CN114181946B - Gene and promoter related to low nitrogen stress resistance of plants and application of gene and promoter - Google Patents
Gene and promoter related to low nitrogen stress resistance of plants and application of gene and promoter Download PDFInfo
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Abstract
The application belongs to the technical field of plant genetic engineering, and relates toNtIAA26Genes, in particular genes related to low nitrogen resistance of flue-cured tobacco and application thereof. The gene related to low nitrogen stress resistance of plants is NtIAA26, and the sequence of the gene is shown in SEQ ID No. 1. The sequence of the promoter for promoting the genes is shown as SEQ ID No. 2. By constructing low nitrogen induced specific promoterNtIAA26The gene creates a new flue-cured tobacco strain resistant to low nitrogen stress, and provides ideas and methods for cultivating new nitrogen efficient varieties. The application constructs an over-expression vector by utilizing the low-nitrogen induced specific promoter IAA26, creates a new low-nitrogen stress resistant flue-cured tobacco strain, and has important application value for reducing nitrogenous fertilizer application, cost and environmental pollution.
Description
Technical Field
The application belongs to the field of plant genetic engineering, and relates toNtIAA26Genes, in particular to genes and promoters related to low nitrogen stress tolerance of plants and application thereof.
Background
Nitrogen is one of the important elements required for plant growth and development, and is an important constituent of proteins, nucleic acids, phospholipids and certain growth hormones in plants. The plant mainly acquires nitrogen from the environment to supply the growth and development of the plant, but the absorption, transportation and assimilation of the nitrogen by the plant are restricted by biological and abiotic factors, the sufficient nitrogen supply is usually ensured by applying nitrogen fertilizer in the agricultural production of China, but the excessive application of the nitrogen fertilizer can cause irreversible damage such as soil, water pollution, eutrophication and the like, the utilization rate of the nitrogen fertilizer by the plant is not increased, the cost is increased, and the resource waste is caused. Therefore, the nitrogen application can be reduced by improving the nitrogen utilization efficiency in agricultural production, and the method has important significance for preventing and controlling the pollution of an ecological system.
The auxin/indole-3-acetic acid (Aux/IAA) protein acts as a key factor in response to auxin signaling and plays an important regulatory role in auxin signaling. The Aux/IAA gene family is a multi-gene family of auxin-induced expression comprising four conserved domains I, II, III and IV. Domain I has transcription repression function. Domain II is an essential element of auxin signal transduction, interacting with the F-box protein TIR1, leading to instability and rapid degradation of Aux/IAA. Domains III and IV can interact with other Aux/IAA or ARF to form homodimers and heterodimers, inhibiting auxin response gene transcription, affecting signal transduction. Aux/IAA family genes have been identified and analyzed in many plants, 29 in Arabidopsis, 31 in rice, 26 in potato, etc. The Aux/IAA family plays an important regulatory role in the growth and development of plants and may increase the tolerance of plants to biotic and abiotic stresses. In recent years, research on the involvement of Aux/IAA family genes in plant low nitrogen and other nutritional stress is gradually reported, but key genes and regulatory mechanisms for regulating growth and development and involved in low nitrogen stress in plants are still unclear.
Tobacco is an important economic crop in China, and can also be used as a mode crop for revealing and controlling the growth and development mechanism of plants. At present, bioinformatic analysis of tobacco genome data shows that at least 77 Aux/IAA members exist in tobacco, but the functions of the members are still unclear. Earlier, we found tobacco auxin repressor genes from transcriptome databasesNtIAA26The expression enhancement of (2) may be related to nitrogen, and thus the tobacco auxin repressor gene under low nitrogen stress was studied intensivelyNtIAA26Is important to improve the nitrogen absorption and utilization efficiency of plants. Can provide theoretical basis for breeding tobacco nitrogen high-efficiency utilization varieties.
Because the over-expression of the 35S strong promoter construction easily causes the influence on certain physiological development of tobacco, the use of the inducible promoter is more beneficial to the growth and development of plants. The tobacco is used as an important cash crop in China, the quality of the tobacco is affected by the use of a large amount of nitrogenous fertilizer, black-cured tobacco leaves are produced, and meanwhile, the cost is increased, the income is reduced, and the environment is affected. Therefore, it is an urgent and significant topic to search for a gene and a promoter that improve the low nitrogen resistance of flue-cured tobacco, and to conduct intensive studies on the gene.
