CN114958906A - Gene related to low potassium stress of tobacco, promoter and application thereof - Google Patents
Gene related to low potassium stress of tobacco, promoter and application thereof Download PDFInfo
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- 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
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- 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
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Abstract
The invention belongs to the technical field of plant genetic engineering, and relates to a gene engineering geneNtIAA13Genes and promoters, in particular to genes and promoters related to low potassium stress of tobacco and application thereof.NtIAA13Application of gene in regulating and controlling growth and development of tobacco under low potassium stress resistance, and the geneNtIAA13The sequence of the gene is shown in SEQ ID No. 1. A promoter for promoting genes related to tobacco growth and development under low potassium stress. The promoter is initiated by constructing a low-potassium induction specific promoter proNtIAA13NtIAA13Expression vector for gene, realizationNtIAA13The gene is specifically and highly expressed under the condition of low potassium stress, so that the potassium content of tobacco leaves is improved on one hand, and the low potassium resistance of tobacco is improved on the other hand. Book (I)The invention can be widely applied to tobaccoIAA13The related theory of gene expression inhibition and the application research of increasing the potassium content of tobacco, resisting low potassium stress and the like.
Description
Technical Field
The invention belongs to the field of plant genetic engineering, and relates to a method for producing a plant proteinNtIAA13The gene, especially the gene related to tobacco low potassium stress, promoter and their application.
Background
Potassium is a mineral nutrient element necessary for plant growth and development, participates in multiple vital activities such as carbon and nitrogen metabolism in plants, is closely related to disease resistance and stress resistance of the plants, and influences the yield and quality of the plants. In order to improve the yield and the quality of crops, a large amount of potash fertilizer is applied in agricultural production, on one hand, because of the shortage of potash ore resources in China and the higher cost of imported potash fertilizer, the use of potash fertilizer increases the production cost; on the other hand, the use of a large amount of potash fertilizers brings environmental pollution and adverse effect on the ecological environment, and is not in accordance with the 'double reduction' policy of agricultural development in China. Therefore, how to improve the utilization efficiency of potassium in crops under the condition of reducing the use of potassium fertilizer is an important research direction in modern agriculture development at present. Patent CN 103031331A discloses a protein for improving the low potassium resistance of rice, which is used for cultivating new varieties of rice with low potassium stress resistance, and patent CN103965308A discloses a gene StWRKY6 with low potassium stress resistance in potatoes; the genes of the low potassium stress resistant channels in different plants are different.
Tobacco is an important economic crop in China and is also an important model plant for researching the molecular mechanism of crops. Bioinformatic analysis of tobacco genome data at present shows that there are at least 77 Aux/IAA members of tobacco, but the functions of these members are still unclear. In the early stage, we found an auxin repressor gene with unknown gene function from tobaccoNtIAA13，The gene is subjected to low potassium stress to enhance expression, so that the deep research on the tobacco under the low potassium stressNtIAA13The regulation and control mechanism is crucial to improving the potassium absorption and utilization efficiency of plants, and can provide theoretical basis for breeding tobacco potassium high-efficiency utilization varieties.
In addition, since overexpression constructed by a strong 35S promoter easily influences certain physiological development of tobacco, the use of an inducible promoter is more beneficial to the growth and development of plants. The tobacco is a potassium-loving crop, potassium can obviously improve the combustibility and safety of the tobacco leaves, enhance the chromaticity and the filling property and improve the quality of the tobacco leaves, and the potassium content of the tobacco leaves is always used as an important index for evaluating the quality of flue-cured tobacco at home and abroad. The potassium content of tobacco leaves in China is generally low, and the difference between the potassium content of the tobacco leaves in China and international high-quality tobacco leaves is large, so that the potassium content of the tobacco leaves in China becomes one of the main factors for limiting the production of the high-quality tobacco leaves in China. Therefore, the research of a gene and a promoter for improving the potassium content and the potassium utilization efficiency of flue-cured tobacco is an urgent and significant subject.
Disclosure of Invention
The invention provides a gene and a promoter related to low potassium stress resistance of tobacco and application thereof, creates a new low potassium stress resistance flue-cured tobacco strain, has important application value for reducing potassium fertilizer application and cost and environmental pollution, and provides ideas and methods for cultivating new high-efficiency potassium varieties.
