CN117187259A - Gene for regulating plant growth and photosynthesis under high-temperature stress condition, and encoding protein and application thereof - Google Patents
Gene for regulating plant growth and photosynthesis under high-temperature stress condition, and encoding protein and application thereof Download PDFInfo
- Publication number
- CN117187259A CN117187259A CN202311157424.4A CN202311157424A CN117187259A CN 117187259 A CN117187259 A CN 117187259A CN 202311157424 A CN202311157424 A CN 202311157424A CN 117187259 A CN117187259 A CN 117187259A
- Authority
- CN
- China
- Prior art keywords
- aghsfa1
- gene
- plants
- plant
- heat
- 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.)
- Granted
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 68
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 18
- 230000029553 photosynthesis Effects 0.000 title claims abstract description 11
- 238000010672 photosynthesis Methods 0.000 title claims abstract description 11
- 230000008635 plant growth Effects 0.000 title abstract description 8
- 230000001105 regulatory effect Effects 0.000 title abstract description 7
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 239000013598 vector Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- 230000003321 amplification Effects 0.000 claims 1
- 230000002708 enhancing effect Effects 0.000 claims 1
- 230000002015 leaf growth Effects 0.000 claims 1
- 238000003199 nucleic acid amplification method Methods 0.000 claims 1
- 230000002786 root growth Effects 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 75
- 240000007087 Apium graveolens Species 0.000 abstract description 21
- 235000015849 Apium graveolens Dulce Group Nutrition 0.000 abstract description 19
- 235000010591 Appio Nutrition 0.000 abstract description 19
- 230000012010 growth Effects 0.000 abstract description 10
- 108091023040 Transcription factor Proteins 0.000 abstract description 7
- 102000040945 Transcription factor Human genes 0.000 abstract description 7
- 230000007246 mechanism Effects 0.000 abstract description 4
- 230000002349 favourable effect Effects 0.000 abstract description 3
- 230000004044 response Effects 0.000 abstract description 3
- 238000012408 PCR amplification Methods 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000010353 genetic engineering Methods 0.000 abstract description 2
- 241000219194 Arabidopsis Species 0.000 description 36
- 230000009261 transgenic effect Effects 0.000 description 19
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 13
- 239000007788 liquid Substances 0.000 description 12
- 230000035882 stress Effects 0.000 description 12
- 229930002875 chlorophyll Natural products 0.000 description 11
- 235000019804 chlorophyll Nutrition 0.000 description 11
- 241000219195 Arabidopsis thaliana Species 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 238000012258 culturing Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 108050008339 Heat Shock Transcription Factor Proteins 0.000 description 5
- 102000000039 Heat Shock Transcription Factor Human genes 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 241000589158 Agrobacterium Species 0.000 description 4
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical group O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 4
- 241001052560 Thallis Species 0.000 description 4
- 230000036579 abiotic stress Effects 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 229940118019 malondialdehyde Drugs 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 239000002689 soil Substances 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 102000040075 HSF family Human genes 0.000 description 3
- 108091069846 HSF family Proteins 0.000 description 3
- 102000003992 Peroxidases Human genes 0.000 description 3
- 102000019197 Superoxide Dismutase Human genes 0.000 description 3
- 108010012715 Superoxide dismutase Proteins 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 2
- 102000002812 Heat-Shock Proteins Human genes 0.000 description 2
- 108010004889 Heat-Shock Proteins Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 210000003484 anatomy Anatomy 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000004790 biotic stress Effects 0.000 description 2
- 229930002868 chlorophyll a Natural products 0.000 description 2
- 229930002869 chlorophyll b Natural products 0.000 description 2
- NSMUHPMZFPKNMZ-VBYMZDBQSA-M chlorophyll b Chemical group C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C=O)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 NSMUHPMZFPKNMZ-VBYMZDBQSA-M 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- LHGVFZTZFXWLCP-UHFFFAOYSA-N guaiacol Chemical compound COC1=CC=CC=C1O LHGVFZTZFXWLCP-UHFFFAOYSA-N 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000019516 nonphotochemical quenching Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 238000004161 plant tissue culture Methods 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
- 230000000171 quenching effect Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- 241000208173 Apiaceae Species 0.000 description 1
- 235000002764 Apium graveolens Nutrition 0.000 description 1
- 241000499895 Bloomeria Species 0.000 description 1
- 244000056139 Brassica cretica Species 0.000 description 1
- 235000003351 Brassica cretica Nutrition 0.000 description 1
- 235000003343 Brassica rupestris Nutrition 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- 108010035563 Chloramphenicol O-acetyltransferase Proteins 0.000 description 1
- 101100364969 Dictyostelium discoideum scai gene Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 101150100310 Hsf gene Proteins 0.000 description 1
- 241000219470 Mirabilis Species 0.000 description 1
