CN116656698B - Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops - Google Patents
Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops Download PDFInfo
- Publication number
- CN116656698B CN116656698B CN202310926763.8A CN202310926763A CN116656698B CN 116656698 B CN116656698 B CN 116656698B CN 202310926763 A CN202310926763 A CN 202310926763A CN 116656698 B CN116656698 B CN 116656698B
- Authority
- CN
- China
- Prior art keywords
- corn
- gene
- drought
- maize
- transgenic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 240000008042 Zea mays Species 0.000 title claims abstract description 101
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 101
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 73
- 235000005822 corn Nutrition 0.000 title claims abstract description 73
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 67
- 230000009261 transgenic effect Effects 0.000 claims abstract description 30
- 230000008641 drought stress Effects 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 15
- 230000006872 improvement Effects 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 238000012214 genetic breeding Methods 0.000 claims abstract description 4
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 28
- 235000009973 maize Nutrition 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 9
- 230000004044 response Effects 0.000 claims description 8
- 239000002773 nucleotide Substances 0.000 claims description 6
- 125000003729 nucleotide group Chemical group 0.000 claims description 6
- 238000012216 screening Methods 0.000 claims description 5
- 230000004069 differentiation Effects 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 3
- 210000001161 mammalian embryo Anatomy 0.000 claims description 3
- 230000001131 transforming effect Effects 0.000 claims description 3
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 2
- 230000006698 induction Effects 0.000 claims description 2
- 210000000056 organ Anatomy 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 35
- 238000010362 genome editing Methods 0.000 abstract description 30
- 230000008723 osmotic stress Effects 0.000 abstract description 24
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract description 13
- 230000002018 overexpression Effects 0.000 abstract description 12
- 230000014509 gene expression Effects 0.000 abstract description 10
- 230000035784 germination Effects 0.000 abstract description 10
- 102000004169 proteins and genes Human genes 0.000 abstract description 9
- 241000219194 Arabidopsis Species 0.000 abstract description 8
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 abstract description 6
- 239000000600 sorbitol Substances 0.000 abstract description 6
- 210000005253 yeast cell Anatomy 0.000 abstract description 6
- 108020004414 DNA Proteins 0.000 description 11
- 239000013598 vector Substances 0.000 description 11
- 239000000047 product Substances 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 241000219195 Arabidopsis thaliana Species 0.000 description 9
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 9
- 229940118019 malondialdehyde Drugs 0.000 description 9
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 230000024346 drought recovery Effects 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 241000589158 Agrobacterium Species 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 108091026890 Coding region Proteins 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- 238000009395 breeding Methods 0.000 description 4
- 230000001488 breeding effect Effects 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 238000010839 reverse transcription Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108091006146 Channels Proteins 0.000 description 3
- 102000034573 Channels Human genes 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 229910000831 Steel Inorganic materials 0.000 description 3
- 108700019146 Transgenes Proteins 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 3
- 230000006978 adaptation Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 238000000227 grinding Methods 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 230000007226 seed germination Effects 0.000 description 3
- 239000010959 steel Substances 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 102000006382 Ribonucleases Human genes 0.000 description 2
- 108010083644 Ribonucleases Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 229930182830 galactose Natural products 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N 2-amino-4-[hydroxy(methyl)phosphoryl]butanoic acid Chemical compound CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108091092584 GDNA Proteins 0.000 description 1
- 239000005561 Glufosinate Substances 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000012520 frozen sample Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000003208 gene overexpression Methods 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000002363 herbicidal effect Effects 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000005257 nucleotidylation Effects 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 1
- 229960001225 rifampicin Drugs 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of plant biology, and discloses application of a corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops, in particular to application in improving drought resistance of corn, and application in germplasm resource improvement, genetic breeding and preparation of corresponding drought resistant transgenic crops. Also disclosed is the expression of the gene in yeast to promote the tolerance of yeast cells to osmotic stress caused by sorbitol, and the overexpression in Arabidopsis to promote germination of transgenic plants under osmotic stress. The invention verifies that the corn gene Zm00001d018037 has the function of improving the drought stress resistance of corn by creating a gene editing mutant, transgenic over-expressing corn genetic material and drought resistance phenotype detection, thereby providing a new technical means for cultivating new strain of drought-resistant corn and creating germplasm resources, and having important agricultural application value and wide practical prospect.
Description
Technical Field
The invention relates to the technical field of biology, in particular to application of a corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops, and especially in improving drought resistance of corn.
