CN110759979B - Transcription factor for improving starch synthesis of wheat grainsbZIP2And uses thereof - Google Patents
Transcription factor for improving starch synthesis of wheat grainsbZIP2And uses thereof Download PDFInfo
- Publication number
- CN110759979B CN110759979B CN201910833620.6A CN201910833620A CN110759979B CN 110759979 B CN110759979 B CN 110759979B CN 201910833620 A CN201910833620 A CN 201910833620A CN 110759979 B CN110759979 B CN 110759979B
- Authority
- CN
- China
- Prior art keywords
- gene
- tubzip2
- tabzip
- wheat
- transcription factor
- 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.)
- Expired - Fee Related
Links
- 229920002472 Starch Polymers 0.000 title claims abstract description 60
- 239000008107 starch Substances 0.000 title claims abstract description 58
- 235000019698 starch Nutrition 0.000 title claims abstract description 58
- 235000021307 Triticum Nutrition 0.000 title claims abstract description 51
- 108091023040 Transcription factor Proteins 0.000 title claims abstract description 39
- 102000040945 Transcription factor Human genes 0.000 title claims abstract description 39
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 20
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 20
- 241000209140 Triticum Species 0.000 title claims abstract 6
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 89
- 230000014509 gene expression Effects 0.000 claims abstract description 44
- 108091026890 Coding region Proteins 0.000 claims abstract description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 5
- 244000098338 Triticum aestivum Species 0.000 claims description 66
- 230000002018 overexpression Effects 0.000 claims description 30
- 230000009261 transgenic effect Effects 0.000 claims description 10
- 239000013604 expression vector Substances 0.000 claims description 7
- 108091033409 CRISPR Proteins 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
- 238000010354 CRISPR gene editing Methods 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 claims description 2
- 238000009825 accumulation Methods 0.000 claims 2
- 230000001737 promoting effect Effects 0.000 claims 1
- 230000001131 transforming effect Effects 0.000 claims 1
- 235000013339 cereals Nutrition 0.000 abstract description 47
- 229920000945 Amylopectin Polymers 0.000 abstract description 25
- 230000002829 reductive effect Effects 0.000 abstract description 14
- 241000196324 Embryophyta Species 0.000 abstract description 12
- 230000008859 change Effects 0.000 abstract description 7
- 102000004169 proteins and genes Human genes 0.000 abstract description 5
- 229920000856 Amylose Polymers 0.000 description 18
- 239000013598 vector Substances 0.000 description 17
- 230000000694 effects Effects 0.000 description 16
- 238000004458 analytical method Methods 0.000 description 12
- 238000003757 reverse transcription PCR Methods 0.000 description 10
- 238000000034 method Methods 0.000 description 9
- 230000033228 biological regulation Effects 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 8
- 238000013518 transcription Methods 0.000 description 8
- 230000007423 decrease Effects 0.000 description 7
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000331 Firefly luciferases Proteins 0.000 description 5
- 108010052090 Renilla Luciferases Proteins 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 239000005089 Luciferase Substances 0.000 description 4
- 108090000848 Ubiquitin Proteins 0.000 description 4
- 102000044159 Ubiquitin Human genes 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000010195 expression analysis Methods 0.000 description 4
- 235000013312 flour Nutrition 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 210000004940 nucleus Anatomy 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 210000001938 protoplast Anatomy 0.000 description 4
- 230000002441 reversible effect Effects 0.000 description 4
- 238000010200 validation analysis Methods 0.000 description 4
- 108700008625 Reporter Genes Proteins 0.000 description 3
- 108010039811 Starch synthase Proteins 0.000 description 3
- 102000039554 bZIP family Human genes 0.000 description 3
- 108091067354 bZIP family Proteins 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000004186 co-expression Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000036961 partial effect Effects 0.000 description 3
- 230000004960 subcellular localization Effects 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 3
- 108090000344 1,4-alpha-Glucan Branching Enzyme Proteins 0.000 description 2
- 102000003925 1,4-alpha-Glucan Branching Enzyme Human genes 0.000 description 2
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 2
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000004568 DNA-binding Effects 0.000 description 2
- 244000077283 Distichlis palmeri Species 0.000 description 2
- 235000005422 Distichlis palmeri Nutrition 0.000 description 2
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 2
- 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 2
- 108060001084 Luciferase Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 238000003559 RNA-seq method Methods 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 230000009418 agronomic effect Effects 0.000 description 2
- 125000003275 alpha amino acid group Chemical group 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 210000005069 ears Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 230000010474 transient expression Effects 0.000 description 2
- 229940100445 wheat starch Drugs 0.000 description 2
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 238000003718 Dual-Luciferase Reporter Assay System Methods 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241001442556 Gloma Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- HXXFSFRBOHSIMQ-VFUOTHLCSA-N alpha-D-glucose 1-phosphate Chemical compound OC[C@H]1O[C@H](OP(O)(O)=O)[C@H](O)[C@@H](O)[C@@H]1O HXXFSFRBOHSIMQ-VFUOTHLCSA-N 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000007036 catalytic synthesis reaction Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 239000004464 cereal grain Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 229950010772 glucose-1-phosphate Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 235000006486 human diet Nutrition 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- PWPJGUXAGUPAHP-UHFFFAOYSA-N lufenuron Chemical compound C1=C(Cl)C(OC(F)(F)C(C(F)(F)F)F)=CC(Cl)=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F PWPJGUXAGUPAHP-UHFFFAOYSA-N 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000003012 network analysis Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000005758 transcription activity Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000000196 viscometry Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
Images
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
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
- C12N15/8245—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Nutrition Science (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Cell Biology (AREA)
- Mycology (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (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 discloses a transcription factor capable of improving the starch synthesis of wheat grainsbZIP2And uses thereof, the transcription factor isTubZIP2A gene, andTubZIP2gene homology gene in common wheatTabZIP2Three copies of (a):TabZIP‑A2、TabZIP‑ B2andTabZIP‑D2the coding region consists of nucleotide sequences shown by SEQ ID No.1, No.2, No.3 and No.4 respectively. In thatTubZIP2In the case of the over-expressed wheat line of (1),AGPL‑cyto、AGPS‑ cytoandSBEIIathe expression of the protein is obviously improved in the middle and later periods of endosperm filling compared with the wild type, while the expression quantity of other Starch Synthesis Related Genes (SSRG) is not changed, and the grain length and thousand kernel weight are also obviously increased. Knocking out genes in common wheatTabZIP2The total starch and amylopectin content of the knockout line is obviously reduced, and the grain length and thousand grain weight are also obviously reduced.TubZIP2Over-expression lines andTabZIP2the plant height, effective tillering, small spike number and grain length of the knockout line have no obvious change.
Description
Technical Field
The invention relates to the field of genetic engineering, in particular to a transcription factor for improving the expression of wheat starch synthesis related genes and application thereof.
Background
Wheat, one of the major crops in the world, not only provides protein, vitamins and dietary fiber for human health, but also is a major source of starch and energy in the human diet. Starch is the main storage component in the endosperm of wheat grains, accounting for 60-70% of the total weight of grains and 70-85% of the total weight of flour.
Starch synthesis in wheat endosperm cells involves a complex series of reactions requiring the involvement of a series of enzymes. At the start of starch synthesis, AGPase catalyzes the conversion of glucose-1-phosphate (G1 c-1-P) and Adenosine Triphosphate (ATP) to adenosine diphosphateAcid glucose (ADP-glucose), is the glucose donor for amylose and amylopectin synthesis. Thereafter, granule-bound starch synthase (GBSS) catalyzes the ADP-glucose to finally form amylose, while Starch Synthase (SS), Starch Branching Enzyme (SBE) and debranching enzyme (DBE) catalyze the ADP-glucose transfer to highly branched glucan branches to finally form amylopectin. The content of the starch of the grains can be effectively improved by regulating and controlling the expression of the starch synthesis related gene, thereby improving the yield of wheat. However, to date, there have been few reports on transcriptional regulation of starch synthesis in cereal grains. WhereinRSR1AndOsbZIP58is a starch synthesis regulation factor identified in rice;ZmNAC36、ZmbZIP91、ZmEREB156and SUSIBA2are transcription factors that regulate starch synthesis identified in maize and barley, respectively. In wheat, except for riceRSR1Homologous geneTaRSR1There is no report on the systematic study of starch synthesis regulation mechanism.