- 101100364971 Mus musculus Scai gene Proteins 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- CGNLCCVKSWNSDG-UHFFFAOYSA-N SYBR Green I Chemical compound CN(C)CCCN(CCC)C1=CC(C=C2N(C3=CC=CC=C3S2)C)=C2C=CC=CC2=[N+]1C1=CC=CC=C1 CGNLCCVKSWNSDG-UHFFFAOYSA-N 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- QKSKPIVNLNLAAV-UHFFFAOYSA-N bis(2-chloroethyl) sulfide Chemical compound ClCCSCCCl QKSKPIVNLNLAAV-UHFFFAOYSA-N 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 208000012839 conversion disease Diseases 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000004148 curcumin Substances 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 229960001867 guaiacol Drugs 0.000 description 1
- 230000008642 heat stress Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 235000010460 mustard Nutrition 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000003938 response to stress Effects 0.000 description 1
- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- MUUHXGOJWVMBDY-UHFFFAOYSA-L tetrazolium blue Chemical compound [Cl-].[Cl-].COC1=CC(C=2C=C(OC)C(=CC=2)[N+]=2N(N=C(N=2)C=2C=CC=CC=2)C=2C=CC=CC=2)=CC=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=CC=C1 MUUHXGOJWVMBDY-UHFFFAOYSA-L 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000005068 transpiration Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000008096 xylene Substances 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a gene for regulating plant growth and photosynthesis under high-temperature stress, and a coding protein and application thereof, belonging to the field of genetic engineering, wherein the nucleotide sequence of the gene is shown as SEQ ID NO.1, and the amino acid sequence of the coding protein is shown as SEQ ID NO. 2. The invention clones a new HSF transcription factor from celery 'Jinnan real celery' by using PCR amplification technology: the heat-resistant gene AgHSFa1-2, the HSF transcription factor can enhance the growth vigor of plant leaves and root systems, promote photosynthesis of plants and promote the tolerance of plants to high temperature; the heat-resistant gene AgHSFa1-2 obtained by the invention is favorable for deeply understanding the response mechanism of celery in a high-temperature environment, and simultaneously lays an important foundation for cultivating high-temperature resistant plants.
Description
Technical Field
The invention relates to the field of genetic engineering, in particular to a gene for regulating plant growth and photosynthesis under high-temperature stress, and a coded protein and application thereof.
Background
Celery (Apium graveolens l.) is an annual or perennial herb of the family umbelliferae, native to the swamp land on the coast of the mediterranean, and is a vegetable crop of great importance, which is widely planted and consumed worldwide. Celery is a semi-cold-resistant vegetable, is not resistant to heat, has an optimal temperature of 15-20 ℃ for seed germination, has an optimal temperature of 15-25 ℃ for plant growth, can grow poorly in an environment above 26 ℃, and is suitable for cultivation and planting in spring and autumn. 'Jian nan Shi Feng' is an excellent local variety bred in Tianjin nan district, has strong stress resistance and fast growth speed, and can be cultivated in the four seasons of the south and north.
Crop growth, development and yield are affected by many biotic and abiotic stresses, such as extreme weather, e.g., drought, low or high temperatures, soil salinization, etc. Plants have developed a variety of defense strategies to relieve abiotic stresses, including morphological mechanisms, plant hormones regulating and regulating the expression of stress response genes.
Previous studies have shown that Transcription Factors (TFs) are key participants in the regulation of the growth and development of several horticultural plants, and can enhance their tolerance to various abiotic and biotic stress conditions. Among them, heat shock transcription factor (HSF) is one of the most important transcription factors of plants. The HSF family can cope with heat stress and other abiotic stresses, including cold, salt, and drought, etc., by binding to the inverted repeat region of HSEs and promoting transcription of Heat Shock Proteins (HSPs). The HSF family can be classified into HSFA, HSFB and HSFC based on the length of the basic amino acid (aa) sequence between the DBD and HR-A/B regions and the number of amino acid residues inserted into the HR-A/B region.
In order to solve the problem of how to improve the stress resistance of celery plants to high temperature, the invention clones a new HSF gene AgHSFa1-2 from celery variety 'Jinnan Shigella', and expounds the function of the gene AgHSFa1-2 in the aspect of plant heat resistance so as to expand the application prospect of HSF family transcription factors.
Disclosure of Invention
The invention aims to provide a gene for regulating plant growth and photosynthesis under high-temperature stress, and encoding protein and application thereof, so as to solve the problems in the prior art, and the obtained heat-resistant gene AgHSFa1-2 can enhance the growth vigor of plant leaves and root systems, promote plant photosynthesis and promote the tolerance of plants to high temperature, and is favorable for deeply knowing the response mechanism of celery under high-temperature environment and laying an important foundation for cultivating high-temperature resistant plants.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a heat-resistant gene AgHSFa1-2, the nucleotide sequence of which is shown as SEQ ID NO. 1.
The invention also provides a protein coded by the heat-resistant gene AgHSFa1-2, and the amino acid sequence of the protein is shown as SEQ ID NO. 2.
The invention also provides a recombinant vector comprising the heat-resistant gene AgHSFa1-2.
The invention also provides a recombinant cell comprising the recombinant vector.
The invention also provides an application of the heat-resistant gene AgHSFa1-2 or the protein or the recombinant vector or the recombinant cell in improving the high temperature stress resistance of plants.
