148 results about "Genetically modified wheat" patented technology

The invention provides a wheat TaAGO4a gene CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats)/-CRISPR-associated protein 9) vector and application thereof and belongs to the field of crop molecular biology. The wheat TaAGO4a gene CRISPR/Cas9 vector and the application thereof have the advantages that gRNA of a third exon of specificity-targeted TaAGO4a is provided firstly, a DNA sequence of the gRNA is shown as SEQ ID NO.1, and the gRNA contains an enzyme cutting site XmnI; subsequently, the CRISPR/Cas9 vector containing the gRNA is provided, and through co-transformation of Cas9 and the specific gRNA into a wheat protoplast as well as enzyme cutting and sequencing technologies, the condition that the gRNA can guide the Cas9 to cut three copies positioned on a chromosome 3A, a chromosome 3B and a chromosome 3D of the TaAGO4a respectively can be detected successfully so as to cause frameshift mutation of the gene and result in afunction or excalation of the gene; the wheat TaAGO4a gene CRISPR/Cas9 vector can be used for preparing TaAGO4a gene-deleted transgenic wheat.

The invention discloses an improved method for detecting and identifying transgenic wheat with high sensitivity and high efficiency. The improved method comprises the steps of: (1) carrying out pretreatment and enzymolysis on tissue with vigorous cell division in a meristematic zone at a root tip of wheat through N2O gas to obtain a metaphase mitotic phase specimen with clear image and good dispersion of chromosomes; (2) marking a fluorescent probe of a target fragment to be detected (for example, a transgenic fragment carried by a transgenic carrier) by utilizing a nick translation method; (3) carrying out fluorescent in-situ hybridization on the metaphase mitotic phase chromosomes of the transgenic wheat obtained in the step (1) by utilizing the fluorescent probe of the target fragment of the transgenic carrier constructed in the step (2); and (4) analyzing fluorescent in-situ hybridization results obtained in the step (3), and comparing the fluorescent in-situ hybridization results with a contrast material to determine the distribution of fluorescent signals in the transgenic wheat to be detected. The improved method provided by the inventions can be applied to chromosomal localization of exogenous transgenic wheat with the sensitivity reaching 2-3kb, and also can be applied in the identification of transgenic signals of other crops.

The invention discloses a method for breeding aphid-resistant transgenic wheat and a special expression vector. A recombinant vector provided by the invention is obtained by inserting gene expression cassettes shown as nucleotides at sites 1 to 3,828 from the tail end 5' in SEQ ID NO (Sequence Identity Number): 1 into multiple cloning sites of a vector pG4AB. Experiments prove that the aphid resistance of the transgenic wheat into which the vector is transferred is obviously higher than that of wild wheat, so that the vector has great significance in the field of genetic breeding of plants.

The invention provides a method for detecting the number of arbuscular mycorrhizal fungi in wheat rhizosphere soil based on real-time fluorescence quantification PCR. The method includes the steps that firstly, sample total DNA of wheat rhizosphere soil is extracted and serves as a template of PCR amplification; then, amplification is conducted with SEQ ID No.1 and SEQ ID No. 2 as primers, and specific fragments of 129 bp are obtained; after bond and conversion are conducted, qPCR amplification is conducted. By means of the method, arbuscular mycorrhiza is authenticated, absolute quantification can be conducted on the arbuscular mycorrhiza accurately, the copy number of the arbuscular mycorrhizal fungi in all periods of duration of transgenic wheat is obtained, and in the growth and development stages of wheat, the arbuscular mycorrhiza copy number tends to be gradually increased as a whole. The rapid, convenient and precise authentication method is built, a theoretical and test basis is provided for subsequent tests, great significance is provided for further evaluation of safety of transgenic crops, and compared with an existing method, the method is high in sensitivity, high in specificity, good in repeatability, convenient to operate and visual in result.

The invention discloses a pollen-specific promoter and an expression vector thereof. Experiments show that the promoter can drive efficient and specific expression of a target gene in the pollen of transgenic rice. The promoter of the invention can be used for functional analysis and identification of genes related to growth and development of plant pollen, as well as creation of plant male sterile lines, prevention of plant transgenic drift, and the like. Especially, the promoter has an important application value in creation of male sterile lines of rice and other gramineous crops and utilization of heterozygous advantages.

The invention provides a method for detecting wheat yellow mosaic virus-resisting transgenic wheat metabolites on the basis of gas chromatography-mass spectrometry technology (GC-MS). The wheat yellow mosaic virus-resisting transgenic wheat metabolites are optimized on the basis of the GC-MS detection method, the type and proportion of an extracted solution, extracting time, volume of the extracted solution, deriving process temperature and time are respectively optimized, the relative peak area is at an optimal detection state under the extracting condition of the extracted solution that the ratio of water to acetonitrile to isopropanol is 2:3:3 when 1mL of extracted solution is added for extracting for 20min, and more metabolites components can be detected. Under the deriving condition that the temperature is 30DEG C and the extracting time is 90min, the relative peak area is large, and the signal intensity is obvious. By adopting the detection method, reliable and effective data can be provided for the study of the metabonomics of the wheat yellow mosaic virus-resisting transgene wheat, and a foundation can be laid for the safety evaluation of the transgenic crop.

