1652 results about "Plant genetic engineering" patented technology

The invention provides a method for deleting a selection marker gene of transgenic rice and belongs to the field of plant gene engineering. The method for efficiently deleting the marker gene of the transgenic rice in a plant level is established by utilizing a genome fixed-point editing technology mediated by a CRISPR/Cas9 system. The whole segment of the marker gene can be effectively deleted by utilizing the method, and only an expression frame of the marker gene is deleted pointedly without change of the expression of other components in the transgenic rice. Thus, the transgenic rice without the selection marker gene can be efficiently bred by utilizing the method, so that the safety doubt of people about the selection marker gene can be completely removed.

The invention belongs to the technical field of plant genetic engineering and specifically relates to a method for directionally knocking out miRNA (micro Ribonucleic Acid) of paddy rice. Aiming at solving the technical problems, a method and a detection means are provided for directionally modifying miRNA sites of plants. The invention discloses a method for obtaining miRNA mutants of the paddy rice; specific operation of the method comprises: knocking out single miRNA, knocking out a plurality of miRNAs or knocking out large fragments of the miRNA by adopting a CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats- associated 9) method. The method provided by the invention is suitable for creation and screening of directionally knocking out the miRNA of the paddy rice.

The invention belongs to the technology field of plant gene engineering, and relates to the field of preparation and application of molecular marker. The molecular marker originates from the ZmGA3ox21 gene section, which locates in the third chromosome of corn and controls the character of plant height of corn. The sequence of the nucleotide, which is used to separate and clone the ZmGA3ox2 gene, is represented by the SEQ ID NO: 1, and the cDNA sequence of the nucleotide is represented by the SEQ ID NO: 2. A molecular marker of corn plant height is obtained, and the nucleotide sequence of the molecular marker is represented by SEQ ID NO: 6-9. It has been proved by utilizing the map-based cloning method that natural variation of the gene can affect the number variation of corn plant height, and provoke the changes of length, expression level, and in-vitro active GA content of corn internodal cells. The natural variation of the gene does not affect yield and flowering phase related characters. It has been proved by the genetic analysis of dwarf mutant d 1-6016 that loss of large sections of the gene can cause dwarfing of corn plants.

The invention relates to the field of plant genetic engineering, and in particular relates to separation, cloning and application of a gene OsPTR9 which can improve nitrogen utilization efficiency and yield of rice. The OsPTR9 is one gene in a rice PTR (peptide transporter family) family, and an encoded protein is a nitrogenous organic matter transporter. In the invention, the OsPTR9 gene is excessively expressed, so that nitrogen fertilizer absorbing efficiency of normal rice is improved, tillering capacity is improved, ear length and thousand seed weight are increased, and functions of the OsPTR9 gene is verified in an OsPTR9 target gene mutant by adopting an RNAi (ribonucleic acid interference) technology. According to the invention, the expression quantity of the OsPTR9 gene is increased or reduced by adopting a genetic engineering technology, which sufficiently shows that the OsPTR9 gene can control the nitrogen absorption, plant tillering, ear length and thousand seed weight andthe like of the rice. The gene has important application values in the aspects of explaining influence of nitrogen on a plant growth and development process, efficiently utilizing nitrogen fertilizerfor the rice and improving the yield.

The invention relates to the technical field of plant genetic engineering, in particular relates to isolating clone and functional verification of a DAN fragment containing a gene OsWRKY45-2 related to rice disease resistance. The gene OsWRKY45-2 codes WRKY proteinoids and can endow rice with resistance to the diseases caused by bacterial pathogenic bacteria, namely the leaf blight bacteria (Xanthomonas oryzae pv.oryzae) and fungal pathogenic bacteria, namely the rice blast bacteria (Magnaporthe grisea). The fragment and the exogenous regulatory sequence thereof are directly transferred into rice, thus the resistance capability of transgenic rice with over-expression OsWRKY45-2 against bacterial blight in and rice blast is improved remarkably.

The invention discloses a DNA fragment separating colony and functional checking of rice tissue special expressing promoter PD54O in plant gene engineering technical domain, which is characterized by the following: the PD54O promoter expresses in rice leaf and leaf sheath and does not express in germ and endosperm; transferring genetic rice plant can resist the invasion of borer with the expression of PD54O regulate and control insect-resisting gene; it does not express Bt protein in seeds of food.