Background
Drought stress seriously affects the growth and development of plants and the yield of crops, and has become an important research topic for stress of plants. Corn is the first crop worldwide, its yield and quality impact human society survival and development, but corn growth and yield are also severely threatened by drought stress. In long-term drought research, components playing important roles in drought stress response of a plurality of plants have been identified, but the research on identification and action pathways of the components is mainly realized by mode plants such as arabidopsis thaliana, and the like, so that the research on important functions and improvement components of drought stress in important crops such as corn, rice and wheat is relatively less, and the identification of the important components and the drought response pathways in response to the drought stress has more theoretical and practical significance.
The protein coded by the corn Zm00001d018037 gene belongs to a NIP channel protein family member, the CDS length of the corn Zm00001d018037 gene is 885 bp, the protein codes for the NIP channel protein with 295 amino acids, and at present, no research report on the gene in the drought stress reaction of corn is seen in the prior art.
Disclosure of Invention
The technical problem to be solved by the invention is how to alleviate the influence of drought stress on monocotyledonous crops, especially on the growth and development of corn and the yield, namely how to improve the drought stress resistance of corn.
The technical scheme of the invention is as follows: the application of the corn gene Zm00001d018037 in improving the drought resistance of monocotyledonous crops is provided, and the application is preferably in improving the drought resistance of corn.
The invention also provides application of the maize gene Zm00001d018037 in germplasm resource improvement, genetic breeding and preparation of corresponding drought-resistant transgenic crops.
Furthermore, the invention also provides a method for improving the drought stress response of monocotyledonous crops, which integrates the corn gene Zm00001d018037 into cells, tissues or organs of the monocotyledonous crops and enables the monocotyledonous crops to be overexpressed, wherein the monocotyledonous crops are preferably corn.
The invention also provides a method for cultivating the high drought-resistant corn, which comprises the following steps: transforming corn gene Zm00001d018037 into corn young embryo, co-culturing, resistance screening, callus induction, subculture and differentiation to obtain T0 generation transgenic corn, and continuous selfing and target gene PCR detection to obtain stable T 3 And (3) obtaining the maize with the excessive expression of Zm00001d018037, namely the maize with high drought resistance, by using the generation homozygous transgenic line.
Since osmotic stress is one of the most important aspects of plants subjected to drought stress, the invention also provides application of the maize gene Zm00001d018037 in improving the osmotic stress tolerance of yeast cells, so that the maize gene Zm00001d018037 is expressed in the yeast, and the osmotic stress tolerance of the yeast cells is improved.
The invention also provides application of the maize gene Zm00001d018037 in improving seed germination capacity under the osmotic stress of arabidopsis, so that the maize gene Zm00001d018037 is expressed in arabidopsis, and the seed germination capacity under the osmotic stress of transgenic arabidopsis is improved.
The prior art predicts that maize gene Zm00001d018037 encodes an NIP channel protein, but no report has been made regarding its role in plant abiotic stress. The inventor discovers that all the gene editing mutant lines cause triplet codon shift by creating a corn gene Zm00001d018037 gene editing mutant line, and reveals the gene editing mutant line of a corn gene Zm00001d018037 through experiments, which shows a drought stress sensitive phenotype, and the corn gene Zm00001d018037 is overexpressed in transgenic corn to promote the enhancement of drought tolerance of the transgenic corn. The gene plays an important role in drought stress response of plants, so that a new technical means is provided for cultivating excellent crop strains with enhanced drought tolerance by using the gene.
The invention has the following beneficial effects: the invention provides the application of the corn gene Zm00001d018037 in improving the drought resistance of monocotyledonous crops, especially in improving the drought resistance of corn, and in germplasm resource improvement, genetic breeding and preparation of corresponding drought-resistant transgenic crops, and also provides the technical scheme that the expression of the gene in yeast promotes the osmotic stress tolerance of yeast cells to sorbitol, and the overexpression in Arabidopsis promotes the germination of transgenic plants under osmotic stress, and provides a new technical means for cultivating new corn varieties with high drought resistance and innovation of excellent corn germplasm resources, so that the invention has important agricultural application value and wide practical prospect.
Drawings
FIG. 1 is an illustration of sequencing and identification of the basic structure, the gene editing target position and the gene editing mutant Zm00001d018037-d1 strain and the Zm00001d018037-d2 strain of the maize gene Zm00001d 018037; wherein A is the description of the basic structure and the gene editing target position of the corn gene Zm00001d018037, B is the description of the sequencing identification of the gene editing type of the corn gene Zm00001d018037-d1 strain, and C is the description of the sequencing identification of the gene editing type of the corn gene Zm00001d018037-d2 strain.