Disclosure of Invention
The invention identifies a transcription factor Co-expressed with Starch Synthesis Related Genes (SSRG) in endosperm filling period by Weighted Gene Co-expression Network Analysis (WGCNA)TubZIP2。TubZIP2And homologous gene thereof in common wheatTabZIP2Can be remarkably improvedAGPL-cyto、AGPS-cytoAndSBEIIathe expression of the gene further improves the total starch and amylopectin content of the wheat grains. At the same time, the user can select the desired position,TubZIP2andTabZIP2has no any adverse effect on other important agronomic traits of wheat.
The invention is helpful for people to understand the regulation and control of wheat starch synthesis,TubZIP2andTabZIP2can also be used as an important target gene for genetic engineering and traditional breeding to culture a new wheat variety with high yield and high quality.
The inventor identifies and obtains the genome data analysis of endosperm of the grain filling stage of the Wulare diagram wheat G1812TubZIP2Has similar expression pattern with partial SSRG gene. The TubZIP2 subcellular location is in nucleus, and 1-128 aa thereof hasTranscriptional activation activity. In the dual-luciferase reporter gene system analysis and the wheat endosperm transient overexpression analysis,TubZIP2promotion ofAGPL-cyto、AGPS-cytoAndSBEIIaactivity of a gene promoter. In thatTubZIP2In the case of the over-expressed wheat line of (1),AGPL-cyto、AGPS-cytoandSBEIIathe expression of (a) is obviously improved in the middle and later stages of endosperm filling compared with the wild type, and the expression quantity of other SSRG genes is not changed. Correspondingly, the total starch content and the amylopectin content of the overexpression line are obviously improved, and the grain length and the thousand grain weight of the overexpression line are also obviously increased. In contrast, knockouts using CRISPR/Cas9 technologyTubZIP2Homologous genes in common wheatTabZIP2The total starch content and amylopectin content of the knockout line are obviously reduced, and the grain length and thousand grain weight are also obviously reduced.TubZIP2Over-expression lines andTabZIP2the plant height, effective tillering, small spike number and grain length of the knockout line have no obvious change. In any case, it is preferable that,TubZIP2andTabZIP2promotes the synthesis of the starch of the wheat grains, and has no negative influence on other important agronomic traits.
The invention aims to provide a transcription factor for regulating and controlling the expression of an SSRG geneTubZIP2The transcription factor promotes starch synthesis.
The transcription factor provided by the inventionTubZIP2Is derived from wheat with a Wulare diagram and has a coding region consisting of a nucleotide sequence shown as SEQ ID No. 1. The DNA shown in SEQ ID No.1 has a length of 1722 bp (excluding stop codon), belongs to the bZIP family, and encodes a protein of 574 amino acid residues.TubZIP2The homologous gene in common wheat isTabZIP2It has three copiesTabZIP-A2、TabZIP-B2AndTabZIP-D2the coding regions are respectively nucleotide sequences shown in SEQ ID No.2, SEQ ID No.3 and SEQ ID No.4, the length of the coding regions is 1722 bp, and 574 protein with amino acid residues is coded.
Meanwhile, the present invention provides a recombinant vector comprising the transcription factor of claim 1.
The invention also provides an expression vector containing the transcription factor, preferably a plant expression vector.
The invention provides an application of the transcription factor in improving the expression of SSRG genes in the filling stage in wheat; the application is to transform the transcription factor into wheat and screen a positive transgenic line over expressing the transcription factor.
The invention also provides a method for over-expressing the transcription factor in endosperm, which is to transform the endosperm of common wheat by using an over-expression vector containing the transcription factor, preferably to transform the endosperm 15 days after the common wheat blossoms by using a gene gun method.
Also, the present invention provides amplificationTubZIP2AndTabZIP2the universal primer pair of the full length of the coding region, wherein the sequence of the forward primer is shown as SEQ ID No.5, and the sequence of the reverse primer is shown as SEQ ID No. 6.
The invention also provides a method for determining and regulating the candidate transcription factor expressed by the SSRG gene by utilizing co-expression analysis, which finds the transcription factor with a similar expression mode to the SSRG gene by analyzing the transcriptome of the endosperm of the wheat in the filling stage of the Ural chart through RNA-Seq.
The invention provides a method for detecting the regulation intensity of a transcription factor on an SSRG gene promoter region, which comprises the steps of co-transforming an overexpression vector of the transcription factor and a report vector of the SSRG gene promoter driving the expression of Firefly luciferase (Firefly luciferase) gene into a wheat protoplast cell, and detecting the intensity of the activity of the transcription factor regulating the SSRG gene promoter region by taking Renilla luciferase (Renilla luciferase) as an internal reference.
The invention provides a method for detecting the regulation and control of SSRG gene expression by transcription factor through endosperm overexpression, which comprises the steps of converting an overexpression vector of the transcription factor into endosperm 15 days after common wheat blossoms by a gene gun method, carrying out dark culture in a hypertonic culture medium for 48 hours, and detecting the change of SSR gene expression in the endosperm by RT-PCR.
The invention provides a Ular pattern wheatTubZIP2Can be used for up-regulating immature seeds of common wheatAGPL-cyto、AGPS-cytoAndSBEIIathe expression of the gene and finally the content of the total starch and the amylopectin of the mature seeds are improved. Its homologous gene in common wheatTabZIP2Has the same function. Due to the fact thatIn this way, the temperature of the molten steel is controlled,bZIP2has important significance for improving the starch quality of wheat.
Drawings
FIG. 1 shows a schematic view of aTubZIP2Evolutionary tree analysis, subcellular localization, transcriptional activation activity and expression pattern analysis. Wherein, A is the structure of TubZIP2, wherein 128-179 amino acids (aa) represent the core region of bZIP family transcription factor; b is an evolutionary tree analysis displayTubZIP2And homologous genes thereof are conserved in wheat families, and have a far relation with homologous genes from other families of the gramineae family; c is the N-terminal (1-127 aa) of TubZIP2 with transcriptional activation activity; d is TubZIP2 located in cell nucleus; e isTubZIP2In thatT. urartuPerforming specific expression in mature endosperm; f isTubZIP2And a part of the SSRG gene inT. urartuCo-expression in the grouting process.
FIG. 2TubZIP2In vitro enhancementAGPL-cyto、AGPS-cytoAndSBEIIatranscription of the gene. A is a schematic diagram of a report vector, an expression vector and an internal reference in a dual-luciferase reporter gene system, wherein REN: renilla luciferase; LUC: firefly luciferase; ubi: ubiquitin; p: a promoter; ter: and (6) terminating. B isTubZIP2Significant improvement in dual luciferase reporter systemsAGPL-cyto、AGPS-cytoAndSBEIIapromoter activity of the gene; c isTubZIP2A schematic representation of the overexpression of the vector in the endosperm; d isTubZIP2Increased overexpression in endospermAGPL-cyto、AGPS-cytoAndSBEIIatranscription of the gene.
FIG. 3TubZIP2Increasing wheat over-expression in linesAGPL-cyto、AGPS-cytoAndSBEIIathe level of transcription of the gene. A and B represent the results of RNA-seq and RT-PCR, respectivelyTubZIP2Overexpression in the immature endosperm of transgenic wheat; C. e and G are in endosperm 7 days, 14 days and 21 days after anthesis in RNA-seq analysis, respectivelyAGPL-cyto、AGPS-cytoAndSBEIIaa change in the level of transcription; D. f and H represent endosperm 7 days, 14 days and 21 days after RT-PCR validationAGPL- cyto、AGPS-cytoAndSBEIIachange in the amount of gene expression.