The invention also provides an application of the heat-resistant gene AgHSFa1-2 or the protein or the recombinant vector or the recombinant cell in improving the growth vigor of plant leaves and root systems.
The invention also provides an application of the heat-resistant gene AgHSFa1-2 or the protein or the recombinant vector or the recombinant cell in improving photosynthesis of plants.
The invention also provides an application of the heat-resistant gene AgHSFa1-2 or the protein or the recombinant vector or the recombinant cell in cultivating and improving high-temperature resistant plants.
The invention also provides a method for improving the high temperature resistance of plants, which transfers the heat-resistant gene AgHSFa1-2 into a receptor plant.
Further, the heat-resistant gene AgHSFa1-2 is amplified by a primer shown as SEQ ID NO.3-4 and then transferred into a receptor plant.
The invention discloses the following technical effects:
the invention clones a new HSF transcription factor from celery 'Jinnan real celery' by using PCR amplification technology: the heat-resistant gene AgHSFa1-2, the HSF transcription factor can enhance the growth vigor of plant leaves and root systems, promote photosynthesis of plants and promote the tolerance of plants to high temperature. The heat-resistant gene AgHSFa1-2 obtained by the invention is favorable for deeply understanding the response mechanism of celery in a high-temperature environment, and simultaneously lays an important foundation for cultivating high-temperature resistant plants.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the growth state of a WT Arabidopsis plant and an AgHSFa1-2 transgenic Arabidopsis T2 generation plant provided in example 1 under high temperature treatment;
FIG. 2 is a graph showing the results of the high temperature treatment of the WT Arabidopsis plant tissue culture seedlings and AgHSFa1-2 transgenic Arabidopsis T2 generation plant tissue culture seedlings provided in example 1; wherein in FIG. 2, WT is the root system of an Arabidopsis wild plant, OE-AgHSFa1-2 is the root system of an Arabidopsis T2 generation plant transformed with AgHSFa1-2 gene;
FIG. 3 is a GUS staining chart of T2 generation plants of Arabidopsis thaliana transformed with AgHSFa1-2 gene provided in example 1; wherein, in FIG. 3, WT represents a wild type control group, OE-AgHSFa1-2 represents an AgHSFa1-2 transgenic Arabidopsis T2 generation plant;
FIG. 4 is a T2 generation PCR detection chart of the transgenic pCAMBIA1301-AgHSFa1-2 gene Arabidopsis thaliana provided in example 1;
FIG. 5 is a graph showing comparison of the results of fluorescent quantitative expression verification of four genes of AtHSPP 8.7, atHSP70-1, atAPX1 and AtGOLS1 after 4-period high-temperature treatment of the WT and AgHSFa1-2 transgenic Arabidopsis provided in example 1, wherein WT represents a wild-type control group, OE-AgHSFa1-2 represents an AgHSFa1-2 transgenic Arabidopsis T2 generation plant; FIG. 5A is AtHSPP 98.7 and B is AtHSP70-1; c is AtAPX1, D is AtGOLS1;
FIG. 6 is a graph showing a comparison of leaf anatomy of WT and AgHSFa1-2 transgenic Arabidopsis T2 generation plants after 4h high temperature treatment provided in example 1, wherein WT represents a wild type control group and OE-AgHSFa1-2 represents AgHSFa1-2 transgenic Arabidopsis T2 generation plants;
FIG. 7 is a graph showing the physiological index comparison of WT and AgHSFa1-2 transgenic Arabidopsis T2 generation plants provided in example 1 after 24h treatment at high temperature; wherein, WT represents a wild type control group, OE-AgHSFa1-2 represents an AgHSFa1-2 transgenic Arabidopsis T2 generation plant; in fig. 7, a is a proline content comparison chart, B is an MDA content comparison chart, C is a CAT enzyme activity comparison chart, D is a SOD enzyme activity comparison chart, and E is a POD enzyme activity comparison chart;
FIG. 8 is a graph showing the chlorophyll content and chlorophyll fluorescence characteristics of the T2 generation plants of Arabidopsis thaliana T2 generation transformed with AgHSFa1-2 gene provided in example 1 after high temperature treatment for 24 hours; wherein, WT represents a wild type control group, OE-AgHSFa1-2 represents an AgHSFa1-2 transgenic Arabidopsis T2 generation plant; in fig. 8, a is a chlorophyll a content comparison chart, B is a chlorophyll B content comparison chart, C is a total chlorophyll content comparison chart, D is a different plant transpiration rate comparison chart, E is a different plant chlorophyll non-photochemical quenching coefficient comparison chart, and F is a different plant chlorophyll photochemical quenching coefficient comparison chart.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Example 1
(1) Extraction of celery total RNA and synthesis of cDNA: total RNA was extracted from celery's Mirabilis Heterophylla' mature leaves using Plant Total RNAIsolation Kit (Chengdu Fuji Biotechnology Co.). By Goldenstar TM RT6 cDNA Synthesis Mix Rnasin selected (Beijing Optimu Corp.) reverse transcribes total RNA extracted into cDNA;
(2) Cloning of celery transcription factor AgHSFa1-2 gene: based on celery transcriptome sequencing information, taking an Arabidopsis thaliana HSF transcription factor family as an information probe, and carrying out search analysis to obtain a celery AgHSFa1-2 gene sequence;
designing a pair of primers, a forward primer, according to the sequence: 5'-ATGGCTTCTACCAACGGC-3' (SEQ ID NO. 3), reverse primer: 5'-TCAGGCCTTTTTGGTGTCTG-3' (SEQ ID NO. 4). Amplifying by using 'jin nan real celery' cDNA as a template, wherein the PCR reaction conditions are as follows: pre-denaturation at 94℃for 5min; denaturation at 94℃for 30s, annealing at 54℃for 30s, extension at 72℃for 60s for 30 cycles; extending at 72℃for 10min. The PCR product was subjected to mass volume fraction 1.0% agarose gel electrophoresis, and then the target band was recovered, and was connected to pUCm-T vector (Shanghai) and transformed into E.coli DH 5. Alpha. And the extracted plasmid was subjected to PCR identification and was then submitted to sequencing by Beijing qingke biosciences Co.
