CN111996200B - Application of TGA7 gene in regulation and control of plant flowering phase - Google Patents
Application of TGA7 gene in regulation and control of plant flowering phase Download PDFInfo
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Abstract
The invention discloses application of a TGA7 gene in regulation and control of plant flowering phase. Arabidopsis TGA7 gene stippled plants delayed flowering compared to wild type plants. Single base mutation in arabidopsis TGA7 gene. The number of rosette leaves of the Arabidopsis plant with the mutated TGA7 gene is more than that of the wild type under both long-day and short-day conditions. In the autonomous pathway mutants, TGA7 was increased in fca-2 and fv-4 and decreased in fld-3 and flk-1. The TGA7 gene expression quantity in wild type WT plants at 16-27 ℃ is obviously increased in sequence, while the TGA7 expression in SVP-41 plants at 23 ℃ is obviously reduced, and the TGA7 expression in SVP plants is obviously increased. The expression levels of key genes FLC, MAF5 and SMZ of the flowering regulation autonomous pathway in an Arabidopsis plant with the mutation TGA7 gene are obviously increased, and the expression level of NF-YC2 is obviously reduced.
Description
Technical Field
The invention relates to the technical field of molecular biology, in particular to application of a TGA7 gene in regulation and control of plant flowering phase.
Background
TGACG-binding (TGA) transcription factor belongs to the bZIP transcription factor family. The TGA family of arabidopsis thaliana has 10 members in total, and plays an important role in plant defense and plant development. These TGAs can interact with the NPR1 gene (inhibitor of non-disease-related gene 1), and are involved in salicylic acid-mediated gene expression (such as PR-1) and disease resistance in Arabidopsis. These TGAs can be associated with cis-regulated tgaccg elements. Studies have shown that this element is present in the promoter of PR1, and that PR1 gene expression requires the promoter to respond to SA and interact with NPR 1. NPR1 was unable to bind directly to the genomic region of PR1, NPR1 first interacted with TGA, mediating the expression of PR 1. Among the 10 TGAs of arabidopsis, NPR1 can interact with 7 TGAs, which in turn can be further divided into 3 sub-branches. TGA1 and TGA4 belong to the first category; TGA2, TGA5, TGA6 constitute the second class; the third category comprises TGA3 and TGA 7. In arabidopsis, only TGA1 and TGA4 interacted with NPR1 in SA-induced leaves, and the remaining TGA interacted with NPR 1. Therefore, all 7 TGAs play important roles in regulating the expression of defense genes and are important components of the plant defense system.
The higher plant flower formation comprises three parts of flower formation induction, flower primordium formation and flower organ formation and development, wherein the flower formation induction directly determines the morning and evening of flower formation. The survival of plants faces a great challenge to the ever changing environment, and many sensing mechanisms of plants have evolved to enable plants to integrate external signals and use them to regulate their metabolism, growth and development. Therefore, the method has important practical significance for the research of the flowering induction path of the plants. The arabidopsis TGA family plays an important role in plant defense, stress response, plant development and the like. Their role in flowering phase regulation has yet to be studied further.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides application of an arabidopsis gene TGA7 in regulating and controlling the flowering period of a plant.
In order to realize the purpose of the invention, the invention adopts the following technical scheme to realize:
the invention provides the function of an arabidopsis gene TGA7 in regulating and controlling the flowering period of arabidopsis.
Further: arabidopsis TGA7 gene stippled plants delayed flowering compared to wild type plants.
Further: the flowering gene causing expression change after the TGA7 gene mutation is FLC, MAF5, SMZ and NF-YC 2.
Further: the number of rosette leaves of the Arabidopsis plant with the mutated TGA7 gene is more than that of the wild type under both long-day and short-day conditions.
Further: the expression quantity of TGA7 in the autonomous pathway mutants fca-2, fve-4, fld-3 and flk-1 is obviously different from that of the wild type, the former two are obviously increased, and the latter two are obviously reduced.
Further: the TGA7 gene is involved in temperature pathway regulation, and the expression quantity of the wild TGA7 gene is obviously increased in sequence in 16-27 ℃. At 23 ℃, compared with wild type, the expression level of the TGA7 gene of SVP-41 and 35S-SVP plants is obviously reduced, and the expression level of the TGA7 gene of the SVP plants is obviously increased.
