants is relieved, and the method has high application values.

The invention discloses an induction method of gulfweed regeneration plant. The induction method of the gulfweed regeneration plant comprises the following steps: preparing a sterile explant, inoculating the explant to an induction culture medium and inducing callus to form at an induction temperature of 15-20 DEG C with a light intensity of 4000-5000Lx and a light period of 8 hours; processing gulfweed rhizoid in 0.2% povidone for 3 minutes, and then sterilizing in a 3% sodium hypochlorite solution for 5 minutes, sterilizing in 0.1% mercury bichloride for 3 minutes so as to obtain a sterile survival explant; adding 0.1-1mg of ZT, 1-2mg of IAA (Indole Acetic Acid), 1-3g of ascorbic acid, 30g of cane sugar and 7g of agar in a 1LPES culture medium, sterilizing at a high temperature and a high pressure for 10-20 minutes to bud the gulfweed.

The invention discloses a BnALS1 mutant gene and protein based on gene editing and application of the BnALS1 mutant gene and protein. The invention also discloses a cytosine base editing vector. The invention reports that single base editing is realized in rape for the first time. A base editing method provided by the invention can directionally improve rape traits. Compared with chemical mutagenesis, the method is faster, more efficient and more accurate. Meanwhile, germplasm resources of the rape resisting herbicides are enriched. Mutant materials R10 and R144 obtained in the invention havestrong tribenuron-methyl herbicide resistance. 20 days after 20 ml of 15 mg ai/L tribenuron-methyl is sprayed to each plant in the 5-6 leaf stage (approximately equivalent to 5 times of a recommendedamount for broadleaf weed prevention and control), the R10 and R144 materials are not affected at all, and transgenic receptor materials of the rape suffer from severe phytotoxicity (new leaves wither, and old leaves stop growing and turn purple).

The invention discloses a Chinese herbal medicine plant cultivation system and device with a hyperlipidemia prevention and treatment effect. The aim is to provide the indoor plant cultivation system for assisting growth of Chinese herbal medicine plants through microorganisms (beneficial flora) and a Chinese herbal medicine plant cultivation device of the system. According to the system, the compatibility of the beneficial flora and Chinese herbal medicine is achieved through special matching, the effect of effectively preventing and treating hyperlipidemia can be achieved, symbiosis of the beneficial flora is achieved, and the promotion effect on growth of the Chinese herbal medicine plants can also be achieved. Meanwhile, the established system can also purify indoor air and beatify an indoor environment, the indoor vertical space is sufficiently used, the cost is low, and the beneficial effect is significant.

The invention relates to a method for obtaining tartary buckwheat seeds easy to shell and belongs to the technical field of genetic seed breeding of crops. According to the provided method, a chemicalmutagenesis method is adopted to obtain the high-quality tartary buckwheat seeds. The adopted technical scheme includes the steps that a, ethylmethane sulfonate is utilized to process 5,000 ethylmethane sulfonate (Heifeng No.1) seeds, and the seeds are soaked in an EMS solution for 8-10 hours and stirred several times during soaking; then, the solution is poured out carefully, sodium thiosulfateis adopted for neutralizing the EMS solution, distilled water is adopted for flushing several times, and the seeds are placed in a fuming cupboard for airing for preparation of sowing; b, the processed seeds are sown in land which is ploughed previously, wherein if the statistic fatality rate of the sown seeds reaches 30-50%, mutagenesis is effective; c, M1-generation mutants are obtained from each single plant in order to harvest potential available mutants; d, after screening and seed baking, it is found out that shells of seeds of three single plants are broken, after continuous planting ofM2-M5 generations, three plant systems with stable gene expression are obtained, and more than 100 single plants are obtained. The method has significant meaning on diversification of tartary buckwheat seed resources, shortening of a traditional seed breeding process and boosting of tartary buckwheat seed breeding work.