Sequencing results show that the nucleotide sequence of the celery AgHSFa1-2 gene is shown as SEQ ID NO.1, and the amino acid sequence of the protein is shown as SEQ ID NO. 2.
SEQ ID NO.1：
ATGGCTTCTACCAACGGCACCAATAATCCGGCCACCGGACGATCTCAATCTCCGGCGATTCCTACTCCGATAACCAACGTAAATGCTCCGCCGCCGTTCCTTGTGAAGACGTACGATATGGTTGATAATCCGGAAATTGATAAGATAGTCTCGTGGAGTGATTCTAACAATAGCTTCATCGTGTGGGATCCACCTGAGTTCGGTAGAGATGTTTTGCCCAAGTATTTTAAACATAACAATTTCTCCAGCTTTGTGCGTCAGCTCAATACTTATGGTTTCAGGAAGGTTGATCCAGACCGCTGGGAGTTTGCTAATGAAGGCTTTTTAAGAGGCCAAAGGCACTTGCTTAGGACTATCGTCCGCCGGAAACCAACTCAGGGCCAAAGCCAACAACCGCATGGACAAAGTTCATCAGTAGGGGCATGTGTTGAAGTGGGGACATTTGGCCTCGAGGAAGAGGTTGAAAGGCTAAAAAGGGACAAGAATGTCCTTATGCAAGAGCTTGTCAGGTTGAGACAGCAACAGCAGACCACTGACCATCATTTACAAACAATGGTGAAGCGCCTTCATGGGATGGAGCAAAGGCAACAGCAGATGATGTCGTTCCTAGCGAAGGCTGTGAATAGCCCTGGCTTTTTGTCACAATTTGTTCAGCAGCAAACTGAGAGTACCAAGCGCTTAACGGAAGGCAATAAGAAACGGAGGATCAAACCTGATGATGGGCAGATTGTCAAGTACCAGCCTAATGTGATTGAAGCTGCAAAAGCATTGCTCAGTCAGATAGCAAAATCAGATGCATCTCCTAAGCTGGAGACCTTTGACCATAGCTCGGATTGTGTTATCCCTAGTGATATCTCCTCACCTTTGACTGCATCAGACCATGGAAGTTCTCCAACTCATAGCTCAGGAGGGACTCTATCCAAGTATCCACCAATTTCTACTGGTTCTTATGTCCCATCAGATTCAGGGGTTATTCTTAGTGGTCCTTCAACCTCCATATCTGATATAAAGGCAGGTGTAACTTCTGATGCCCTTATAACCGCACAACTTCCTGATGTAAGCTCATTGATCGGGGAACATGAGTTGCCGTCTGTCAGTCTTCCTCCAGATGACATTAATATCCTTGATATCTCCAAGTTTGAAGAAATGATTTCAGGAAGCAATGTCAGTAATCCTAGTGGAAGTAATGGTGCATTCCTAGATCCAGCATCATTGGAGGCAGATCAAACCATGTCCTTGGACATTGGCGATCTCTCTCCTGATATGGGTATCGACTGGGATAATAACTCTTACTTCGATGAGATGCAGGTACTTAATTCGGTTGAACCCGCTTGGGAACAGTTTCTCGAACAGAGCTTGGTTCAGGCAGACGGTGAGGCAATGGATTCTACTTTAGAGGATTCTACACAGAAGAATGAAGTGAAAACTTTGGAGAATGAATGGTCTAAGGCTCAGATGCAAAGGCTTACAGAACAGATGGATCATCTCTCATCAGACACCAAAAAGGCCTGA；
SEQ ID NO.2：
MASTNGTNNPATGRSQSPAIPTPITNVNAPPPFLVKTYDMVDNPEIDKIVSWSDSNNSFIVWDPPEFGRDVLPKYFKHNNFSSFVRQLNTYGFRKVDPDRWEFANEGFLRGQRHLLRTIVRRKPTQGQSQQPHGQSSSVGACVEVGTFGLEEEVERLKRDKNVLMQELVRLRQQQQTTDHHLQTMVKRLHGMEQRQQQMMSFLAKAVNSPGFLSQFVQQQTESTKRLTEGNKKRRIKPDDGQIVKYQPNVIEAAKALLSQIAKSDASPKLETFDHSSDCVIPSDISSPLTASDHGSSPTHSSGGTLSKYPPISTGSYVPSDSGVILSGPSTSISDIKAGVTSDALITAQLPDVSSLIGEHELPSVSLPPDDINILDISKFEEMISGSNVSNPSGSNGAFLDPASLEADQTMSLDIGDLSPDMGIDWDNNSYFDEMQVLNSVEPAWEQFLEQSLVQADGEAMDSTLEDSTQKNEVKTLENEWSKAQMQRLTEQMDHLSSDTKKA*。
(3) Construction of a recombinant expression vector of AgHSFa1-2 gene:
1) Firstly, obtaining a linearization vector of pCAMBIA1301 by using a double enzyme digestion BamHI and ScaI (Thermo Scientific) method, and then purifying by agarose gel electrophoresis and a gel recovery kit (Shanghai) to obtain the linearization vector of pCAMBIA1301 with high purity;
2) The fragment of interest DNA and linearization vector pCAMBIA1301 at 3:1 into a centrifuge tube with the volume of 1.5mL, connecting the centrifuge tube for about 30min at room temperature after uniform mixing, adding 10 mu l of reaction solution into 50 mu l of DH5 alpha competent cells, slightly mixing the mixture by a pipette, incubating the mixture on ice for 30min, and rapidly placing the mixture on ice for cooling after heat shock in a water bath with the temperature of 42 ℃ for 45 seconds;