FIG. 4TubZIP2The total starch content and the amylopectin content of the overexpression system are improved. A isTubZIP2Remarkably improving the total starch content of the over-expression line planted in the 2017-plus 2018 Beijing, the dike and the Zhao county; b, finding that the number of A-type starch granules in the over-expression line seeds planted in Zhao county is more than that of wild type grains in scanning comparison by an electron microscope, wherein A and B respectively represent A-type and B-type starch granules; C. d and E are the amylopectin content, the amylose content and the ratio of amylose to amylopectin of the three-site overexpression lines, respectively; f is the higher peak viscosity of the overexpression line of Beijing in RVA analysis compared with the wild type.
FIG. 5 TubZIP2The thousand kernel weight and the grain width of the over-expression line grains are improved. A and B are respectively wild type and overexpression line plant and spike morphology; C. d and E are respectively the plant height, effective tillering and the number of spikelets per spike of the wild type and the overexpression line; f is the grain length and grain width shape of wild type and over-expression line grains; g isTubZIP2No influence on the grain length of the transgenic line; h and I are eachTubZIP2The grain width and thousand grain weight of the over-expression line are improved.
FIG. 6 TabZIP2In vitro enhancementAGPL-cyto、AGPS-cytoAndSBEIIatranscription of the gene. A isTabZIP2Specific expression in common wheat endosperm; b isTabZIP2AndAGPL-cyto、AGPS-cytoandSBEIIaco-expression; c isTabZIP2A schematic representation of the overexpression of the vector in the endosperm; d isTabZIP2Increased overexpression in endospermAGPL-cyto、AGPS- cytoAndSBEIIatranscription of the gene.
FIG. 7TabZIP2The total starch content and amylopectin content of the knockout line were reduced. A is Beijing, on the dike and Zhao countyTabZIP2The total starch content of the knockout system is obviously reduced; b, finding that the number of A-type starch grains in the knock-out seeds planted in Zhao county is less than that of wild type grains in electron microscope scanning comparison, wherein A and B respectively represent A-type and B-type starch grains; c is the amylopectin content of the three-site knockout line; d is the lower peak viscosity of the Beijing-planted knockout line compared to the wild type in the Rapid Viscometry (RVA) analysis; e and F show a significant reduction in grain width and thousand grain weight, respectively, for the knockout line.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 Ural wheatTubZIP2And the evolutionary tree analysis, the subcellular localization, the transcriptional activation activity and the co-expression analysis of the SSR genes, and specifically comprises the following steps:
(1) wheat (Triticum aestivum L.)bZIP2Cloning of the genes: according toTRIUR3_00571The 5 'end and the 3' end of (3) and designed amplificationTubZIP2The primer of the coding region (the sequence of the forward primer is shown as SEQ ID No.5, the sequence of the reverse primer is shown as SEQ ID No. 6), and the cDNA of endosperm 15 days after the wheat blossom of the Ural chart is used as a template to clone to obtain the geneTubZIP2And the sequence is shown as SEQ ID No. 1. According toTubZIP2Sequence search IWGSC BLAST (https:// urgi. versalles. inra. fr/BLAST _ IWGSC @
dbgroup=wheat_iwgsc_refseq_v1_chromosomes&program = blastn) and WheatExp BLAST (https:// was. pw. usda. gov/WheatExp /), usingTubZIP2The full-length primer takes the cDNA of the endosperm 15 days after the spring bloom of common wheat as a template,TubZIP2the full-length primer (the sequence of the forward primer is shown as SEQ ID No.5, and the sequence of the reverse primer is shown as SEQ ID No. 6) is subjected to PCR amplification and monoclonal sequencing to obtain the common wheatTabZIP2Three copies ofTabZIP-A2、TabZIP-B2AndTabZIP-D2the sequences of the two are respectively shown as SEQ ID No.2, SEQ ID No.3 and SEQ ID No. 4.
(2) And (3) analyzing the evolutionary tree: will be provided withTubZIP2AndTabZIP2and the amino acid sequences of genes with more than 80% sequence similarity in GenBank are compared by Clustal Omega (http:// www.ebi.ac.uk/Tools/msa/clustalo /) software, and then a evolutionary tree is constructed by Mega 5.05 (http:// www.megasoftware.net /), and the results are shown in FIG. 1B,TubZIP2and the homologous genes in the wheat family are independently gathered into a branch in the evolutionary tree analysis and are far away from the homologous genes from other families, which indicates thatbZIP2The genes are conserved in the wheat family.
(3) Detection of transcriptional activation Activity:TubZIP2the amino acid sequence was determined by NCBI Protein BLAST (https:// blast.ncbi.nlm.nih.gov/blast.cgi)
PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK _ LOC = blastome), and analysis shows that TubZIP2 is a transcription factor of bZIP family, and comprises a basic leucoine zipper (bZIP) structural domain (128-179 aa), and the result is shown in FIG. 1A, wherein 128-179 aa of TubZIP2 is a DNA binding domain (bZIP domain). Recombinant plasmids were constructed by combining the full-length of TubZIP2 and the coding region sequences corresponding to fragments 1-127 aa, 128-179 aa and 180-574 aa, respectively, with pGBKT7 (Clontech, California, USA). Transformation of yeast strains AH109 (Clontech, California, USA) with the recombinant plasmids the results are shown in FIG. 1C, all strains grow normally on solid medium (SD/-Trp), but only strains transformed with the complete gene and the recombinant plasmid for the transcriptional activation region can turn X-gal blue, whereas strains transformed with the recombinant plasmid for the DNA binding region cannot turn blue. This indicates that TubZIP2 has transcriptional activation activity, and its N-terminal 1-127 aa functions for transcriptional activation activity.
(4) Subcellular localization: will be provided withTubZIP2The recombinant plasmid pJIT163-UBI-TubZIP2-hGFP fused with GFP is constructed by connecting to a pJIT163-UBI-hGFP vector, the protoplast cells of wheat leaves are transformed, and the expression of TubZIP2 fusion is realized under the drive of a ubitin promoter. The results are shown in FIG. 1D, TubZIP2 localized to the nucleus of protoplast cells.TubZIP2Localization in the nucleus indicates that it functions in the nucleus, consistent with the characteristics of transcription factors.
(5) Tissue-specific expression analysis: extracting total RNA of root, stem, leaf and endosperm 15 days after wheat blossom of Ural chart by Trizol methodTubZIP2Specific primers, their tissue-specific expression was verified by RT-PCR, the results are shown in FIG. 1E,TubZIP2expressed in large amounts in the endosperm.
(6) Wheat (Triticum aestivum L.)bZIP2Verification of Gene and SSRG Gene expression patterns: in the endosperm of the wheat in the filling stage of the Ural chart, the use ofTubZIP2And specific primers of partial SSRG genes, and the expression patterns of the two genes are verified by RT-PCR. The results are shown in figure 1F of the drawings,TubZIP2and expression of the selected SSRG gene peaked 5 days after the flower and then gradually decreased, which is consistent with the results of RNA-Seq.
Example 2 common wheatTabZIP2And tissue-specific expression and in vitro promotion of the SSRG GeneAGPL-cyto、AGPS-cytoAndSBEIIatranscription of (4).
(1) Tissue-specific expression analysis: extracting total RNA of root, stem, flag leaf, ear and endosperm of plant 15 days after spring bloom of common wheat TabZIP-A2、TabZIP-B2AndTabZIP-D2the specific primers of (1) were verified for their tissue-specific expression by RT-PCR, and the results are shown in FIG. 6A, in roots, stems, flag leaves, ears 15 days after flowering,TabZIP-B2there is a small amount of expression of the gene,TabZIP-A2andTabZIP-D2trace expression is present; in the endosperm 15 days after the flower,TabZIP-A2、TabZIP-B2andTabZIP-D2are abundantly expressed and are expressed inTabZIP-B2The expression level of (2) is highest.
（2）TabZIP2Validation of gene and SSRG gene expression patterns: RT-PCR is utilized to detect the endosperm of common wheat in the China spring filling stageTabZIP2And the expression pattern of the SSRG gene, as shown in FIG. 6B,TabZIP-A2、 TabZIP-B2andTabZIP-D2has the same expression pattern with partial SSRG gene, namely starting from 5 days after the flower and reaching the peak value 10 days after the flower, and then the expression quantity begins to decline; this also suggests that these three copies are involved in the regulation of the expression of the SSRG gene in Triticum aestivum.