Further: the expression levels of key genes FLC, MAF5 and SMZ of the flowering regulation autonomous pathway in an Arabidopsis plant with the mutation TGA7 gene are obviously increased, and the expression level of NF-YC2 is obviously reduced.
The invention has the advantages and beneficial effects that:
1. by utilizing the existing plant biotechnology, the invention screens and obtains the homozygous plant of the single base mutant of the Arabidopsis thaliana mutant TGA7, and compares the homozygous plant with the flowering time and the rosette leaf number of a wild plant growing under the same condition, and finds that the expression change of flowering related genes (FLC, MAF5, SMZ and NF-YC2) in the plant caused by the mutated TGA7 gene is delayed compared with the flowering phase of the wild plant.
2. On the basis, the regulation mechanism is further researched. Five flowering regulation pathways are processed, and finally the expression of the TGA7 is regulated by an autonomous pathway and a heat sensation pathway. In the autonomous pathway mutants, TGA7 was increased in fca-2 and fv-4 and decreased in fld-3 and flk-1, suggesting that the autonomous pathway may affect the expression of TGA 7. In the ambient temperature range, TGA7 expression increases with temperature. Furthermore, TGA7 expression was higher in 35S: SVP and lower in SVP-41 at 23 ℃ in wild type WT, SVP-41 and 35S: SVP plants, suggesting that the temperature pathway may modulate the expression of TGA7 at ambient temperature.
3. After identification of key flowering-time related differences, it was found that there are 4 differentially expressed genes involved in the flowering-time pathway, which are related to flowering-time, play an important role in the flowering process, and are important turning points for plants from vegetative growth to reproductive growth. Compared with wild type WT seedlings, the expression levels of FLC, MAF5 and SMZ are up-regulated in the TGA7 mutant, while the expression level of NF-YC2 is down-regulated. The technical scheme of the invention has important significance on how to accurately regulate and control the flowering time of the TGA7 gene.
Drawings
FIGS. 1(a) - (b) are a gene schematic and homozygote identification electropherogram of the TGA7 mutant, respectively, wherein the black and gray boxes in the locus of the TGA7 mutant in (a) represent exons and untranslated regions, respectively, and the black bars represent introns; tga7 is a genomic point mutation, and the EcoRV enzyme cutting site C in the wild type is mutated into T; (b) the middle M channel is a reference group, the 1 channel is a wild type, and the 2 channel is a TGA7 mutant.
FIGS. 2(a) - (c) are statistics of the flowering phenotype and number of rosette leaves at flowering under Long Day (LD) conditions and the number of rosette leaves under Short Day (SD) conditions for the TGA7 mutant, respectively (.: P < 0.05).
FIG. 3 shows that the expression level of TGA7 gene is controlled in 6 flowering pathways by fluorescent quantitative PCR. Wherein (a) is the detection of the TGA7 gene expression level in the photoperiod pathway related mutants gi-1, co-9, ft-10; (b) TGA7 gene expression level detection after three and five weeks of gibberellin (GA3) treatment; (c) detecting the expression quantity of the TGA7 gene after the vernalization treatment; (d) the TGA7 gene expression quantity in the autonomous pathway related mutants of fpa-7, fca-2, fld-3, fve-4 and flk-1 is detected; (e) the expression quantity of the TGA7 gene is changed at 16 ℃, 23 ℃ and 27 ℃ under different temperature conditions; (f) the TGA7 gene was expressed in WT, SVP-41 and 35S: SVP at 16 ℃ and 23 ℃ (P < 0.05).
FIG. 4 shows the expression levels of genes related to the flowering time in wild-type WT and tga7 (.: P <0.05) by fluorescent quantitative PCR, wherein (a) is FLC, (b) is MAF5, (c) is SMZ, and (d) is NF-YC 2.
Detailed Description
The technical solution of the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.
Reagents used in the above experiments and the like were purchased from TAKARA, Roche, TIANGEN, CWBiO and the like.
Reagents and drug instructions used in the experiments: see molecular cloning, third edition.