The technical scheme of the invention is realized as follows:
NtIAA13application of gene in regulating and controlling growth and development of tobacco under low-potassium stress resistance, and the geneNtIAA13The sequence of the gene is shown in SEQ ID No. 1.
A promoter for promoting genes related to tobacco growth and development under low potassium stress.
Furthermore, the sequence of the promoter is shown as SEQ ID No. 2.
Furthermore, the gene sequence is shown as SEQ ID No. 1.
A recombinant vector containing the aboveNtIAA13Genes and promoters as described above.
The recombinant vector is applied to the regulation and control of the growth and development of tobacco under the low potassium stress resistance.
The recombinant vector is applied to culturing new flue-cured tobacco varieties which efficiently utilize potassium.
Further, the method comprises the following steps: and (3) genetically transforming the recombinant vector into the tobacco to be modified, and culturing to obtain the superior tobacco with the high-efficiency utilization of the potassium.
Preferably, the tobacco to be modified is a flue-cured tobacco variety.
Preferably, the tobacco to be modified is flue-cured tobacco variety K326.
The invention has the following beneficial effects:
1. the invention utilizes agrobacterium transformation method to obtainNtIAA13Over-expressing the plants, found theNtIAA13Under the low potassium stress, the gene improves the potassium content and the potassium utilization efficiency of tobacco plants by regulating and controlling the expression of related genes of a high-affinity potassium transporter NtHAK gene family, further regulates and controls the growth and development of the tobacco plants under the low potassium stress, improves the low potassium stress tolerance of the tobacco plants, and can provide theoretical basis for cultivating new low potassium-resistant flue-cured tobacco varieties.
2. The present application findsNtIAA13Expression was highest in roots, second in stems, and lowest in leaves (FIG. 4A). Further determines that the whole tobacco seedling cultured for 21 days has no N, P, K, Ca and Mg under different stresses,NtIAA13relative expression level of genes. The results show that low potassium stress is up-regulated in tobaccoNtIAA13Expression of the gene (FIG. 4B). These findings indicate that it is possible to identify,NtIAA13the gene participates in the growth and development of tobacco and can regulate and control the low potassium stressNtIAA13Gene expression; by constructing low-potassium induction specific promoter to startNtIAA13Genes create a new flue-cured tobacco strain resistant to low potassium stress, and provide ideas and methods for cultivating new high-efficiency potassium varieties. The over-expression vector is constructed by utilizing the low-potassium induced specific promoter proNtIAA13, so that a new low-potassium stress resistant flue-cured tobacco strain is created, and the method has important application value for reducing potassium fertilizer application, cost and environmental pollution.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a drawing ofNtIAA13Electrophorogram of gene clones.
FIG. 2 isNtIAA13Subcellular localization maps in tobacco epidermal cells.
FIG. 3 is a schematic diagram of an embodiment of the present inventionNtIAA13Expression patterns in different parts of tobacco and under different nutrient deficiency stresses.
FIG. 4 shows the phenotypic characteristics biomass, chlorophyll content and antioxidant capacity of the transgenic material NtIAA13 under different potassium supply conditions for wild type tobacco (WT) and proNtIAA13 provided by the present invention.
FIG. 5 shows the potassium content and potassium accumulation of wild type tobacco (WT) and proNtIAA13 transgenic material NtIAA 13.
FIG. 6 shows the expression of high affinity potassium transporter gene of the transgenic material NtIAA13 in wild type tobacco (WT) and proNtIAA 13.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art based on the embodiments of the present invention without inventive step, are within the scope of the present invention.
NtIAA13Application of gene in regulating and controlling growth and development of tobacco under low-potassium stress resistance, and the geneNtIAA13The sequence of the gene is shown in SEQ ID No. 1.
A promoter for promoting a gene related to the growth and development of tobacco under low potassium stress.
Furthermore, the sequence of the promoter is shown as SEQ ID No. 2.
Furthermore, the gene sequence is shown as SEQ ID No. 1.
A recombinant vector containing the aboveNtIAA13Genes and promoters as described above.
The recombinant vector is applied to the regulation and control of the growth and development of tobacco under low potassium stress resistance.
The recombinant vector is applied to culturing new flue-cured tobacco varieties which efficiently utilize potassium.
