72 results about "Coleoptile" patented technology

Plant cell expansion is regulated by wall relaxation and yielding, which is thought to be catalyzed by elusive "wall loosening" enzymes. By employing a reconstitution approach, we initially found that a crude protein extract from the cell walls of growing cucumber seedlings possessed the ability to induce the extension of isolated cell walls. This activity was restricted to the growing region of the stem and could induce the extension of isolated cell walls from various dicots and monocots, but was less effective on grass coleoptile walls. Sequential HPLC fractionation of the active wall extract revealed two proteins with molecular masses of 29 and 30 kD, as measured by SDS-PAGE, associated with such activity. Each protein, by itself, could induce wall extension without detectable hydrolytic breakdown of the wall. We proposed the name "expansins" for this class of proteins. Expansins have been isolated from various plant sources including oat, cucumber, broccoli, celery, tomato, cotton, cabbage, and corn, and also from snail and its feces. These proteins weaken the intermolecular bonds between plant wall polysaccharides. They decrease the mechanical strength of commercial products made from polysaccharides, such as paper, and therefore present a novel approach in developing new technologies in industries which make use of such polysaccharides, such as in the paper industry, in the applications of polysaccharide gums and related products. These proteins moreover present a novel approach in the control of plant growth.

The invention discloses a novel wheat gene TaMYB7D capable of adjusting and controlling the synthesis and the metabolism of anthocyanin, and belongs to the field of plant genetic engineering, wherein the novel wheat gene TaMYB7D is obtained through separation of the purple-grain wheat variety highland 115. The TaMYB7D is located on a wheat chromosome 7D, and a protein product subcell is located in a cell nucleus. A deduced amino acid sequence shows that TaMYB3 encodes an MYB transcription factor capable of adjusting and controlling the synthesis and the metabolism of anthocyanin. The expression quantity of the TaMYB7D in the coleoptiles of a purple wheat variety highland 115 grain can be increased under the induction of light. During transient expression, the TaMYB7D and the pericarpic cell of the highland 115 grain synthetize anthocyanin. Moreover, coseparation occurs between the color of the coleoptiles of the highland 115 grain and the expression of the TaMYB-7D, that is, the TaMYB7D has transcriptional activity used for adjusting and controlling the synthesis and the metabolism of anthocyanin, and is the major gene of the purple coleoptiles of the TaMYB7D highland 115 grain.

The invention provides a method for doubling cone haploids. The method comprises the following steps: 1, cutting off the tips of coleoptiles of buds of corn haploids; and 2, soaking embryos of the buds of the corn haploids without the tips of coleoptiles in mixed liquid of colchicines and dimethyl sulfoxide, so as to obtain corn buts with doubled chromosome. The method for doubling cone haploids, which is provided by the invention, can remarkably improve the survival rate and maturing rate of the corn haploids, wherein the survival rate is higher than 70%, which is far higher than the survival rate of a control group which is generally lower than 50%, the maturing rate is higher than 10%, which is far higher than the maturing rate of the control group which is generally about 5%; moreover, the use amount of colchicines is 5-6 times. When applied to a haploid breeding system, the method can effectively promote the reform of the corn breeding technology to realize large-scale and engineering haploid breeding.

The invention relates to a monocotyledon transgenic method for invading growing points of seed buds minimally and fully. According to the technical key point, the monocotyledon transgenic method comprises the following steps of: germinating seeds for 1 to 2 days, and when buds extend to be 0.2 to 2 centimeters long, removing coleoptile to expose the growing points; pricking and brushing the growing points by using a minimally-invasive brush of which the diameter of a single brush hair is 4 to 20 micrometers, the exposed length is 0.5 to 3 millimeters and the number of the brush hairs is 100 to 5,000 and which is dipped with agrobacterium mediated conversion liquid to perform full minimally-invasive conversion; after performing co-culture, developing further to form seedlings, promoting the development of spikes and grains, and harvesting in a plant division mode; and performing identification on T1 generation. The monocotyledon transgenic method has the advantages that tissue culture, the limit of genotypes and resistance screening are avoided, the method is easy and convenient to operate and easy to realize in large scale and is suitable for all monocotyledons which bear seeds. By the monocotyledon transgenic method, the transformation effects that the genetic transformation rate of wheat is 49 percent, the genetic transformation rate of paddy is 66.3 percent and the genetic transformation rate of corn is 100 percent are achieved.

