CN112280788B - Salicomia Herbacea HgS5 gene and application thereof - Google Patents
Salicomia Herbacea HgS5 gene and application thereof Download PDFInfo
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Abstract
The invention relates to a halogeton drought-resistant gene HgS5 and application thereof, belonging to the technical field of plant bioengineering. The invention clones drought-resistant gene HgS5 from salt-tolerant plant salina, the nucleotide sequence of the gene cDNA is shown as SEQ ID NO.1, and the molecular weight is 1738bp. The drought resistance of the arabidopsis plants can be obviously improved by converting the HgS5 gene. The drought-resistant gene HgS5 provided by the invention provides gene resources for cultivating new varieties of drought-resistant plants (crops).
Description
Technical Field
The invention belongs to the technical field of plant bioengineering, and particularly relates to a halophyte drought-resistant gene HgS5 and application thereof.
Background
Drought is a major natural disaster faced by people from ancient times, even today with developed agricultural technologies, water stress caused by drought and water shortage is still a major factor for limiting high and stable yield of plants (crops), along with economic development and population growth of people, water resource shortage phenomenon is more and more serious, and the expansion of arid areas and the aggravation of drought degree are directly or indirectly caused, so that drought trend has become a focus of global attention. Especially in northwest regions of China, drought has more serious influence on agricultural production.
Development of water-saving agriculture in arid regions is imperative to realize sustainable development of agriculture in northwest arid regions in China. Besides greatly developing water conservancy facilities and improving the crop growth environment, the method has great significance in culturing new varieties of crops with strong drought resistance. The cultivation of new varieties of drought-resistant crops is mainly realized through two ways of traditional crossbreeding and biotechnology breeding, but the biotechnology breeding has the advantages of strong pertinence, high breeding speed, high breeding efficiency and the like, and is valued and promoted by breeders, but most of crops have poor drought resistance, narrow genetic foundation and development of excellent drought-resistant genes as key limiting factors of drought-resistant breeding success and failure of crops in terms of traditional crops. A large number of drought-tolerant rural wild plants, such as cistanche salsa, broom cypress, ammopiptanthus mongolicus, halogeton, and the like, are distributed in northwest arid regions of China, contain abundant drought-resistant genes, discover the resources of the native drought-resistant genes, and have important value for accelerating the cultivation process of new varieties of drought-resistant crops in China.
The salina (Halogeton glomeratus) belongs to annual herbs of chenopodiaceae, is a drought-resistant and salt-tolerant pioneer plant widely distributed in northwest arid regions of China, has strong drought resistance and salt tolerance, and a salt tolerance mechanism mainly comprises two aspects of isolation of salt vacuole and limiting absorption of root salt, has obvious advantages in the development of drought-resistant and salt-tolerant genes, but has serious lag in research on the development of drought-resistant and salt-tolerant genes in the salina, and needs to be enhanced urgently.
Problems of the prior art: the whole genome of the halogeton has not been published, and the application of the halogeton HgS5 gene and drought resistance thereof is not seen in the prior art.
Disclosure of Invention
In view of the defects of the prior art, the invention provides a halogeton HgS5 gene and application thereof in drought resistance.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
1. the full-length sequence of the halogeton HgS5 gene is shown in a sequence table, and the cDNA coding sequence of the halogeton HgS5 gene is the nucleotide sequence from 298 th to 1407 th positions in SEQ ID NO.1, and the halogeton HgS5 gene comprises 1110bp. The coded amino acid sequence consists of 370 amino acids, as shown in SEQ ID NO.2.
2. The cloning and obtaining method of the halogeton HgS5 gene comprises the following steps: (1) sequence information acquisition; (2) cloning of HgS5 gene; (3) HgS5-pMD19-T transformed E.coli competent cells; (4) screening for bluish white spots and sequencing; (5) the bacterial liquid suitable for sequencing is stored in glycerol.