Disclosure of Invention
The application provides a gene and a promoter related to low nitrogen stress resistance of plants and application thereof, and the low nitrogen induced specific promoter is constructed to startNtIAA26The gene creates a new flue-cured tobacco strain resistant to low nitrogen stress, and provides ideas and methods for cultivating new nitrogen efficient varieties. The application constructs an over-expression vector by utilizing the low-nitrogen induced specific promoter IAA26, creates a new low-nitrogen stress resistant flue-cured tobacco strain, and has important application value for reducing nitrogenous fertilizer application, cost and environmental pollution.
The technical scheme of the application is realized as follows:
the gene related to low nitrogen stress resistance of plants is NtIAA26, and the sequence of the gene is shown in SEQ ID No. 1.
The sequence of the promoter for promoting the genes is shown as SEQ ID No. 2.
An over-expression vector comprising the above-described gene, said over-expression vector being controlled by the above-described promoter.
The over-expression vector is applied to improving the low nitrogen stress resistance of tobacco.
The over-expression vector is applied to cultivation of new varieties of flue-cured tobacco with high utilization rate of nitrogen.
The application comprises the following steps: the over-expression vector of the gene related to low nitrogen stress resistance of the plant is constructed by using the promoter, and then the over-expression vector is genetically transformed into a tobacco variety K326 which is not low nitrogen resistant, so that a new variety of flue-cured tobacco with high efficiency utilization of nitrogen is cultivated.
The application has the following beneficial effects:
1. the application utilizes agrobacterium transformation method to obtainNtIAA26Over-expression vector, foundNtIAA26Under low nitrogen stress, the gene can improve the nitrogen accumulation amount and the nitrogen utilization efficiency of the tobacco plant by regulating and controlling the expression of the antioxidant related genes and the nitrate transporter NRT1 and NRT2 gene family related genes, so as to regulate and control the growth and development of the tobacco plant under low nitrogen stress, improve the tolerance of the tobacco plant to low nitrogen stress, and provide theoretical basis for cultivating new varieties of low nitrogen resistant flue-cured tobacco.
2. The application discovers thatNtIAA26The highest expression was in the stems, next to the old leaves, the lowest in the new leaves (fig. 4A). Further determining that the whole tobacco seedlings cultured for 21 days are under different stresses (N, P, K, ca and Mg),NtIAA26relative expression levels of genes. The results show that low nitrogen stress upregulates tobaccoNtIAA26Expression of the gene (FIG. 4B). These findings indicate that the number of the active cells,NtIAA26the gene participates in the growth and development of plants and can regulate and control the low nitrogen stressNtIAA26Gene expression.
Drawings
In order to more clearly illustrate the embodiments of the application or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic view ofNtIAA26Subcellular localization maps in tobacco epidermal cells.
FIG. 2 shows an embodiment of the present applicationNtIAA26Expression patterns at different sites in tobacco and under different stress of lack of elements.
FIG. 3 shows the phenotypic characteristics of biomass, chlorophyll and antioxidant capacity of wild type tobacco (WT) and proIAA26 transgenic materials under different nitrogen supply conditions provided in the examples of the present application.
FIG. 4 shows the nitrogen uptake rate and nitrogen utilization efficiency of wild-type tobacco (WT) and proIAA26 transgenic materials provided by examples of the present application.
FIG. 5 shows the expression of nitrate transporter genes in wild-type tobacco (WT) and proIAA26 transgenic materials provided by the examples of the present application.
Detailed Description
The technical solutions of the present application will be clearly and completely described in conjunction with the embodiments of the present application, and it is apparent that the described embodiments are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without any inventive effort, are intended to be within the scope of the application.
The gene related to low nitrogen stress resistance of plants is NtIAA26, and the sequence of the gene is shown in SEQ ID No. 1.
The sequence of the promoter for promoting the genes is shown as SEQ ID No. 2.
An over-expression vector comprising the above-described gene, said over-expression vector being controlled by the above-described promoter.
The over-expression vector is applied to improving the low nitrogen stress resistance of tobacco.
The over-expression vector is applied to cultivation of new varieties of flue-cured tobacco with high utilization rate of nitrogen.
The application comprises the following steps: the over-expression vector of the gene related to low nitrogen stress resistance of the plant is constructed by using the promoter, and then the over-expression vector is genetically transformed into a tobacco variety K326 which is not low nitrogen resistant, so that a new variety of flue-cured tobacco with high efficiency utilization of nitrogen is cultivated.