A method for artificially transplanting endophytic fungi from the natural grass to the lawn grass comprises the steps as follows: inoculating the seeds which are determined to be aseptic onto the surface of a water-agar culture medium under the aseptic condition, and slightly pressing; sealing the culture dish, placing the culture dish into a constant-temperature incubator, and carrying out the dark culture to obtain aseptic coleoptile; longitudinally cutting a small opening at the upper part of the coleoptile with a depth that the coleoptiles is not penetrated, injecting mycelia into the opening, inoculating, placing into a constant-temperature incubator and carrying out the dark culture for 5 days; culturing in the dark and in the light alternately for 4 days, transplanting into the aseptic vermiculite, sealing the flower pot opening by fresh-keeping film to increase the humidity of ambient air for one week; removing the fresh keeping film, culturing at the room temperature for one month, and detecting the infection rate. The method has a homologous transplanting success rate of 93% and a heterogenous transplanting success rate of 76%, has the advantages of rapidness, convenience, low cost, high success rate, etc., and overcomes the defect of low success rate in the prior art.

The invention relates to a rice seedling promoter for dry direct seeding and an application method, in particular to a seedling promoter which can improve the sprouting up capacity of rice coleoptile, increase the emergence rate and reduce pest and disease damage in a rice seedling stage under a dry direct seeding condition of rice as well as an application method, and solves the problems of low sprouting up capacity of the rice coleoptile, low field emergence rate, poor seedling quality, serious pest and disease damage threat and the like for dry direct seeding of the rice. Links including trace fertilizer application, acid regulation, pest and disease damage control and the like are finished through one-step seed dressing before seeding, the labor intensity is reduced, and the production efficiency is improved. The rice seedling promoter for dry direct seeding comprises micronutrients, an acid regulator, a chelating agent, an insecticide and a fungicide. The application method comprises the following steps: 1, 30 ml of the rice seedling promoter for dry direct seeding is weighed and added with 30 ml of clear water, and the materials are mixed uniformly; 2, 15 kg of rice seeds are weighed and mixed and stirred uniformly with a rice seedling promoter solution, and a mixture is air-dried; 3, dry direct seeding of the treated rice seeds is performed. The rice seedling promoter for dry direct seeding has few residues and long efficacy, and a culture effect on strong rice seedlings is good.

The invention relates to a pesticide preparation and a preparation method thereof, in particular to a trifluralin emulsion in water and a preparation method thereof. The trifluralin emulsion in water comprises the following components in part by mass: 0.5 to 48 parts of trifluralin, 1 to 20 parts of solvent, 1 to 16 parts of emulsifier, 0.1 to 5 parts of thickening agent, 0 to 8 parts of antifreezing agent, 0.1 to 1.6 parts of defoaming agent, 0 to 3 parts of antimicrobial agent, a proper amount of pH value regulator, and the balance of deionized water. The trifluralin emulsion in water is a selective pre-emergence soil treatment agent, and is mainly absorbed through coleoptiles and hypocotyls of weeds. The trifluralin emulsion in water has the effect on gramineous and partial small-seed broadleaf weeds, and is applicable to cotton, soybeans, rape, peanuts, potatoes and the like. The trifluralin emulsion in water does not contain or rarely contains an organic solvent, so the manufacturing cost and use cost are reduced, and the toxicity produced to operators, storers and transporters, and users is reduced in the process of production, storage and transportation; and the trifluralin emulsion in water is a product which is environmental-friendly and has high safety.

The invention discloses a fast and efficient agrobacterium-mediated rice stem tip genetic transformation method. The method comprises the following major steps: directly soaking mature rice seeds in tap water, accelerating germination, then transversely cutting the coleoptiles to expose stem tips as transformation receptors, performing genetic transformation by virtue of agrobacterium mediation without sterilization treatment, putting the transformed materials under an appropriate environment to allow plants to grow, and carrying out transgenic screening and identification to obtain the transgenic rice plants. According to the fast and efficient agrobacterium-mediated rice stem tip genetic transformation method, callus induction and a series of subsequent tissue culture processes are avoided, the defects of materials affected by bacteria, difficult regeneration, vitrification, browning, low survival rate of transplanted tissue cultured seedlings and the like in the tissue culture process are avoided, the rice genetic transformation cost is reduced, the restriction of the rice genotype to the transformation efficiency is effectively overcome, and a large quantity of transgenic rice plants can be obtained in short time; the fast and efficient agrobacterium-mediated rice stem tip genetic transformation method has a significant value in good-quality rice culture and rice genetic transformation research.