3) 300 μl of LB liquid medium was added and incubated at 37℃for 45-60min. Centrifuging at 5,000rpm for 2min, collecting thallus, discarding part of supernatant, re-suspending the thallus with the rest culture medium, lightly coating with sterile coating rod on LB solid culture medium containing Kan resistance, and culturing in 37 deg.C incubator for 16-24 hr;
4) Selecting a plurality of clones on a recombinant reaction conversion plate for colony PCR identification, identifying positive colonies, selecting corresponding single colonies, culturing overnight in a liquid LB culture medium containing Kan antibiotics at 37 ℃ and 200rpm incubator, extracting plasmids or directly sequencing bacterial liquid to identify the accuracy of the vector;
5) After successful identification, the recombinant plasmid pCAMBIA1301-AgHSFa1-2 was stored.
(4) Transfer of the recombinant vector into Agrobacterium GV3101: (1) 2 mug of recombinant plasmid pCAMBIA1301-AgHSFa1-2 is added into each 100 mug of GV3101 agrobacterium competent cells, the mixture is stirred by hands to the bottom of a tube and mixed uniformly, and then the mixture is placed on ice for 5min, liquid nitrogen for 5min, water bath at 37 ℃ for 5min and ice bath for 5min. Adding 700 μl of LB liquid medium without antibiotics, and shake culturing at 28deg.C for 2 hr; (2) the cells were collected by centrifugation at 6000rpm for 1min, 100. Mu.l of the supernatant was left, resuspended cells were gently blown, and uniformly spread on 30ml of YEB solid medium containing 15. Mu.l Kan and 15. Mu.l Rif, and placed upside down in a 28℃incubator for 2 days, and the results were easily verified by colony PCR by picking up a number of positive clones.
(5) Cultivation of Arabidopsis thaliana: (1) according to the experiment, a certain amount of Arabidopsis seeds are taken and filled in a sterile 1.5mL centrifuge tube, 1mL of 75% ethanol is added, the mixture is mixed up and down, the supernatant is discarded, and the process is repeated for 3 times. 1mL of ultrapure water was added, the seeds were washed, the supernatant was discarded, and the procedure was repeated 3 times. Seed on the prepared MS plate with a 1mL pipette on an ultra clean bench; (2) sealing the flat plate, pouring the flat plate into a 4 ℃ for purification for 72 hours under the dark condition, and after the end, vertically culturing the flat plate in an illumination incubator, wherein transplanting can be performed after one week of seedling emergence; (3) the seedlings are planted into the soil of the small basin by forceps, firstly, the seedlings are moisturized for 24 hours by using a preservative film, and the seedlings are placed in a plant growth room for cultivation until the arabidopsis grows and is bolting (30 days) for transformation experiments.