Example 3 wheatbZIP2Function of the Gene
1. Wheat (Triticum aestivum L.)bZIP2Direct regulation of the SSRG Gene
(1) Dual luciferase reporter gene system detectionTubZIP2Regulation of SSR gene promoter: driven by the ubiquitin promoterTubZIP2(Ubi P:: TubZIP 2) in large scale, and constructionTubZIP2The expression vector of (1); promoter using SSRG Gene (-2000 bp to-1 bp)) Drives the expression of Firefly luciferase (Firefly luciferase) gene, constructs an expression vector (SSRG P:: LUC), and uses the vector as a reporter vector. The two recombinant vectors are co-transformed into wheat protoplast cells, a Renilla luciferase (Renilla luciferase) overexpression vector (Ubi P:: REN) is used as a control, the vector schematic diagram is shown in 2A, and LUC/REN represents the expression activity of an SSR gene promoter. Fluorescence intensity was measured using the Dual-Luciferase Reporter Assay System kit (Promega, Madison, USA) and Gloma 20/20 Luminometer (Promega, Madison, USA). The results are shown in figure 2B which shows,TubZIP2significantly improves the SSRG geneAGPL-cyto、AGPS-cytoAndSBEIIathe promoter activity of (1).
（2）bZIP2Endosperm over-expression of (a): the pJIT163-UBI-hGFP vector (ampicillin resistance), supplied by Cabernet Gaussler, institute of genetics and developmental biology, of the Chinese academy of sciences, driven the expression of GFP by the ubiquitin promoter. Will be provided withTubZIP2、TabZIP-A2、TabZIP-B2AndTabZIP-D2respectively recombined in pJIT163-UBI-hGFP vector, and the ubiquitin promoter in the vector is drivenbZIP2For high-level expression, the vector schemes are shown in FIGS. 2C and 6C, respectively.
Disinfecting seeds of common wheat 15 days after spring flowering in China with 70% alcohol, bombarding endosperm with a gene gun to realize transient expression of the recombinant plasmid in the endosperm, and then culturing the recombinant plasmid in a hypertonic culture medium for 48 hours in a dark place; extracting total RNA of endosperm, and performing reverse transcription to obtain cDNA; RT-PCR validationTubZIP2、TabZIP-A2、TabZIP-B2AndTabZIP-D2the transient expression and its effect on the expression of the SSRG gene are shown in FIG. 2D: (TubZIP2) It is shown that, compared with the wild-type endosperm introduced with the empty plasmid,TubZIP2can be obviously improvedAGPL-cyto、AGPS-cytoAndSBEIIa the promoter activity of (a);TabZIP2the results of (a) are shown in figure 6D,TabZIP-A2、TabZIP-B2andTabZIP-D2can both up-regulate the expression of the SSRG gene, in whichTabZIP-B2The strength of the up-regulation of (a) is the maximum,TabZIP-D2the effect is not obvious.
Wheat (Triticum aestivum L.)TubZIP2Overexpression line RNA-seq and phenotype identification (1) wheat transgenic plant screening:construction and utilizationGlu-1Bx14Promoter driveTubZIP2The over-expression carrier makes the transcription factor specifically express in kernel endosperm, and makes gene gun transformation on the transgenic platform of institute of genetics and developmental biology of Chinese academy of sciences, the obtained transformed seedling is planted in greenhouse, and according to the conventional method, its genome DNA is slightly extracted, 100 ng of genome DNA is taken as template, and the primer pair (shown by SEQ ID No.5 and SEQ ID No. 6) is used, and utilizes the wild type wheat genome DNA as control to make conventional PCR amplification so as to further detect positive plant. Homozygous selfing to T2Transgenic wheat of the generations was used for subsequent experiments.
(2) And (3) taking materials of endosperm 7 days, 14 days and 21 days after the flowering respectively, quickly freezing and storing the materials by using liquid nitrogen, placing the materials in a refrigerator at the temperature of-80 ℃, extracting RNA by using Trizol, and performing transcriptome sequencing (RNA-seq) on three repeated samples. Simultaneously reverse transcribing the RNA to cDNATa4045(ubiquinol-cytochrome C reaction enzyme ron-sulfurr subunit) as a control, and the expression of the SSRG gene was verified by RT-PCR using a primer specific to the SSRG gene. The results are shown in FIG. 3A and FIG. 3B, and the results of RNA-seq and RT-PCR validation both showTubZIP2Expression was significantly upregulated in all three phases; FIGS. 3C and 3D showTubZIP2No obvious regulation effect is realized 7 days after the flowers bloom; as shown in figures 3E and 3F,TubZIP2can be remarkably increased at 14 days after the flower blossomAGPL-cyto、AGPS-cytoAndSBEIIaexpression of the gene; FIGS. 3G and 3H show that the gene has reduced regulation and is mainly up-regulated 21 days after floweringSBEIIaExpression of the gene.
(3) And (3) determining the total starch content of the mature seeds: total starch content was determined by means of a Total starch kit (K-TSTA; Megazyme). The results are shown in fig. 4A, where the total starch content of wheat over-expressed lines was significantly increased in three locations, beijing, hebeibei embankment and zhao county, compared to wild type, with an average increase of about 2.88%, 4.93% and 4.38% for three lines OE82, OE85 and OE87, respectively.AGPS-cytoAndAGPL-cytois the rate-limiting enzyme in the starch biosynthesis pathway, and the increase in total starch content is associated with an increase in the transcription level of both transgenic lines.
(4) Scanning mature seeds by an electron microscope: and transversely cutting the mature seeds of the wild wheat and the over-expression wheat by using a blade, and scanning the transverse section by using an electron microscope. As shown in fig. 4B, the over-expressed line had more type a starch granules, which is consistent with an increase in total starch content.
(5) And (3) determining the amylose content of the mature seeds: and (3) measuring the amylose content of the over-expression system and wild flour by using a DPCZ-II type amylose analyzer, wherein the difference value of the total starch content and the amylose content is the amylopectin content. As shown in FIG. 4C, the amylopectin content of the three site-overexpressed lines was significantly higher than that of the wild type, with an average increase of 4.79% for OE82 and 8.06% and 7.50% for OE85 and OE87, respectively, which is associated with the resultsSBEIIaThe catalytic synthesis of the amylopectin is consistent. There was no difference in amylose content on the hebei embankment and the zhao county (9.57% reduction in OE87 in zhao county), but the amylose content of OE82, OE85 and OE87 in the beijing area was reduced by 7.15%, 5.02% and 5.60%, respectively, as shown in fig. 4D. The values for three over-expressed amylose/amylopectin from three sites were significantly reduced due to the increased amylopectin content and the steady and even reduced amylose content (no significant change in amylose/amylopectin from OE82 on embankment and zhao county), as shown in fig. 4E.
(6) Overexpression lines mature seed RVA characteristics: rapid Viscometers (RVA) are widely used to evaluate the gelatinization properties of starch, and the peak viscosity produced by RVA is inversely related to the amylose to amylopectin ratio, consistent with a decrease in the straight-to-branched ratio, as shown in fig. 4F, with a significant increase in the peak viscosity for both over-expressed lines of wheat grown in beijing. .
(7) Plant phenotype identification of over-expressed line wheat: the characteristics of transgenic wheat in three places are investigated, and as shown in fig. 5A, 5B and 5F, the characteristics of plants, ears and grains of wild type and overexpression lines are respectively represented; FIGS. 5C, 5D and 5E show no significant differences in plant height, effective tillering and number of spikelets per plant and wild type, respectively, for the over-expressed lines; fig. 5G, 5H and 5I show grain length, grain width and thousand kernel weight, respectively, of the kernel, where there was no significant difference in grain length, and both grain width and thousand kernel weight of the over-expressed lines were higher than wild type. Wherein the grain width increase amplitude of OE85 is maximum and is as high as 6.62%, and the increase amplitude of OE82 is minimum and is only 3.72%. The increase in grain width caused a change in thousand kernel weight of the kernels, with an average increase in the thousand kernel weight of OE85 of the most, up to 13.50%, and an average increase in OE82 and OE87 of 6.98% and 11.46%.