FIG. 2 is an illustration of the detection of drought tolerance, detection of moisture content of plant tissue before and after severe drought treatment, and detection of accumulation of membranous oxidation products (MDA) of plant tissue before and after severe drought treatment for maize gene Zm00001d018037 gene editing mutants (ZmNIP 2a-d1 and ZmNIP2a-d 2) after severe drought treatment; wherein A is the description of drought tolerance detection of maize gene Zm00001d018037 gene editing mutants (ZmNIP 2a-d1 and ZmNIP2a-d 2) after severe drought treatment, B is the description of water content detection of plant tissues before and after severe drought treatment, and C is the description of accumulation detection of membranous oxidation products (MDA) of plant tissues before and after severe drought treatment.
FIG. 3 is a schematic representation of maize genesZm00001d018037Description of PCR detection of transcript levels in overexpressed maize lines.
FIG. 4 is a schematic representation of maize genesZm00001d018037And (5) gene editing and identification of transgenic super-expression genetic material seedling drought resistance phenotype.
FIG. 5 is a diagram showing that expression of ZmNIP2a in yeast promotes osmotic stress tolerance of yeast.
FIG. 6 is an illustration of germination rate statistics after 4 days of osmotic stress tolerance during germination of ZmNIP2a gene-promoted mode plant Arabidopsis thaliana, and overexpression of ZmNIP2a gene; wherein A is an illustration of osmotic stress tolerance during germination of Arabidopsis thaliana which is a ZmNIP2a gene promotion mode, and B is an illustration of germination rate statistics after 4 days of osmotic stress of Arabidopsis thaliana over-expressed by the ZmNIP2a gene.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The methods used in the examples are conventional methods known to those skilled in the art, and the reagents used, if not specified, are commercially available products. The corn gene editing mutant and the transgenic super-expression material are derived from a corn molecular breeding and gene editing platform in a national key laboratory of the stress adaptation and improvement of co-constructed crops of Henan university, and corn B73-329 is taken as an experimental base material.
For the convenience of expression and simplicity and clarity of experimental records, the corn Zm00001d018037 gene is simply expressed as ZmNIP2a, and the nucleotide sequence is shown as NO.1 in the ST.26 sequence table; the single nucleotide addition (+ bp) gene editing mutant of the maize Zm00001d018037 gene is expressed asZmNIP2a-d1，The nucleotide sequence is shown as NO.2 in a ST.26 sequence table; the deletion (-7 bp) gene editing mutant of the maize Zm00001d018037 gene is expressed asZmNIP2a-d2，The nucleotide sequence is shown as NO.3 in the ST.26 sequence table.
Creating gene editing mutant and transgenic super-expression corn genetic material
(1) According to the gene coding sequence of the corn Zm00001d018037 gene, selecting a target (double target, target 1 sequence: ACGCCGTCGGACACATCTC, target 2 sequence: GAAGATGTCCGCCAGCGAC, wherein target 1 is the main component), designing primers according to the target sequence (four primer sequences are Zm00001d018037MT1T2-F: AATAATGGTCTCAGGCGACGCCGTCGGACACATCTC, zm00001d018037MT1T2-F0: GACGCCGTCGGACACATCTCGTTTTAGAGCTAGAAATAGC, zm00001d018037MT1T2-R0: GTCGCTGGCGGACATCTTCCGCTTCTTGGTGCC, zm00001d018037MT1T2-R: ATTATTGGTCTCTAAACGTCGCTGGCGGACATCTTC) respectively, carrying out four-primer PCR amplification by taking an intermediate vector pCBC-MT1T2 as a template, and purifying a PCR amplified fragment. The amplified fragment was subjected to cleavage ligation with the gene editing final vector pBUE411 in the following manner:
taking 5 mu L of the liquid, enzyme cutting, connecting products to convert competent cells of escherichia coli, culturing overnight on an antibiotic LB culture medium containing kanamycin, and screening positive clones. The positive transformants were cultured overnight (37 ℃ C., 220 rpm) on liquid LB medium. And extracting the successfully connected recombinant plasmid DNA, and sequencing to confirm the existence of the designed target sequence in the recombinant vector. And (3) transferring the successfully constructed gene editing vector into an agrobacterium EHA105 strain, submitting the constructed crop adversity adaptation of the Henan university province and improving a national key laboratory corn molecular breeding and gene editing platform to perform genetic material creation work. And breeding the created gene editing T0 generation material to obtain T1 generation gene editing germplasm. And (5) planting a gene editing strain, taking corn leaves of the strain, and extracting DNA.
