CN110760524A - Specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance - Google Patents
Specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance Download PDFInfo
- Publication number
- CN110760524A CN110760524A CN201910957559.6A CN201910957559A CN110760524A CN 110760524 A CN110760524 A CN 110760524A CN 201910957559 A CN201910957559 A CN 201910957559A CN 110760524 A CN110760524 A CN 110760524A
- Authority
- CN
- China
- Prior art keywords
- specific dna
- com58276
- plant
- dna molecule
- arabidopsis thaliana
- 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
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
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
Abstract
The invention discloses a specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance. The nucleotide sequence of the specific DNA fragment com58276 is shown as SEQ ID NO: 1 is shown. Experiments prove that the specific DNA fragment com58276 is introduced into the wild type Arabidopsis thaliana Columbia-0 subtype to obtain transgenic Arabidopsis thaliana; compared with the wild type Arabidopsis thaliana Columbia-0 subtype, the drought resistance of the transgenic Arabidopsis thaliana is improved, and the improvement of the drought resistance is reflected by the reduction of the malonaldehyde content, the increase of the soluble sugar content, the reduction of the water loss rate and the improvement of the survival rate. Therefore, the specific DNA fragment com58276 can regulate the plant stress resistance. The invention has important application value.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a specific DNA fragment com58276 and application thereof in regulation and control of plant stress resistance.
Background
Caragana microphylla is a leguminous shrub mainly growing in sandy land of desert or semi-desert regions, and has a highly developed root system and extremely strong drought and cold resistance. Caragana microphylla is not only important pasture and industrial raw material, but also can maintain water and soil, prevent soil desertification and have important economic and Ecological value (Fang X W, Li J H, Xiong Y C, Xu D H, Fan X W, Li F M. Responses of Caraganaakorshiki Kom. to shoot removal: mechanisms undersiding growth [ J ]. Ecological Research, 2007, 23 (5): 863-871.).
At present, the researches on caragana korshinskii mainly focus on the aspects of biological characteristics, physiological changes, anatomical structures and the like, and the researches on stress resistance related genes and stress resistance related fragments are few. The CkWRKY1 gene is isolated from caragana microphylla, and the encoded CkWRKY1 protein belongs to WRKY transcription factor family. Research shows that CkWRKY1 protein is an anti-adversity related protein and plays an important role in the caragana microphylla to cope with adversity stress (Yang Q, Yin J J, Li G, Qi L W, Yang F Y, Wang R G, Li G J. reference gene selection for qRT-PCR in Caragana akkorshinski Kom. under differential stress conditions [ J ]. Molecular biology reports, 2014, 41(4): 2325-2334.).
Disclosure of Invention
The invention aims to improve the stress resistance, such as drought resistance, of plants.
The invention firstly protects a specific DNA molecule, and the nucleotide sequence of the specific DNA molecule can be shown as SEQ ID NO: 1 is shown.
Expression cassettes containing said specific DNA molecules also belong to the scope of protection of the present invention.
The expression cassette (from 5 'to 3') may include a promoter region, a transcription initiation region, a gene region of interest (containing the specific DNA molecule), a transcription termination region, and optionally a translation termination region. The promoter region and the gene region of interest may be native/analogous to the host, or the promoter region and the gene region of interest may be native/analogous to each other, or the promoter region and/or the gene region of interest may be heterologous to the host or to each other. As used herein, "heterologous" refers to a sequence that is a sequence derived from a foreign species, or, if from the same species, is substantially modified in composition and/or genomic locus in nature by deliberate human intervention. Optionally containing a transcription termination region homologous to the transcription initiation region, to the operably linked gene region of interest, and to the plant host; or; the target gene region and the host are exogenous or heterologous. The transcription termination region may be derived from the Ti-plasmid of Agrobacterium tumefaciens, such as the octopine synthase and nopaline synthase termination regions.
The expression cassette may also include a 5' leader sequence. The 5' leader sequence may enhance translation.
In preparing the expression cassette, adapters or linkers may be used to ligate specific DNA molecules, or other manipulations may be involved to provide appropriate restriction sites, remove excess DNA, remove restriction sites, etc. To achieve this, in vitro mutagenesis, primer repair, restriction, annealing, re-substitution, such as transitions and transversions, may be performed.