Wheat (Triticum aestivum L.)TabZIP2Phenotypic characterization of knockout lines
(1) Determining the total starch content of mature seeds of the knockout line: 2017 and 2018 wheat planted in Beijing, Hebei dyke and Zhao countyTabZIP2Knock-out line T2The passage line was used for subsequent assays. Total starch content was determined by means of a Total starch kit (K-TSTA; Megazyme). As shown in FIG. 7A, the total starch content of the knockout lines in Beijing, Dingshang and Zhao county locations was significantly lower than that of the wild type, bZIP2kn-ab 、bZIP2kn-abd1 and bZIP2knAn average reduction of 4.18%, 5.69% and 5.75% for-abd 2.
(2) Electron microscopy scanning of knockout line mature seeds: mature seeds of wild wheat and knockout wheat are transversely cut by a blade, the transverse section is subjected to electron microscope scanning, and the A, B type starch grain distribution of the seeds is changed along with the reduction of total starch. As shown in fig. 7B, the knockout line contained fewer a-type starch grains.
(3) Determining the amylose content of mature seeds of the knockout line: measuring amylose content of wild type and knockout line flour with DPCZ-II type amylose analyzer, wherein the difference between total starch content and amylose content is amylopectin content, as shown in FIG. 7C, the amylopectin content of knockout line at three positions except bZIP2 on dikeknNo apparent ab change, three lines in the other three sites showed a decrease, bZIP2kn-ab average decrease 3.42%, bZIP2kn-abd1 and bZIP2knThe decrease of-abd 2 was 5.57% and 10.10%, respectively. Reduction of Total starch content and amylopectin content and knock-out linesTabZIP2To pairTaAGPL-cyto、TaAGPS-cytoAndTaSBEIIathe transcriptional activity was consistently decreased. (4) RVA characteristics of knockout line mature seeds: analysis of the flour viscosity by RVA, as shown in figure 7D, the peak viscosity of the knockout line was significantly lower than the wild type. A decrease in amylopectin content in the knockout line causes a significant decrease in the peak viscosity of the RVA.
(5) Knockout line mature seed kernelShape: as shown in FIGS. 7E and 7F, the grain width and thousand grain weight of the three spot knockout lines were significantly lower than the wild type, where bZIP2kn-ab、bZIP2kn-abd1 and bZIP2knThe average grain width of abd2 decreased by 3.79%, 5.40% and 5.78%, corresponding to an average reduction in thousand kernel weight of 9.19%, 14.78% and 15.30%.TabZIP2In knockout linesTaAGPL-cyto、TaAGPS-cytoAndTaSBEIIathe transcription activity of the compound enzyme is reduced, so that the total starch content and the amylopectin content of mature grains are reduced, the grain width of the grains is obviously reduced, and the thousand grain weight of the grains is further obviously reduced compared with the wild type.
<110> institute of genetics and developmental biology of Chinese academy of sciences
<120> transcription factor bZIP2 for improving total starch content and amylopectin content of wheat grains and application thereof
<160> 6
<210> 1
<211> 1722
<212> CDS
<213> TubZIP2, Triticum aestivum L.)
<400> 1
ATGGCCATCG CGGAGGCGTC CCCCTTCGCG GACCTGCCCT TCCCCGACGA CCTGCCGGAG TTCCCGCACG GCCCCGTCGG GGACGACGAC GCCTTCGCCC TGGACGGCTT CGATCTCGAC GATCTGGACA TCGACTTCGA CTTCGACCTC GACCTCGACC TCCTCCCCAC CGACGACGTG CGGCTCCCGT CGCCGCCGCC GCCGCTCGCC ACGTCCTCCT CCTCGGCCGG GTCGCCGGGC GGGGCAGGGG ACTCCTCCTC CGGCTCCGGT GGCGGAGCGG ACGGCGGCCT GAAGAACGAC GAGTCCTCGG AGACGTCTTC GAGGAGCGCG AGCGCTGGGA GCGACGGCAA GGCTAAGGAG GGGGAGGGTG AGGACGACAA GCGGCGGGCG CGGCTGGTGC GGAACCGGGA GAGCGCGCAC CTGTCGCGGC AGAGGAAGAA GCAGTACGTG GAGGAGCTGG AGGGGAAGGT CAAGGCCATG CAGGCCACCA TCGCCGACCT CTCCACCAGG ATCTCCTGCG TCACTGCTGA GAACGCCGCT CTCAAGCAGC AACTGGCCGG TGCTGGTGGA GCAGGCGTCC CGCCGCCGCT GCCGATGTAC CCGGGATTGT ACCCTTTGCC GCTGCCATGG ATGCACCCGG CTTATGCGAT GGGAGCGCGC GGCTCCCAAG TGCCGCTCAT GCCGATACCT CGGCTGAAAA CCAAGCAGCC TGCGTCGGCT GCCGCAGAGC CACCGGCCAA GAAGTCTAGG AAGACCAAGA AGGTTGCCAG TGTTAGCCTC CTTGGATTGC TGTTCCTGAC GATGCTCTGT GGGTGTTTGG TTCCTGCGGT AAATCGGATG TATGGAGCAG TTGATTCCCG AGAAGGAATT GTGCTTGGTC CATCACAATC ACGTCATGGG AGGGTTCTGG CTGTTGACGG GCCTCGAGAT GGTGTCCCGG AAGGTGTTGA TTCGAAGTTG CCACATAATT CAAGTGAGAC GCTCCCGGCC TTGTTGTATA TTCCAAGGAA TGGGAAGCAT GTCAAGATCA ATGGAAATCT TGTTATCCAG TCTGTTGTTG CGAGTGAGAA AGCTTCTTCG CGCATGTGTC ACTCTGATGG GAAGACTTCA TGTAACCAAG GACAAGAAGA TACTAGTTTG GCAATTCCTG GCCATGTTGC TCAGTTGAAT TCTGGAGAAG TCATGGAATC TGCTAAAGCA ATAAAAAATA AACTGATGGC TTTACCTCCT GGAGATGGAA GCATATACAG AGACGATGAT GAATTACTGC CACAATGGTT TAGTGAAGCA ATGTCAGGTC CTATGTTGAG TTCCGGAATG TGCACCGAAG TGTTCCAGTT TGATATATCG CCGACCACCA TTGTTCCTGT CTACTCCAGT GGTATGCACA ATGCATCACA TAACTCCACG GAGAACCTCC CCTCCAGTCA GTCCCATAAG GTCAAGAACA GAAGGATTTT ACATACCATG GCCGTTCCCC TAAAAGGTTC AACGTCCAAC CACACCGATC ACCTCAAAGC GCACCCCAAG AACGAGAGCT TTGCTGGAAA GAAACCGGCT TCATCGGTGG TGGTCTCTGT CCTGGCTGAC CCTAGAGTGG ATGCTGATGG AAGGATCTCT TCGAAGTCAT TGTCGCGTAT ATTTGTTGTA GTCCTCGTTG ACAGTGTAAA GTATGTCACT TACTCTTGCG TCCTGCCGTT CAAAACCCAT AGCCCTCATC TG
<210> 2
<211> 1722
<212> CDS
<213> TabZIP-A2, Triticum aestivum L.)