EXAMPLE 1 obtaining of homozygous mutants
1. Obtaining of mutants
The TGA7 mutant seeds used were purchased from the Arabidopsis thaliana Resource Center (ABRC, the Arabidopsis Biological Resource Center, http:// www.arabidopsis.org /). Wherein the mutant is constructed by point mutation as shown in figure 1(a) under the number CS89835 of tga 7; c in the EcoRV enzyme cutting site of the wild genome is mutated into T, so that the EcoRV enzyme cutting site at the position of the genome of the tga7 is deleted, and the homozygous mutant can be obtained by enzyme cutting identification after PCR reaction. the full-length gene Coding (CDS) sequence of the tga7 gene is shown in SEQ ID NO. 1.
2. DNA extraction from Arabidopsis thaliana plant leaves
Taking a proper amount of seeds in wet filter paper, placing the filter paper in a refrigerator at 4 ℃ for dark treatment for 48 hours, finishing dormancy breaking of the seeds, transferring the seeds into soil, placing the seeds in a greenhouse for culture (16 hours of illumination/8 hours of dark, 23 ℃, 10000Lux), dropping five seeds in one pot, covering the seeds with a preservative film to keep stable germination conditions, continuing culturing the seeds for 2 days until 11 days, and taking a proper amount of leaves to extract genomes.
The experiment adopts self-prepared TPS extract, and the specific operation is as follows:
firstly, collecting a sample into a 1.5ml centrifuge tube, adding a proper amount of TPS and small steel balls, grinding the sample by using a grinder at 55Hz for 2min, and shaking uniformly.
② centrifuging for 10min at 13000rpm, taking out the supernatant and transferring to a new 1.5ml centrifuge tube, adding equal amount of isopropanol.
③ 13000rpm centrifugation for 5min, abandoning the supernatant, adding 500. mu.L 75% ethanol, and shaking up and down for several times. Centrifuging at 13000rpm for 5min, discarding the supernatant, and repeating the previous step.
13000rpm for 2min, sucking the liquid in the tube as much as possible by a pipette, drying in an oven at 37 ℃, adding 50 mu L of warm ddH2O, and dissolving completely.
TPS extracting solution formula
TPS(100ml)： 1M Tris-HCl(pH8.0) 10ml
0.5M EDTA(pH8.0) 40ml
2M KCl 50ml
PCR identification of homozygotes of the TGA7 mutant
(1) Designing an identifying primer in a forward direction TAAAGTTATCGCAGTTAGAGC as shown in SEQ ID NO. 2; reverse direction CCGCATCAATCACAATG as shown in SEQ ID NO. 3. After PCR amplification by using the primer, a 689bp mutant or wild type TGA7 fragment is obtained.
(2) And (3) PCR reaction system:
(3) PCR reaction procedure:
detecting by 1% agar gel electrophoresis and 5000bp Marker. The fragments were cut into 1.5ml centrifuge tubes.
(4) Product recovery (kit)
The product recovery kit of Roche company is adopted in the experiment, and the specific operation is as follows:
putting the glue to be recovered into a 1.5ml centrifuge tube, adding a proper amount of Binding Buffer, standing for a few minutes at 60 ℃ until the glue is completely melted, and shaking the glue by hand for a few times during the process to accelerate the melting.
② transferring the mixed solution to an adsorption column, and centrifuging the mixture for 1min at 8000 rpm.
③ adding 600 mul Washing Buffer after abandoning the waste liquid, centrifuging the waste liquid for 1min by 13000rpm, abandoning the waste liquid, repeating the operation once
And fourthly, the mixture is separated for 1min, and the waste liquid is discarded.
Fifthly, placing the adsorption column in a new 1.5ml centrifuge tube, standing at room temperature until the column head membrane is dried, then dripping 30 mul warm eluent, centrifuging the eluent for 2min at 13000rpm, and collecting the liquid, namely the pure PCR product.
(5) Enzyme digestion
Detecting by 1% agar gel electrophoresis and 5000bp Marker. The wild type genome is the EcoRV enzyme cutting site, after C is mutated into T, the EcoRV enzyme cutting site of the genome of tga7 is deleted, the wild type is cut by enzyme to obtain 389bp and 300bp fragments, but the mutant can not be cut and still is 689 bp. The electrophoresis results are shown in FIG. 1(b), and the tga7 mutant is a homozygous mutant.