Further, the method comprises the following steps: and (3) genetically transforming the recombinant vector into the tobacco to be modified, and culturing to obtain the superior tobacco with the high-efficiency utilization of the potassium.
Preferably, the tobacco to be modified is a tobacco variety.
Preferably, the tobacco to be modified is flue-cured tobacco variety K326.
Example 1
Aiming at the prior artNtIAA13Cloning and expressing, the invention clones from tobacco K326NtIAA13Gene and promoter sequences, constructionNtIAA13Promoter initiationNtIAA13Overexpression vector of gene and genetic transformation to obtainNtIAA13Overexpressing transgenic plants using wild-type tobacco K326 andNtIAA13the biomass and the oxidation resistance of the plant under the low potassium stress are analyzed by the overexpression transgenic plant,The potassium content, the accumulated potassium content and the expression level of the related gene of the high-affinity potassium transport protein are found by analysisNtIAA13The over-expression of the gene can regulate the growth and development of tobacco plants under low potassium stress, improve the tolerance of the tobacco plants to the low potassium stress,
tobacco auxin suppressor geneNtIAA13The sequence of (A) is shown as SEQ ID No.1, and the sequence of the promoter is shown as SEQ ID No. 2.
Tobacco auxin response factorNtIAA13The cloning method of the gene and the promoter specifically comprises the following steps:
the first step is as follows: extracting total RNA of tobacco leaves; taking K326 root system as material, extracting Total RNA by using a method provided by Eastep Super Total RNA Extraction Kit (Shanghai Promega biological products Co., Ltd.), determining the OD260/280 value of the extracted Total RNA, and detecting the integrity of the extracted Total RNA by using 1.2% agarose gel electrophoresis;
the second step is that: obtaining total cDNA and total genome DNA of tobacco root systems; taking 1ug of K326 young leaf total RNA sample as a template, and carrying out reverse transcription by using a HiScript III 1st Strand cDNA Synthesis Kit (+ gDNA wiper) Kit (Nanjing Nozanza biotechnology, Inc.) to obtain cDNA; extracting total DNA of the tobacco genome by adopting EasyPure Plant Genomic DNA Kit of Beijing all-purpose gold biotechnology limited according to a reference instruction;
in a preferred embodiment of the invention, the reverse transcription conditions are: 40min at 42 ℃, 30min at 50 ℃, 5min at 99 ℃ and 5min at 5 ℃.
The third step: according to predictions on NCBINtIAA13Design of Gene and promoter sequences in tobaccoNtIAA13The gene and promoter cloning primer, the primer sequence constructed by eukaryotic expression vector is shown in table 1:
TABLE 1 primer sequences
In a preferred embodiment of the invention, the eukaryotic expression vector is proNtIAA13CAMBIA1305 engineered vector driving the objective replacement of the 35S constitutive promoter.
The fourth step:tobaccoNtIAA13PCR amplification of genes and promoters. Respectively taking 1 mu L of cDNA and genomic DNA of tobacco root system as templates, adopting a standard 50 mu L PCR reaction system, and amplifying by Phusion ultra-fidelity DNA polymeraseNtIAA13Full-length CDS sequence and promoter sequence of gene;
in a preferred embodiment of the invention, the parameters for PCR amplification of the fragment of interest are as follows: the sequence of NtIAA13 CDS (pre-denaturation at 94 ℃ for 4min, 35 cycles, and heat preservation at 72 ℃ for 10 min. the cycle process is denaturation at 94 ℃ for 1min, annealing at 58 ℃ for 1min, and extension at 72 ℃ for 1 min);NtIAA13a gene promoter with about 1500bp sequence (pre-denaturation at 94 ℃ for 4min, 35 cycles, heat preservation at 72 ℃ for 10 min. the cycle process is denaturation at 94 ℃ for 1min, annealing at 58 ℃ for 1min, and extension at 72 ℃ for 1.5 min);
the construction method of the over-expression vector provided by the embodiment of the invention specifically comprises the following steps:
and (3) constructing and testing an overexpression vector. For constructing an overexpression vector proNtIAA13pCAMBIA1305, first, using tobacco genomic DNA as a template and amplifying it with primersNtIAA13The entire promoter sequence of (a) was constructedNtIAA13Promoter-driven CAMBIA1305 intermediate vector, followed by cloningNtIAA13Full length CDS sequence (FIG. 1), constructed by enzymatic cleavageNtIAA13Promoter initiationNtIAA13The expression vector of (1); selecting positive clones to carry out PCR identification on bacteria liquid, sending positive clones to sequence, verifying that the sequence is correct, and successfully constructing the vector;
subcellular localization of NtIAA13 in tobacco epidermal cells
For analysis of subcellular localization, one would have no terminal codonNtIAA13The cDNA was fused to the N-terminus of the Green Fluorescent Protein (GFP) gene in the pBWA (V) HS vector. Subcellular localization was transiently expressed in leaf epidermal cells of Nicotiana benthamiana and control cells were similarly transformed with 35S:: GFP. The transformed tobacco skin samples were then stored at 25 ℃ for 16 hours in the dark. Transient expression of GFP was monitored by confocal laser scanning microscopy (Nikon C2-ER). As shown in FIG. 2, NtIAA13 is localized in the nucleus and is a regulatory factor.