The expression cassette may also include a selectable marker gene for screening transformed cells or tissues. Selectable marker genes can be used to screen transformed cells or tissues. Marker genes include genes encoding antibiotic resistance, such as the genes encoding neomycin phosphotransferase II (NEO), Hygromycin Phosphotransferase (HPT), genes providing resistance to herbicidal compounds (e.g., glufosinate, 2, 4-D). Other selectable markers include phenotypic markers, such as fluorescent proteins. The selection markers listed above are not limiting. Any selectable marker gene may be used in the present invention.
Recombinant plasmids containing the specific DNA molecules also belong to the protection scope of the invention. The recombinant plasmid can be obtained by inserting the specific DNA molecule into a starting plasmid. The recombinant plasmid can be specifically a recombinant plasmid obtained by inserting the specific DNA molecule into a multiple cloning site of a starting plasmid.
The starting plasmid may include a selectable marker gene for selection of transformed cells. Selectable marker genes can be used to screen transformed cells or tissues. Marker genes include genes encoding antibiotic resistance, such as the genes encoding neomycin phosphotransferase II (NEO), Hygromycin Phosphotransferase (HPT), genes providing resistance to herbicidal compounds (e.g., glufosinate, 2, 4-D). Other selectable markers include phenotypic markers such as fluorescent proteins. The selection markers listed above are not limiting. Any selectable marker gene may be used in the present invention.
The recombinant plasmid may comprise any of the expression cassettes described above containing the specific DNA molecule.
The recombinant plasmid can be specifically a recombinant plasmid pBinGlyRed3-com 58276. The recombinant plasmid pBinGlyRed3-com58276 is a plasmid which replaces a small DNA fragment between recognition sequences of restriction enzymes EcoRI and XmaI of a pBinGlyRed3 vector with a nucleotide sequence shown as SEQ ID NO: 1 (the DNA molecule shown in SEQ ID NO: 1, namely the specific DNA fragment com58276) to obtain the recombinant plasmid.
Recombinant microorganisms containing said specific DNA molecules also belong to the scope of protection of the present invention.
The recombinant microorganism can be obtained by introducing the recombinant plasmid into the starting microorganism.
The starting microorganism may be a bacterium, yeast, algae or fungus. The bacteria may be gram positive or gram negative bacteria. The gram-negative bacterium may be Agrobacterium tumefaciens (Agrobacterium tumefaciens). The Agrobacterium tumefaciens (Agrobacterium tumefaciens) may specifically be Agrobacterium tumefaciens EHA 105.
The recombinant microorganism may specifically be EHA105/pBinGlyRed3-com 58276. EHA105/pBinGlyRed3-com5827 is obtained by introducing the recombinant plasmid pBinGlyRed3-com58276 into Agrobacterium tumefaciens EHA 105.
Transgenic cell lines containing the specific DNA molecules also belong to the protection scope of the invention.
None of the transgenic cell lines containing the specific DNA molecule comprises propagation material. The transgenic plants are understood to comprise not only the first generation of transgenic plants obtained by transforming the recipient plant with the specific DNA molecule, but also the progeny thereof. For transgenic plants, the gene can be propagated in that species, and can also be transferred into other varieties of the same species, including particularly commercial varieties, using conventional breeding techniques. The transgenic plants include seeds, callus, whole plants and cells.
The invention also protects the application of the specific DNA molecule, which can be A1) or A2):
A1) regulating and controlling the stress resistance of the plants;
A2) cultivating the transgenic plant with the changed stress resistance.
In the application, the regulation and control of the plant stress resistance can be the improvement of the plant stress resistance.
In the application, the transgenic plant with the changed stress resistance can be cultured to be a transgenic plant with improved stress resistance.
The invention also provides a method for cultivating transgenic plants, which comprises the following steps: introducing the specific DNA molecules into the starting plant to obtain a transgenic plant; the transgenic plants have improved stress resistance compared to the starting plants.
In the above method, the "introducing a specific DNA molecule into a starting plant" may be introducing an expression cassette containing the specific DNA molecule or a recombinant plasmid containing the specific DNA molecule into the starting plant.
Any of the plants described above may be any of the following c1) to c 5): c1) a dicotyledonous plant; c2) a monocot plant; c3) a cruciferous plant; c4) arabidopsis thaliana; c5) the wild type Arabidopsis thaliana Columbia-0 subtype.