Example 2 statistical analysis of flowering time and Lotus throne number
Growing under the condition of 23 ℃ and under the condition of long illumination (16h/8h, illumination/dark) or short illumination (8h/16h, illumination/dark) 10000Lux illumination. And selecting 20 plants of a normally growing arabidopsis wild type plant WT and a tga7 plant respectively, and counting the number of rosette leaves after the plants bloom.
Statistical results are shown in FIG. 2(a-c), where the time of flowering is later than that of the wild type in TGA7 plants, and the number of rosette leaves is greater than that of the wild type, indicating that the plants delayed flowering after TGA gene mutation in Arabidopsis thaliana. the TGA7 mutant exhibited delayed flowering under both long and short light conditions, suggesting that TGA7 may not be involved in the photoperiodic pathway.
Example 3 analysis of expression of five genes involved in the manipulation of the flowering regulatory pathways by TGA mutants and wild type
1. Five flowering regulation approaches
(1) Vernalization: and (3) plant: col, FRI
Processing a group: the non-resistant culture medium 1/2MS is inoculated at 4 deg.C for 2 days to break dormancy, placed at 4 days in a greenhouse, placed at 4 deg.C for 6weeks, placed at 5 days in the greenhouse, and then planted in soil (LD/SD).
② comparison: after the treatment group started to break dormancy for 36 days, the non-resistant medium 1/2MS was seeded at 4 deg.C for 2 days to break dormancy, placed in the greenhouse for 9 days, and then planted in the soil (LD/SD).
And thirdly, sampling at 9d after seedling transplanting, and quantitatively analyzing the TGA7 gene expression quantity.
(2) Gibberellin: and (3) plant: col, tga7
Processing a group: SD 100. mu.M GA3, sprayed once per week from day 7 of germination.
② comparison group: SD 0.1% ethanol, sprayed once a week from day 7 of spontaneous germination.
③ ZT8 Arabidopsis thaliana leaves were taken and stored at-80 ℃.
(3) The photoperiod: and (3) plant: col, tga7, gi-1, co-9, ft-10
And (3) treatment: sampling 9d seedlings after LD germination
(4) And (3) autonomous: and (3) plant: col, tga7, fld-3, flk-1, fve-4, fpa-7, fca-2, ld-1
And (3) treatment: sampling 9d seedlings after LD germination
(5) Temperature: and (3) plant: col + svp-41+35s-svp
And (3) treatment: 9d seedlings were sampled after LD germination at 16 deg.C, 23 deg.C, 27 deg.C.
2. Extraction of Total RNA of Arabidopsis thaliana (RNA kit of Tiangen Co.)
The 75% ethanol is used for cleaning and disinfecting the operation table when the glove is worn so as to prevent the pollution in the environment; in order to prevent degradation of RNA enzyme in saliva, the mask is worn in the whole process; in order to reduce the degradation degree of RNA, extraction should be carried out at low temperature as much as possible; furthermore, it should be ensured that all samples directly contacted during the process are RNase-Free. The experiment adopts an RNA kit of Tiangen company, and the specific operation is as follows:
firstly, shearing 2 pieces of arabidopsis thaliana rosette leaves at ZT16 time point by using clean scissors, placing the pieces in a 1.5ml centrifuge tube, adding a small steel ball, quickly placing the pieces in liquid nitrogen for freezing, and grinding the pieces into powder by using an automatic grinder for 35Hz and 2 min.
Preparing RL with 1 percent of beta-mercaptoethanol, adding a proper amount of the RL into the powder, shaking and uniformly mixing, transferring to a CS column, and centrifuging at a high speed for 5min at a low temperature to obtain clear liquid.
③ transferring the supernatant to a new 1.5ml centrifuge tube which is added with absolute ethyl alcohol with 0.5 times volume in advance, uniformly mixing and transferring to a CR3 column, centrifuging at low temperature and high speed for 1min, and discarding the waste liquid.
And fourthly, adding 350 mu l of RW1, centrifuging at low temperature for 1min, adding 80 mu l of DNaseI working solution after discarding the waste liquid, standing at 28 ℃ for 30min, and adding 500 mu l of RW.
Low-temperature centrifuging for 1min, and discarding the waste liquid.
And sixthly, carrying out air separation for 2min, and discarding the waste liquid.
Seventhly, placing the CR3 column head in a new 1.5ml centrifuge tube, standing at room temperature until the column head membrane is dried, adding 30 mu l of warm eluent or sterilized ddH2O on the membrane, and centrifuging at high speed for 2min at normal temperature to obtain the total RNA.