<400> 2
ATGGCCATCG CGGAGGCGTC CCCCTTCGCG GACCTGCCCT TCCCCGACGA CCTGCCGGAG TTCCCGCACG GCCCCGTCGG GGACGACGAC GCCTTCGCCC TGGACGGCTT CGATCTCGAC GATCTGGACA TCGACTTCGA CTTCGACCTC GACCTCGACC TCCTCCCCAC CGACGACGTG CAGCTCCCGT CGCCGCCGCC GCCGCTCGCC ACGTCCTCCT CCTCGGCCGG GTCGCCGGGC GGGGCAGGGG ACTCCTCCTC CGGCTCCGGT GCCGGAGCGG ACGGCGGCCT GAAGAACGAC GAGTCCTCGG AGACGTCTTC CAGGAGCGCG AGCGCTGGAA GCGACGGCAA GGCTATGAAC GGGGAGGGTG AGGACGACAA GCGGCGGGCG CGGCTGGTGC GGAACAGGGA GAGCGCACAC CTGTCGCGGC AGAGGAAGAA GCAGTACGTG GAGGAGCTGG AGGGGAAGGT CAAGGCCATG CAGGCCACCA TCGCCGACCT CTCCACCAGG ATCTCCTGCG TCACGGCTGA GAACGCCGCT CTCAAGCAGC AACTGGCCGG TGCCGGTGGC GCAGGCGTCC CGCCGCCGCT GCCGATGTAC CCTGGATTGT ACCCTTTGCC GCTGCCATGG ATGCACCCGG CTTATGCGAT GGGAGCGCAC GGCTCCCAAG TGCCGCTCAT GCCGATACCT CGGCTGAAAA CCAAGCAGCC TGCGTCGGCT GCCGCAGAGC CACCGGCCAA GAAGTCTAGG AAGACCAAGA AGGTTGCTAG TGTTAGCCTC CTTGGATTGC TGTTCCTGAT GATGCTCTGT GGGTGTTTGG TTCCTGCGGT AAATCGGATG TATGGAGCAG TTGATTCCCG AGAAGGAATT GTGCTTGGTC CATCACAATC ACGTCATGGG AGGGTTCTGG CTGTTGACGG GCCTCGAGAT GGTGTCTTGG AAGGTGTTGA TTCGAAGTTG CCGCATAATT CAAGTGAGAC GCTCCCGGCG TTGTTGTATA TTCCTAGGAA TGGGAAGCAT GTCAAGATCA ATGGAAATTT GGTTATCCGG TCTGTTGTTG CGAGTGAGAA AGCCTCTTCG CGCATGTGTC ACTCTGATGG GAAGACTTCA TGTAACCAAG GACAAGAAGA TACTAGTTTG GCAATTCCTG GCCATGTTGC TCAGTTGAAT TCTGGAGAAG TCATGGAATC TGCTAAAGCA ATAAAAAATA AACTGATGGC TTTACCTCCT GGAGATGGAA GCATATACAG AGACGATGAT GAATTACTGC CACAATGGTT TAGTGAAGCA ATGTCAGGTC CTATGTTGAG CTCCGGAATG TGCACCGAAG TGTTCCAGTT TGATATATCG CCGACCACCA TTGTTCCTGT CTACTCCAGT GGTATGCACA ACGCATCACA TAACTCCACG GAGAACCTCA CCTCCAGTCA GTCCGATAAG GTCAAGAACA GAAGGATTTT ACATTCCATG GCCGTTCCCC TAAAAGGTTC AACGTCCAAC CACACCGATC ACCTCAAAGC GCACCCCAAG AACGAGAGCT TTGCTGGAAA CAAACCGGCT TCATCAGTGG TGGTCTCTGT CCTGGCTGAC CCGAGAGTGG ACGCTGATGG AAGGATCTCT TCGAAGTCAT TGTCGCGTAT ATTTGTTGTA GTCCTTGTTG ACAGTGTAAA GTATGTCACT TACTCTTGCG TCCTGCCGTT CAAAACCCAT AGCCCTCATC TG
<210> 3
<211> 1722
<212> CDS
<213> TabZIP-B2, Triticum aestivum L.)
<400>3
ATGGCCCTCG CGGAGGCGTC CCCCTTCGCG GACCTGCCCT TCCCCGACGA CCTGCCGGAG TTCCCGCACG GCCCCGTCGG GGACGACGAC GCCTTCGCCC TGGACGGCTT CGATCTCGAC GATCTGGACA TCGACTTCGA CTTCGACCTC GACCTCGACC TCCTCCCCAC CGACGACGTG CAGCTCCCGT CGCCCCCACC GCCGCTCGCC ACGTCCTCGT CCTCGGCCGG GTCGCCGGGC GGGGCAGGGG ACTCCTCCTC CGGCTCAGGT GGCGGAGCGG ACGGCGCCCT GAAGAACGAC GAGTCCTCGG AGACGTCTTC CAGGAGCGCG AGCGCTGGGA GCGACGGCAA GGCTAAGGAC GGGGAGGGTG AGGACGACAA GCGGCGGGCG CGGCTGGTGC GGAACCGGGA GAGCGCGCAT CTGTCGCGGC AGAGGAAGAA GCAGTACCTC GAGGAGCTGG AGGGGAAGGT CAAAGCCATG CAGGCCACCA TCGCCGACCT CTCCACCAGG ATCTCCTGCG TCACTGCCGA GAACGCTGCT CTCAAGCAGC AGCTGGCTGG CGCCGGTGGC GCAGGCGTCC CCCCGCCGCT GCCGATGTAC CCCGGATTGT ACCCTTTGCC ACCGCCATGG ATGCACCCTG CTTATGCGAT GGGAGCGCGC GGCTCCCAAG TGCCGCTCAT GCCCATACCT CGGCTGAAAA CCAAGCAGCC TGCGTCGGCT GCCGCAGAGC CACCGGCCAA GAAGTCTAGG AAGACCAAGA AGGTTGCCAG TGTTAGCCTC CTTGGATTGC TGTTCCTGAT GATGCTCTGC GGGTGTTTGG TTCCTGCAGT AAATCGGATG TATGGACCAG TTGATTCCCG AGAAGGAATT GTGCTTGGTC CTTCACAATC ACGTCATGGG AGGGTTCTGG CTGTTGATGG GCCTCGAGAT GGTGTCTCGG AAGGTGTTGA TTCGAAGTTG CCACATAATT CAAGTGAGAC GCTCCCGGCA TTGTTGTATA TTCCGAGGAA TGGGAAACAT GTCAAGATCA ATGGTAATCT TGTTATCCAG TCTGTTGTTG CGAGTGAGAA AGCTTCTTCA CGCATGTGTC ACTCTGATGG GAAGACTTCA TGTAACCAAG GACAAGAAGA TACTAGTTTG GCAATTCCTG GCCACGTTGC TCAGTTGAAT TCTGGAGAAG TCATGGAATC TGCTAAAGCA ATAAAAAATA AACTGATGGC TTTACCTCCT GGAGATGGAA GCATATACAG AGAGGATGAT GAATTACTGC CACAATGGTT TAGTGAAGCA ATGTCAGGTC CTATGTTGAG CTCTGGAATG TGCACCGAAG TGTTCCAGTT TGATATATCG CCAACCACCA TTGTTCCTGT CTACTCCAGT GGTATGCACA ACGCATCACA TAACTCCACG GAGAACCTCC CCTCCAGTCA GTCCCATAAG GTCAAGAACA GAAGGATTTT ACATTCCATG GCCATTCCCC TAAAAGGTTC AACGTCCAAC CACACCGATC ACCTCAAAGC GCACCCCAAG AACGAGAGCT TTGCTGGAAA CAAACCGGCT TCATCGGTGG TGGTCTCTGT CCTGGCTGAC CCTAGAGTGG ATGCTGATGG AAGGATCTCT TCGAAGTCAT TGTCGCGTAT ATTTGTTGTA GTCCTTGTTG ACAGTGTAAA GTATGTCACT TACTCTTGCG TCCTGCCGTT CAAAACCCAT AGCCCTCATC TG
<210> 4
<211> 1722
<212> CDS
<213> TabZIP-D2, Triticum aestivum L.)