Any of the above stress resistance may be drought resistance.
As noted above, an increase in drought resistance may be manifested in at least one of a decrease in malondialdehyde content, an increase in soluble sugar content, a decrease in water loss rate, and an increase in survival rate.
In one embodiment of the invention, a recombinant plasmid pBinGlyRed3-com58276 (containing a specific DNA molecule with a nucleotide sequence shown as SEQ ID NO: 1) is introduced into a Columbia-0 subtype of wild type Arabidopsis thaliana to obtain transgenic Arabidopsis thaliana; compared with the wild type Arabidopsis thaliana Columbia-0 subtype, the drought resistance of the transgenic Arabidopsis thaliana is improved, and the improvement of the drought resistance is reflected by the reduction of the malonaldehyde content, the increase of the soluble sugar content, the reduction of the water loss rate and the improvement of the survival rate. As can be seen, SEQ ID NO: 1 can regulate and control the stress resistance of plants. The invention has important application value.
Drawings
FIG. 1 shows the growth state of Arabidopsis seedlings to be tested before natural drought, after natural drought and after 3 days of rehydration.
FIG. 2 is a statistical result of the survival rate of Arabidopsis seedlings.
FIG. 3 shows the statistical result of the water loss rate of the isolated leaf of Arabidopsis thaliana.
FIG. 4 is a statistical result of the soluble sugar content in Arabidopsis thaliana leaves.
FIG. 5 is a statistical result of the malondialdehyde content in Arabidopsis thaliana leaves.
FIG. 6 shows the growth state of Arabidopsis thaliana on MS solid medium containing 180mM mannitol.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention.
The experimental procedures in the following examples are conventional unless otherwise specified.
The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
In the quantitative tests in the following examples, three replicates were set up and the results averaged.
Caragana microphylla seeds were collected from a Denseian desert botanical garden in Gansu, by inventor (010-.
Wild type Arabidopsis thaliana (Arabidopsis thaliana) (Columbia-0 subtype) is described in the following references: kim H, Hyun Y, Park J, Park M, Kim M, Kim H, Lee M, Moon J, Lee I, Kim J.A geneticin between column responses and flowing time through FVE in Arabidopsis thaliana. Nature genetics.2004, 36: 167-.
Indicates a very significant difference, P < 0.01. Indicates significant difference, P < 0.05.
Example 1 obtaining of transgenic com58276 Arabidopsis thaliana and identification of drought resistance
First, obtaining the specific DNA fragment com58276
1. Sowing caragana microphylla seeds in sandy soil, and performing normal growth management to obtain caragana microphylla plants which grow for 1 month; and stopping watering for 20 days to obtain the caragana plants subjected to drought treatment.
The caragana plant subjected to drought treatment has developed root system, white leaves, thick epidermal hair and strong drought resistance.
2. And (2) taking the leaves of the caragana plant subjected to drought treatment obtained in the step (1), extracting total RNA (ribonucleic acid) firstly, and then carrying out reverse transcription to obtain the cDNA (complementary deoxyribonucleic acid) of the caragana.
3. After the step 2 is completed, taking the cDNA of caragana as a template, and adopting a primer F: 5'-TCGCTTTCTCCTTAACCGTA-3' and primer R: 5'-AAACCTTAAATTGAGATACT-3' and recovering PCR amplification product of about 322 bp.
4. Sequencing the PCR amplification product recovered in the step 3.
The sequencing result shows that the nucleotide sequence of the PCR amplification product recovered in the step 3 is shown as SEQ ID NO: 1 is shown.
Converting SEQ ID NO: 1 is named as specific DNA fragment com 58276.
II, obtaining the recombinant plasmid pBinGlyRed3-com58276
1. The pBinGlyRed3 vector (Zhang C, Iskandarov U, Klotz E T, et al. Athrastochytrid diacetyleglycerol acyltransferase 2 with branched specificity primers oly acid content in engineered Arabidopsis thaliana genes sections. journal of Experimental Botany, 2013, 64(11):3189 and 3200.) was digested with restriction endonucleases EcoRI and XmaI, and the vector backbone of about 9kb was recovered.