2. Reverse transcription (Tiangen reverse transcription kit)
The experiment used a reverse transcription kit from Tiangen corporation: FastQuantRT Kit (with gDNase), the specific procedure was as follows:
(ii) 0.2ml RNase-Free EP tube, the following ingredients were added:
② after mixing evenly, standing for 3min at 42 ℃; then placed on ice for 5 min.
③ adding the following components into the RNase-Free EP tube after the reaction:
fourthly, after being mixed evenly, the mixture is placed at 42 ℃ for standing for 30 min; then transferring to 95 ℃ and standing for 3min to obtain the cDNA, and storing the cDNA at-20 ℃.
3. Analysis of Gene expression level
CFX96 real-time polymerase chain reaction was performed using UltraSYBR mix (Beijing, China). The obtained wild-type and tga7 mutant cDNAs were subjected to fluorescent quantitative PCR detection, and the expression level of the gene was analyzed by performing three biological replicates using the expression level of TUB2 as a control.
Design of primer
Primers for quantitative real-time PCR
(ii) PCR reaction system:
③ qPCR reaction program:
the above amplification cycles were 45 for quantitative detection. Five flowering control pathways were performed, and statistical results are shown in fig. 3, and it was found that the expression of TGA7 was not changed by vernalization in both wild type and FRI type FLC plants, indicating that neither GA nor vernalization pathways regulate TGA 7. It finally follows that the expression of TGA7 is regulated by an autonomous pathway and a temperature pathway. In the autonomous pathway mutants, TGA7 was increased in fca-2 and fv-4 and decreased in fld-3 and flk-1, suggesting that the autonomous pathway may affect the expression of TGA 7.
SVP plays a key role in the temperature pathway, and SVP-41 mutants showed a temperature insensitive flowering phenotype in previous studies. We further analyzed whether the expression of TGA7 is also affected by ambient temperature. In the ambient temperature range, TGA7 expression increases with increasing temperature. In addition, TGA7 was more stably expressed in 16 ℃ wild type, SVP-41 and 35S: SVP plants, while TGA7 was more highly expressed in 35S: SVP plants and TGA7 was less expressed in SVP-41 plants at 23 ℃, suggesting that a temperature pathway might control the expression of TGA7 under ambient temperature.
FIG. 3 fluorescent quantitative PCR to determine whether the expression level of TGA7 gene is regulated in 6 flowering pathways. (a) The TGA7 gene expression quantity in the photoperiod pathway related mutant gi-1, co-9, ft-10 is detected; (b) TGA7 gene expression level detection after three and five weeks of gibberellin (GA3) treatment; (c) detecting the expression quantity of the TGA7 gene after the vernalization treatment; (d) the TGA7 gene expression quantity in the autonomous pathway related mutants of fpa-7, fca-2, fld-3, fve-4 and flk-1 is detected; (e) the expression quantity of the TGA7 gene is changed at 16 ℃, 23 ℃ and 27 ℃ under different temperature conditions; (f) the TGA7 gene was expressed in WT, SVP-41 and 35S: SVP at 16 ℃ and 23 ℃ (P < 0.05).
Example 4 analysis of expression of the major 4 flowering-related genes in TGA mutants and wild type
1. Total RNA extraction from Arabidopsis thaliana
The method is as shown in example 3
2. Reverse transcription
The method is as shown in example 3
TGA mutant and wild type flowering Gene expression analysis
The method is as shown in example 3
FIG. 4(a-d) shows that the expression levels of FLC, MAF5 and SMZ in the tga7 mutant are obviously increased and the expression level of NF-YC2 is obviously reduced by the fluorescent quantitative PCR analysis of Arabidopsis seedlings which grow for 9 days after sprouting under the long-day condition. This is consistent with the late-flowering phenotype of the tga7 mutant.
The above evidence demonstrates that by mutating the TGA gene, the expression of a part of the flowering-related gene can be influenced, thereby altering the flowering phase of the plant.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; such modifications and substitutions do not materially depart from the spirit and scope of the invention as claimed.