An object of the present invention is to discover an endogenous wheat sequence satisfying the conditions of: a) it is universally present in varieties of wheat, b) the amount present (detected amount) is not affected depending on the wheat variety, c) even if other grains are present, only wheat can be detected without cross-reactivity, and d) it is amplified quantitatively by the PCR reaction. A further object of the present invention is to provide a method of accurately detecting and quantitating endogenous wheat DNA in a test sample by the polymerase chain reaction. The present invention provides a method of detecting or quantitating endogenous wheat DNA in a test sample by the polymerase chain reaction, the method comprising: a step of using a nucleic acid in the test sample or a nucleic acid extracted from the test sample as a template to amplify the nucleic acid of a region consisting of the base sequence identified as SEQ ID NO: 2 or a partial sequence thereof with a primer pair capable of amplifying that region; and a step of detecting or quantitating the amplified nucleic acid.

The invention discloses an einkorn wheat low molecular weight glutenin subunit, its encoding gene and use, wherein the einkorn wheat low molecular weight glutenin subunit is the protein of amino acid residue No.2 sequence in the sequence table, or SEQ ID No.2 derived protein by the substitution, deletion or addition of one or several amino acid residual radicals to the SEQ ID No.2 amino acid residual radical sequence, and having the same activity with the SEQ ID No.2 amino acid residual radical sequence. The encoding gene of the einkorn wheat low molecular weight glutenin subunit is one of the following nucleic acid sequences, (1) DNA sequence of SEQ ID No.1 in the sequence table, (2) the polynucleotide of SEQ ID No.2 in the sequence table, (3) DNA sequence having over 90% homology with DNA sequences defined by SEQ ID No.1 in the sequence table and encoding the same functional protein. The gene can be used in the cultivation of transgenic wheats.

The invention discloses a breeding method for a genetically modified wheat capable of resisting both sheath blight and root rot and a related biological material thereof. The method for breeding the disease-resistant genetically modified wheat comprises the following step: the coding gene of a protein with an amino acid sequence shown as SEQ ID No. 2 is introduced into a receptor wheat, so that the genetically modified wheat, the disease resistance of which is higher than the disease resistance of the receptor wheat, is obtained; and the disease resistance is resistance to sheath blight and/orresistance to root rot. A transgenic experiment in which TaOCS is introduced into wheat proves that compared with the resistance of the receptor wheat, the resistance of the genetically modified wheat overexpressed by TaOCS to sheath blight and root rot is remarkably enhanced, indicating that TaOCS is a protein related to the sheath blight and root rot of plants, and TaOCS and the coding gene thereof can be used for enhancing the resistance of plants to the sheath blight and/or root rot.

The invention discloses a coded gene sequence and amino acid sequence with high-expressive activity macromolecular glutelin subunit with sequence as SEQ ID No.1 and SEQ ID No.2 in the biological gene engineering technical domain, which is characterized by the following: providing astopic endosperm promoter with high-expressive activity macromolecular glutelin subunit with sequence as SEQ ID No.3; constructing gene-transmitting carrier; utilizing gene shooting method, agricillin dielectric method and pollen tube channel method to transmit gene to common wheat; culturing superior wheat.

The invention discloses a plant yellow dwarf resistance-associated protein TiDPK1, a coding gene and application thereof. The protein is a protein 1) or 2) as follows: 1) the protein consisting of an amino acid sequence shown by a sequence 2 in a sequence table; and 2) the protein which is derived from 1) by performing substitution and/or deletion and/or addition on the amino acid sequence of the sequence 2 in the sequence table and is related to stress resistance of plants. The coding gene TiDPK1 of the protein, transgenic lines and a method also belong to the protection of the invention. Experiment proves that the gene is specifically expressed in yellow dwarf resistant wheat (such as YW642) and is regulated up by the induction of BYDV; TiDPK1 is a membrane protein; BYDV concentration is raised and susceptibility index rises in silent original yellow dwarf resistant wheat YW642 of the TiDPK1; over-expressed transgenic wheat improves resistance to yellow dwarf; and the TiDPK1 gene is shown to be an important gene resistant to yellow dwarf.

The present invention discloses a method for obtaining transgene wheat by using a gene gun, and a special culture medium. According to the present invention, wheat immature embryo is adopted as explant, a hypertonic culture medium provided by the present invention is directly used to carry out a hypertonic treatment without pre-culture, gene gun bombardment is performed, and the special culture medium provided by the present invention is adopted to carry out a hypertonic treatment, callus induction, differentiation and rooting culture after bombardment, wherein a screening agent is used during the differentiation and rooting stages and is not used during the callus induction state so as to significantly improve the wheat gene transformation efficiency, wherein the wheat gene transformation efficiency can be up to 36%; the method has advantages of short transformation period (the time from immature embryo taking to obtaining of the regenerated plant having root, stem and leaf is only 8-10 weeks), high transformation stability and high repeatability; and the effective approach is provided for wheat genetic improvement industrialization, and the broad application prospects are provided.