2. Taking the cDNA of caragana microphylla obtained in the step one (2) as a template, and adopting a primer FF: 5' -ACGGGGGACTGA ATTCGCTTTCTCCTTAACCGTA-3' (recognition site for the restriction enzyme EcoRI is underlined) and primer FR: 5' -CCGCCTCGAGCCCGGGAAACCTTAAATTGAGATACT-3' (recognition sites for restriction enzyme XmaI are underlined) was subjected to PCR amplification, and a PCR amplification product of about 354bp was recovered.
3. And (3) carrying out homologous recombination on the vector skeleton recovered in the step (1) and the PCR amplification product recovered in the step (2) to obtain a recombinant plasmid pBinGlyRed3-com 58276.
The recombinant plasmid pBinGlyRed3-com58276 was sequenced. According to the sequencing results, the recombinant plasmid pBinGlyRed3-com58276 was structurally described as follows: the DNA small fragment between the restriction enzymes EcoRI and XmaI recognition sequences of the pBinGlyRed3 vector was replaced with a DNA fragment having the nucleotide sequence of SEQ ID NO: 1 (the DNA molecule shown in SEQ ID NO: 1 is the specific DNA fragment com58276) to obtain the recombinant plasmid.
Thirdly, obtaining of recombinant Agrobacterium
The recombinant plasmid pBinGlyRed3-com58276 is introduced into Agrobacterium tumefaciens EHA105 to obtain recombinant Agrobacterium, which is named as EHA105/pBinGlyRed3-com 58276.
Fourth, obtaining of transgenic com58276 Arabidopsis thaliana
1. The T.thaliana inflorescence floral dip transformation method (described in Clough, S.J., and Bent, A.F. Floraldip: expressed method for Agrobacterium-mediated transformation of Arabidopsis thaliana plant J. (1998)16, 735-743.) was used to transform EHA105/pBinGlyRed3-com58276 into wild type Arabidopsis thaliana to obtain T.thaliana1The com58276 Arabidopsis seeds were transferred.
2. Observation of T under Green light1The illumination condition of com58276 Arabidopsis seeds is transferred, and then seeds capable of emitting red light are planted to obtain T1The coms 58276 positive seedlings are transferred. T is1The seeds received by the com58276 positive seedlings are T2The com58276 Arabidopsis seeds were transferred.
3. Observing the T of the different strains screened in the step 2 under a green light lamp2The illumination of com58276 Arabidopsis seeds was transferred. If the ratio of the number of red-emitting seeds to the number of red-emitting seeds in a line is 3: 1, the strain is a strain in which a copy of the specific DNA fragment com58276 is inserted, and the seeds received in the strain are T3The com58276 Arabidopsis seeds were transferred.
4. Observing the T screened in the step 3 under a green light lamp3Transgenic com58276 Arabidopsis thaliana seedsThe plants capable of emitting red light seeds are T3The transgenic com58276 Arabidopsis thaliana was homozygous. 2 of them are T3The generation homozygous transgenic com58276 Arabidopsis lines were named line3 and line7, respectively, and the subsequent experiments were performed.
Fifth, molecular identification
The Arabidopsis seed to be tested is T of line33T of line7, generation seed3Generation seed or wild type arabidopsis seed.
(1) Taking an arabidopsis seed to be detected, soaking the arabidopsis seed in 70% (v/v) ethanol water solution for 30s, and washing the arabidopsis seed with sterile water for 3 times; then spread on MS solid medium and vernalize for 2 days at 4 ℃.
(2) And (3) after the step (1) is finished, taking the arabidopsis thaliana seeds to be detected, and carrying out light-dark alternate culture (16h illumination culture/8 h dark culture) at the temperature of 22 ℃ for 28 days to obtain the arabidopsis thaliana seedlings to be detected.
(3) Extracting genome DNA of the arabidopsis seedling to be detected, taking the genome DNA as a template, and performing PCR amplification by adopting a primer pair consisting of 5'-GGACTCTTGACCATGG-3' and 5'-ATTCGAGCTGGTCACC-3' to obtain a PCR amplification product; then, the following judgment is made: if some PCR amplification product contains DNA segment of about 500bp, the Arabidopsis seedling to be tested corresponding to the PCR amplification product is positive seedling.
The results showed that T of line33T of generation seed and line73The seedlings obtained by the generation of seeds are all positive seedlings.
Sixth, drought resistance identification of trans com58276 Arabidopsis thaliana
The Arabidopsis seed to be tested is T of line33T of line7, generation seed3Generation seed or wild type arabidopsis seed.