Example 1
Is not aligned with the prior artNtIAA26Cloning and expression of the application from tobacco K326NtIAA26Gene, constructionNtIAA26Over-expression vector of gene and genetic transformation are carried out to obtainNtIAA26Over-expressing transgenic plants using wild type tobacco K326 andNtIAA26the over-expression transgenic plant analyzes the biomass, the antioxidation capability, the nitrogen accumulation and the utilization efficiency of the plant under the low nitrogen stress and the expression level of the nitrate transporter related genes, and the analysis shows thatNtIAA26The gene overexpression can regulate the growth and development of the tobacco strain under the low nitrogen stress, improve the tolerance of the tobacco strain to the low nitrogen stress,
tobacco auxin inhibitor geneNtIAA26The sequence of (2) is shown as SEQ ID No.1, and the sequence of (2) is shown as SEQ ID No. 2.
Tobacco auxin response geneNtIAA26The cloning method of (2) specifically comprises:
the first step: extracting total RNA of tobacco leaves; using K326 young leaves as a material, extracting total RNA by using a method provided by Eastep Super Total RNA Extraction Kit (Shanghai Probex Bioproduct Co., ltd.) kit, measuring OD260/280 value of the extracted total RNA, and detecting the integrity by using 1.2% agarose gel electrophoresis;
and a second step of: obtaining total cDNA and total genome DNA of tobacco leaves; reverse transcription was performed using 1ug of K326 young leaf total RNA sample as a template and HiScript type III 1st Strand cDNA Synthesis Kit (+gDNA wind) kit (Nanjinouzan Biotechnology Co., ltd.) to obtain cDNA; extracting total DNA of tobacco genome by adopting EasyPure Plant Genomic DNA Kit of Beijing full gold biotechnology Co., ltd and referring to the instruction book;
in a preferred embodiment of the application, the reverse transcription conditions are: 42 ℃ 40min,50 ℃ 30min,99 ℃ 5min,5 ℃ 5min.
And a third step of: tobacco leafNtIAA26Designing a gene cloning primer; through tobacco Aux/IAA genehomePhylogenetic analysis of familiesNtIAA26The coding region sequence and genome DNA sequence of the gene are compared with each other by DNAMAN V6.0 software to select the polypeptide sequenceNtIAA26Primer design is carried out on the specific locus of the gene to designNtIAA26The sequence of the primer NtIAA26-F constructed by amplifying the full-length cDNA and the genome DNA sequence of the gene and constructing a eukaryotic expression vector is shown in table 1; the NtIAA26-R sequence is shown in Table 1:
TABLE 1
In a preferred embodiment of the application, the eukaryotic expression vector is proNtIAA26The target gene CAMBIA1305 is started to modify the vector.
Fourth step: tobacco leafNtIAA26PCR amplification of genes. Using 1 mu L of first strand of tobacco young leaf cDNA and total genome DNA as templates, adopting a standard 50 uLPCR reaction system, and amplifying by using Phusion super-fidelity DNA polymeraseNtIAA26Full-length cDNA and genomic DNA sequences of the gene;
in a preferred embodiment of the application, the parameters for PCR amplification of cDNA are as follows: pre-denatured at 94 ℃ for 4min,35 cycles, and heat-preserving at 72 ℃ for 10min. When the genome DNA is amplified, the extension time is increased to 3min;
the cycle was run at 94℃for 1min denaturation, 58℃for 1min annealing, and 72℃for 2min extension.
The method for constructing the over-expression vector provided by the embodiment of the application specifically comprises the following steps:
and (5) constructing and checking an over-expression vector. To construct the overexpression vector proNtIAA26pCAMBIA1305, first, amplified using primersNtIAA26Construction of a 35S promoterNtIAA26Is then replaced by a vector promoter, and is constructed by enzyme digestionNtIAA26Promoter initiationNtIAA26Is a vector of (a) an expression vector of (b); selecting positive clones for bacterial liquid PCR identification, sequencing positive clones, verifying correct sequences and successfully constructing a vector;
RNA extraction and cDNA synthesis of tobacco (Nicotianatabacum cv, K326) leaves
Tobacco K326 leaf RNA was extracted with TRIZOL Plus and 1. Mu.g of total RNA was reverse transcribed into cDNA template. Using tobacco cDNA as a template, and performing PCR amplification by using a specific primer to obtainNtIAA26The fragment of the gene,
subcellular localization of NtIAA26 in tobacco epidermal cells
To analyze subcellular localization, the NtIAA26 cDNA without terminal codons was fused to the N-terminus of the Green Fluorescent Protein (GFP) gene in pCAM35s vector. Subcellular localization was transiently expressed in leaf epidermal cells of Nicotiana benthamiana and the same transformation was performed with 35S:: GFP on control cells. 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). The results are shown in fig. 1, with NtIAA26 localized in the nucleus.