The invention belongs to the technical field of plant gene engineering, disclosing a separated and cloned major gene GS5 capable of controlling the width and the weight of a rice grain and the DNA sequence of the allelic gene of the major gene GS5. The DNA sequence is shown as SEQ ID NO.1 (Zhenshan 97B) and SEQ ID NO.3 (H94) and contains 10 exons. The amino acid sequence of the major gene and the amino acid sequence of the allelic gene are shown as SEQ ID NO.2 and SEQ ID NO.4. By using two large-grain rice varieties and two small-grain rice varieties for comparative sequencing, in an approximately 6.1kb range, 22 common base differences exist between the large-grain variety and the small-grain variety, wherein 18 mutations are in a promoter area, 4 mutations are in a code area and 5 amino acids are caused to be changed. By using a transgenic technology, GS5 transgenic rice plants are obtained and express that the width and the weight of the rice grain are obviously improved when being compared with the control width and the control weight of the rice grain. The character changes are quite coincident with the two genotype expressions of a Zhenshan 97 near-isogenic line and a GS5 near-isogenic line. The invention additionally discloses a method of near-isogenic line breeding, gene cloning and gene transfer and application thereof.

The invention belongs to the technical field of plant gene engineering, and discloses a screening method for real-time fluorescent quantitative PCR reference genes of Hibiscus hamabo Sieb.et Zucc. Thescreening method utilizes a transcriptome database of Hibiscus hamabo Sieb.et Zucc, selects 10 candidate reference genes, and takes 10 candidate reference gene sequences as templates to design real-time fluorescent quantitative PCR specific primers, selects different tissues of Hibiscus hamabo Sieb.et Zucc and Hibiscus hamabo Sieb.et Zucc leaves under various abiotic stresses and hormone treatments as experimental materials for performing a fluorescent quantitative PCR experiment, and finally, performs data analysis by applying geNorm, NormFinder and BestKeeper software. The most stable reference genes in the screening method are ACT and SKIP. The screening method for real-time fluorescent quantitative PCR reference genes of Hibiscus hamabo Sieb.et Zucc establishes a screening system forstable reference genes of hibiscus hamabo, and provides stable reference genes for developing gene function research of Hibiscus hamabo Sieb.et Zucc.

The invention relates to the technical field of plant genetic engineering, and discloses an agrobacterium-mediated sugarcane genetic transformation method with vacuum infiltration assistance. The agrobacterium-mediated sugarcane genetic transformation method includes: (1) induction and multiplication of embryonic callus of sugarcane core leaves, (2) activation and cultivation on agrobacterium tumefaciens with target genes, (3) agrobacterium infestation with vacuum infiltration assistance on embryonic callus of sugarcane core leaves, (4) resistant material screening and germination, and (5) resistant material verification. With the agrobacterium-mediated sugarcane genetic transformation method, the agrobacterium can pass through the gaps among callus cells and enter deep cells on vacuum, and tiny wounds can be generated on the surfaces of the cells, so that phenolic substances are secreted by plants to activate shear and transfer of T-DNA (transferred deoxyribonucleic acid) in Ti plasmids. The agrobacterium-mediated sugarcane genetic transformation system is perfected. Genetic transformation rate of sugarcane is remarkably improved as compared with that of the prior art.

The invention belongs to the technical field of plant gene engineering, in particular to a modified glyphosate-resistant gene and a glyphosate-resistant rice cultivation method and comprises modification of glyphosate genes, building of plant expression vectors of the glyphosate-resistant gene and a glyphosate-resistant transgenosis rice cultivation method. Artificially-synthesized modified glyphosate-resistant gene serves as a target gene, the sequence of the gene is as shown in SEQ ID NO: 1, and the target gene is led into rice receptors by an agrobacterium tumefaciens mediated transformation method to obtain high glyphosate-resistant transgenosis rice GT28. the glyphosate-resistant gene of GT28 is recombined by the aid of sexual hybridization and somatic hybridization technique to obtain new rice germplasm, and new glyphosate-resistant varieties (strains) of rice are cultivated.

The invention relates to the field of plant genetic engineering, and provides a rice nitrogen transporter gene OsPTR10, wherein the protein sequence of the rice nitrogen transporter gene OsPTR10 is shown as SEQ ID No.1, and the cDNA sequence of the rice nitrogen transporter gene OsPTR10 is shown as SEQ ID No.2. According to the rice nitrogen transporter gene OsPTR10, through the excessive expression of the OsPTR10 gene, the normal efficiency in absorbing nitrogenous fertilizer of the rice is improved, the content of soluble protein of plants is increased, further, the tiller number is increased, the ear length and filling grain number are increased, and finally, the yield is improved; through the RNAi technology, the expression quantity of the OsPTR 10 gene is reduced, and the functions of the gene are verified in the OsPTR10 target gene mutant. The gene has the significant application values on the aspects of the statement on the influences of the nitrogen element on the growth and development of the plants and on the aspects of the efficient utilization for the nitrogen fertilizer of the rice and the improvement on the yield of the rice.

The invention relates to the technical field of the plant genetic engineering, in particular to the isolation and cloning and functional verification of the DNA fragment of oryza officinalis anti-Xanthomonas oryzae major gene Xa3/Xa26-2. The Xa3/Xa26-2 gene is used to code the leucine-rich protein kinase protein. The Xa3/Xa26-2 gene ensures that rice can resist diseases caused by bacterial pathogen-Xanthomonas oryzae pv. oryzae. The DNA fragment and the regulatory sequence thereof are directly transferred in rice, and the resistance capability to Xanthomonas oryzae of the transgenic rice carrying Xa3/Xa26-2 is significantly enhanced.

The invention, relating to the field of plant genetic engineering, particularly relates to a nucleotide sequence of a gene from an allied specie Aegilops variabilis of wheat for resistance to Cereal cyst nematode, and an application on improving plant resistance to insects based on the gene through the method of biotechnology. The nucleotide sequence of the gene for resistance to Cereal cyst nematode disclosed in the invention is a complete encoding sequence of the gene for resistance to Cereal cyst nematode with the width of 3586bp, comprising 3 exons and 2 introns, and can code 934 amino acids, and is used in improving plant resistance.

The invention discloses application of a nitrate radical transporter gene OsNRT1.8 in rice breeding and belongs to the field of plant genetic engineering. The protein encoding amino acid sequence of the gene OsNRT1.8 is shown as SEQ ID NO.1, and a cDNA sequence is shown as SEQ ID NO.2. By establishing rice gene OsNRT1.8 overexpression plant and a gene OsNRT1.8 interference plant, it is found that normal rice tiller number and spike number of each plant can be increased by improving gene OsNRT1.8 expression. Therefore, the gene OsNRT1.8 can be used for increasing rice yield in rice breeding. The gene OsNRT1.8 has an important application value on the aspects of elaboration about nitrogen influence on plant growth and development and improvement of rice plant type.

The invention belongs to the technical field of gene engineering, in particular relates to application of Chilo suppressalis (Walker) endogenesis small RNA in rice inset resistance improvement, and aims to verify the function and study the application of an obtained Chilo suppressalis endogenesis small RNA sequence. By means of high-flux small RNA sequencing and bioinformatics analysis, a DNA sequence of Chilo suppressalis endogenesis small RNA csu-15, and the nucleotide sequence of the DNA sequence is as shown in SEQ ID NO:1. The sequence of csu-15 is adopted to establish an artificial microRNA (amiRNA) expression carrier and convert rice. The in-vitro stalk insect inoculation experiment shows that Chilo suppressalis growth can be remarkably inhibited in transgenic rice.

The invention discloses a method for building a high-efficiency regeneration system of superior corn self-bred line agriculture line 531, belonging to the field of plant genetic engineering and transgenosis breeding. The invention takes an agriculture line 531 rataria as an explant, induces in a callus induction medium and produces an II-type embryonic callus; the II-type embryonic callus is subjected to embryoid induction under light in an embryoid induction medium to produce a green embryoid; then, the green embryoid is transported to a regeneration medium and is cultured into a regeneration plant under light; root induction is carried out in a rooting medium, and acclimatization is carried out in Hogland nutrient solution to ensure that a new thick root grows on the root of the regeneration plant; the root is transplanted to nutritional soil for rejuvenation culture; and finally, the root is transplanted to a land for growing field crops to normally grow and seed. The regeneration technology is suitable for the superior corn self-bred line agriculture line 531 with high application value, can ensure that the superior quality of agriculture line 531 corn can be inherited in the corn transgenosis breeding process, and has an important meaning for functional genome group research.

The invention relates to the technical field of plant genetic engineering, and more particularly relates to separated clone and functional verification of DNA segments of paddy disease-resistant related gene OsEDR1. By utilizing transgenic technology based on RNA interference (RNAi) principle, part of the DNA segments of the OsEDR1 gene are connected with a vector expressing double-strand RNA to be converted into rice varieties, and the expression of the OsEDR1 gene in the rice varieties can be inhibited. Genetic transformation rice with obviously reduced OsEDR1 gene expression quantity has the remarkably improved resistibility for bacterial blight and bacterial streak disease, so as to prove that the OsEDR1 gene is negative regulatory factor in the reaction of resisting bacterial blight and bacterial streak disease of rice, thus the disease resistance of the rice can be improved by inhibiting the expression of the OsEDR1 gene.