3. The method for verifying the HgS5 gene function of the salina comprises the following steps: (1) constructing a gene HgS5 overexpression vector; (2) Agrobacterium-mediated transformation and cultivation of Arabidopsis thaliana; (3) identifying the drought resistance of the arabidopsis through HgS5 gene transfer.
4. Application of halogeton HgS5 gene in improving drought resistance of plants.
The beneficial effects are that: the invention provides an HgS5 gene sequence, which is cloned from drought-resistant and salt-tolerant plant salina to a new drought-resistant gene, and further transformed Arabidopsis thaliana is subjected to drought resistance identification, and the result proves that the drought resistance of the Arabidopsis thaliana strain transformed with the HgS5 gene is greatly improved. The molecular weight of the HgS5 is less than 2000bp, the gene is small, the genetic transformation operation is convenient, and the acquisition of the drought-resistant gene HgS5 provides a precious gene resource for the cultivation of drought-resistant crops and new plant varieties.
Drawings
Fig. 1: gene HgS5 cDNA amplification map; wherein: m is D2000 Marker;1-2 is the PCR product of the HgS5 gene.
Fig. 2: constructing a colony PCR electrophoresis chart by the vector; wherein, M is DL 2000DNA Marker; the arrows 1-5 are all colony PCR products which can be successfully amplified to the target genes after being connected with an over-expression vector.
Fig. 3: double enzyme digestion verification diagrams of the over-expression vector; wherein, the enzyme cutting sites are Kpn I and BamH I.
Fig. 4: PCR detection of resistance plants; wherein, the numbers "1" to "12" are all the numbers of the resistant seedlings, "-" is the pure water blank control, "+" is the plasmid control, "WT" is the wild plant control, and "M" is the DL 2000DNA Marker.
Fig. 5: verifying drought resistance of the T1 generation; the transgenic arabidopsis thaliana and the wild type arabidopsis thaliana are subjected to drought resistance treatment, and are photographed in treatments of 0d, 3d, 6d, 9d and 12d respectively, wherein the upper row is wild type arabidopsis thaliana WT, and the next row is transgenic arabidopsis thaliana with HgS5 gene.
Fig. 6: verifying drought resistance of the T2 generation; the transgenic arabidopsis thaliana and the wild type arabidopsis thaliana are subjected to drought resistance treatment, and are photographed in treatments of 0d, 3d, 6d, 9d and 12d respectively, wherein the upper row is wild type arabidopsis thaliana WT, and the next row is transgenic arabidopsis thaliana with HgS5 gene.
Fig. 7: PCR detection electrophoresis patterns of T2 generation resistant plants; wherein, M is DL 15000+2000DNA Marker; WT is wild type arabidopsis as a control.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the following detailed description of the embodiments of the present invention will be given with reference to the accompanying drawings. Examples of these preferred embodiments are illustrated in the accompanying drawings. The embodiments of the invention shown in the drawings and described in accordance with the drawings are merely exemplary and the invention is not limited to these embodiments.
It should be noted here that, in order to avoid obscuring the technical solution of the present invention due to unnecessary details, only the structures and/or processing steps closely related to the solution according to the present invention are shown in the drawings, while other details having little relation are omitted.
Example 1
The embodiment provides a halogeton HgS5 gene, the full-length sequence of the gene is shown in a sequence table, and the cDNA coding sequence of the gene is the nucleotide sequence from 298 th to 1407 th positions in SEQ ID NO.1, and the gene comprises 1110bp. The coded amino acid sequence consists of 370 amino acids, as shown in SEQ ID NO.2.
Example 2
The embodiment provides a cloning and obtaining method of a salina HgS5 gene, comprising the following steps:
1. sequence information acquisition: the salt stress-induced expression gene HgS5 was identified based on the results of transcriptomic sequencing of salt stress responses of Salicomia Herbacea in 2015 (Transcriptomic profiling ofthe salt-stress response in the halophyte Halogeton glomeratus [ J ]. BMCgenomics,2015, 16 (1): 169).