Determination ofNtIAA26Is regulated by low nitrogen
Wild (WT) tobacco seeds were sterilized, uniformly spread in a sponge seedling tray, and placed in a constant temperature and humidity culture chamber (28 ℃ in the daytime, 14 ℃ in the evening, 22 ℃ in the evening, 10 hours, every 24 hours of circulation; humidity set at 60%). To detect different tobacco tissuesNtIAA26Tissue-specific expression of the gene, sampling root, stem, new leaf and old leaf of tobacco cultured for 10 weeks (new leaf is the second leaf from top to bottom, old leaf is the seventh leaf from top to bottom) and measuring by qRT-PCR method, the result shows that,NtIAA26the highest expression was in the stems, next to the old leaves, the lowest in the new leaves (fig. 2A). Further determining that the whole tobacco seedlings cultured for 21 days are under different stresses (N, P, K, ca and Mg),NtIAA26relative expression levels of genes. The results show that low nitrogen stress upregulates tobaccoNtIAA26Expression of the gene (FIG. 4B). These findings indicate that the number of the active cells,NtIAA26the gene participates in the growth and development of plants and can regulate and control the low nitrogen stressNtIAA26Gene expression.
ObtainingNtIAA26Overexpression transgenic tobacco plants
Amplification from tobacco leaves with the specific primers in Table 1NtIAA26For CDS of (2)NtIAA26Is over-expressed by (a). The PCR product was ligated into pCAMBIA1305 vector driven by the Cocois mosaic Virus (CaMV) 35S promoter and nopaline synthase terminator. The construct was transferred by electrophoresis to the Agrobacterium tumefaciens strain EHA105 and transformed into tobacco (Nicotianatabacum cv, K326). By introduction ofNtIAA26Over-expressing the construct, 10 transgenic tobacco lines were obtained. Analysis identified two transgenic lines (designated OX1 and OX 2) with better overexpression was confirmed and used in this study.
VerificationNtIAA26Tolerance of overexpressed transgenic tobacco to low nitrogen stress. By introduction ofNtIAA26Overexpression constructs, wild-type tobacco (WT) seeds andNtIAA26-OXthe transgenic tobacco seeds (T2 generation) are sterilized, uniformly spread in a sponge seedling tray, and placed in a constant temperature and humidity culture room (28 ℃ in the daytime, 14 hours at 22 ℃ in the evening, 10 hours in each 24 hours of circulation, and the humidity is set to be 60%). When the seeds germinated, two leaves were grown, 1/4 Hoagland nutrient solution was initially added for 4 days, then 1/2 Hoagland nutrient solution was added for 4 days, then Hoagland nutrient solution was added for 7 days. Tobacco seedlings for about 21 days were subjected to NaCl (150 mmol/L) stress treatment for 7 days, and tobacco seedlings were harvested, and leaves and roots were rinsed with deionized water.
Example of embodiment effect analysis
The following studies were performed on wild-type and transgenic strain samples:
1. study of phenotypic characteristics and biomass of tobacco (fig. 3a, b); indicating that under low nitrogen stress conditions,NtIAA26-OXthe growth vigor and biomass of the transgenic plants are significantly better than that of the WT plants.
2. Chlorophyll content was determined (fig. 3C); indicating that under low nitrogen stress conditions,NtIAA26-OXthe chlorophyll content of the transgenic plants was significantly higher than that of the control WT.
3. The activities of antioxidant enzyme SOD and POD were measured (FIG. 3D, E); under the condition of low nitrogen stress,NtIAA26-OXthe activity of SOD and POD of the transgenic tobacco is obviously higher than that of wild type tobacco. This indicatesNtIAA26The over-expression of (2) can enhance the antioxidant capacity of tobacco.
4. Measuring the nitrogen accumulation amount and the nitrogen utilization efficiency (FIG. 4); at 24h of low nitrogen treatment, both nitrogen accumulation and nitrogen utilization efficiency of the NtIA26 overexpressing transgenic lines OX1, OX2 were significantly higher than the control WT. The low-nitrogen treatment reduces the absorption and utilization of nitrogen by the tobacco strain,NtIAA26overexpression can enhance accumulation and utilization of nitrogen in tobacco.