The invention discloses SNP (single nucleotide polymorphism) molecular markers related to salt tolerance and hypoxia tolerance of rice and application of the SNP molecular markers. The SNP molecular markers provided by the invention are located at the 26th, 874th and 154th sites of rice chromosome 1, and the polymorphism is A or C. The SNP molecular markers are significantly correlated with the length of coleoptiles in rice under hypoxia and high salt germination conditions. The relative length of coleoptiles in a rice variety with a genotype being AA at the sites is significantly greater thanthat of a rice variety with the genotype being CC under hypoxia and high salt germination conditions. The invention also designs PCR primers based on the SNP sites, and the nucleotide sequences of the primers are shown in sequence 1 and sequence 2. The SNP molecular markers provided by the invention are verified in natural populations that the markers can effectively predict the hypoxia and highsalt germination of rice, and thus can be used for marker-assisted breeding of salt tolerance, hypoxia tolerance and direct seeding molecular breeding in rice.

The invention discloses a gene OsBEAR1 for adjusting and controlling growth and development of coleoptiles of oryza sativa, and an application of the gene OsBEAR1. The MSU ID of the gene is ''LOC_Os11g15210'', and the gene is used for coding an amino acid sequence as shown in SEQ ID NO:3. By a gene editing method, the gene mutant is knocked out in Oryza sativa L. cv. Nipponbare, growth and development of the coleoptiles of oryza sativa seedlings after OsBEAR1 knockout are accelerated, and the length of the coleoptiles of the oryza sativa seedlings after OsBEAR1 knockout is notably longer thanthat of coleoptiles of receptor oryza sativa seedlings. The elongation of the coleoptiles facilitates sprouting of oryza sativa and direct sowing in the fields. According to the gene OsBEAR1 disclosedby the invention, a base is provided for reformation and breeding of the varieties of oryza sativa suitable for direct sowing in the fields.

The invention discloses a method for identifying salt tolerance of different wheat varieties in a germination period and belongs to the field of application of agricultural technologies. According tothe method for identifying salt tolerance of different wheat varieties in the germination period, the germination rate, germination potential, root length, first leaf length, coleoptile length, bud dry weight, seedling fresh weight, root dry weight and dry substance transport rate of wheat seeds with four different concentrations are measured, the relative value and the weighted average are calculated, the weighted average of all indexes is imported into a comprehensive index calculation formula, and a comprehensive salt tolerant index is calculated. The method for identifying the salt tolerance of different wheat varieties in the germination period is high in systematicness, simple and convenient to operate, wide in applicability and high in comprehensiveness, and is more accurate in evaluation of the tolerance of the wheat in the germination period and has more reliable results than a salt-tolerant membership function value method.

The invention provides a method for evaluating the maximum elongation characteristics of the mesocotyl and the coleoptile of Zea mays. The method comprises the following steps of: (1) carefully choosing Zea mays seeds; (2) carrying out field planting of Zea mays materials; (3) analyzing the physical characteristics of Zea mays selfing line seeds; (4) analyzing the nutrient quality of the Zea mays selfing line seeds; (5) observing the maximum elongation characteristics of the mesocotyl and the coleoptile in the Zea mays selfing line; and (6) analyzing a result. The method disclosed by the invention takes the Zea mays selfing line seeds with different deep-sowing tolerance characteristics as a test material, the maximum elongation characteristics, which perform a main decisive effect for the deep-sowing tolerance characteristics of the Zea mays, of the mesocotyl / coleoptile / seedling length in the corresponding Zea mays selfing line are observed so as to obtain the 12-day-old accumulated mesocotyl length / coleoptile length / sum of the mesocotyl and the coleoptile / ratio of the mesocotyl to the coleoptile of the Zea mays selfing line as well as 22-day-old accumulated seedling length of the Zea mays selfing line during the maximum elongation characteristics of the mesocotyl / coleoptile / seedling length, optimal multiple regression equations between the physical characteristics and the nutrient quality characteristics of the seed and the above 12-day-old accumulated mesocotyl length / coleoptile length / sum of the mesocotyl and the coleoptile / ratio of the mesocotyl to the coleoptile of the Zea mays selfing line as well as 22-day-old accumulated seedling length are established, so that the potential maximum elongation characteristics of the mesocotyl, the coleoptilea and the seedling length of different Zea mays selfing lines can be quickly, nondestructively, objectively and accurately predicted, the Zea mays selfing line with good deep-sowing tolerance characteristics can be screened, and a technical reference is provided for quick screening and breeding application of the germplasm resources of the Zea mays selfing line with the good deep-sowing tolerance characteristics.