HgS5 Gene cloning: the applicant uses the leaf of halogeton as material to isolate the HgS5 gene.
The preparation method of the halogeton drought-resistant gene HgS5 is mainly characterized by comprising the following steps:
(1) Leaves of 3-7d salt-grown grass seedlings are stressed by 200-500mM NaCl and used as materials, total RNA is extracted by a Trizol method, a cDNA synthesis kit (Dalianbao bioengineering Co., ltd.) is adopted, a cDNA first strand is synthesized by reverse transcription, and the gene fragment is amplified. Two enzyme cutting sites of Kpn I and BamH I are respectively added on the upstream and downstream primers of the salina drought-resistant gene HgS5.
The upstream primer F1 is 5')GGGGTACCGACGTTGTTTCCAGCCCACTTC-3’
The downstream primer R1 is 5')CGGGATCCAGAATGTTAAACACTTTACAATAC-3’
The PCR reaction system is as follows:
the amplification procedure was:
the electrophoresis of the PCR amplification products is shown in FIG. 1.
(2) The PCR product was recovered by using a gel recovery kit (Dalianbao bioengineering Co., ltd.; gel Extraction Mini Kit), and then the recovered product was connected to a pMD19-T (Dalianbao bioengineering Co., ltd.) carrier, and the specific operation steps were that 1. Mu.L of the pMD19-T Vector (Simple) carrier, 4. Mu.L of the recovered product, and 5. Mu.L of Solution I were slowly and uniformly mixed in a small PCR tube, lightly centrifuged for 3sec, left in a 16℃water bath, and then connected overnight, and the next day was taken to 4℃for storage, thereby obtaining a connecting Solution.
Transformation of competent cells of E.coli with HgS5-pMD19-T
1) LBA medium plates (containing 100 mug/mL ampicillin Amp) were prepared, and after 20 mug IPTG and 80 mug L X-gal were mixed evenly, each plate was evenly coated and dried in the dark for use;
2) Taking out DH5 alpha competent cells from-80 ℃, immediately placing into an ice box, and melting on ice;
3) Adding 5 μl overnight connection solution into 50 μl competent cells, mixing with the bottom of the flick tube, and placing in ice box for 30min;
4) Heat shock is carried out for 90 seconds in a water bath kettle at 42 ℃, the water bath kettle is quickly put into an ice box, and the ice bath is carried out for 3 minutes, so that the operation is stable during the period;
5) 400 mu L of LB liquid medium is added into the centrifuge tube, and the culture is carried out for 1 to 1.5 hours at 200rpm at 37 ℃ of a shaking table;
6) The bacterial liquid in the centrifuge tube is sucked, evenly coated on the flat plate prepared in the first step (200 mu L of bacterial liquid is coated on one flat plate), slightly dried, sealed and placed in a dark environment at 37 ℃ for culture overnight.
4. Screening for blue and white spots and sequencing (sequencing result is shown in sequence table)
1) 10mL of LB medium (containing 100. Mu.g/mL of ampicillin) was added to a 50mL centrifuge tube (sterilized);
2) Randomly picking a plurality of white single colonies on a conversion plate by using toothpicks (sterilized), respectively placing the white single colonies into a centrifuge tube, sealing a tube orifice, and culturing overnight under the dark condition of 200rpm at 37 ℃;
3) Sucking out 1 mu L of bacterial liquid as a template for PCR detection;
4) Sequencing the bacterial liquid with correct PCR detection.
5. The bacterial liquid suitable for sequencing is stored in glycerol.
Example 3
The embodiment provides application of a halogeton HgS5 gene, which comprises the following steps:
1. construction of Gene HgS5 overexpression vector
1. Sucking 5. Mu.L of the glycerol bacteria finally obtained in example 2, culturing overnight in 10mL of LB liquid medium (containing 100. Mu.g/mL ampicillin Amp) at 37 ℃ under 200rpm in the dark for 12 hours, and extracting plasmids;
2. meanwhile, extracting a plasmid of an over-expression vector pCambia2301, and carrying out double enzyme digestion;
the extracted plasmid was double digested with two endonucleases KpnI and BamHI.