DeterminingNtIAA13Is regulated by low potassium signal
Wild Type (WT) tobacco seeds are sterilized, uniformly scattered in a sponge seedling tray, and placed in a constant temperature and humidity culture room (28 ℃ in the daytime, 14 hours in the daytime, 22 ℃ in the evening, 10 hours in 24-hour circulation, and the humidity is set to be 60%). For detecting different tobacco tissuesNtIAA13The tissue specificity expression of the gene, the root, the stem and the leaf of the tobacco cultured for 10 weeks are sampled in different parts and are measured by a qRT-PCR method (the adopted primers are shown in Table 1), and the result shows that,NtIAA13expression was highest in roots, second in stems and lowest in leaves (FIG. 3A). Further determines that the whole tobacco seedling cultured for 21 days has no N, P, K, Ca and Mg under different stresses,NtIAA13relative expression levels of the genes. The results show that low potassium stress is up-regulated in tobaccoNtIAA13Expression of the gene (FIG. 3B). These findings indicate that it is possible to identify,NtIAA13the gene participates in the growth and development of plants and can regulate and control the low potassium stressNtIAA13And (4) expressing the gene.
To obtainNtIAA13Overexpression of transgenic tobacco plants
The specific primers in Table 1 were amplified from tobacco leavesNtIAA13CDS sequence ofNtIAA13Over-expression of (a). The PCR product was ligated into pCAMBIA1305 vector driven by proNtIAA13 promoter and nopaline synthase terminator. The construct was transferred to Agrobacterium tumefaciens strain EHA105 by electrophoresis and transformed into tobacco (Nicotiana tabacum cv, K326). By introduction ofNtIAA13The construct was overexpressed, and 16 transgenic tobacco lines were obtained. The analysis identified two transgenic lines with better overexpression (named OX1 and OX 2) that were confirmed and used in this study.
AuthenticationNtIAA13Tolerance of transgenic tobacco to low potassium stress is overexpressed. By introduction ofNtIAA13Over-expression constructs, wild-type (WT) tobacco seeds andNtIAA13-OXthe transgenic tobacco seeds (T2 generation) were sterilized, uniformly scattered in a sponge seedling tray, and placed in a constant temperature and humidity culture room (28 ℃ in the daytime, 14 hours, 22 ℃ in the evening, 10 hours, cycle every 24 hours; humidity setting is 60%). When the seeds germinated, they grew into two leaves, 1/4 Hoagland nutrient solution was added for 4 days, and then 1/2 Hoagland nutrient solution was addedThe solution was incubated for 4 days, then Hoagland's nutrient solution was added for 14 days. About 30 days of tobacco seedlings were subjected to normal potassium (2 mmol/L) low potassium (0.1mmol/L) stress treatment for 7 days, tobacco seedlings were harvested and leaves and roots were rinsed with deionized water.
Examples of effects analysis
The following studies were performed on wild type and transgenic line samples:
1. study of tobacco biomass (fig. 4A); indicating that under low potassium stress conditions,NtIAA13-OXthe biomass of the transgenic plant is higher than that of Wild Type (WT) tobacco, and the growth vigor is relatively good.
2. Measuring chlorophyll content (fig. 4B); indicating that under low potassium stress conditions,NtIAA13-OXthe chlorophyll content of the transgenic plants was significantly higher than the control WT.