5. Determination of nitrate transporter genesNtNRT1.1、NtNRT1.5、NtNRT1.7、NtNRT2.5Expression level of (a) (fig. 5); under the condition of low nitrogen stress,NtIAA26-OXtransgenic tobaccoNtNRT1.1、NtNRT1.5、NtNRT2.5The gene expression level is obviously higher than that of the wild typeNtNRT1.7The gene expression level is significantly lower than that of the wild type.
The above results indicate thatNtIAA26The over-expression of the gene can regulate the growth and development of the tobacco plant under low nitrogen stress, improve the tolerance of the tobacco plant to the low nitrogen stress, and provide basis for the salt-tolerant tobacco cultivation technology and the comprehensive utilization of tobacco leaves.
The foregoing description of the preferred embodiments of the application is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the application.
<110> Sichuan province, henan agricultural university, china tobacco Total
<120> Gene, promoter related to plant low nitrogen stress tolerance and use thereof
<141> 2021-11-22
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1521
<212> DNA
<213> Nicotiana tabacum L.
<400> 1
ataactctcc ctcttatatt tttagcttcc ctttatttac ttctccttct gttctctctc 60
tccattaact ctattcttca ctcttgaata ctctcaagat aaaacccact ctcactctca 120
gaaaatatat tagtcatcac catatataat tgctcttcct cctttttctt gacctatttt 180
tctttgtctt tgtggggttc ctttgaggtt ttatatcttc tggattcgac atatatacct 240
cttgtattag atcatctgat cagagaactt atacccagtt tttttttctc tatcaatatg 300
tgggtttctg tcacttgaag ctttggttga aactcaagga agttacttag gtagatatca 360
aacggaggtt ggtcttttat ttaattatta ttgacgttga atacataaga aagttaaaag 420
aaatggaggg ttattcaaga aacgaggaga aaaagctgga gttaaggctt ggtccgccaa 480
acggagactg gtctagagaa aaggatgaag cccttttccc ttttggttac aatatcagca 540
ccaacaatgg aagccaagtt cagcagcaaa agttttcttc atttcttcag cttcaaacaa 600
cgccacagaa acagagtgtg atggcacagg aatcattaca gacagctggt tgcattaaag 660
cagtaaatga tgcagaaaag aaggcatttt caccagctac aaaaacaggt cctgtctccc 720
acagtgctca gaaaaggact gcaccagcac cagtggtggg ttggcctcca attcgttcat 780
ttaggaagaa tattgcaagt agcagcagct ctttgaaacc tgtgactgcg tcacaaaatg 840
tggtcccaaa taagagtgat gccaacaaaa aaccaatgga aatttgtcag aaaggattat 900
ttgttaaaat caatatggat ggtattccta ttggaaggaa agtagatctc aaagcttatg 960
acagctatga taaactctcc tctgctgttg atgaactttt tagaggcctt cttgcagctc 1020
aaagtgatca atctggtggt gggaaagaga agaaggaaga aggagagaaa gcgataactg 1080
gattattgga cggaagtggc gaatatacgc ttgtttatga ggataatgaa ggtgacagaa 1140
tgcttgttgg ggatgtccct tggcatatgt ttgtgtctac tgtgaaaagg ctgcgtgtgt 1200
tgaaaagctc agacctctca acacttactc gtggaaataa gctgacctga tgatctgatc 1260
taacgcgcaa tcttgagaat caacaagaga atgaatgact tgttttgcat ggaattgaaa 1320
agtttttcat tattgaagta gtagaagcaa aaagagtttt tgtaaattat gaatattgct 1380
gcactgctaa tatttgtgtt tggaaggcaa aaaatgaata ttgctttctt tgttttgtcc 1440
ctttttgtat ccataggatg tgtgctatct ccagttaatt tctccaagtg tataaaatta 1500
aaatgtaaac cgttcgttct c 1571
<210> 2
<211> 1746
<212> DNA/RNA
<213> Nicotiana tabacum L.