The enzyme digestion system is as follows:
a total volume of 20. Mu.L was digested at 37℃for 3 hours, and the digested product was purified.
3. Then, the purified and recovered vector and the target fragment were ligated with T4 DNA ligase.
The connection system is as follows:
10×T4 DNA Ligase 1ul
7ul of target fragment HgS5 gene
Vector pCambia2301 DNA 1ul
The total volume was 9. Mu.L. Keeping the temperature in a water bath at 65 ℃ for 3min, and then carrying out ice bath for 1-2min, and then adding T4 DNA Ligase 1ul and connecting at 16 ℃ for 12-16h.
4. The ligation mixture pCambia2301-HgS5 was transformed into E.coli DH 5. Alpha. Competent cells.
1) LBA medium plates (containing 100 mug/mL ampicillin Amp) were prepared, and after 20 mug IPTG and 80 mug L X-gal were mixed evenly, each plate was evenly coated and dried in the dark for use;
2) Taking out DH5 alpha competent cells from-80 ℃, immediately placing into an ice box, and melting on ice;
3) Adding 5 μl overnight connection solution into 50 μl competent cells, mixing with the bottom of the flick tube, placing in ice box for 30min,
4) Heat shock is carried out for 90 seconds in a water bath kettle at 42 ℃, the water bath kettle is quickly put into an ice box, and the ice bath is carried out for 3 minutes, so that the operation is stable during the period;
5) 400 mu L of LB liquid medium is added into the centrifuge tube, and the culture is carried out for 1 to 1.5 hours at 200rpm at 37 ℃ of a shaking table;
6) The bacterial liquid in the centrifuge tube is sucked, evenly coated on the flat plate prepared in the first step (200 mu L of bacterial liquid is coated on one flat plate), slightly dried, sealed and placed in a dark environment at 37 ℃ for culture overnight.
5. Screening blue and white spots, PCR detection of bacterial liquid (shown in figure 2), and preserving glycerol bacteria.
1) 10mL of LB medium (containing 100. Mu.g/mL of ampicillin) was added to a 50mL centrifuge tube (sterilized);
2) Randomly picking a plurality of white single colonies on a conversion plate by using toothpicks (sterilized), respectively placing the white single colonies into a centrifuge tube, sealing a tube orifice, and culturing overnight under the dark condition of 200rpm at 37 ℃;
3) Sucking out 1 mu L of bacterial liquid as a template for PCR detection;
4) The bacterial liquid suitable for detection is stored in glycerin.
6. mu.L of glycerol bacteria was aspirated, and the resultant was cultured overnight in 10mL of LB liquid medium (containing 100. Mu.g/mL ampicillin Amp) at 37℃under 200rpm in the dark for 12 hours to extract a recombinant plasmid.
7. The recombinant plasmid was digested at 37℃for 3 hours.
The double enzyme digestion system comprises:
the total volume was 20. Mu.L. And the double enzyme digestion products are subjected to electrophoresis detection (shown in figure 3).
The gene is constructed into an over-expression vector and is transformed into model plant Arabidopsis thaliana. The gene is verified to have the capability of obviously improving drought tolerance of plants. The gene HgS5 of the invention is utilized to construct various plant expression vectors, and can be widely applied to cultivation of new drought-resistant varieties of transgenic plants (crops).
2. Agrobacterium-mediated transformation and culture method for arabidopsis thaliana
1. LBA4404 Agrobacterium was prepared and activated, and recombinant plasmids pCambia2301-HgS5 were transferred into competent Agrobacterium to prepare an Agrobacterium dip.
2. The method comprises the steps of infecting arabidopsis inflorescences by a flower dipping method, covering plastic films on infected plants, culturing for 24 hours in a dark place, transferring to a greenhouse, maintaining at 22 ℃ and humidity of about 70% under the illumination condition for 16 hours, and harvesting T0 generation seeds by individual plants after the arabidopsis is mature.