The invention discloses a planting method of sugary enhanced sweet corn. The planting method comprises following steps: seed soaking and germination accelerating, seedling raising in a seedling raising plate, soil preparation, seedling transplanting, field management, harvesting and the like. Compared with the prior art, the planting method has the advantages as follows: seeds are soaked in a traditional Chinese medicine liquid, and the problem of low seedling emergence rate caused by high sugar content and soft coleoptile of the sugary enhanced sweet corn is solved; seedlings are raised with a specific seedling raising substrate, management is enhanced in a seedling raising period, and the survival rate of the raised seedlings is increased; according to the characteristics that weeds are prone to running riot in a sugary enhanced sweet corn field and the sugary enhanced sweet corn is intolerant of drought in the earlier stage and is intolerant of waterlogging in the later stage, corresponding soil preparation and fertilizer and water application are performed after seedling transplanting, so that the planted sweet corn has good palatability, good marketability and obvious economic benefits.

The invention relates to an anniversary large-scale maize transformation method and belongs to the field of agricultural biotechnology and crop breeding. The anniversary large-scale maize transformation method mainly comprises the following steps of: (1) sowing maize acceptor materials in the maize growing reason in batches at an interval of 4-7 days; (2) selfing or carrying out sister cross, and controlling the number of plants pollinated each day to be approximately identical; (3) after pollinating for 9-12 days, stripping young embryos to carry out agrobacterium tumefaciens mediation conversion; (4) germinating maize seeds so as to obtain seedlings when the young embryos cannot be obtained; (5) intercepting coleoptile knots to induce calluses and subculturing; and (6) transforming calluses by adopting agrobacterium tumefaciens mediation conversion. The anniversary large-scale maize transformation method has the advantage of effectively overcoming the shortcomings of shortage of acceptor materials for maize transformation and high cost, anniversary large-scale transformation is easy to realize, and thus a large number of transgenosis seedlings are obtained.

The invention relates to a method for rapidly identifying wheat fusarium stem rot. The method comprises the following steps: 1, preparation of fusarium graminearum spore suspension; 2, germinating wheat seeds at a room temperature till the coleoptile length is 1.4-1.6 cm; 3, cutting 2-4 mm from the top end of coleoptile, taking the fusarium graminearum spore suspension of 2-4 microliters and inoculating at a coleoptile wound; 4, culturing in an illumination incubator for 7 days and measuring the coleoptile lesion length. The method provided by the invention has the beneficial effects that culture conditions are simple; the wheat fusarium stem rot inoculation is simple and rapid, the absorption is more rapid due to the existence of the coleoptile wound, the resistance level of the wheat fusarium stem rot can be identified within a shorter time, expensive equipment is not required; a large number of repetitions can be performed at a time, and the statistical method is high in accuracy, low in experiment cost, labor and force-saving, good in identification effect on wheat fusarium stem rot resistance and suitable for popularization and application.

The invention discloses a callus induction method taking paris polyphylla coleoptiles as explants. The method comprises the following steps: firstly sterilizing and pre-culturing latent buds to be germinated on a paris polyphylla rootstock; and selecting the white inner-layer coleoptiles of the buds the volume of which is obviously increased after preculture as the explants, inoculating the explants into a callus induction culture medium with the concentration of MS+6-BA of being 0.5-2.0mg / L<-1>, the concentration of NAA of being 0.5-4mg / L<-1>, the concentration of coconut juice of being 20-80mg / L<-1>, the concentration of sucrose of being 30-50g / L<-1> and the concentration of agar of being 6.0-7.0g / L<-1>, and culturing for 50-60 days to obtain white or yellow-white compact calluses formed on the surfaces of the explants. The method disclosed by the invention better has the beneficial effects of better solving the problems of the traditional paris polyphylla production method such as long growth time, high cost and the like, and effectively alleviating the shortage phenomenon of the current paris polyphylla medicinal material.