<400> 2
aagtttgcag caaaggaaaa attgttgtgg gacttggccg gagaagattg ctgtaattaa 60
aaggaatttc agtccatttt aagcataata aacaacgggc ttgaagaact aaagttatgg 120
agagtctttg ttgggtttca acacgatgag cttttggtct ctgcttttaa tgttgttctt 180
gctgatgggt cttcttttcg tgttaatata tgtcaaggct tgtcactgtc gttttccttg 240
cttctaatgt tgttcttgct gatgtatttt catgtatttg ttgtattcat tgtccttttt 300
tcattgtaat tcaatgtatt ttgttgtatt ccatgtattt cattgtattc actgtcttct 360
tttctcattg tatttcaatg tatcccgttg tattttatgt atttcattgt attcactatc 420
tcgctatata ccatgaatat attcataagt tttttttaat taatataatt tatgtattca 480
gatgtattat ataatttctc tgaagattgc tatatttttg gggtattttt cgattgagaa 540
tcttttttat aactgaaaat acaaaatttg tgtgttataa ttgagtttgt tgagttatat 600
taggagtcta ttatgttaat tgattcattt tcccttttaa aaacaatgtg atcccctatt 660
tcacgccgta aatacaatcg aatacaataa tctgtccagt tgtaatccca tgtttctctc 720
catgaataca gtcgaataca ctcgaataca ataactgatt agctggactt ccctgattca 780
cgcctatatt tgctacagta ttcatgaata caataactta aatacatcaa atacatctta 840
taactatata aaacgtatct ataatctgta atatagcaaa tgatatctat aaataactaa 900
ttactgctaa aaatagtact ttatgaaaat ttcccttgtt ttatatgttt tatgaatagg 960
accggcccct aaagtgtgtg tatgtgagtc tatatgagag agggaaaagg tcacaaaaat 1020
aaaataaaat aaaataaaaa aggtagtcag cacagcacgc tttgcttaat gaaccaaaca 1080
aaatacaaca aaaaaagttt tgggggtttg tggttgcgtg agttttgcta caattccgtt 1140
tgctgttttt aggattgggt ccgttccttt tgagtgaaga gaatctcccc ctgcccccta 1200
atccctatat acataactct ccctcttata tttttagctt ccctttattt acttctcctt 1260
ctgttctctc tctccattaa ctctattctt cactcttgaa tactctcaag ataaaaccca 1320
ctctcactct cagaaaatat attagtcatc accatatata attgctcttc ctcctttttc 1380
ttgacctatt tttctttgtc tttgtggggt tcctttgagg ttttatatct tctggattcg 1440
acatatatac ctcttgtatt agatcatctg atcagagaac ttatacccag tttttttttc 1500
tctatcaata tgtgggtttc tgtcacttga agctttggtt gaaactcaag gtatttagag 1560
caacgatctt caactttcca tgaccttttg ccatgttttt aaccatagct tatgcttaaa 1620
cgtaaatttc ttgtgaacat gatgttgtct tcttttttgc aggaagttac ttaggtagat 1680
atcaaacgga ggttggtctt ttatttaatt attattgacg ttgaatacat aagaaagtta 1740
aaagaa 1746
Claims (2)
1. The application of the gene related to low nitrogen stress resistance of plants in the cultivation of new varieties of flue-cured tobacco with high utilization efficiency of nitrogen is characterized in that: the gene is NtIAA26, and the sequence is shown in SEQ ID No. 1; the promoter sequence for starting the NtIAA26 gene is shown in SEQ ID No. 2.
2. The use according to claim 1, characterized in that: the method comprises the following steps: the over-expression vector of the NtIAA26 gene started by the promoter is constructed, and then the over-expression vector is genetically transformed into a low nitrogen intolerant tobacco variety K326, so that a new variety of flue-cured tobacco with high efficiency utilization of nitrogen is cultivated.
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Non-Patent Citations (4)
| Title |
|---|
| NCBI Reference Sequence: XM_016632242.1.PREDICTED: Nicotiana tabacum auxin-responsive protein IAA26-like (LOC107807809), mRNA.NCBI Reference Sequence: XM_016632242.1.2016,全文. * |
| PREDICTED: Nicotiana tabacum auxin-responsive protein IAA26-like (LOC107807809), mRNA;NCBI Reference Sequence: XM_016632242.1;NCBI Reference Sequence: XM_016632242.1;全文 * |
| 低氮胁迫对烟草生物学性状、生长素及NtPINs基因家族的影响;王立志;魏跃伟;黄明月;朱彦铭;丁永乐;贾宏;;中国烟草学报(01);全文 * |
| 水稻品种"Lemont"响应低氮培养及共培稗草的上调表达基因分析;方长旬;许铁城;黄力坤;王清水;何海斌;林文雄;;中国生态农业学报(09);第1188页以及1189页右栏第1段 * |
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