3. And (3) carrying out low-temperature treatment and sterilization on the harvested transgenic arabidopsis thaliana T0 generation seeds, then broadcasting the seeds in a 1/2MS solid medium containing an antibiotic Kan (40 ug/ml), screening resistant seedlings, transplanting and culturing, randomly cutting plant leaves to extract DNA after the seedlings grow up, amplifying target genes by conventional PCR, carrying out electrophoresis on amplified products (shown in figure 4), and comparing and confirming the amplified products with carrier plasmids containing the target genes.
PCR detection primer:
an upstream primer: 35s-F: GACGCACAATCCCACTATCC
A downstream primer: hgS5-R CGGGATCCAGAATGTTAAACACTTTACAATAC
PCR amplification system:
total volume 25. Mu.L
PCR amplification procedure:
3. identification of drought resistance of Arabidopsis through HgS5 gene transfer
1. The obtained 11T 0 generation positive seedlings are routinely cultivated until the seeds are harvested, the T1 generation HgS5 transgenic arabidopsis seeds and the wild type arabidopsis (WT) seeds are sown in a culture medium (turf: vermiculite: perlite is mixed in a volume ratio of 1:3:0.5), the seedlings are cultivated in an artificial greenhouse, after the seedlings grow for 1 month, the watering of nutrient solution is stopped, natural drought treatment is carried out, the growth condition of the seedlings is observed (as shown in the attached drawing 5), and as a result, the growth speed of the transgenic arabidopsis is obviously faster than that of the wild type arabidopsis, the wild type arabidopsis plants begin to wilt when the drought treatment is carried out for 6 days, the transgenic arabidopsis normally grows, the wild type arabidopsis seriously wilt when the drought treatment is carried out for 9 days, the transgenic arabidopsis only partially wilt, the wild type arabidopsis completely withers when the seedling is 11 days, the transgenic arabidopsis aggravates, and is not completely withered.
2. Sowing the harvested transgenic arabidopsis T2 generation seeds and wild arabidopsis (WT) seeds into a culture medium (turf: vermiculite: perlite is mixed in a volume ratio of 1:3:0.5), culturing in an artificial greenhouse, randomly cutting plant leaves to extract DNA after seedlings grow up, amplifying target genes by conventional PCR, comparing with the wild arabidopsis, carrying out electrophoresis on amplified products (shown in figure 7), and carrying out PCR detection primers:
an upstream primer: 35s-F: GACGCACAATCCCACTATCC
A downstream primer: hgS5-R CGGGATCCAGAATGTTAAACACTTTACAATAC
PCR amplification system:
total volume 25. Mu.L
PCR amplification procedure:
after the seedlings grow for 1 month, the natural drought treatment is stopped, and the growth condition of the seedlings is observed (as shown in the attached figure 6), and the result shows that the growth speed of the transgenic Arabidopsis thaliana HgS5 gene is obviously faster than that of the wild Arabidopsis thaliana, the wild Arabidopsis thaliana plant starts to wilt when the drought treatment is 6d, the transgenic Arabidopsis thaliana grows normally, the wild Arabidopsis thaliana seriously wilt when the transgenic Arabidopsis thaliana is 9d, only part of the transgenic Arabidopsis thaliana wilts, the wild Arabidopsis thaliana completely withers when the transgenic Arabidopsis thaliana reaches 11d, the wilt degree of the transgenic Arabidopsis thaliana is aggravated, and the transgenic Arabidopsis thaliana is not completely withered.
The foregoing is merely exemplary of the application and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the application and are intended to be comprehended within the scope of the application.