The SLS extraction method of the strain corn DNA is as follows: taking 0.1 g of the maize leaves of the strain, placing the maize leaves in a 2 mL centrifuge tube which is sterilized in advance and is provided with small steel balls, and quick-freezing the maize leaves in liquid nitrogen; the quick frozen sample was shaken in a mill at 60 Hz frequency for 1 min, 750 μl of SLS DNA extract (1% SLS, 0.02M EDTA (ph=8.0), 0.1M Tris-HCl (ph=8.0), 0.1M NaCl) was added to the sample, and vigorously shaken to mix it thoroughly. Adding 750 mu L of phenol: chloroform: isoamyl alcohol (25:24:1) and fully shaking, standing for 5 min at room temperature, and centrifuging at high speed for 10 min at 4 ℃ and 12,000 rpm; transferring the supernatant to a centrifuge tube of 1.5 mL, adding equal volume of isopropanol, mixing upside down, and standing in a refrigerator at 4deg.C for 30 min; high-speed centrifugation at 12,000 rpm at 4℃for 10 min to precipitate DNA; washing the DNA with 75% ethanol for 2 times; and (3) dissolving DNA by adding 100 mu L of sterilized ultrapure water, and using the DNA as a template. The amplified DNA fragments were sequenced and aligned by specific primer amplification (forward primer: GCAGTGCGTAGGAAGGAACT; reverse primer: GGTGTGCGTTGTTATACGCC), and the gene editing type of the created gene editing strain was confirmed (as shown in FIG. 1B).
(2) Gene editing mutant drought tolerance detection:
wild type B73-329 andZmNIP2a-d1、ZmNIP2a-d2the mutants are respectively planted in nutrition bowls, the weight of nutrition soil in each nutrition bowl is the same, and the mutants are cultivated in a greenhouse. After 10 days of growth, watering was stopped, drought treatment was performed, and drought phenotype during the treatment was observed. After 5 days of drought treatment, both gene-edited lines exhibited a more severe leaf wilting and curling phenotype, while no leaf curl was apparent in response to controls B73-329 (fig. 2A). The relative water content of the leaf of the plant at this time was examined (as follows), and the relative water content of the leaf of the two gene editing mutants was significantly lower than that of B73-329 (FIG. 2B). Active oxygen accumulation and oxidative damage are one of the important hazards of drought to plants. Detection of B73-329 and after drought treatmentZmNIP2a-d1、 ZmNIP2a-d2The content of Malondialdehyde (MDA) which is a membrane oxidation product in the mutant shows that the MDA content of two gene editing strain tissues after drought treatment is obviously higher than that of a control B73-329 (figure 2C), and the damage of the surface by drought stress is more serious.
The relative water content detection method comprises the following steps: the fresh weight of the plant tissue is weighed and recorded as W1, then the plant tissue is placed in an oven for full drying at 65 ℃, the dry weight is obtained by weighing, W2 is obtained by recording, and the relative water content of the plant tissue can be obtained by using a formula of [ relative water content= (W1-W2)/W1 x 100% ].
MDA (membrane oxidation product malondialdehyde) is measured by weighing 0.2 g experimental material into a centrifuge tube with steel balls in advance, immediately placing into a high-throughput tissue grinder after quick freezing by liquid nitrogen, vibrating and grinding for 1 min at 8,000 rpm, then adding 2 mL of 10% TCA (trichloroacetic acid) solution at 4 ℃ and 4,000 rpm, centrifuging for 10 min, sucking 1 mL supernatant, adding 1 mL of 0.5% TBA solution, uniformly mixing, reacting in a boiling water bath for 15 min, cooling, 4 ℃ and 10,000 rpm, centrifuging for 10 min, transferring 200 mu L of supernatant into an ELISA plate after centrifuging, measuring the absorbance at three positions of 532 nm, 600 nm and 450 nm in the ELISA meter, and calculating MDA content (mu mol/g) =MDA concentration (mu mol/L) multiplied by the volume of extract (mL)/weight of sample (g) according to the following formula.
(3) Transgenic overexpression genetic germplasm creation and expression detection:
using the cDNA of corn wild type B73-329 as a template, a specific primer was designed (primer sequence is as follows:
F: gcgcgccatttaaatactagtATGTCGACCAACTCGAGGTCCAACTC；
r catggtggatcccatactagtCACTTGGATGTGGTCGAGCTCGTCGTC, wherein the lower case letters are the joint sequences on the transgenic super-expression vector, the upper case letters are the primers in the coding sequence of the Zm00001d018037 gene), and PCR amplification and purification are carried out to recover amplified fragments. The amplified fragment recovered by purification was ligated with the transgenic overexpressing empty vector pcmcia 3300 digested with restriction enzymes (single step cloning ligation of novzan). The ligation product was transformed into E.coli competent cells and cultured overnight on a solid LB medium containing kanamycin, thereby selecting positive transformants. And extracting the successfully connected recombinant plasmid DNA and sequencing. And (3) transforming the recombinant vector into an agrobacterium EHA105 strain, submitting the constructed crop adversity adaptation of the Henan university province and improving a national key laboratory corn molecular breeding and gene editing platform to perform transgenic overexpression genetic germplasm creation work.