The invention provides a purpose of gamma-aminobutyric acid for improving corn seed germination and root system growth under salt stress, relates to the purpose of the gamma-aminobutyric acid for the growth and development of corn, and aims to provide a new purpose of the gamma-aminobutyric acid. The gamma-aminobutyric acid has a promoting effect on the corn seed germination and seedling growth, has a promoting effect on growth of corn seed radicles and coleoptiles under the salt stress, and is higher in promoting effect on the growth of the radicles under the salt stress than that the coleoptiles. The restraining effect of the salt stress on the activity of alpha-amylases can be relieved, so that the restraining effect of the salt stress on the heights of corn seedlings is relieved. Accumulation of the fresh weight and the dry weight of stems and leaves under the salt stress is restored. The gamma-aminobutyric acid has the promoting effect on the growth of the seedling root systems under the salt stress, specifically relieves the restraining effect of NaCl on root fresh weight, root dwt, root lengths, root surface areas, root volume, root average diameters and root tip number, and is used in the field of breeding of corn.

The invention discloses an embryoid induction method taking a polyphylla bud axis as an explant. The method comprises the following steps: firstly, carrying out disinfection and pre-cultivation on an un-germinated bud on polyphylla rhizome, and then cutting off coleoptile of the bud of which the volume obviously expands after pre-cultivation; inserting the bud axis into a callus-inducing medium to cultivate from the center after cutting in length, and then cultivating the callus tissue generated by the bud axis to obtain polyphylla embryonic tissues; finally carrying out induction cultivation on the embryonic tissues, so as to generate white spherical embryoids. By adopting the embryoid induction method, a new path is supplied for cultivation of excellent variety of the polyphylla and massive and rapid propagation of tissue culture seedlings; mass production of polyphylla plants at low cost within a short period of time can be achieved.

The invention discloses a method for culturing corn haploid coleoptile section tissue and a specific culture medium thereof. The differentiation culture medium provided by the invention is the culture medium used in the differentiation and shoot formation of corn haploid coleoptile section induced callus, and is prepared by adding the following substances into MS basic culture medium: proline, casein hydrolyzate, carbon and gel, wherein the final concentration of the proline is 0.7g / L, and the final concentration of the casein hydrolyzat is 0.5g / L. The method of the invention adopts corn coleoptile section as the explant induced callus, and then differentiates the callus into a haploid strain to establish a stable and high-efficiency tissue culture system. The invention can be used for differentiation rate genetic research, material selection for tissue culture, and the like.

The present invention is a method of shoot apical meristem transformation for monocot plant via sufficient and micro wounding (SMW). The technical process includes: expose the apical meristem by removing the coleoptile away when the shoot grows to 0.2-2 cm after 1-2 days of seed germination; make sufficient and micro wounding transformation to the apical meristem by stabbing and brushing for 2-3 times using the SMW brush having 100-5000 bristles which is 4-20 μm in diameter for each one and 0.5-3 mm in exposed length, and dipped with the Agrobacterium tumefaciens containing binary vector harboring exogenous genes; develop the treated meristems directly to normal plants after co-cultivation; promote the plants to develop big spikes and set more seeds; harvest the seeds of T0 plants separately; detect and identify the transformation results in T1 generation which is bred from each individual T0 plant. The advantages of the invention are independent of tissue culture, unlimited in genotype, unnecessary to carry resistant marker, simple and large scale to perform, and applicable to all monocot plants which can set seeds. The transformation efficiencies for wheat, rice and maize using this method are 49%, 66.3%, and 100%, respectively.

The present invention provides a method for transforming exogenous gene by coleoptile of germinating seed, which belongs to the technical field of plant genetic engineering. The invention mainly comprises the following steps: a) germinating seeds in conventional method, carrying out transformation when coleoptiles grow to an appropriate length; b) absorbing the activated and cultivated agrobacterium liquid with a micro-syringe; c) vertically penetrating the syringe needle into the growing points of damaged plumules through the top of coleoptiles, or injecting the bacterium liquid to the growing points after coleoptiles are crosscut with a blade above the growing point of plumules on the base of coleoptiles and conducting transforming and cocultivating; d), spraying herbicide when the seeds grow to seedlings, taking herbicide resistant seedlings as transgenic plants of primary selection because of the gene of herbicide resistant being the marker gene of transformation; e) growing the plants of primary selection in the field and conducting gene identification to the seeds obtained. This invention is characterized by simple, rapid, efficient and large-scale transformation, and can be applied to gramineous plants of which the germinated seeds have coleoptiles.