<400> 4
ATGGCCATCG CGGAGGCGTC CCCCTTCGCG GACCTGCCCT TCCCCGACGA CCTGCCGGAG TTCCCGCACG GCCCCGTCGG GGACGACGAC GCGTTCGCCC TGGACGGCTT CGATCTCGAC GATCTGGACA TCGACTTCGA CTTCGACCTC GACCTCGACC TCCTCCCCAC CGACGACGTG CAGCTCCCGT CGCCGCCGCC GCCGCTCGCC ACGTCCTCGT CCTCGGCCGG GTCGCCGGGC GGGGCAGGGG ACTCCTCCTC CGGCTCCGGT GGCGGAGCGG ACGGCGGCCT GAAGAACGAC GAGTCCTCGG AGACGTCTTC CAGGAGCGCG AGCGCTGGGA GCGACGACAA GGCTAGGGAT GGGGAGGGTG AGGACGCCAA GCGGCGCGCG CGGCTGGTGC GGAACAGGGA GAGCGCGCAC CTGTCGCGGC AGAGGAAGAA GCAGTACGTG GAGGAGCTGG AGGGGAAGGT CAAAGCCATG CAGGCCACCA TCGCCGATCT GTCCACCAGG ATCTCCTGTG TCACCGCCGA GAACGCTGCT CTCAAGCAGC AACTGGCTGG CGCAGGTGGT GCAGGGGTCC CGCCGCCGCT TCCTATGTAC CCAGGATTGT ACCCTTTGCC ACCGCCATGG ATGCACCCGG CTTATGCGAT GGGAGCGCGC GGCTCCCAAG TGCCGCTCAT GCCGATACCT CGGCTGAAAA CCAAGCAGCC TGCGTCGGCT GCCGCAGAGC CACCGGCCAA GAAGTCCAGG AAGACCAAGA AGGTTGCGAG TGTCAGCCTC CTTGGATTGT TGTTACTGAT GATGCTCTGC GGGTGTTTGG TTCCTGCGGT AAATCGGATG TATGGAGCAG TTGATACTCG AGAAGGAATT GTGCTTGGTC CATCACAATC ACGTCATGGG AGGGTTCTGG CTGTTGATGG GCCTCGAGAT GGTGTCTCGG AAGGTGTTGA TTCGAAGCTG CCACATAATT CAAGTGAAAA GCTCCCGGCG TTGTTGTATA TTCCAAGGAA TGGGAAGCAT GTCAAGATCA ATGGAAATCT TGTTATCCAG TCTGTTGTTG CGAGTGAGGA AGCTTCTTCG CGCATGTGTC ACTCTGATGG GAAGACTTCA TGTAACCAAG GGCAAGAAGA TACTAGTTTG GCAATTCCTG GCCATGTTGC TCAGTTGAAT TCTGGAGAAG TCATGGAATC TGCCAAAGCA ATAAAAAACA AACTGATGGC TTTACCTCCT GGAGATGGAA GCATATACAG AGACGATGAT GAATTACTGC CACAATGGTT TAGTGAAGCA ATGTCAGGTC CTATGTTGAG CTCCGGAATG TGCACCGAAG TGTTCCAGTT CGATATATCA CCGACCACCA TTGTTCCTGT CTACTCCAGT GGTATGCACA ACGCATCACA TAACTCCACG GAGAACCTCC CCTCCAGTCA GTCCCATAAG GTCAAGAACA GAAGGATTTT ACATTCCATG GCCATTCCCC TAAAAAGTTC AACGTCCAAC CACACCGATA ACCTCAAAGC GCACCCCAAG AACGAGAGCT TTGCTGGAAA CAAACCGGCT TCATCGGTGG TGGTCTCTGT CCTGGCTGAC CCTAGAGTGG ATGCTGATGG AAGGATCTCT TCGAAGTCAT TGTCGCGTAT ATTTGTTGTG GTCCTTGTTG ACAGTGTAAA GTATGTCACT TACTCTTGCG TCCTGCCGTT CAAAACCCAT AGCCCTCATC TG
<210> 5
<211> 20
<212> DNA
<213> artificially synthesized sequence
<400> 5
ATGGCAGACCACCTTCAAGT
<210> 6
<211> 22
<212> DNA
<213> artificially synthesized sequence
<400> 6
TCAGTACTTCCACATGCCATCC
Claims (5)
1. The application of the transcription factor for improving the synthesis of the seed starch in improving the expression of the SSRG gene in the filling stage in wheat is characterized in that: the transcription factor isTubZIP2A gene or isTubZIP2Gene homology gene in common wheatTabZIP2Two copies of (a):TabZIP-A2、TabZIP-B2saidTubZIP2Gene, gene,TabZIP-A2Gene, gene,TabZIP-B2The coding region of the gene consists of nucleotide sequences shown by SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3 respectively.
2. The use of claim 1, wherein: transforming the transcription factor into wheat, and screening a positive transgenic line over expressing the transcription factor.
3. The application of the transcription factor for improving the synthesis of the seed starch in improving the expression of the SSRG gene and promoting the accumulation of the starch in the wheat is characterized in that: converting the over-expression vector of the transcription factor into common wheat, and screening to obtain a positive transgenic line with increased starch content; the transcription factor isTubZIP2A gene or isTubZIP2Gene homology gene in common wheatTabZIP2Two copies of (a):TabZIP-A2、TabZIP-B2saidTubZIP2Gene, gene,TabZIP-A2Gene, gene,TabZIP-B2The coding region of the gene consists of nucleotide sequences shown by SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3 respectively.
4. Use according to claim 3, characterized in that: the transcription factor isTubZIP2。
5. The application of the transcription factor for improving the synthesis of the seed starch in reducing the expression of the SSRG gene and the starch accumulation in common wheat is characterized in that: the transcription factor isTubZIP2A gene or isTubZIP2Gene homology gene in common wheatTabZIP2Two copies of (a):TabZIP-A2、TabZIP-B2saidTubZIP2Gene, gene,TabZIP-A2Gene, gene,TabZIP-B2The coding regions of the genes respectively consist of nucleotide sequences shown in SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3, and the genes are knocked out by using CRISPR/Cas9 technologyTabZIP2Two copies of (a):TabZIP-A2、TabZIP-B2and screening out positive transgenic lines.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910833620.6A CN110759979B (en) | 2019-09-04 | 2019-09-04 | Transcription factor for improving starch synthesis of wheat grainsbZIP2And uses thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910833620.6A CN110759979B (en) | 2019-09-04 | 2019-09-04 | Transcription factor for improving starch synthesis of wheat grainsbZIP2And uses thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110759979A CN110759979A (en) | 2020-02-07 |
| CN110759979B true CN110759979B (en) | 2021-09-03 |
Family
ID=69329420
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910833620.6A Expired - Fee Related CN110759979B (en) | 2019-09-04 | 2019-09-04 | Transcription factor for improving starch synthesis of wheat grainsbZIP2And uses thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110759979B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114807163B (en) * | 2022-03-28 | 2023-08-25 | 河南农业大学 | Application of TaSBE I Gene in Promoting Wheat Starch Synthesis |
| CN117866983B (en) * | 2024-03-11 | 2024-05-07 | 浙江大学海南研究院 | Application of OsbZIP10 gene in regulation of amylose content of rice grains |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1860231A (en) * | 2003-06-06 | 2006-11-08 | 阿博根有限公司 | Transcription factors |
| CN101563461A (en) * | 2006-08-02 | 2009-10-21 | 克罗普迪塞恩股份有限公司 | Plants having improved characteristics and a method for making the same |
| CN101792773A (en) * | 2009-05-07 | 2010-08-04 | 山东大学 | Application of sh2 and bt2 genes in enhancing starch content in corn seeds and increasing weight of seeds |
| CN102471374A (en) * | 2009-07-07 | 2012-05-23 | 巴斯夫植物科学有限公司 | Plants having modulated carbon partitioning and a method for making the same |
| CN103160541A (en) * | 2011-12-09 | 2013-06-19 | 中国科学院上海生命科学研究院 | Transcription factor for regulating physical and chemical properties of plant seed starch |
| CN103619164A (en) * | 2011-04-22 | 2014-03-05 | 巴斯夫植物科学有限公司 | Plants having enhanced yield-related traits and method for making the same |
| CN104725493A (en) * | 2013-12-19 | 2015-06-24 | 中国科学院上海生命科学研究院 | Endosperm starch synthesis related transcription factor and application thereof |
| CN105753952A (en) * | 2016-01-29 | 2016-07-13 | 北京市农林科学院 | Plant drought tolerance related protein Tabzip174 as well as coding gene and application thereof |
| WO2017117633A1 (en) * | 2016-01-07 | 2017-07-13 | Commonwealth Scientific And Industrial Research Organisation | Plants with modified traits |
| CN107058343A (en) * | 2017-06-21 | 2017-08-18 | 四川农业大学 | Applications of the transcription factor VP1 in regulation and control crop plant height and Grain starch content |