Recombinant Agrobacterium single colonies were inoculated into 2-3 mL liquid medium containing 100. Mu.g/mL kanamycin and 50. Mu.g/mL rifampicin, and shake-cultured overnight at 28 ℃. The next day was transferred to 50 mL liquid medium containing antibiotics for shake culture and resuspended to an OD600 of between 0.8-1.0 to obtain recombinant Agrobacterium suspension. And (3) infecting the maize immature embryo of B73-329 by adopting the obtained recombinant agrobacterium suspension, and then sequentially carrying out co-culture, resistance screening (the resistance screening adopts herbicide glufosinate), pre-differentiation, differentiation and rooting to obtain a T0 generation regenerated plant. Selfing the transgenic plants in the generation T0 to obtain generation T1 seeds, wherein the plants grown from the generation T1 seeds are generation T1 plants; the plants are selfed until T3 generation seeds are obtained, namely, the T3 generation seeds of the over-expression transgenic plants are obtained, the T3 generation lines obtained after the selfing of different T0 generation regenerated plants are respectively named as OE-1 and OE-2, and in the process, corn B73-329 is used as a control plant, namely, wild WT.
The genetic materials such as wild corn B73-329, transgene over-expression and the like are respectively 4 to 6 strains (OE-1 and OE-2), a few leaves are cut, and total RNA is extracted.
The extraction method of the specific corn RNA comprises the following steps: preparing a 1.5 mL RNase-free centrifuge tube with small steel balls, taking about 0.5 g fresh plant leaves, placing the fresh plant leaves in the centrifuge tube, covering the centrifuge tube with a cover, quick-freezing the fresh plant leaves in liquid nitrogen for 5-10 seconds, immediately placing the fresh plant leaves in a high-throughput tissue grinding instrument, and vibrating and grinding the fresh plant leaves at 8,000 rpm for 1 min to enable the leaves to fully crush powder (wearing masks and gloves in the process to prevent pollution of exogenous RNase); adding 1 mL pre-cooled Trizol on ice into the centrifuge tube, quickly mixing the mixture upside down, and standing the mixture at room temperature for 5 min; adding 200 μl of chloroform, shaking vigorously for 15 s, standing at room temperature for about 5 min, centrifuging at 1, 2000 rpm for 15 min at 4deg.C; separating the mixture into three layers after centrifugation, transferring the upper transparent water phase containing RNA into a 1.5 mL centrifuge tube without RNase, slowly sucking to avoid sucking into the middle and lower organic phase, and discarding the precipitate; adding isopropanol with the same volume, slightly reversing and uniformly mixing, standing for 10-20 min at room temperature, and precipitating RNA; centrifuging at 12,000 rpm and 4 ℃ for 10 min, obtaining RNA by a small amount of white precipitate at the bottom of the centrifuge tube, and discarding the supernatant; adding 1 mL of 75% ethanol, blowing, rinsing and precipitating, centrifuging at 12,000 rpm and 4 ℃ for 5 min, discarding the supernatant, and rinsing twice repeatedly; pouring the centrifuge tube into an ultra-clean bench after uncapping, naturally airing for 10 min, and fully volatilizing alcohol; 50 μl DEPC treated water was added to the centrifuge tube, RNA was fully dissolved, and the concentration was measured for reverse transcription to synthesize cDNA.
Preparation of cDNA: the extracted RNA was mixed in an RNase-free centrifuge tube using the Norpran HiScript 3 RT SuperMix for qPCR kit as follows
Genomic DNA removal system
| System component | Volume of |
| RNase-free ddH2O | to 16 μL |
| 4×gDNA wiper buffer | 4 μL |
| Template RNA | 1 pg-1μg |
After gently stirring and mixing, the mixture was reacted for 2 min at 42℃using a PCR instrument. After the completion of the reaction, 5X HIScript qRT SuperMix was directly added to the reaction tube.
Reverse transcription reaction system
| System component | Volume of |
| 5×HIScript qRT SuperMix | 4 μL |
| Reaction solution of the first step | 16 μL |
After mixing by gentle blowing, the mixture was reacted at 37℃for 15 minutes using a PCR instrument, and reacted at 85℃for 5 s. The product can be used for qPCR reaction immediately after measuring the concentration, or stored at-20 ℃.