The invention discloses a rapid and efficient genetic transformation method for agrobacterium tumefaciens-mediated wheat stem tips. The method mainly comprises the steps of carrying out water seed soaking, accelerating germination and cold treatment on mature wheat seeds, transversely cutting coleoptiles of the wheat seeds until exposing stem tips, taking the cut wheat seeds as a transformation acceptor material, carrying out genetic transformation by virtue of vacuum treatment mediation on agrobacterium tumefaciens, placing the transformed material in a suitable growth environment, and carrying out transgenic identification after the plants grow, so as to obtain transgenic wheat plants. By virtue of the rapid and efficient genetic transformation method, the culture of callus tissues of the wheat is omitted, so that the influences caused by various factors to the genetic transformation of the wheat are reduced, and the problems of vitrification, browning, cell mutation and the like in the tissue culture process are avoided; the genetic transformation conditions of the wheat are optimized, so that the economic cost of the genetic transformation of the wheat is lowered, the restriction of the genotype of the wheat to the transformation efficiency is effectively overcome, and a large amount of transgenic wheat plants can be rapidly obtained; and the rapid and efficient genetic transformation method has significant values to the culture of high-quality new-variety wheat and the research of the genetic transformation of the wheat.

The invention relates to the technical field of plant breeding, in particular to a bud stress resistance detection device and a bud stress resistance detection method. According to the invention, a beverage bottle is subjected to waste use and is used for bud stress resistance detection. The device is simple in structure and is capable of providing a growing environment which more approaches a natural state for buds, the growth of coleoptile and radicle is not restricted, and the solution concentration change caused by water evaporation is avoided. Therefore, a result of an experiment carried out by adopting the device is more accurate. The method is capable of fully utilizing the device, computer software is combined for processing, the result of the experiment is rapider and more accurate to process, the experiment error caused by long-term exposure of a plant root system is avoided, and the result of the experiment is more accurate and reliable.

The invention relates to a method for transforming an exogenous gene by using maize coleoptile joint induced calluses, which belongs to the field of agricultural biotechnology and crop breeding. The method mainly comprises the following steps of: (1) placing sterilized maize seeds in a sprouting culture medium for cultivation, and germinating to obtain seedlings; (2) cutting off the coleoptile joints from the seedlings to induce the calluses; (3) carrying out subculture propagation on the calluses, and introducing the exogenous gene by adopting an agrobacterium tumefaciens mediated method; (4) obtaining kanamycin-resistant calluses by 3-4 times of screening; (5) respectively carrying out embryoid induction and differentiation to obtain transgenosis regeneration seedlings; and (6) carrying out PCR (Polymerase Chain Reaction) detection on the regeneration seedlings to check whether the transgenosis is introduced. According to the method, acceptor materials are free of limitation of seasons and convenient to obtain and have strong regeneration capacity; and the large-scale genetic transformation of the maize is easy to realize.

The invention discloses a method for rapidly evaluating rice SRBSDV variety resistance through detecting a viral multiplication speed in coleoptile. The method comprises the following steps: (1) selecting an identifying scaleplate; (2) performing hypoxia induction on the rice to be tested and identifying the coleoptile of the scaleplate variety; (3) inoculating the toxic sogatella furcifera on the coleoptile and performing dark culture; (4) performing fluorogenic quantitative PCR to obtain a relative transcript level (RQ value) of an SRBSDV virus S9 and a reference gene in each rice variety period time; (5) drawing an SRBSDV proliferation index trend line; and (6) evaluating the resistance level and the resistance grade of the rice variety to be tested on the Southern rice black-streaked dwarf virus according to the SRBSDV proliferation index trend line formula. By detecting the viral multiplication speed in the coleoptiles and rapidly evaluating the resistance of the rice SRBSDV germplasm resistance, the resistance level and the resistance grade of the rice variety on the SRBSDV from the molecular level in the rice germination stage are determined, the identification period is short and the reliability is high, and the method has big breeding application value.