3. Measuring the activity of antioxidant enzyme SOD and MDA content (figure 4C, D); under the condition of low potassium stress, the method can reduce the stress of potassium,NtIAA13-OXthe MDA content of the transgenic tobacco is obviously lower than that of the wild type, and the activity of SOD is obviously higher than that of the wild type. This indicates thatNtIAA13Can enhance the antioxidant capacity of tobacco.
4. Measuring potassium content and potassium accumulation (FIG. 5); at 7 days of low potassium treatment, compared to control WT,NtIAA13the potassium content and potassium accumulation of over-expressed transgenic lines OX1 and OX2 are both significantly higher than those of wild type. The low-potassium treatment reduces the absorption and utilization of potassium by tobacco plants,NtIAA13over-expression can enhance the absorption and accumulation of potassium by tobacco.
5. Determination of nitrate transporter genesNtHAK1、NtHAK5、NtHAK6、NtNKT1Expression level of (2) (fig. 6); under the condition of low potassium stress, the method can reduce the stress of potassium,NtIAA13-OXof transgenic tobaccoNtHAK1、NtHAK5The gene expression level is obviously higher than that of the wild type, andNtHAK6、NtNKT1the difference between the gene expression level and the wild type is not great.
The above results show thatNtIAA13The over-expression of the gene can regulate and control the growth and development of tobacco plants under low potassium stress, improve the low potassium stress tolerance of the tobacco plants and provide a basis for the comprehensive utilization of salt-tolerant tobacco cultivation technology and tobacco leaves.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
<110> Bijie City of tobacco Co, Guizhou province
Henan university of agriculture
<120> gene related to low potassium stress of tobacco, promoter and application thereof
<130> sequence Listing
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 879
<212> DNA
<213> Nacobbus aberrans
<400> 1
atggaaccaa cacttggttt acttgacact ggtgaaaaag gttgttcaaa tatggccaaa 60
gtggaacaag attacaatat ggacatgtgt tctgaggaag aaacagagct tgagttgggg 120
ttaggactta gtctaaacag tggtggtggg gtggtggcaa aagctaagaa atcagcatgg 180
ggtgattatg gtagaatttt aactgctaaa gattttccta atggattttc agctgcaggg 240
agatctatta ttaataatag tggtgtttct tctggtacta aaagagctgc tgattttgtt 300
ggttctaata ctgatgttgg atctcctcct actggttcca gtcaggttgt gggatggcca 360
cccataaggg catacagaat gaacagcttg gttaatcaat caaaggttct aaatgctgaa 420
gaagacaagg gagttggcgg gaacgataag aaggagcatt caaagaagaa aatcaatcat 480
ggaaatgcca aggacgacgc gacttctatc aaagaaaaag ggcatcttgg ttttgtaaag 540
gtgaatatgg atggtttgcc tattggaaga aaggtggatt tgaatgctca cacttgctat 600
gaatccttag cagaaacctt agaggatatg ttctgcaaat caactaaaag tggtgaaaag 660
gaacaaacaa caaagtcttt taagctcttg gatggatcat ctgaatttgt gctcacatat 720
gaggataaag aaggagactg gatgcttgtt ggagatgttc catgggagat gtttgtcaac 780
agtgtgaaaa ggctaagaat tatgaggact tctgaggcta atggacttgg tccaagaatc 840
cctcagaagc aggagagaca aaaagggaaa ccaatctaa 879
<210> 2
<211> 1507
<212> DNA
<213> Nacobbus aberrans
<400> 2
ggaataaaac ttaaaatatt attttgagtt tgaaattact ttctgcagta gtaaaatcag 60
tacttactta tgcagaaatc attttatgcg ggattgacta tgaaataaaa tttcttatat 120
tgttgtcatt taatgtataa taatttttac attattgttc atccgtatat caatcaatgc 180
attaaagtta cattactagt ccatgaccta aatacacaac aaggtggagt aaaacgagaa 240
aaattacttc atactccata caataacaaa taacttttgt aaaaatgcct cttttcattt 300
catctaatta gctgtcctac tttgcatgca tgagactgag aatgatatgc acgagaatta 360
ttagttgaaa ggacaatcat tttaatgttg taaaacaaat ttctattttt attgaattac 420
acatatttat attgtttctg attttttaca acaagattga tttaaatgat aaatcctgtg 480
ataatatttc aaactaccct gttgcggtat tatttattgg atattacatt cataaaatat 540