<110> Gansu agricultural university
<120> Salicomia Herbacea HgS5 gene and application thereof
<160>2
<210> 1
<211>1738
<212> DNA
<213> Salicomia Herbacea (Halogeton glomeratus)
<400> 1
1 GACGTTGTTT CCAGCCCACT TCAGGAGAAC TATCTGATTT CATTCCACTA TTGCGTGAAG
61 TTTTTGGGCA AAAATCGGAC AAATTTCAAA GAAATTCTGC TGTCATTCTG ACTTGTGGTT
121 CTGTTTATCG AATAGCACTT GCTTTGTTTA ATCAACACTG CAAAAATGTG GCTAGCTTGT
181 TTTTAGTAAG ATGAAGACCT CACCGCTGCT GTCATTCTGA CTTGTGGTTC TGTTTATCGT
241 ATAGCACTTG CTTTGTTTAA TCAACACTGC AAAAATGTGG CTAGCTTGTT TTTAGTAAGA
301 TGAAGACCTC ACCGGACCCT CAGACTACTC CTGAAAGACA GTCCACCGAT CAAGTTTCCC
361 ATGAGTCCCA AAGTGGTCAA CAGAATAACA ATCAAGAAGC TCCAGTCCCT GACTCAGGTT
421 CAGCTGCAGT TTCAAGCAAT GATGGACGCA AAGTTTCTCG AGAAGATATT GAACTTGTCC
481 AAAATTTGAT AGAACGGTGC TTGCAACTGT ATATGAATAA AGATGAGGTG GTCAAAACCC
541 TCTTGAATCG TGCAAGGATT GATCCTGGAT TTACTACTTT AGTATGGCAG AAGTTGGAGG
601 AAGAAAATTC TGATTTTTTC AGAGCTTATT ATATAAGGCT CAAATTGAAG AAGCAGATTG
661 TTTTATTCAA TCATCTTCTT GAACACCAAT ATAATTTGAT GAAATATCCT GTTCCTCCGA
721 AAGTTCCTTT GGCTCCAATC CAGAATGGAA TACATCCTAT GCCTGTTAAC AACTTGCCTA
781 TGGGATACCC TGTCCTACAA CCACCTCCAA TGCCAGCCAC TGGTCAACCT CATCTTGATG
841 CTATGAGCCA TGGAATGTCT AGCTGCCATG TGGTTAATGG AGTCCCTGCT CCCAGTAATT
901 ATCAACCCAT GAGAATGAAT TCTGGGAATG ATATGGTGAT AGAAAGTAGC GCAGCTGAAG
961 CTGCACCGGC AGTTCCTCCC AGCACTGGCA TGACATCCAT GTCAGACATT CCCATCAGTC
1021 CAACTTCAGT GGCATCCAGT GGACATTTCC CATTCACTGC TTCAGAAATA CCTGGGATGG
1081 GAGTAGATAC CTCAGCACTA GATTCGGCAT TTCAATCTGA TGTGGGAAGT TCAATTGGAT
1141 TGCAGCTTCC ATCTGATGGA GGAGCTGGAA ATCCTAAAGA TTCTCTTCGT TCTTTGGCGC
1201 AGATTCCTTG GAATTTCAGT CTTTCTGACC TAACGGCAGA CTTATCAAAC TTGGGAGATC
1261 TTGGGCCTTT GGGAAATTAT CCTGGTTCAC CATTTCTGCC TTCCGATTCT GATATCTTGC
1321 TGGATTCGCC AGATAATGAG GATATAGTGG AGGAGTTCTT TGTCGATTCA GTCCCTGGGC
1381 AGCCACCACA ATCTGATGAG GAGAAGCCAT GAGCTAAGTT CTACCCTGTG CATTCTCCAA
1441 ATTCGATAGG CTGAACCTAA GCTCTGGAGA TTAGACCAGA GAATTATTCT GTAGGAAGGA
1501 GAAAGGAGTC AAGGACTGGC ATGAATTATC ATTTACATAG ACACATCGGG GATTGGATGA
1561 CGTCATGTAA TAGCCTATCG TCAGATGTAA ACAAGATTTT GCTGATGATT GTTCGGTTAT
1621 CGTTAGACTT TGTAATCCCA TCTTGTAGAT GCCATGCTAA TATGCCATAA CATTATGAGA
1681 CATATCATTC TTGATTCATT TTTCCTGGAA TTCAGTATTG TAAAGTGTTT AACATTCT
<210> 2
<211>370
<212> protein
<213> Salicomia Herbacea (Halogeton glomeratus)
<400> 2
1 MKTSPDPQTT PERQSTDQVS HESQSGQQNN NQEAPVPDSG SAAVSSNDGR KVSREDIELV
61 QNLIERCLQL YMNKDEVVKT LLNRARIDPG FTTLVWQKLE EENSDFFRAY YIRLKLKKQI