The culture conditions were: 22 ℃; 12h light/12 h dark; the light intensity was 12000 Lx.
First, natural drought resistance identification
The experiment was repeated three times to obtain an average, and the procedure for each repetition was as follows:
1. taking 3 culture pots, planting the seeds of arabidopsis to be tested in each pot, and culturing for 3 weeks; then, each pot keeps 2 arabidopsis seedlings to be detected with basically consistent growth vigor.
2. And (4) after the step 1 is finished, naturally drying for two weeks.
3. And (3) after the step 2 is finished, rehydrating, and counting the survival rate of the arabidopsis seedlings to be detected after 3 days.
Before natural drought, after natural drought and 3 days after rehydration, the growth state of the arabidopsis seedlings to be tested is shown in figure 1(WT is wild arabidopsis). The results show that, before natural drought, wild type Arabidopsis thaliana and 2T were used3The phenotype of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) has no significant difference; after natural drought for 2 weeks, the leaves of wild type Arabidopsis were all yellow and curled, and even the plants died directly, while 2T3The leaves of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) are partially yellow, partially curled and bolting; after 3 days of rehydration, leaves of wild type Arabidopsis died, plants died, and 2T' s3The generation-homozygous transgenic com58276 Arabidopsis lines (line3 and line7) all survived.
The survival rate statistics of the Arabidopsis seedlings to be tested are shown in FIG. 2(WT is wild type Arabidopsis). The results showed 2T compared to wild type Arabidopsis thaliana3The survival rate of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) is significantly improved.
Second, measuring the water loss rate of the in vitro blade
The experiment was repeated three times to obtain an average, and the procedure for each repetition was as follows:
1. planting the seeds of the arabidopsis to be tested, and culturing for 3 weeks to obtain the seedlings of the arabidopsis to be tested.
2. And (3) after the step (1) is finished, taking the arabidopsis thaliana seedlings to be detected with basically consistent growth vigor, picking the leaves, and weighing by a one-ten-thousandth balance to obtain the fresh weight.
3. And (3) after the step 2 is finished, placing the leaves in a culture dish, placing the leaves at room temperature for 2h, 4h, 6h, 8h, 10h or 12h, and weighing the leaves by using a ten-thousandth balance, namely weighing the leaves after water loss.
4. And (4) after the step 3 is completed, calculating the water loss rate of the in vitro blade.
Water loss (%) - (fresh weight (g) — weight after water loss (g))/(fresh weight (g)) × 100%
And drawing a water loss curve by taking the leaf placing time as an abscissa and the corresponding water loss rate as an ordinate. The water loss curve is shown in FIG. 3(WT is wild type Arabidopsis thaliana). The results showed 2T compared to wild type Arabidopsis thaliana3The water loss rate of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) is significantly reduced.
Thirdly, measuring the content of soluble sugar
The experiment was repeated three times to obtain an average, and the procedure for each repetition was as follows: and 2, collecting leaves of the arabidopsis seedlings to be detected, and detecting the content of soluble sugar by using a plant soluble sugar content detection kit (a product of Solarbio company).
The results of the detection are shown in FIG. 4(WT is wild type Arabidopsis thaliana). The results showed 2T compared to wild type Arabidopsis thaliana3The soluble sugar content of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) was significantly increased.
Determination of Malondialdehyde (MDA)
The experiment was repeated three times to obtain an average, and the procedure for each repetition was as follows: and 2, collecting leaves of the arabidopsis thaliana seedlings to be detected, and detecting the content of malondialdehyde by using a Malondialdehyde (MDA) content detection kit (product of Solarbio).
The results of the assay are shown in FIG. 5(WT is wild type Arabidopsis thaliana). The results showed 2T compared to wild type Arabidopsis thaliana3The malondialdehyde content of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) was significantly reduced.
Fifth, identification of drought resistance under mannitol stress conditions (i.e., drought simulation by addition of mannitol)
The experiment was repeated three times to obtain an average, and the procedure for each repetition was as follows:
1. 20 seeds of Arabidopsis thaliana to be tested are taken, treated with 70% (v/v) ethanol water for 5min, sterilized with 2.6% (v/v) sodium hypochlorite water for 10min, and then washed with sterilized spitting water for 5 times.