agttaggaga tcaaactcat tatttatgca ctttctatgg tataactaat tcgtactaat 600
taagcactgt ttattatttt aaataggtac aaatcaagca agtggtttct gaattccgga 660
aactaaatgg ttaaaacaga aaattaaggg gtcgtttggt ttgaagacat agttatattg 720
tgattaatta tgttgatatt agatataatg agattagtta tgttgagatt aattattctg 780
ctattatttc ttattgacta tttggtatgt tgtattaatt ctacgattgt caattttatt 840
actttatctt aggataactt atcttgtgat tactattcca atctttgaca gataaaaatt 900
atccttatac tatttttaat ctggattagt aaccaaataa aaattaattt tttctaaatt 960
ttaataatac taaatttttc ttccaaaatt atttttattt atctatccta ccaaactaca 1020
ccaaaaaaac actatctctt ccttcaaacc tcttatattt ttttaaggta agaaaagtag 1080
tagtactaaa atactatact ttctaaagac attttagaca tttacgaaac tcgagaagaa 1140
aaaaatgatt cccatactaa tatttgcttc caacatttgg acaaaagcaa aggggacaac 1200
ccaaaaacac tttcaagtag taatatcaaa agcatggcaa aatacaagcc aaaccaaaaa 1260
ccaaaaatca tgagctcgag aaattgaaat gaatacaata aaagaaactt gaggaagaag 1320
aattttgaca gtgcttgttg cactagcaga catatgggtc ccataatcat cactagctcc 1380
accatacatc cccaccccca cccccacccc ccaccccata ataaaccact atacattctc 1440
ttcattattc tctcaaatcc attttcttga aatcttactc acaaaatctt gaaacccctt 1500
ttgaatg 1507
Claims (10)
1.NtIAA13The application of the gene in regulating and controlling the growth and development of the tobacco under the low potassium stress resistance is characterized in that: the above-mentionedNtIAA13The sequence of the gene is shown in SEQ ID No. 1.
2. A promoter, characterized in that: used for starting genes related to the growth and development of tobacco under low potassium stress.
3. The promoter according to claim 2, wherein: the sequence of the promoter is shown as SEQ ID No. 2.
4. The promoter according to claim 3, wherein: the gene sequence is shown in SEQ ID No. 1.
5. A recombinant vector characterized by: the recombinant vector comprising the recombinant vector of claim 1NtIAA13A gene and the promoter according to claim 2 or 3.
6. The use of the recombinant vector of claim 5 for regulating the growth and development of tobacco resistant to low potassium stress.
7. The use of the recombinant vector of claim 5 in the cultivation of a new flue-cured tobacco variety with high potassium utilization efficiency.
8. Use according to claim 7, characterized by the steps of: and (3) genetically transforming the recombinant vector into the tobacco to be modified, and culturing to obtain the superior tobacco with the high-efficiency utilization of the potassium.
9. Use according to claim 8, characterized in that: the tobacco to be modified is a tobacco variety.
10. Use according to claim 9, characterized in that: the tobacco to be modified is flue-cured tobacco variety K326.
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| CN115725643A (en) * | 2022-10-19 | 2023-03-03 | 河南农业大学 | Application of NtMYB35 transcription factor in tobacco black shank resistance |
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| CN104450777A (en) * | 2014-10-13 | 2015-03-25 | 南京农业大学 | Method for improving potassium absorption efficiency of plant and resisting against potassium deficiency stress and recombinant expression vector used therein |
| CN106831967A (en) * | 2015-12-03 | 2017-06-13 | 北京大学 | Reduce IAA10 albumen and its encoding gene expression is improving plant to the application in fractilinea oryzae resistance |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115725643A (en) * | 2022-10-19 | 2023-03-03 | 河南农业大学 | Application of NtMYB35 transcription factor in tobacco black shank resistance |
| CN115725643B (en) * | 2022-10-19 | 2023-08-15 | 河南农业大学 | Application of NtMYB35 transcription factor in tobacco black shank resistance |
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| CN114958906B (en) | 2023-08-08 |
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