(6) Genetic transformation of Arabidopsis thaliana: (1) agrobacterium activation: respectively adding 10 mu LRif and 20 mu L Kan (Sigma Co.) into 20mL LB liquid medium, shaking, inoculating, and oscillating at 28 deg.C and 220rpm for 8-10 hr to obtain activated bacterial liquid of Agrobacterium; (2) and (3) culturing agrobacterium tumefaciens in an enlarged manner: respectively adding 100 mu L of Rif and 200 mu L of Kan into 200mL of YEB liquid culture medium, adding 5-10mL of activated bacterial liquid, performing shake culture at 28 ℃/220rpm for 14-16h until the OD value is 1.6-2.0, centrifuging at 4500rpm for 10min, discarding supernatant of the precipitate bacterial body, and naturally airing; (3) adding 100mL of 5% sucrose and 20 mu L of SILWETL-77 surfactant solution into the precipitated thalli to resuspend the thalli, and blowing the thalli uniformly by a liquid transfer device to resuspend the thalli; (4) adding the bacterial liquid in the centrifugal bottle into a plate, folding the arabidopsis inflorescence, immersing the plate, gently shaking for 15s, sleeving the plant with a black bag after the transformation, keeping moisture for 24h in a dark place, and repeatedly transforming once again after a week.
(7) T1 generation positive plant screening: planting the seeds harvested in the T0 generation of arabidopsis thaliana, sterilizing the T0 generation of seeds, inoculating an MS screening culture medium containing 30mg/L hygromycin (25 mg/L cephalosporin is added for bacteriostasis), culturing for 7-10 days at 22 ℃ by illumination, screening to obtain T1 generation positive plants (plants with seedlings and root systems which grow normally), transplanting the positive seedlings into nutrient soil, covering the positive seedlings with a preservative film for 2-3 days, removing the film, and then growing normally. And (3) extracting DNA from the leaves of the screened T1 generation positive plants, identifying that the leaves contain AgHSFa1-2 genes by using a PCR method, carrying out molecular verification of target genes of transgenic plants, and finally confirming that the genes have been transferred into the T1 generation positive plants.
(8) Positive detection of transgenic plants in T2 generation: (1) collecting T1 generation positive plants to obtain T1 generation seeds, continuously screening hygromycin to obtain T2 generation positive plants, taking out the T2 generation positive plants with forceps when the plants grow to 10d, and staining the T2 generation positive plants and the WT control by using a GUS staining kit (Beijing cool pacing technology Co., ltd.) to obtain the results shown in figure 3; (2) the obtained positive plants are transplanted and grown, the genomic DNA of the leaves is extracted for PCR molecular identification, and the T2 generation positive plants are determined, as shown in figure 4.
(9) Phenotype observation of Arabidopsis positive plants: (1) sowing wild type WT Arabidopsis and T1 generation seeds in sterilized nutrient soil at the same time, purifying at 4 ℃ for 72 hours, transferring into a plant growth room, and performing high temperature stress treatment when Arabidopsis leaves develop to 5cm (20 d); (2) WT and OE-AgHSFa1-2 plants were simultaneously placed in a 38℃light incubator and kept at a humidity of about 60% for 24h, as shown in FIG. 1.
(10) Root system activity determination of arabidopsis positive plants: (1) sterilizing wild WT Arabidopsis and T1 generation seeds with 75% ethanol for 10s, repeating for 3 times, cleaning with sterile water, inoculating on MS plate, purifying at 4deg.C for 72 hr, transferring into illumination incubator, vertically culturing, and culturing at 25deg.C until root system grows obviously; (2) when the root systems of the arabidopsis are grown for 10d, the temperature of the illumination incubator is adjusted to 38 ℃, the humidity is kept at about 60%, and the treatment is carried out for 24h, as shown in figure 2.
(11) Quantitative expression verification of arabidopsis positive plants: (1) high temperature treatment of WT and OE-AgHSFa1-2 plants: treating at 38deg.C for 0h, 4h, 12h, and 24h respectively, taking leaf samples, immediately quick-freezing with liquid nitrogen, and storing in-80deg.C refrigerator to obtain RNA and cDNA according to the method in (1); (2) selection of 4 genes of Arabidopsis thaliana and Arabidopsis thalianaThe transcription expression level of Actin in mustard is used for designing an expression detection primer for an internal reference sequence, as shown in table 1; (3) real-time quantitative PCR was performed using Beijing engine biotechnology Co.Ltd 2×TSINGKE TM Master qPCR Mix (SYBR Green I) kit is carried out according to the operation instructions; (4) the calculation formula of the relative transcription expression level of the target gene is 2 -ΔΔCt Δct= (Ct target gene-ctaction) treatment group- (Ct target gene-ctaction) control group; (5) the data were analyzed and plotted to verify the relative expression level as shown in fig. 5.
(12) Leaf structural characteristic determination of arabidopsis positive plants
Wild type WT Arabidopsis plants and transgenic plants AgHSFa1-2 were subjected to a high temperature treatment at 38℃for 4 hours, leaf samples were carefully taken with forceps, fixed with FAA fixative, dehydrated with ethanol, treated with xylene and absolute ethanol, paraffin sections were prepared, double counterstained with safranine and green, sealed, photographed with a Nikon Eclipse E100 optical microscope, and each sample was repeated 3 times as shown in FIG. 6.
(13) Determining the physiological index of the arabidopsis positive plant:
high temperature treatment of wild type WT Arabidopsis plants and transgenic plants AgHSFa 1-2: treating at 38deg.C for 24h, and determining proline content by ninhydrin method; measuring MDA content by using a Malondialdehyde (MDA) detection kit; measuring the activity of hydrogen peroxide (CAT) enzyme by an ultraviolet spectrophotometry; measuring the activity of superoxide dismutase (SOD) by using a nitrogen blue tetrazolium photochemical reduction method; the guaiacol method determines Peroxidase (POD) activity.