The invention discloses a culture method of virus-free seedlings of curcuma wenyujin. The method comprises the following steps of selecting main rhizomes of curcuma wenyujin for accelerating germination until robust buds are obtained on the main rhizomes; cutting the robust buds, performing cleaning, stripping outer-layer coleoptiles, selecting growing points, and inoculating the growing points into an induced germination differentiation culture medium for culture; transferring the obtained virus-free seedlings to a proliferation culture medium for proliferation culture and repeated culture; and transferring single plantlet obtained by culture to a rooting and seedling strengthening culture medium for culture until a seedling which can be planted out of a bottle is obtained. By adopting the method disclosed by the invention, a large number of high-quality seedlings with the same genetic background and consistent growth vigor can be produced in a relatively short period, and the methodis particularly suitable for industrialized seedling production of the virus-free seedlings of the curcuma wenyujin.

The invention discloses a fast and efficient agrobacterium-mediated wheat stem tip genetic transformation method. The method mainly comprises the following steps: taking mature wheat seeds, soaking the seeds in water, performing accelerating germination, performing cold treatment, transversely cutting coleoptile, exposing stem tips to serve as transformation receptor materials, performing vacuum treatment mediation on agrobacterium, performing genetic transformation, placing the transformed materials in a suitable growth environment, and performing transgenic identification on grown plants, thereby obtaining the transgenic wheat plants. According to the method disclosed by the invention, wheat callus does not need to be cultured, influences of various factors on genetic transformation of wheat are reduced, the problems such as vitrification, browning, cell mutation and the like in the tissue culture process are solved, the conditions of genetic transformation of wheat are optimized, the economic cost of genetic transformation of wheat is reduced, restriction of wheat genotype on transformation efficiency is effectively overcome, lots of transgenic wheat plants can be rapidly obtained, and the method has significant values for cultivating high-quality new wheat varieties and performing study on genetic transformation of wheat.

The invention belongs to the technical field of agricultural production, and provides a method for synchronously identifying the drought resistance of rice in the germination period and the later period. The method comprises the following steps: sterilizing seeds of a to-be-detected rice variety, putting into a culture dish, adding distilled water, and performing germination culture at 30 DEG C in a constant temperature incubator; picking seeds with uniform and consistent exposed buds, and performing abscisic acid culture and distilled water contrast culture; performing dark culture at 30 DEG C in the constant temperature incubator for 5 days; and measuring the dry weight of coleoptiles, calculating the inhibition ratio and identifying the drought resistance according to the inhibition ratio. By the method for identifying the drought resistance of indoor rice, abscisic acid replaces a hypertonic solution such as ethylene glycol, a problem that the conventional method for identifying the drought resistance of the rice in the early period cannot accurately reflect the drought resistance of the rice in the later period is solved, the drought resistance of the rice in the germination period and the drought resistance in the later period are synchronously identified, and the method has high promotion and application value.

This invention relates to a new class of proteins called expansins, and methods related thereto are presented. This class of proteins can be characterized as catalysts of the extension of plant cell walls and the weakening of the hydrogen bonds in pure cellulose. Two proteins have been isolated from washed wall fragments of cucumber hypocotyls, referred to as “cucumber expansin-29” and “cucumber expansin-30”. Moreover, three peptide fragments from the purified cEx-29 protein were sequenced, then oligonucleotide primers were designed to amplify a portion of the expansin cDNA using polymerase chain reaction with a cDNA template derived from cucumber seedlings, and then the PCR fragment was used to screen a cDNA library to identify full length clones. Another expansin protein has been isolated from oat coleoptiles (oat expansin oEx-29), while three additional expansin sequences have been identified in Arabidopsis and an additional two in rice.

The invention relates to an application of an OsARD1 gene to improving of the flooding tolerance of rice. The nucleotide sequence of the OsARD1 gene is SEQ ID No.1, and the encoded amino acid sequence of the OsARD1 gene is SEQ ID No.2. The experiment results show that the coleoptile and the lamina of an overexpressed transgenic plant are both longer than those of a wild type control plant, so that the OsARD1 gene overexpressed transgenic plant is more rapidly contacted with air and shine through more rapid elongation of the coleoptile and the lamina under the flooding condition, then the normal aerobic metabolism and the photosynthesis of the plant are recovered, and enough nutrients are guaranteed for the growth of the part flooded by water. Further experiments prove that overexpression of the OsARD1 gene strengthens the flooding tolerance of rice to a certain degree. By improving the flooding tolerance of the rice and cultivating flooding tolerant rice varieties, the loss of the rice in the flood disaster is reduced, and the application is of great significance for guaranteeing food security of people and the agricultural sustainable development.