Sequence listing
<110> university of teachers in Hangzhou
Application of <120> TGA7 gene in regulation and control of plant flowering phase
<160> 17
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1107
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
atgatgagtt cttcttctcc aacacaactt gcatctttaa gagacatggg aatctatgag 60
ccatttcaac aaattgtcgg ttggggaaat gttttcaaat ctgatatcaa tgatcatagt 120
cccaatactg ctacttcctc tattattcag gttgatccta gaattgatga tcacaacaac 180
aacatcaaga taaattatga ttcttctcat aaccagatcg aagcagaaca accttctagt 240
aatgataatc aagatgatga tggcaggatt catgataaga tgaaacggcg tttagcgcag 300
aaccgagaag cggctcgcaa aagtcgtttg agaaagaagg cttatgttca gcagttagag 360
gaaagccggt taaagttatc gcagttagag caagaactcg aaaaggttaa gcagcagggc 420
catttaggac catctgggag tattaacaca gggattgcat catttgagat ggaatattca 480
cactggctac aagaacaaag cagaagagtt agcgaactac gaacagcgct tcaatctcat 540
ataagcgaca tagaactcaa gatgctagta gagagttgct tgaaccatta cgctaatctt 600
ttccaaatga aatccgatgc agcaaaagcc gatgttttct acttgatatc gggaatgtgg 660
cgaacttcaa ccgaaagatt cttccaatgg attggagggt ttcgtccatc cgaactttta 720
aacgttgtga tgccttatct tcaaccatta acggatcaac aaatcttgga agtgagaaac 780
ctccaacaat catcacaaca agcagaggat gctctgtctc aagggattga taaacttcaa 840
cagagtttag ctgaaagcat tgtgattgat gcggttatcg agtccacgca ttatcccact 900
cacatggctg cagctataga gaatcttcaa gcattagaag gatttgtgaa tcaagcagat 960
catctgaggc aacaaacttt gcaacaaatg gcgaagatct taacgacaag acaatcggct 1020
cgaggtttac tagctttagg agagtatctt catagacttc gtgctcttag ttctctttgg 1080
gcagctcgtc cacaagaacc aacttaa 1107
<210> 2
<211> 21
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
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taaagttatc gcagttagag c 21
<210> 3
<211> 17
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 3
ccgcatcaat cacaatg 17
<210> 4
<211> 21
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 4
atcatagtcc caatactgct a 21
<210> 5
<211> 19
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 5
atcctgccat catcatctt 19
<210> 6
<211> 26
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 6
gatggagctt gtgaagaacc ttcagg 26
<210> 7
<211> 27
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 7
cagccgttga tgattggtgg ttacttg 27
<210> 8
<211> 27
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 8
ctagccagat ggagaataat catcatg 27
<210> 9
<211> 27
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 9
ttaaggtggc taattaagta gtgggag 27
<210> 10
<211> 27
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 10
agggagaagg agccatgaag tttggtg 27
<210> 11
<211> 27
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 11
gtcttcagag gtttcatggt tgccatg 27
<210> 12
<211> 21
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 12
ccctgctaca actggaacaa c 21
<210> 13
<211> 19
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 13
caccctggtg catacactg 19
<210> 14
<211> 21
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 14
cttcagatgt tctgggctaa c 21
<210> 15
<211> 18
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 15
ctccgcagag atcatcct 18
<210> 16
<211> 21
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 16
atccgtgaag agtacccaga t 21
<210> 17
<211> 21
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 17
aagaaccatg cactcatcag c 21
Claims (2)
1. The application of the Arabidopsis TGA7 gene in regulating and controlling the flowering phase of plants is characterized in that the Arabidopsis TGA7 gene sequence is shown as SEQ ID NO. 1.
2. The use of an Arabidopsis TGA7 gene according to claim 1 for regulating the flowering phase of a plant, wherein the plant having a point mutation in the Arabidopsis TGA7 gene has a delayed flowering compared to a wild-type plant; wherein the EcoRV enzyme cutting site C in the Arabidopsis thaliana TGA7 gene is mutated into T.
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| Characterization of genes associated with TGA7 during the floral transition;Xiaorui Xu et al.;《BMC Plant Biology》;20210811;第21卷;第1-11页 * |
| 转录因子TGA7参与拟南芥响应干旱胁迫的分子机制研究;段军叶;《中国博士学位论文全文数据库 基础科学辑》;20190415(第4期);A006-22 * |
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