(14) Determination of chlorophyll content and chlorophyll fluorescence characteristics of leaves of arabidopsis positive plants
High temperature treatment of wild type WT Arabidopsis plants and transgenic plants AgHSFa 1-2: treating at 38deg.C for 24h by using absolute ethanol and absolute acetone 1:1, measuring chlorophyll a, chlorophyll b and total chlorophyll content by a mixed liquid leaching method; light energy conversion efficiency (Fv/Fm), photochemical quenching (qP) and non-photochemical quenching (qN) of PS II were measured by using a chlorophyll fluorescence meter (PAM-2500), 3 plants were selected for each strain, 3 leaves were selected for each plant, and 3 times were measured for each leaf. The above experiments were repeated 3 times as shown in fig. 8.
TABLE 1 primer sequences
(15) Test results
1) The growth state and phenotype of the T2 generation positive plant obtained by transferring the celery AgHSFa1-2 gene obtained by cloning into an Arabidopsis plant are obviously superior to those of the WT plant under the high-temperature stress of 38 ℃;
2) Root system measurement results show that the root system length of the transgenic plant is obviously longer than that of a wild arabidopsis control, and the expression of the AgHSFa1-2 gene promotes the growth of the arabidopsis root system under high temperature stress, as shown in figure 2;
3) The fluorescent quantitative PCR results show that 4 genes in the OE-AgHSFa1-2 plant respond to high temperature stress, the expression level also rises with the increase of the treatment time, and the expression level rises rapidly after 12 hours and is kept at a higher level, and the detail is shown in figure 5;
4) Leaf anatomy showed that the number and extent of stomatal opening of transgenic plants was significantly higher than that of wild type arabidopsis controls, indicating that expression of AgHSFa1-2 gene contributed to the stomatal opening of recipient plants at high temperature (fig. 6);
5) The physiological index analysis result can prove that the celery AgHSFa1-2 gene can significantly improve the stress resistance of a receptor plant to high temperature (figure 7);
6) The leaf chlorophyll content and chlorophyll fluorescence characteristic results show that the photosynthesis intensity of the transgenic AgHSFa1-2 plant under high temperature stress is obviously stronger than that of the WT wild Arabidopsis variety (figure 8).
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (10)
1. A heat-resistant gene AgHSFa1-2 is characterized in that the nucleotide sequence is shown as SEQ ID NO. 1.
2. A protein encoded by the heat-resistant gene AgHSFa1-2 according to claim 1, wherein the amino acid sequence is shown in SEQ ID No. 2.
3. A recombinant vector comprising the heat-resistant gene AgHSFa1-2 of claim 1.
4. A recombinant cell comprising the recombinant vector of claim 3.
5. Use of a thermotolerant gene AgHSFa1-2 according to claim 1 or a protein according to claim 2 or a recombinant vector according to claim 3 or a recombinant cell according to claim 4 for increasing the tolerance of a plant to high temperature stress.
6. Use of the heat-resistant gene AgHSFa1-2 according to claim 1 or the protein according to claim 2 or the recombinant vector according to claim 3 or the recombinant cell according to claim 4 for improving plant leaf and root growth vigor.
7. Use of a thermo-resistant gene AgHSFa1-2 according to claim 1 or a protein according to claim 2 or a recombinant vector according to claim 3 or a recombinant cell according to claim 4 for enhancing photosynthesis in plants.
8. Use of the thermotolerant gene AgHSFa1-2 according to claim 1 or the protein according to claim 2 or the recombinant vector according to claim 3 or the recombinant cell according to claim 4 for the cultivation and improvement of plants with high temperature resistance.
9. A method for improving the high temperature resistance of plants, which is characterized in that the heat-resistant gene AgHSFa1-2 as defined in claim 1 is transferred into a recipient plant.