121 VLFNHLLEHQ YNLMKYPVPP KVPLAPIQNG IHPMPVNNLP MGYPVLQPPP MPATGQPHLD
181 AMSHGMSSCH VVNGVPAPSN YQPMRMNSGN DMVIESSAAE AAPAVPPSTG MTSMSDIPIS
241 PTSVASSGHF PFTASEIPGM GVDTSALDSA FQSDVGSSIG LQLPSDGGAG NPKDSLRSLA
301 QIPWNFSLSD LTADLSNLGD LGPLGNYPGS PFLPSDSDIL LDSPDNEDIV EEFFVDSVPG
361 QPPQSDEEKP
Claims (2)
1. The salina HgS5 gene is characterized in that the full-length sequence of the HgS5 gene is shown in SEQ ID NO:1, wherein the cDNA coding sequence is the nucleotide sequence from 298 th to 1407 th positions in SEQ ID NO.1, and comprises 1110bp; the coded amino acid sequence consists of 370 amino acids and is shown as SEQ ID NO.2.
2. Use of a salicorne HgS5 gene according to claim 1 for increasing drought resistance in arabidopsis thaliana, characterized in that said use is achieved by overexpression of the HgS5 gene.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001053323A2 (en) * | 2000-01-24 | 2001-07-26 | Ramot University Authority For Applied Research & Industrial Development Ltd. | Plants tolerant of environmental stress conditions, methods of generating same and novel polynucleotide sequence utilized thereby |
| CN105705643A (en) * | 2013-07-08 | 2016-06-22 | 路易斯安那州州立大学及农业机械学院管理委员会 | Abiotic stress resistance |
| CN106939314A (en) * | 2017-04-24 | 2017-07-11 | 甘肃农业大学 | Salt sward anti-drought gene Hg.S4 and its application |
| CN107022552A (en) * | 2017-04-24 | 2017-08-08 | 甘肃农业大学 | Salt sward resistant gene of salt HgS2 and its application |
| CN107287212A (en) * | 2017-07-20 | 2017-10-24 | 甘肃农业大学 | Salt sward resistant gene of salt HgS3 and its application |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001053323A2 (en) * | 2000-01-24 | 2001-07-26 | Ramot University Authority For Applied Research & Industrial Development Ltd. | Plants tolerant of environmental stress conditions, methods of generating same and novel polynucleotide sequence utilized thereby |
| CN105705643A (en) * | 2013-07-08 | 2016-06-22 | 路易斯安那州州立大学及农业机械学院管理委员会 | Abiotic stress resistance |
| CN106939314A (en) * | 2017-04-24 | 2017-07-11 | 甘肃农业大学 | Salt sward anti-drought gene Hg.S4 and its application |
| CN107022552A (en) * | 2017-04-24 | 2017-08-08 | 甘肃农业大学 | Salt sward resistant gene of salt HgS2 and its application |
| CN107287212A (en) * | 2017-07-20 | 2017-10-24 | 甘肃农业大学 | Salt sward resistant gene of salt HgS3 and its application |
Non-Patent Citations (1)
| Title |
|---|
| 盐生草盐胁迫响应基因HgS5功能研究;彭亚萍;中国优秀硕士学位论文全文数据库;全文 * |
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