| CN107586324A (en) * | 2016-07-06 | 2018-01-16 | 中国农业科学院作物科学研究所 | TabZIP15 albumen and its encoding gene and application |
| CN108424918A (en) * | 2018-04-04 | 2018-08-21 | 河北省农林科学院粮油作物研究所 | It is a kind of inhibit wheaten starch synthesis transcription factor WSR1 genes and its application |
-
2019
- 2019-09-04 CN CN201910833620.6A patent/CN110759979B/en not_active Expired - Fee Related
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1860231A (en) * | 2003-06-06 | 2006-11-08 | 阿博根有限公司 | Transcription factors |
| CN101563461A (en) * | 2006-08-02 | 2009-10-21 | 克罗普迪塞恩股份有限公司 | Plants having improved characteristics and a method for making the same |
| CN101792773A (en) * | 2009-05-07 | 2010-08-04 | 山东大学 | Application of sh2 and bt2 genes in enhancing starch content in corn seeds and increasing weight of seeds |
| CN102471374A (en) * | 2009-07-07 | 2012-05-23 | 巴斯夫植物科学有限公司 | Plants having modulated carbon partitioning and a method for making the same |
| CN103619164A (en) * | 2011-04-22 | 2014-03-05 | 巴斯夫植物科学有限公司 | Plants having enhanced yield-related traits and method for making the same |
| CN103160541A (en) * | 2011-12-09 | 2013-06-19 | 中国科学院上海生命科学研究院 | Transcription factor for regulating physical and chemical properties of plant seed starch |
| CN104725493A (en) * | 2013-12-19 | 2015-06-24 | 中国科学院上海生命科学研究院 | Endosperm starch synthesis related transcription factor and application thereof |
| WO2017117633A1 (en) * | 2016-01-07 | 2017-07-13 | Commonwealth Scientific And Industrial Research Organisation | Plants with modified traits |
| CN105753952A (en) * | 2016-01-29 | 2016-07-13 | 北京市农林科学院 | Plant drought tolerance related protein Tabzip174 as well as coding gene and application thereof |
| CN107586324A (en) * | 2016-07-06 | 2018-01-16 | 中国农业科学院作物科学研究所 | TabZIP15 albumen and its encoding gene and application |
| CN107058343A (en) * | 2017-06-21 | 2017-08-18 | 四川农业大学 | Applications of the transcription factor VP1 in regulation and control crop plant height and Grain starch content |
| CN108424918A (en) * | 2018-04-04 | 2018-08-21 | 河北省农林科学院粮油作物研究所 | It is a kind of inhibit wheaten starch synthesis transcription factor WSR1 genes and its application |
Non-Patent Citations (10)
| Title |
|---|
| bZIP转录因子在植物生长发育及非生物逆境响应的作用;刘慧洁等;《浙江农业学报》;20190624;第31卷(第7期);1205-1214 * |
| Pivotal role of bZIPs in amylose biosynthesis by genome survey and transcriptome analysis in wheat (Triticum aestivum L.) mutants;Pankaj Kumar等;《Scientific Reports》;20181122;第8卷(第1期);1-15 * |
| PREDICTED: Aegilops tauschii subsp. tauschii bZIP transcription factor 60-like (LOC1 09734334), mRNA;Genbank Database;《Genbank》;20170224;XM_020293548 * |
| Triticum aestivum bZIP family transcription factor bZIP-A2 (bZIP-A2) mRNA, complete cds;Luo, G.等;《Genbank》;20200122;MN022880 * |
| Triticum aestivum bZIP family transcription factor bZIP-B2 (bZIP-B2) mRNA, complete cds;Luo,G.等;《Genbank》;20200122;MN022881 * |
| Triticum aestivum bZIP family transcription factor bZIP-D2 (bZIP-D2) mRNA, complete cds;Luo,G.等;《Genbank》;20200122;MN022882 * |
| Triticum aestivum bZIP transcription factor mRNA, complete cds;Geng,X.L.等;《Genbank》;20160301;KT753298 * |
| Triticum urartu cultivar G1 81 2 unplaced genomic scaffold scaffold45505, whole genome shotgun sequence;Ling,H.Q.等;《Genbank》;20150320;KD034294 * |
| TubZIP28, a novel bZIP family transcription factor from Triticum urartu, and TabZIP28, its homologue from Triticum aestivum, enhance starch synthesis in wheat;Yanhong Song等;《New Phytologist》;20200118;第226卷(第5期);1384-1398 * |
| 小麦淀粉合成酶的研究进展;廖金花等;《贵州农业科学》;20141231;第42卷(第2期);18-22 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110759979A (en) | 2020-02-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wei et al. | GRAIN INCOMPLETE FILLING 2 regulates grain filling and starch synthesis during rice caryopsis development | |
| Robson et al. | Leaf senescence is delayed in maize expressing the Agrobacterium IPT gene under the control of a novel maize senescence‐enhanced promoter | |
| Sun et al. | Characterization and ectopic expression of CoWRI1, an AP2/EREBP domain-containing transcription factor from coconut (Cocos nucifera L.) endosperm, changes the seeds oil content in transgenic Arabidopsis thaliana and rice (Oryza sativa L.) | |
| Chávez-Bárcenas et al. | Tissue-specific and developmental pattern of expression of the rice sps1 gene | |
| Li et al. | Enhancement of grain number per spike by RNA interference of cytokinin oxidase 2 gene in bread wheat | |
| CN101970668A (en) | Plants with modified starch metabolism | |
| JP2009509509A (en) | Nucleic acids and proteins associated with sucrose degradation in coffee | |
| CN110759979B (en) | Transcription factor for improving starch synthesis of wheat grainsbZIP2And uses thereof | |
| CN107937411A (en) | Chinese white poplar PtoWRKY40 genes, its expression vector and construction method and application | |
| Gallagher et al. | Cloning and characterization of a putative fructosyltransferase and two putative invertase genes from the temperate grass Lolium temulentum L. | |
| Yang et al. | PvNAC1 and PvNAC2 are associated with leaf senescence and nitrogen use efficiency in switchgrass | |
| Wang et al. | A plastidic ATP/ADP transporter gene, IbAATP, increases starch and amylose contents and alters starch structure in transgenic sweetpotato | |
| CN105647940B (en) | The method and its application of OsGRF6 gene raising rice yield | |
| CN113862265A (en) | Method for improving rice grain shape and appearance quality | |
| CN103172715A (en) | Plant epidermal hair controlling gene and application thereof | |
| CN110777150B (en) | Application of protein GmPLATZ in regulation and control of plant seed yield | |
| CN116218876A (en) | Gene OsB12D3 for regulating rice chalkiness, encoding protein and application thereof | |
| CN102373224A (en) | Auxin transport protein gene of paddy rice and application thereof | |
| CN104805100B (en) | Paddy gene OsS μ 2 applications in plant leaf blade aging is delayed of BP | |
| CN105636974A (en) | Expression of thermostable starch synthase genes improves the yield in heat stress | |
| CN108165557A (en) | Application of the wheat TaZCCT2 genes in the flowering of plant time is regulated and controled | |
| CN108341857A (en) | A kind of and rice yield GAP-associated protein GAP and its encoding gene and application | |
| US11124802B2 (en) | Modulating plant abiotic stress responses using the Kanghan gene family | |
| CN104774853A (en) | Rice root regulation function and application of E3 ubiquitin ligase gene OsPIW | |
| CN115216455B (en) | Application of ENB1 gene and encoding protein thereof in regulation and control of plant grain size and grain weight |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210903 |