Performing reverse transcription to obtain cDNA, performing PCR amplification with specific primer with the sequence of F5'-GTACGTACGTGTGCTAGCTAG-3' and R5'-GATGTCATGGATCTCGTTGTTG-3' to obtain amplified product, detecting the amplified product to obtain the relative expression of transgenic overexpressed strain gene,
the cDNA prepared by the reverse transcription kit is used as a template, and PCR amplification (F: 5'-GTACGTACGTGTGCTAGCTAG-3'; R: 5'-GATGTCATGGATCTCGTTGTTG-3') is performed by using specific primers to perform the identification work of the transcript level of the transgenic overexpressed strain.
The NIP2a transcripts in the overexpressing lines OE-1 and OE-2 were far higher than in the corresponding controls (B73-329), as shown in FIG. 2: the genetic materials such as transgene over-expression and the like reach about 20 times and 25 times higher than the relative expression quantity of wild genes of B73-329 respectively in corn strains OE-1 and OE-2 (figure 3).
Under the condition of the greenhouse, the genetic materials such as the earth cultivated wild corn (B73-329, namely wild WT), corn Zm00001d018037 transgene overexpression and the like reach 4 to 6 strains of corn strains (OE-1 and OE-2) respectively, and for comparison, corresponding gene editing mutant strains are planted at the same timeZmNIP2a-d1；ZmNIP2a-d2). Normally growing for 10 days, then stopping water supply, and performing drought treatment. After 5 days of drought treatment, compared with the wild nature (B73-329, wild nature WT) of the corresponding control material, the corn Zm00001d018037 gene edits nucleotide deletion mutationZmNIP2a-d1The method comprises the steps of carrying out a first treatment on the surface of the Coding sequence shift mutant of corn Zm00001d018037 geneZmNIP2a-d2) Shows a sensitive phenotype with significantly reduced drought tolerance, whereas transgenic overexpressing maize lines (OE-1And OE-2) showed a significant increase in drought tolerance (as shown in FIG. 4), and the above experiment was repeated 8 times with completely consistent results.
The studies of the inventors demonstrate that Zm00001d018037 gene improves drought stress response and promotes osmotic stress tolerance of plants. The functional identification of the gene under drought and osmotic stress of plants shows the innovation of the invention. These achievements have important theoretical and production significance for promoting drought stress tolerance of plants and the like.
Since osmotic stress is one of the most important aspects of drought stress, the present invention also investigated the expression of the maize Zm00001d018037 gene in yeast and promoting the growth of yeast under osmotic stress conditions.
Constructing a ZmNIP2a yeast expression fusion vector (pYES 2) vector system in which the ZmNIP2a gene is driven by GAL promoter and glucose is the promoter inhibitor, so that in a medium containing glucose,ZmNIP2agenes are not normally expressed, and in other glycogen media, such as in galactose-containing media,ZmNIP2acan be expressed, thereby causing the difference of growth vigor under the osmotic stress of yeast.
HandleZmNIP2aConstruction of the Gene coding sequence topYES2In the fusion vector formed, the constructed fusion vector is then transformed into a yeast (INVSV 1) strain. On glucose-containing medium, there was no significant difference in the vigor of sorbitol-treated and untreated transformants, whereas on sorbitol-supplemented galactose yeast medium, along with dilution of the yeast concentration gradient, the vigor of the yeast cells carrying the ZmNIP2a recombinant transformants was significantly better than that of the yeast cells transformed with the pYES2 empty vector, i.e. ZmNIP2a promoted yeast growth under osmotic stress caused by sorbitol. (as shown in fig. 5).
Since osmotic stress is one of the most important aspects of drought stress, the invention also researches the germination condition of the arabidopsis seeds under the osmotic stress condition promoted by the Zm00001d018037 gene of corn.
Method for creating transformation by adopting agrobacterium to infect arabidopsis thalianaZmNIP2aGene overexpression arabidopsis thaliana, and detection of transgenic overexpression arabidopsis thaliana strainPhenotype under osmotic stress. Normal arabidopsis seeds (wild type and transgenic overexpression) were inoculated on MS and ms+sorbitol medium, the phenotype of the seeds germinated for 6 days (a), and germination rate statistics at 4 days (B). The above experimental results show that overexpression in ZmNIP2a arabidopsis thaliana promotes germination under osmotic stress. "x" indicates that the difference is extremely significant.
Compared with wild type Arabidopsis thaliana (Col-0), the transgenic overexpressing Arabidopsis thaliana strain (OE-4, OE-5) exhibited enhanced tolerance to osmotic stress (MS+150 mM sorbitol) in the seed germination stage, and the germination rate of the transgenic overexpressing strain and the like was significantly higher than that of WT when grown for 4 days (FIG. 6).
The foregoing is merely a preferred embodiment of the invention, and it should be noted that modifications could be made by those skilled in the art without departing from the principles of the invention.