Compositions and methods are provided for the production and selection of transgenic plants and plant parts, for increasing the transformation frequency of a plant or plant part, and for regulating the expression of a transgene, such as a herbicide tolerance polynucleotide. The methods and compositions allow for the delay in the expression of herbicide tolerance polynucleotides until a point in development during which herbicide selection is more efficient. Compositions comprise polynucleotide constructs comprising an excision cassette that separates a transgene, such as a herbicide tolerance polynucleotide, from its promoter and host cells comprising the same. The excision cassette comprises a polynucleotide encoding a site-specific recombinase operably linked to an inducible promoter and expression of the recombinase leads to excision of the excision cassette and expression of the transgene.

The invention discloses a C4 type phosphoenolpyruvate carboxylase (PEPC) gene of corn and an application thereof in wheat. The novel PEPC gene provided by the invention can be stably expressed and inherited in wheat, and the photosynthetic efficiency is remarkably improved compared with that of an acceptor material, thereby providing an important gene source and parent material support for improving the photosynthetic efficiency of C3 crop by means of a C4 type high photosynthetic efficiency gene and breeding high photosynthetic efficiency transgenetic wheat variety with the level of output greatly improved.

The invention discloses a glyphosate-resistant gene, a special expression vector and an application of the glyphosate-resistant gene to preparation of glyphosate-resistant transgenic wheat. The invention provides a protein which is (a) or (b): (a) a protein consists of an amino acid sequence shown in a sequence 3 in a sequence table; and (b) a protein which is derived from the amino acid sequence shown in the sequence 3 in the sequence table through the substitution and/or deficiency and/or addition of one or more amino acid residues and derived from a sequence 2 related to glyphosate resistance. Experiments prove that artificial design is carried out on wheat by utilizing the condon preference of the wheat and the chloroplast targeting sequence from wheat RubisCO small subunit is added at N segment to obtain a modified gene through an artificial synthesis method; the modified gene is applied as a selection marker gene in wheat transformation so as to obtain the glyphosate-resistant transgenic wheat; and researches prove that the resistance of the transgenic wheat is higher than the transgenic resistance of the wheat before optimization.

The invention discloses a method for cultivation of transgenic wheat resisting take-all and sharp eyespot and related biological materials thereof. The method for cultivation of transgenic wheat of disease resistance includes: importing an related gene which has a code sequence composed of 7th-405th nucleotides in the sequence 1 of a sequence table into recipient wheat so as to obtain the transgenic wheat of disease resistance with disease resistance higher than that of the recipient wheat. The transgenic wheat of disease resistance is the wheat of a1 or a2: a1. transgenic wheat resisting take-all and sharp eyespot, with the disease resistance being resistance to take-all and sharp eyespot; and a2. anti-take-all transgenic wheat, with the disease resistance being resistance to take-all. Experiments prove that, compared with Yangmai 18, the anti-take-all effect of the transgenic wheat with the imported anti-take-all related gene is improved by 60%-80%, and the anti-sharp eyespot effect is improved by 20%-33%.

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 growing controversy over GM/novel-trait crops and food products that possess novel industrial or nutritional traits, has resulted in an increased need for technologies that allow the rapid and reliable identification of GM/novel-trait crops so that they can be identified, and segregated if necessary, at the various stages of production and processing. A visual seed identity system is described that permits the identification and tracing of each individual seed through the upstream value chain, from field increase to the processing plant. These traits are genetically controlled, transmitted to progeny by either pollen or ovule and expressed in developing seed, showing a xenia effect. That novelty of this proposed IP system will facilitate early mitigation measures if out-crossing or admixtures occur and exceed a threshold limit set by regulatory agencies.

The invention discloses an agrobacterium tumefaciens-medicated genetic transformation method of wheat. The agrobacterium tumefaciens-medicated genetic transformation method comprises the following steps of: firstly, inoculating recombinant Agrobacterium tumefaciens containing a target DNA (Deoxyribonucleic Acid) into wheat immature embryos; and then co-culturing the inoculated wheat immature embryos for enabling the T-DNA of the recombinant Agrobacterium tumefaciens to be integrated with a chromosome of the wheat; and secondly, directly inoculating the wheat immature embryos co-cultured in the first step in a long-seedling culture medium for culturing to obtain transgenic wheat seedlings with roots, stems and leaves. The method disclosed by the invention has the advantages that the applicability is wide, so that the method is suitable for transforming various wheat species; the method is simple in operation, and thus a transgenic plant can be obtained without inducing a tissue culture and regenerating the plant; and the transformation period is short, i.e. the process from the infection to the obtaining of the transgenic wheat seedlings with the roots, the stems and the leaves only requires 11-14 days. The method has very important significance for genetic improvement on the wheat species and the obtaining of new species of the transgenic wheat.