2. After completion of step 1, the Arabidopsis seeds to be tested were sown in a medium (MS solid medium, MS solid medium containing 150mM mannitol, MS solid medium containing 180mM mannitol, or MS solid medium containing 200mM mannitol) and vernalized at 4 ℃ for 3 days.
3. The medium which completed step 2 was vertically cultured for 6 days, and the growth state of Arabidopsis thaliana was observed.
The growth state of Arabidopsis thaliana on MS solid medium containing 180mM mannitol is shown in FIG. 6(WT is wild type Arabidopsis thaliana). The results showed that wild type Arabidopsis thaliana and 2T were present on MS solid medium3The phenotype of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) has no significant difference; 2T compared with wild type Arabidopsis thaliana on MS solid medium containing mannitol3The survival rate of the generation-homozygous com58276 Arabidopsis lines (line3 and line7) is increased to a certain extent.
The results show that the drought resistance of the arabidopsis can be improved by introducing the specific DNA fragment com58276 into the wild arabidopsis. The improvement in drought resistance is manifested by a reduction in malondialdehyde content, an increase in soluble sugar content, a reduction in water loss rate, and an improvement in survival rate.
<110> institute of biotechnology of Chinese academy of agricultural sciences
<120> specific DNA fragment com58276 and application thereof in regulation and control of plant stress resistance
<160>1
<170>PatentIn version 3.5
<210>1
<211>322
<212>DNA
<213>Caragana korshinskii Kom.
<400>1
tcgctttctc cttaaccgta gctagtcgtc ttggccggaa tcacagatgg ccgcgcgcca 60
tctctcccct ctcgcgctcc ttcccgttcg cgcatcaccc ctctccctct cgatctctcc 120
ttttcgcgcc tctccccttt tttcctctcg cgacttcact gcagtcattc gtaatagcca 180
cgcacggcca aggatggcgc gccggtcacc ttctccccac cttcttctct atcacgaccg 240
ccctcccttt tctctcctcc tgatgcactt tcctatcttt ctagctgctg ctgctactta 300
gcagtatctc aatttaaggt tt 322
Claims (10)
1. The nucleotide sequence of the specific DNA molecule is shown as SEQ ID NO: 1 is shown.
2. An expression cassette comprising a specific DNA molecule according to claim 1.
3. A recombinant plasmid comprising the specific DNA molecule of claim 1.
4. A recombinant microorganism comprising a specific DNA molecule according to claim 1.
5. A transgenic cell line comprising the specific DNA molecule of claim 1.
6. The use of the specific DNA molecule according to claim 1, which is A1) or A2):
A1) regulating and controlling the stress resistance of the plants;
A2) cultivating the transgenic plant with the changed stress resistance.
7. A method of breeding a transgenic plant comprising the steps of: introducing a specific DNA molecule according to claim 1 into a starting plant to obtain a transgenic plant; the transgenic plants have improved stress resistance compared to the starting plants.
8. The method of claim 7, wherein: the expression "introducing a specific DNA molecule into a starting plant" refers to introducing an expression cassette containing the specific DNA molecule or a recombinant plasmid containing the specific DNA molecule into the starting plant.
9. The use of claim 6, or the method of claim 7 or 8, wherein: the plant is any one of the following c1) to c 5): c1) a dicotyledonous plant; c2) a monocot plant; c3) a cruciferous plant; c4) arabidopsis thaliana; c5) the wild type Arabidopsis thaliana Columbia-0 subtype.