10. The method according to claim 9, wherein the thermo-resistant gene AgHSFa1-2 is transferred into a recipient plant after amplification by a primer shown in SEQ ID NO. 3-4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311157424.4A CN117187259B (en) | 2023-09-08 | 2023-09-08 | Gene for regulating plant growth and photosynthesis under high-temperature stress condition, and encoding protein and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311157424.4A CN117187259B (en) | 2023-09-08 | 2023-09-08 | Gene for regulating plant growth and photosynthesis under high-temperature stress condition, and encoding protein and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117187259A true CN117187259A (en) | 2023-12-08 |
| CN117187259B CN117187259B (en) | 2024-03-01 |
Family
ID=89004734
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311157424.4A Active CN117187259B (en) | 2023-09-08 | 2023-09-08 | Gene for regulating plant growth and photosynthesis under high-temperature stress condition, and encoding protein and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117187259B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110131680A1 (en) * | 2009-09-15 | 2011-06-02 | Stephen Bowley | Method of enhancing gene expression in plants |
| EP2840141A1 (en) * | 2013-08-21 | 2015-02-25 | Industry-Academic Cooperation Foundation, Yonsei University | Gene implicated in abiotic stress tolerance and growth accelerating and use thereof |
| CN107674873A (en) * | 2017-10-19 | 2018-02-09 | 河北省农林科学院遗传生理研究所 | Wheat thermal excited transcryption factor gene TaHsfA2i and its encoding proteins and application |
| CN114480432A (en) * | 2022-03-31 | 2022-05-13 | 四川农业大学 | Celery heat-resistant gene AgHSFA6a-1 and application thereof |
-
2023
- 2023-09-08 CN CN202311157424.4A patent/CN117187259B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110131680A1 (en) * | 2009-09-15 | 2011-06-02 | Stephen Bowley | Method of enhancing gene expression in plants |
| EP2840141A1 (en) * | 2013-08-21 | 2015-02-25 | Industry-Academic Cooperation Foundation, Yonsei University | Gene implicated in abiotic stress tolerance and growth accelerating and use thereof |
| CN107674873A (en) * | 2017-10-19 | 2018-02-09 | 河北省农林科学院遗传生理研究所 | Wheat thermal excited transcryption factor gene TaHsfA2i and its encoding proteins and application |
| CN114480432A (en) * | 2022-03-31 | 2022-05-13 | 四川农业大学 | Celery heat-resistant gene AgHSFA6a-1 and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| JIE LI ET AL.: "The miR165/166–PHABULOSA module promotes thermotolerance by transcriptionally and posttranslationally regulating HSFA1", 《THE PLANT CELL》, vol. 35, 3 May 2023 (2023-05-03), pages 2952 - 2971 * |
| 周瑾等: "芹菜TCP基因家族全基因组鉴定、进化分析和表达研究", 《四川农业大学学报》, vol. 40, no. 3, 30 April 2022 (2022-04-30), pages 145 - 155 * |
| 李岩等: "芹菜热激转录因子基因AgHSFB2的克隆及不同温度处理下的表达响应", 《芹菜TCP 基因家族全基因组鉴定、进化分析和表达研究》, vol. 38, no. 3, 31 December 2015 (2015-12-31), pages 360 - 368 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117187259B (en) | 2024-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108841841B (en) | Cloning of tomato transcription factor SlbZIP6 and application thereof in high temperature stress resistance | |
| CN111593058B (en) | Bna-miR169n gene and application thereof in controlling drought resistance of brassica napus | |
| CN114480432B (en) | Celery heat-resistant gene AgHSFA6a-1 and application thereof | |
| WO2022247591A1 (en) | Heat shock-related gene zmhsf11 and application thereof in regulating plant heat tolerance | |
| CN113621625B (en) | Application of sesame SiERF103 gene in enhancing plant resistance | |
| CN114591972B (en) | Celery heat-resistant gene AgDREBA6c and application thereof | |
| CN117187259B (en) | Gene for regulating plant growth and photosynthesis under high-temperature stress condition, and encoding protein and application thereof | |
| CN115772212A (en) | Alfalfa chloroplast MsSAP22 gene and application thereof in improving drought resistance of plants | |
| CN116479007B (en) | Celery AgDREBA6a gene and application thereof in improving high-temperature stress resistance of plants | |
| CN111423500B (en) | SiMYB56 protein and application of encoding gene thereof in regulation and control of plant drought resistance | |
| CN113307854A (en) | Salt-tolerant protein, gene encoding same and application | |
| CN116621959B (en) | Soybean GmMADS5 gene and application thereof in plant flowering phase regulation | |
| CN115011631B (en) | Protein for regulating drought resistance of corn at seedling stage, and coding gene and application thereof | |
| CN114591971B (en) | Drought-resistant VvCCD7 gene of grape as well as amino acid sequence and application thereof | |
| CN111875686B (en) | Application of sesamin SiLLR in regulation and control of plant root system development | |
| CN114807174B (en) | Genetic locus for reversely regulating resistance of rice to rice blast bacteria and application thereof | |
| CN113652434B (en) | Gorgon fruit DNA molecule with function of promoting rice grain enlargement and application thereof | |
| CN111363019B (en) | Application of SiMYB56 protein and coding gene thereof in regulation and control of low nitrogen tolerance of plants | |
| CN114805513B (en) | Tobacco NtOEE1 gene and application thereof in regulation of stem and leaf included angle and plant height | |
| CN116143892B (en) | Application of OsGN11 gene in improving rice grain number per ear character | |
| CN110922459B (en) | Application of SlSNAT1 protein and related biological material thereof in regulation and control of plant seed aging resistance | |
| CN107226849B (en) | Application of rice GW5 gene in cultivating grain type-changed transgenic plant | |
| CN116396370A (en) | Plant coding gene and protein with functions of improving cadmium resistance and promoting biomass improvement and transgenic strain | |
| CN116836993A (en) | Duke pear bHLH transcription factor gene PbebHLH155 and application thereof | |
| CN116254294A (en) | Cultivation method for enhancing drought resistance of tomatoes |
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 |