Claims (6)
1. The application of the corn gene Zm00001d018037 in improving drought resistance of crops is characterized in that the nucleotide sequence of the corn gene Zm00001d018037 is shown in a sequence table SEQ ID NO.1, the crop corn variety is corn B73-329, and the corn gene Zm00001d018037 is excessively expressed in the corn B73-329.
2. The use according to claim 1, in the improvement of maize germplasm resources.
3. The use according to claim 1, in maize genetic breeding.
4. The use according to claim 1, for the preparation of corresponding drought-resistant transgenic crop maize.
5. A method for improving the drought stress response of crop maize B73-329, characterized in that the maize gene Zm00001d018037 according to claim 1 is integrated into the cells, tissues or organs of crop maize B73-329 and overexpressed.
6. A method for cultivating high drought resistant corn, comprising the steps of: transforming corn gene Zm00001d018037 into corn B73-329 young embryo, co-culturing, resistance screening, callus induction, subculturing and differentiation to obtain T 0 Transgenic corn with stable T is obtained through continuous selfing and target gene PCR detection 3 The generation of homozygous transgenic lines obtains corn which is over-expressed with a corn gene Zm00001d018037, namely the high drought resistant corn, and the nucleotide sequence of the corn gene Zm00001d018037 is shown in a sequence table SEQ ID NO. 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310926763.8A CN116656698B (en) | 2023-07-27 | 2023-07-27 | Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310926763.8A CN116656698B (en) | 2023-07-27 | 2023-07-27 | Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116656698A CN116656698A (en) | 2023-08-29 |
| CN116656698B true CN116656698B (en) | 2023-11-03 |
Family
ID=87724491
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310926763.8A Active CN116656698B (en) | 2023-07-27 | 2023-07-27 | Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116656698B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118562873A (en) * | 2024-08-01 | 2024-08-30 | 河南大学三亚研究院 | Application of corn gene Zm00001d053828 in improving drought resistance of crop corn |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105586349A (en) * | 2016-03-14 | 2016-05-18 | 中国科学院遗传与发育生物学研究所 | Drought-resistant salt-tolerant gene TaPIP1A of wheat and encoded protein and application of TaPIP1A |
| CN105713078A (en) * | 2016-04-28 | 2016-06-29 | 中国农业科学院作物科学研究所 | Application of drought-tolerance-related protein in regulating drought tolerance of plants |
| CN106191076A (en) * | 2016-07-26 | 2016-12-07 | 江苏省农业科学院 | Plant PIP1；10 genes and application thereof |
| AR110831A2 (en) * | 2018-01-16 | 2019-05-08 | Evogene Ltd | METHOD FOR IMPROVING THE CONDITIONS OF A PLANT SUBMITTED TO ADVERSE FACTORS, ISOLATED MOLECULE THAT INCLUDES A CONSTRUCTION OF NUCLEIC ACID AND ISOLATED VEGETABLE CELL THAT INCLUDES AN EXOGEN POLYETHYPE |
| CN112322654A (en) * | 2020-10-16 | 2021-02-05 | 山东大学 | Application of corn transcription factor ZmMYB42 gene in plant drought-resistant breeding |
| CN112795552A (en) * | 2021-03-10 | 2021-05-14 | 河南大学 | Application of Zm0001d024568 gene and encoding protein thereof in drought stress resistance of corn |
| CN115700282A (en) * | 2022-12-21 | 2023-02-07 | 河南大学三亚研究院 | Corn drought-resistant gene Zm00001d030678 and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7589257B2 (en) * | 2006-02-09 | 2009-09-15 | Pioneer Hi-Bred International Inc. | Genes for enhancing nitrogen utilization efficiency in crop plants |
-
2023
- 2023-07-27 CN CN202310926763.8A patent/CN116656698B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105586349A (en) * | 2016-03-14 | 2016-05-18 | 中国科学院遗传与发育生物学研究所 | Drought-resistant salt-tolerant gene TaPIP1A of wheat and encoded protein and application of TaPIP1A |
| CN105713078A (en) * | 2016-04-28 | 2016-06-29 | 中国农业科学院作物科学研究所 | Application of drought-tolerance-related protein in regulating drought tolerance of plants |
| CN106191076A (en) * | 2016-07-26 | 2016-12-07 | 江苏省农业科学院 | Plant PIP1；10 genes and application thereof |
| AR110831A2 (en) * | 2018-01-16 | 2019-05-08 | Evogene Ltd | METHOD FOR IMPROVING THE CONDITIONS OF A PLANT SUBMITTED TO ADVERSE FACTORS, ISOLATED MOLECULE THAT INCLUDES A CONSTRUCTION OF NUCLEIC ACID AND ISOLATED VEGETABLE CELL THAT INCLUDES AN EXOGEN POLYETHYPE |