The invention relates to the technical field of plant genetic engineering and discloses clone and application of semi-dominant gene qGL3 capable of controlling grain length and grain weight of rice kernel. In the invention, a main-effective QTL (quantitative trait loci) SEQ ID NO:1 which is capable of simultaneously controlling the grain length and grain weight of rice kernel and a semi-dominant allelic gene SEQ ID NO:3 having advantageous function are separated and cloned; the two genes have the capacities of modifying the yield and appearance quality of rice; and at the same time, the two homologous genes of the two genes in rice are cloned and a fact that the two homologous genes have regulation effect on the length of rice kernel as well is proved. Thus, the gene qGL3 and the advantageous allelic gene qGL3-D thereof as well as the two homologous genes of the gene qGL3 all can be applied to crop genetic modification.

The invention belongs to the technical field of plant genetic engineering, and specifically relates to cloning and an application of a Wo gene which is related to tomato hair generation. The cDNA (complementary deoxyribonucleic acid) sequence of the cloned Wo gene is as shown in SEQ ID NO.1(sequence identity number 1) in a sequence table, the corresponding amino acid sequence is as shown in SEQ ID NO.2, and the Wo gene encodes 731 amino acids. The invention further relates to a promoter of the Wo gene and cloning of two allelic genes of the Wo gene. The Wo gene is over-expressed in tomatoes and can improve the amount of epidermal hair of the tomatoes, significantly enhance the insect resistance of the tomatoes and enhance the anti-virus capability.

The invention relates to the technical field of plant genetic engineering, in particular to a gene for regulating and controlling the corn drought stress resistance. The gene is a ZmPP2CA10 deletion mutation gene; the deletion segment is a continuous base sequence with a distance being 301bp away from the upstream of the ZmPP2CA10 initial codon ATG; the continuous basic group sequence is shown as SEQ ID NO:1; the nucleotide sequence of the mutated ZmPP2CA10 promoter is shown as SEQ ID NO:2. The deletion mutation gene can be used for corn drought resistance relevant molecular marker auxiliary breeding and corn drought resistance auxiliary identification; the operation is convenient; the accuracy rate is high.

A method for using the promotor LEAP of rice's drought inducting gene to improve the resistance of plant to drought features that the promotor LEAP of rice OsLEAl gene is cloned. Its nucleotide sequence is shown by SEQ ID No.1 and is used to code a drought induced expression protein. Two cis-acting elements ABRE contained in its promotor region can be specifically response to drought and ABA. Its inductive expression carrier can stongly induce the expression of target gene when the drought stress is applied to a plant. The method for culturing the drought-resistant plant is also disclosed.

The invention relates to the technical field of plant genetic engineering, and particularly discloses an application of a corn CIPK42 protein and a coding gene of the corn CIPK42 protein in regulationand control of salt stress tolerance of plants. The corn ZmCIPK42 gene is found to be able to positively regulate and control the salt tolerance of the plants; and the salt tolerance of the plants can be effectively improved by increasing of the expression quantity of the ZmCIPK42 gene. According to the invention, transgenic corn and arabidopsis thaliana plants with ZmCIPK42 overexpression are constructed; and compared with a non-transgenic wild type, the transgenic corn and arabidopsis thaliana plants are significantly improved in salt tolerance and growth. Discovery of the salt-tolerant function of the ZmCIPK42 gene provides a novel gene target and resource for cultivation of salt-tolerant plant varieties, is of great significance to research of a salt-tolerant molecular mechanism of the plants, and lays a certain theoretical foundation for research of a salt stress response mechanism of the plants and a molecular mechanism of resisting adverse environments.

The invention relates to the technical field of plant gene engineering and provides an inducible promoter for pathogenic bacteria of rice. The promoter can be activated by rice bacterial leaf blight bacterium xanthomonas PXO99 and PXO61, a rice baeterial leaf streak bacterium RS105 and a corn banded sclerotial blight bacterium YWK-196, and can also be activated by abscisic acid, gibberellic acid, salicylic acid and sodium chloride. Truncation of the promoter verifies that two sections from -513bp to -411bp and from -411bp to -309bp are critical areas induced by the pathogenic bacteria. Various plant expression vectors can be constructed with the adoption of the promoter, and the promoter can be used for improving plant quality and other beneficial production traits through a plant gene engineering technology.

The invention relates to the field of plant genetic engineering and specifically relates to isolation and cloning and application of OsELF 3 gene capable of controlling the heading stage of paddy rice. According to results of functional complementation experiments of the gene, the OsELF 3 gene has the function of controlling the heading stage of paddy rice, the heading stage of plants can be regulated through improving or weakening the expression level of the OsELF 3 gene by using genetic engineering technology, and therefore, the heading stage of a variety is improved to be adapted to the variety to different areas and seasons. The gene cloned in the invention and coded protein thereof can be extensively used for regulating flowering characteristics of plants and improving growth stages of plant varieties and have critical application values.