10. The use of claim 6 or 9, or the method of any one of claims 7 to 9, wherein: the stress resistance is drought resistance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910957559.6A CN110760524B (en) | 2019-10-10 | 2019-10-10 | Specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910957559.6A CN110760524B (en) | 2019-10-10 | 2019-10-10 | Specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110760524A true CN110760524A (en) | 2020-02-07 |
| CN110760524B CN110760524B (en) | 2020-08-07 |
Family
ID=69331705
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910957559.6A Active CN110760524B (en) | 2019-10-10 | 2019-10-10 | Specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110760524B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101078015A (en) * | 2006-05-25 | 2007-11-28 | 中国科学院遗传与发育生物学研究所 | Korshinsk peashrub transcription factor CkAREB and application of the same in adversity resistant plant cultivation |
| CN104086635A (en) * | 2014-04-10 | 2014-10-08 | 内蒙古农业大学 | New drought resistant gene CkDHN1 in Caragana korshinskii Kom. |
| CN106674338A (en) * | 2015-11-09 | 2017-05-17 | 中国科学院植物研究所 | Application of stress resistance-related protein to regulation and control on stress resistance of plants |
| WO2019154346A1 (en) * | 2018-02-11 | 2019-08-15 | 北京紫光英力化工技术有限公司 | Use of bio-based sulfonate as nutrient source and conditioning agent |
-
2019
- 2019-10-10 CN CN201910957559.6A patent/CN110760524B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101078015A (en) * | 2006-05-25 | 2007-11-28 | 中国科学院遗传与发育生物学研究所 | Korshinsk peashrub transcription factor CkAREB and application of the same in adversity resistant plant cultivation |
| CN104086635A (en) * | 2014-04-10 | 2014-10-08 | 内蒙古农业大学 | New drought resistant gene CkDHN1 in Caragana korshinskii Kom. |
| CN106674338A (en) * | 2015-11-09 | 2017-05-17 | 中国科学院植物研究所 | Application of stress resistance-related protein to regulation and control on stress resistance of plants |
| WO2019154346A1 (en) * | 2018-02-11 | 2019-08-15 | 北京紫光英力化工技术有限公司 | Use of bio-based sulfonate as nutrient source and conditioning agent |
Non-Patent Citations (2)
| Title |
|---|
| YAN LONG等: ""Identification of drought response genes by digital gene expression (DGE)"", 《GENE》 * |
| 符娜: ""柠条锦鸡儿植物抗旱基因的筛选"", 《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110760524B (en) | 2020-08-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113337520B (en) | Upland cotton GhA0749 and GhD0744 transcription factors and application thereof in flowering regulation | |
| WO2022135246A1 (en) | R gene for controlling matching of soybean-rhizobium, protein and use thereof | |
| CN110872598B (en) | Cotton drought-resistant related gene GhDT1 and application thereof | |
| CN109837296B (en) | New salt-tolerant drought-tolerant function of corn gene ZmNAC77 and application thereof | |
| CN110804090B (en) | Protein CkWRKY33 and coding gene and application thereof | |
| CN110358772B (en) | OsEBP89 gene for improving abiotic stress resistance of rice, and preparation method and application thereof | |
| CN113621625B (en) | Application of sesame SiERF103 gene in enhancing plant resistance | |
| CN109180791B (en) | Gene related to plant drought tolerance, and coding protein and application thereof | |
| CN112626084B (en) | Strawberry MYB transcription factor FvMYB24 gene, expression protein and application | |
| CN110760524B (en) | Specific DNA fragment com58276 and application thereof in regulating and controlling plant stress resistance | |
| CN104673803B (en) | Application of gene methylation in regulation of gene expression | |
| CN106892973A (en) | Plant adversity resistance related protein GhMYB4 and encoding gene and application | |
| WO2013010368A1 (en) | Use of rice aerenchyma formation key gene oslsd2 | |
| CN101824080B (en) | Picea wilsonii transcription factor PwHAP5 and coding gene and application thereof | |
| CN105732785B (en) | Application of protein GhDHN1 in regulation and control of plant stress resistance | |
| Zhao et al. | Transformation of modified cowpea trypsin inhibitor gene and anti-bacterial peptide gene in Brassica pekinensis protoplasts mediated by Agrobacterium tumefaciens | |
| CN110760522B (en) | AK209 gene and its coded protein and application in resisting stress and increasing yield | |
| CN112375766B (en) | Rice antioxidant capacity related gene BRHIS1 and application thereof | |
| CN114958866B (en) | Gene for regulating soybean branch number and application thereof | |
| CN116536286B (en) | Rice OsCTK1 protein and application of encoding gene thereof | |
| CN114908106B (en) | Rose salt tolerance gene RrLBD40 and application thereof | |
| CN116121269B (en) | Gene TrMYB118 for regulating synthesis of plant anthocyanin and application thereof | |
| CN114835787B (en) | Application of Quercus suberectus QsSRO1 gene and encoding protein thereof in plant stress resistance | |
| CN117305266B (en) | Gene OsBDG1 related to rice stress resistance and application of coded protein thereof | |
| KR101190255B1 (en) | Antibiotics marker free creeping bentgrass having resistance against two herbicides |
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 |