| CN112322654A (en) * | 2020-10-16 | 2021-02-05 | 山东大学 | Application of corn transcription factor ZmMYB42 gene in plant drought-resistant breeding |
| CN112795552A (en) * | 2021-03-10 | 2021-05-14 | 河南大学 | Application of Zm0001d024568 gene and encoding protein thereof in drought stress resistance of corn |
| CN115700282A (en) * | 2022-12-21 | 2023-02-07 | 河南大学三亚研究院 | Corn drought-resistant gene Zm00001d030678 and application thereof |
Non-Patent Citations (9)
| Title |
|---|
| 1, ZmNIP2 ; 4 and ZmTIP4 ; 4 involved in urea transport.BMB reports.2012,96-101. * |
| A Nucleoporin NUP58 modulates responses to drought and salt stress in maize (Zea mays L.);Zhixue Liu等;Plant Science;1-12 * |
| Enhanced Drought Stress Tolerance by the Arbuscular Mycorrhizal Symbiosis in a Drought-Sensitive Maize Cultivar Is Related to a Broader and Differential Regulation of Host Plant Aquaporins than in a Drought-Tolerant Cultivar;Gabriela Quiroga等;Frontiers in Plant Science;1-15 * |
| Functional divergence of the NIP III subgroup proteins involved altered selective constraints and positive selection;Qingpo Liu等;BMC Plant Biology;1-12 * |
| Riliang Gu等.Isolation and characterization of three maize aquaporin genes, ZmNIP2 * |
| Targeted mining of drought stress-responsive genes from EST resources in Cleistogenes songorica;Jiyu Zhang等;Journal of Plant Physiology;1844-1851 * |
| The function of high-affinity urea transporters in nitrogen-deficient conditions;Marcel P. Beier等;Physiologia Plantarum;802-808 * |
| Using the Maize Nested Association Mapping (NAM) Population to Partition Arbuscular Mycorrhizal Effects on Drought Stress Tolerance into Hormonal and Hydraulic Components;Juan Manuel Ruiz-Lozano等;Int. J. Mol. Sci.;1-25 * |
| Zea mays NOD26-like membrane integral protein ZmNIP2-1 (LOC542643), mRNA;Gu R等;Genbank;FEATURES和ORIGIN部分 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116656698A (en) | 2023-08-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107541520B (en) | OsSAUR11 gene related to rice root development and stress resistance, coding protein and application | |
| CN116656698B (en) | Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops | |
| CN111826381B (en) | Centipede grass root promoting gene EoSINAT5, plant expression vector and application thereof | |
| CN108715852B (en) | Tomato fruit mature gene Sl0658 and application thereof | |
| CN111424037B (en) | Cymbidium CgWRKY70 gene and application thereof | |
| CN110004173B (en) | Method for obtaining non-transgenic storable fresh-eating Chinese wolfberry | |
| CN116064572B (en) | MdWOX11 gene and protein for promoting adventitious root development and application thereof | |
| CN113234720B (en) | Wheat long-chain non-coding RNAlncR156 and application thereof in regulation and control of wheat response to drought stress | |
| CN113337522B (en) | Application of cotton GhNFYC4 gene in promoting plant flowering | |
| CN111454966B (en) | Cymbidium CgWRKY4 gene and application thereof | |
| CN111304222B (en) | Cymbidium CgWRKY11 gene and application thereof | |
| CN111304220B (en) | Cymbidium CgWRKY3 gene and application thereof | |
| CN111304223B (en) | Cymbidium CgWRKY24 gene and application thereof | |
| CN109628468A (en) | A kind of Chunlan CgWRKY53 gene and its application | |
| CN111424040B (en) | Cymbidium CgWRKY21 gene and application thereof | |
| CN114644701B (en) | Use of proteins derived from corn and related biomaterials | |
| CN111424039B (en) | Cymbidium CgWRKY65 gene and application thereof | |
| CN114672498B (en) | Dragonfly pineapple AfCAL gene, cloning method, expression vector and application | |
| CN111424041B (en) | Cymbidium CgWRKY49 gene and application thereof | |
| CN111424038B (en) | Cymbidium CgWRKY40 gene and application thereof | |
| CN116640769B (en) | Peanut AhGATA gene and application thereof in improving stress resistance of plants | |
| CN111500624B (en) | Use of CrSMT genes to increase resistance of plants to biotic and abiotic stress | |
| CN113173981B (en) | Phyllostachys pubescens PeDREB3 gene and application thereof in plant cold resistance regulation | |
| CN111304221B (en) | Cymbidium CgWRKY31 gene and application thereof | |
| CN110982921B (en) | Application of cymbidium miR159a in accelerating plant life cycle |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |