CN106754953A - Application of the arabidopsis SSCD1 gene mutations in jasmonic synthesis in regulation and control plant - Google Patents
Application of the arabidopsis SSCD1 gene mutations in jasmonic synthesis in regulation and control plant Download PDFInfo
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Abstract
A kind of application of arabidopsis SSCD1 gene mutations in jasmonic synthesis in regulation and control plant, the SSCD1 mutant gene sequences are as shown in SEQ ID No.2.Expression by contrasting jasmonic synthetic gene in short-day condition Wildtype Arabidopsis thaliana under Col 0 and SSCD1 genic mutation type arabidopsis sscd1 bodies of the invention, the content of jasmonic, the expression of jasmonic defense pathway downstream responses gene, it was found that the mutation of the SSCD1 genes can influence the synthesis of jasmonic in plant, change the content of jasmonic in plant, activation jasmonic defense pathway, induce the expression of jasmonic defense pathway downstream responses gene, transformed by the gene, the jasmine acid content in plant can be improved, cultivate the plant with jasmine acid content high.
Description
Technical field
The invention belongs to genetic engineering applied technical field, arabidopsis SSCD1 gene mutations are related generally in short-day bar
The synthesis of jasmonic can be regulated and controled under part, so as to influence the system of defense that jasmonic is participated in and mediated in plant.
Background technology
To the March of Second Year, China's most area belongs to short-day period to annual September.And this period is exactly
In the season of crop harvesting, in order to ensure the yield and quality of crops, agricultural producer reduces disease pest by spraying pesticide
Harmful generation, but with the improvement of people ' s living standards, the demand to nuisanceless, pure natural green agricultural product increasingly increases,
Spraying pesticide is increasingly difficult to achieve the requirement of people using the agricultural product of chemical fertilizer.Therefore, it is necessary to change biography in agricultural production
The method that the dependence agricultural chemicals of system carrys out prevention and elimination of disease and pests.
Jasmonic is a kind of novel hormonal being widely distributed in plant, with going deep into for studying it, it has been found that
Jasmonic to plant grow and plant defence play the role of it is important.There are some researches show jasmonic can be notable
Suppress the growth of plant, adjust the development (Kazan and Manners.2012) of pollen, promote the aging of blade, fruit into
It is ripe, induce the formation (Creelman and Mullet.1995) of plant tuber.On the other hand, jasmonic can participate in regulation plant
The defense reaction of thing, enhancing plant is to pest and disease damage, the resistance (Yang et al.2012) of adverse circumstance environment.In agricultural production, just
Having spray jasmonic by external source and prevent and treat the method for diseases and pests of agronomic crop, but is influenceed by external environment, this method
Effect and unstable.
Arabidopsis SSCD1 genes are a key genes for participating in internal tyrosine degradation pathway, and the gene is undergone mutation
A kind of phenotype (Han et al.2013) for simulating scab can be formed under the conditions of short-day.Substantial amounts of research shows to produce simulation
The defense pathway that the plant of scab phenotype, its internal jasmonic or other hormones are mediated often is changed significantly.But
It is that current research and application on the gene in terms of jasmine acid content in lifting plant has not been reported.
The content of the invention
It is an object of the invention to provide a kind of arabidopsis SSCD1 gene mutations in jasmonic synthesis in regulation and control plant
Application, arabidopsis SSCD1 gene mutations can promote in arabidopsis body jasmonic synthesis under the conditions of short-day, improve jasmonic
Content.To carry out the prevention and control of plant diseases, pest control by means of jasmonic in agricultural production and regulating crop growth provides certain reference and side
Help, for the plant of jasmine acid content high under the conditions of cultivation short-day lays the foundation.
It is that, up to above-mentioned purpose, the technical solution adopted by the present invention is：A kind of arabidopsis SSCD1 mutators are in short-day bar
Application under part in jasmonic synthesis in regulation and control plant, the SSCD1 mutant gene sequences are as shown in SEQ ID No.2.
Under the conditions of short-day, SSCD1 mutators can induce the expression of jasmonic synthetic gene in plant, lifting body
Interior jasmine acid content, and induce the expression of jasmonic defense pathway downstream responses gene, the i.e. mutation of the SSCD1 genes can to adjust
Control the synthesis of jasmonic, it is possible to the defense pathway that jasmonic is mediated in activated plant body.
It is of the invention mainly to pass through technique for gene engineering means, make the SSCD1 genes in arabidopsis body that point mutation to occur, it is impossible to
Normal encoding fumarylacetoacetase (FAH), interrupts internal tyrosine degradation pathway.Under the conditions of short-day, SSCD1
Gene undergo mutation after Arabidopsis plant body in jasmonic synthetic gene LOX2 and AOS expression substantially raise, jasmonic contains
Amount is obvious to be raised, while jasmonic defense pathway is activated, responsive genes expression quantity is significantly raised downstream.
In this way, the present invention intends south by contrasting short-day condition Wildtype Arabidopsis thaliana under Col-0 and SSCD1 genic mutation type
The expression of jasmonic synthetic gene, the content of jasmonic, the table of jasmonic defense pathway downstream responses gene in mustard sscd1 bodies
Reach, finding the mutation of the gene can influence the synthesis of jasmonic in plant, change the content of jasmonic in plant, activate jasmine
Jasmine acid defense pathway, induces the expression of jasmonic defense pathway downstream responses gene, is transformed by the gene, can improve
Jasmine acid content in plant, cultivates the plant with jasmine acid content high.
Brief description of the drawings
Fig. 1 is the wildtype Arabidopsis thaliana Col- of growth 3 days under the conditions of being transferred to short-day after being grown under long-day conditions 2 weeks
The expression of jasmonic biosynthesis gene LOX2, AOS in 0 and SSCD1 gene mutation arabidopsis sscd1 material bodies.
Wherein, A：LOX2 genes, B：AOS genes, C：Wildtype Arabidopsis thaliana Col-0, S：SSCD1 gene mutation arabidopsis
Sscd1 materials.
Fig. 2 is the wildtype Arabidopsis thaliana of growth 2 to 3 days under the conditions of being transferred to short-day after being grown under long-day conditions 2 weeks
Jasmonic content detection result in Col-0 and SSCD1 gene mutation arabidopsis sscd1 material bodies.
Fig. 3 is the wildtype Arabidopsis thaliana Col- of growth 3 days under the conditions of being transferred to short-day after being grown under long-day conditions 2 weeks
Jasmonic defense pathway downstream responses gene PDF1.2 in 0 and SSCD1 gene mutation arabidopsis sscd1 material bodies, VSP2,
The expression of THI2.1.
Wherein, A：PDF1.2 genes, B：VSP2 genes, C：THI2.1 genes, C (slogan banner)：Wildtype Arabidopsis thaliana Col-0,
S：SSCD1 gene mutation arabidopsis sscd1 materials.
Specific embodiment
Following arabidopsis SSCD1 gene mutation bodies seed (sscd1 mutant seeds) for referring to are to intend south by EMS mutagenesis
Mustard wild type (Col-0) and obtain, referring to the mutagenesis processing procedure described in CN102911946B, the sscd1 mutant
Middle SSCD1 genes (SSCD1 gene C DS sequences are as shown in SEQ ID No.1) are undergone mutation (CDS after the SSCD1 gene mutations
Sequence is as shown in SEQ ID No.2), prevent it from normal encoding related protein.
Under the conditions of the short-day of embodiment 1, SSCD1 mutators make jasmonic synthetic gene LOX2 and AOS in mutant
Expression is substantially raised
(A) culture of material：Wildtype Arabidopsis thaliana Col-0 seeds and sscd1 mutant seeds (are contained with thimerosal respectively
Have 20% 84 thimerosals and 0.01% TritonX-100) sterilization 10 minutes, it is layered on MS after sterile water wash 3-5 times and (contains
Have 1% sucrose) solid medium on, vernalization under 4 DEG C of cryogenic conditions.It is placed in long-day culturing room after 3 days to be cultivated, specifically
Condition of culture is：Light dark cycles are 16h/8h, 22 DEG C of temperature, intensity of illumination 100umolm-2s-1, relative humidity 70%.Growth
After 7 days, seedling is moved on on new aseptic MS culture mediums, process of transplanting seedlings it is noted that do not damage seedling, while to reserve enough
Space ensure seedling can normal growth, the seedling of transplanting is continued to be placed in long-day culturing room and is cultivated 7 days, be then transferred to short day
Cultivated 3 days according to culturing room, short-day condition of culture is：Light dark cycles are 8h/16h, 22 DEG C of temperature, intensity of illumination 100umolm-
2s-1, relative humidity 70% respectively obtains wildtype Arabidopsis thaliana Col-0 seedling material and sscd1 mutant seedlings materials.
(B) extraction of arabidopsis Col-0 and sscd1 mutant seedlings materials RNA and the acquisition of cDNA：By short-day condition
2 kinds of Arabidopsis thaliana Seedlings materials (100-200mg) of lower growth 3 days are respectively placed in mortar, add the rapid grind into powder of liquid nitrogen,
Then move into 1.5ml centrifuge tubes, add 1ml Trizol Reagent (Invitrogen, Carlsbad, CA, USA), then
Acutely vibration, makes plant powder fully be mixed with RNA extract solutions, is stored at room temperature 5min；To adding 200 μ l chloroforms in centrifuge tube,
Acutely vibration 1min, is then stored at room temperature 5min；4 DEG C, 12000g centrifugations 15min；Supernatant is transferred in new centrifuge tube, plus
Enter the isopropanol isometric with supernatant and high level salt solution mixed solution (high level salt solution with 1.2M sodium chloride, 0.8M Chinese catalpas lemon acid sodium and
1% (V/V) DEPC water is prepared), fully mix, precipitation at room temperature l0min；4 DEG C, 12000g centrifugations 10min；Supernatant is removed, is added
The ethanol of 1ml 75%, turns upside down, fully cleaning precipitation, 4 DEG C, 8000g, 5min centrifugations；Supernatant is removed, remaining ethanol is blotted,
Room temperature Xia Liao do to ethanol and volatilize completely, add 10-30 μ l0.1%DEPC water, RNA is fully dissolved, respectively obtain 2 kinds of materials
RNA.1 μ l RNA are taken respectively, detect OD260/OD230, OD260/OD280 value and concentration.
The Rever Tra Ace qPCR RT Master Mix with that (TOYOBO) company produces are spun using Japan
GDNA Remove kits (FSQ-301) synthesize cDNA：RNA concentration of the first step according to measured by calculates good required RNA volumes,
The RNA for making each system contain 1000ng, and supplied to 14 μ l with ddH2O, 65 DEG C are processed 5 minutes, and taking-up is immediately placed on ice；
(the gDNA Remover containing 1/50 volume are molten to the 4x DN Master Mix solution for adding 3 μ l in each system for second step
Liquid), 37 DEG C are processed 5 minutes, and taking-up is immediately placed on ice；3rd step is to the 5xRT Master that 3 μ l are added in each reaction system
The solution of Mix II, 37 DEG C for the treatment of 15min, 50 DEG C for the treatment of 5min, 98 DEG C for the treatment of 5min, cDNA synthesis are completed, and 2 kinds of materials are taken respectively
1 μ l cDNA, detect OD260/OD230, OD260/OD280 value and concentration.
(C) QRT-PCR detects 2 kinds of expressions of the gene of material respectively：Calculate every according to measured cDNA concentration
CDNA volumes needed for individual reaction system, cDNA contained by each system is 200ng, mends the μ l of ddH2O to 2, then sequentially adds 2.2 μ l
DdH2O, the forward primer of 0.4 μ l, the reverse primer of 0.4 μ l, Roche (Roche) SYBR fluorescence dye liquors of 5 μ l, amount to 10 μ l
Reaction volume, be internal reference to intend southern Jie ACTIN2 genes.QRT-PCR instruments are the CFX of BIO-RAD companies production
Connect Real-Time System, specific response procedures are：95 DEG C of 10min of the first step；Second step 95 DEG C of 15s, 55 DEG C
1min, 72 DEG C of 30s are circulated 40 times.The data obtained uses 2-ΔΔCtCalculated.
This experiment the primer sequence is as follows：
ACTIN2-FP：5-AGCACTTGCACCAAGCAGCATG-3 (SEQ ID No.3),
ACTIN2-RP:5-ACGATTCCTGGACCTGCCTCATC-3(SEQ ID No.4)；
LOX2-FP:5-TGCTCCCCAGAATCATCAAA-3 (SEQ ID No.5),
LOX2-RP:5-AAACTCGTCGTCTCGTAACCAT-3(SEQ ID No.6)；
AOS-FP:5-TTTTACGACCAAGGAGCTGAAG-3 (SEQ ID No.7),
AOS-RP:5-CGGTGGCATGTTGACTCTGTA-3(SEQ ID No.8)。
Result as shown in figure 1, under the conditions of short-day grow 3 days, in the sscd1 material bodies that SSCD1 genes are undergone mutation
The expression quantity of jasmonic synthetic gene LOX2 and AOS will significantly be higher than wild type Col-0 materials.Show：Short-day condition
Under, SSCD1 mutators can induce the expression of jasmonic synthetic gene.
Under the conditions of the short-day of embodiment 2, SSCD1 mutators can lift jasmine acid content in plant
(A) culture of material：Wildtype Arabidopsis thaliana Col-0 seeds and sscd1 mutant seeds (are contained with thimerosal respectively
Have 20% 84 thimerosals and 0.01% TritonX-100) sterilization 10 minutes, it is layered on MS after sterile water wash 3-5 times and (contains
Have 1% sucrose) solid medium on, vernalization under 4 DEG C of cryogenic conditions.It is placed in long-day culturing room after 3 days to be cultivated, specifically
Condition of culture is：Light dark cycles are 16h/8h, 22 DEG C of temperature, intensity of illumination 100umolm-2s-1, relative humidity 70%.Growth
After 7 days, seedling is moved on on new aseptic MS culture mediums, process of transplanting seedlings it is noted that do not damage seedling, while to reserve enough
Space ensure seedling can normal growth, the seedling of transplanting is continued to be placed in long-day culturing room and is cultivated 7 days, be then transferred to short day
Cultivated 3 days according to culturing room, short-day condition of culture is：Light dark cycles are 8h/16h, 22 DEG C of temperature, intensity of illumination 100umolm-
2s-1, relative humidity 70%.
(B) jasmonic content detection：2 kinds of fresh arabidopsis material 2g are taken respectively, is shredded, add 4 DEG C of precoolings of 15ml
80% methyl alcohol is extracted 12 hours, and homogenate is ground to form on ice, and 1400rpm/min is centrifuged 15 minutes, takes supernatant, uses 5ml
The Na of 0.2mol/L2HPO4-H3PO4Buffer solution adds appropriate petroleum ether extraction, discards upper strata ether phase, retains water phase, and repetition is extracted to
Upper strata it is limpid it is colourless untill, plus Na2HPO4Regulation pH value is 8.0.Again plus 5ml ethyl acetate hybrid extractions, 6000rpm/min from
The heart 5 minutes, takes ester phase, repeats 1-2 times.Ester is concentrated under 20 DEG C of reduced pressures and is drained, be settled to 505 Chromatographic Pure Methanols
1ml, is then determined with high performance liquid chromatography.
Result as shown in Fig. 2 by detect under the conditions of short-day the wildtype Arabidopsis thaliana Col-0 materials of growth 2 to 3 days and
Jasmine acid content in SSCD1 gene mutation arabidopsis sscd1 materials, it is found that under short-day growth conditions, SSCD1 genes are dashed forward
Jasmine acid content in the arabidopsis sscd1 material bodies of change will be considerably higher than wildtype Arabidopsis thaliana Col-0 materials, with short day
According to the increase of growth time, the jasmine acid content in sscd1 material bodies is also increased as.Show：Under the conditions of short-day,
SSCD1 mutators can lift the jasmine acid content in plant, and this lifting is protected always with continuing for short-day
Hold.
Under the conditions of the short-day of embodiment 3, SSCD1 mutators induction jasmonic defense pathway downstream responses gene
The expression of PDF1.2, VSP2, THI2.1
(A) culture of material：Wildtype Arabidopsis thaliana Col-0 seeds and sscd1 mutant seeds (are contained with thimerosal respectively
Have 20% 84 thimerosals and 0.01% TritonX-100) sterilization 10 minutes, it is layered on MS after sterile water wash 3-5 times and (contains
Have 1% sucrose) solid medium on, vernalization under 4 DEG C of cryogenic conditions.It is placed in long-day culturing room after 3 days to be cultivated, specifically
Condition of culture is：Light dark cycles are 16h/8h, 22 DEG C of temperature, intensity of illumination 100umolm-2s-1, relative humidity 70%.Growth
After 7 days, seedling is moved on on new aseptic MS culture mediums, process of transplanting seedlings it is noted that do not damage seedling, while to reserve enough
Space ensure seedling can normal growth, the seedling of transplanting is continued to be placed in long-day culturing room and is cultivated 7 days, be then transferred to short day
Cultivated 3 days according to culturing room, short-day condition of culture is：Light dark cycles are 8h/16h, 22 DEG C of temperature, intensity of illumination 100umolm-
2s-1, relative humidity 70% respectively obtains wildtype Arabidopsis thaliana Col-0 seedling material and sscd1 mutant seedlings materials.
(B) extraction of arabidopsis Col-0 and sscd1 seedling materials RNA and the acquisition of cDNA：To be grown under the conditions of short-day
2 kinds of Arabidopsis thaliana Seedlings materials (100-200mg) of 3 days are respectively placed in mortar, add the rapid grind into powder of liquid nitrogen, then will
It is moved into 1.5ml centrifuge tubes, adds 1ml Trizol Reagent (Invitrogen, Carlsbad, CA, USA), then
Acutely vibration, makes plant powder fully be mixed with RNA extract solutions, is stored at room temperature 5min；To adding 200 μ l chloroforms in centrifuge tube,
Acutely vibration 1min, is then stored at room temperature 5min；4 DEG C, 12000g centrifugations 15min；Supernatant is transferred in new centrifuge tube, plus
(high level salt solution is with 1.2M sodium chloride, 0.8M Chinese catalpas lemon acid sodium to enter the isopropanol isometric with supernatant and high level salt solution mixed solution
And 1% (V/V) DEPC water prepare), fully mix, precipitation at room temperature l0min；4 DEG C, 12000g centrifugation 10min, remove supernatant, plus
Enter the ethanol of 1ml 75%, turn upside down, fully cleaning precipitation, 4 DEG C, 8000g, 5min centrifugations；Supernatant is removed, remaining second is blotted
Alcohol, room temperature Xia Liao do to ethanol and volatilize completely, add 10-30 μ l0.1%DEPC water, RNA is fully dissolved, respectively obtain 2 kinds
The RNA of material；1 μ l RNA are taken respectively, detect OD260/OD230, OD260/OD280 value and concentration.
The Rever Tra Ace qPCR RT Master Mix with that (TOYOBO) company produces are spun using Japan
GDNA Remove kits (FSQ-301) synthesize cDNA：RNA concentration of the first step according to measured by calculates good required RNA volumes,
The RNA for making each system contain 1000ng, and supplied to 14 μ l with ddH2O, 65 DEG C are processed 5 minutes, and taking-up is immediately placed on ice；
(the gDNA Remover containing 1/50 volume are molten to the 4x DN Master Mix solution for adding 3 μ l in each system for second step
Liquid), 37 DEG C are processed 5 minutes, and taking-up is immediately placed on ice；3rd step is to the 5xRT Master that 3 μ l are added in each reaction system
The solution of Mix II, 37 DEG C for the treatment of 15min, 50 DEG C for the treatment of 5min, 98 DEG C for the treatment of 5min, cDNA synthesis are completed, and respectively obtain 2 kinds of materials
The cDNA of material.1 μ l cDNA are taken respectively, detect OD260/OD230, OD260/OD280 value and concentration.
(C) QRT-PCR detects the expression of gene：According to needed for measured cDNA concentration calculates each reaction system
CDNA volumes, cDNA contained by each system is 200ng, mends the μ l of ddH2O to 2, then sequentially adds ddH2O, the 0.4 μ l of 2.2 μ l
Forward primer, the reverse primer of 0.4 μ l, Roche (Roche) SYBR fluorescence dye liquors of 5 μ l, amount to 10 μ l reaction volume, with
It is internal reference to intend south Jie ACTIN2 genes.QRT-PCR instruments are the CFX Connect Real- of BIO-RAD companies production
Time System, specific response procedures are：95 DEG C of 10min of the first step；Second step 95 DEG C of 15s, 55 DEG C of 1min, 72 DEG C of 30s circulations
40 times.The data obtained uses 2-ΔΔCtCalculated.
This experiment the primer sequence is as follows：
ACTIN2-FP：5-AGCACTTGCACCAAGCAGCATG-3 (SEQ ID No.3),
ACTIN2-RP:5-ACGATTCCTGGACCTGCCTCATC-3(SEQ ID No.4)；
PDF1.2-FP:5-GCTTCCATCATCACCCTTATC-3 (SEQ ID No.9),
PDF1.2-RP:5-TTGGCTTCTCGCACAACTT-3(SEQ ID No.10)；
THI2.1-FP:5-GGTTGGGTAAACGCCATTCT-3 (SEQ ID No.11),
THI2.1-RP:5-CATTGTTCCGACGCTCCATT-3(SEQ ID No.12)；
VSP2-FP:5-GGATTGAACCCATCATACTCAG-3 (SEQ ID No.13),
VSP2-RP:5-CACGAGACTCTTCCTCACCTTT-3(SEQ ID No.14)。
Result by contrasting short-day condition Wildtype Arabidopsis thaliana under Col-0 and SSCD1 gene mutation as shown in figure 3, intended
The expression of jasmonic defense pathway downstream responses gene PDF1.2, VSP2, THI2.1, as a result sends out in southern mustard sscd1 materials
After existing SSCD1 gene mutations, under the conditions of short-day, the expression quantity of its internal PDF1.2, VSP2, THI2.1 gene is significantly
Raise, its expression quantity has compared with Col-0 and significantly lifted.Show：Under the conditions of short-day, SSCD1 mutators can cause
The expression of jasmonic defense pathway downstream responses gene, the defense pathway that jasmonic is mediated in regulation and control plant.
In sum, by above-mentioned experiment, it was demonstrated that SSCD1 mutators can induce jasmonic biological under the conditions of short-day
The expression of synthetic gene, promotes to produce the accumulation of jasmonic in plant, and cause jasmonic defense pathway downstream responses gene
Expression.The present invention is to improve plant by lifting plant endogenous jasmine acid content under the conditions of short-day to resist pest and disease damage
Property provide theoretical foundation, and provide practical basis to cultivate jasmine acid content plant high.
Bibliography：
1.Kazan k,Manner JM(2012).JAZ repressors and the orchestration of
phytohormone crosstalk.TRENDS IN PLANT SCIENCE.17:22-31
2.Creelman,R.A.,and Mullet,J.E(1995).Jasmonic acid distribution and
action in plants-regulation during development and response to biotic and
abiotic stress.Proceedings of the National Academy of Sciences of the United
States of America 92:4114-4119
3.Yang,Dong-Lei,Yao,Jian,Chuan-ShengMei,et al(2012).Plant hormone
jasmonate prioritizes defense over growth by interfering with gibberellin
signaling cascade.Proceedings of the National Academy of Sciences of the
United States of America.19:1192-1200
4.Han CY,Ren CM,Zhi TT,Zhou Z,Liu Y,Chen F,Peng W,Xie DX(2013)
Disruption of fumarylacetoacetate hydrolase causes spontaneous celldeath
under short-day conditions in Arabidopsis.Plant Physiol.162:1956-1964。
SEQUENCE LISTING
<110>Agricultural University Of Hunan
<120>Application of the arabidopsis SSCD1 gene mutations in jasmonic synthesis in regulation and control plant
<130> 012
<160> 14
<170> PatentIn version 3.3
<210> 1
<211> 1266
<212> DNA
<213>The CDS sequences of SSD1 genes
<400> 1
atggcgttgc tgaagtcttt catcgatgtt ggctcagact cgcacttccc tatccagaat 60
ctcccttatg gtgtcttcaa accggaatcg aactcaactc ctcgtcctgc cgtcgctatc 120
ggcgatttgg ttctggacct ctccgctatc tctgaagctg ggcttttcga tggtctgatc 180
cttaaggacg cagattgctt tcttcagcct aatttgaata agttcttggc catgggacgg 240
cctgcgtgga aggaagcgcg ttctacgctg caaagaatct tgtcatctaa tgaacctatc 300
ttgcgagaca atgatgtttt gaggagaaaa tcattccatc agatgagtaa agtggaaatg 360
attgttccta tggtgattgg ggactataca gacttctttg catctatgca tcacgcgaag 420
aactgcggac ttatgttccg tgggcctgag aatgcgataa acccaaattg gtttcgtctt 480
cccattgcat atcatggacg ggcatcatct attgtcatct ctgggactga cattattcga 540
ccaagaggtc agggccatcc acaaggaaac tctgaaccat attttggacc ttcgaagaaa 600
cttgattttg agcttgagat ggctgctgtg gttggtccag gaaatgaatt gggaaagcct 660
attgacgtga ataatgcagc cgatcatata tttggtctat tactgatgaa tgactggagt 720
gctagggata ttcaggcgtg ggagtatgta cctcttggtc ctttcctggg gaagagtttt 780
gggactacta tatccccttg gattgttacc ttggatgcgc ttgagccttt tggttgtcaa 840
gctcccaagc aggatccacc tccattgcca tatttggctg agaaagagtc tgtaaattac 900
gatatctcct tggaggttca acttaaacct tctggcagag atgattcttg tgtaataaca 960
aagagcaact tccaaaactt atattggacc ataacgcagc agctagcaca ccataccgtt 1020
aacggttgca atttgaggcc tggtgatctc cttggcacag gaaccataag cggaccggag 1080
ccagattcat atgggtgcct acttgagttg acatggaatg gacagaaacc tctatcactc 1140
aatggaacaa ctcagacgtt tctcgaagac ggagaccaag tcaccttctc aggtgtatgc 1200
aagggagatg gttacaatgt tgggtttgga acatgcacag ggaaaattgt tccttcaccg 1260
ccttga 1266
<210> 2
<211> 1266
<212> DNA
<213>The CDS sequences of SSD1 genes in ssd1 mutant
<400> 2
atggcgttgc tgaagtcttt catcgatgtt ggctcagact cgcacttccc tatccagaat 60
ctcccttatg gtgtcttcaa accggaatcg aactcaactc ctcgtcctgc cgtcgctatc 120
ggcgatttgg ttctggacct ctccgctatc tctgaagctg ggcttttcga tggtctgatc 180
cttaaggacg cagattgctt tcttcagcct aatttgaata agttcttggc catgggacgg 240
cctgcgtgga aggaagcgcg ttctacgctg caaagaatct tgtcatctaa tgaacctatc 300
ttgcgagaca atgatgtttt gaggagaaaa tcattccatc agatgagtaa agtggaaatg 360
attgttccta tggtgattgg ggactataca gacttctttg catctatgca tcacgcgaag 420
aactgcggac ttatgttccg tgggcctgag aatgcgataa acccaaatta gtttcgtctt 480
cccattgcat atcatggacg ggcatcatct attgtcatct ctgggactga cattattcga 540
ccaagaggtc agggccatcc acaaggaaac tctgaaccat attttggacc ttcgaagaaa 600
cttgattttg agcttgagat ggctgctgtg gttggtccag gaaatgaatt gggaaagcct 660
attgacgtga ataatgcagc cgatcatata tttggtctat tactgatgaa tgactggagt 720
gctagggata ttcaggcgtg ggagtatgta cctcttggtc ctttcctggg gaagagtttt 780
gggactacta tatccccttg gattgttacc ttggatgcgc ttgagccttt tggttgtcaa 840
gctcccaagc aggatccacc tccattgcca tatttggctg agaaagagtc tgtaaattac 900
gatatctcct tggaggttca acttaaacct tctggcagag atgattcttg tgtaataaca 960
aagagcaact tccaaaactt atattggacc ataacgcagc agctagcaca ccataccgtt 1020
aacggttgca atttgaggcc tggtgatctc cttggcacag gaaccataag cggaccggag 1080
ccagattcat atgggtgcct acttgagttg acatggaatg gacagaaacc tctatcactc 1140
aatggaacaa ctcagacgtt tctcgaagac ggagaccaag tcaccttctc aggtgtatgc 1200
aagggagatg gttacaatgt tgggtttgga acatgcacag ggaaaattgt tccttcaccg 1260
ccttga 1266
<210> 3
<211> 22
<212> DNA
<213>Synthesis
<400> 3
agcacttgca ccaagcagca tg 22
<210> 4
<211> 23
<212> DNA
<213>Synthesis
<400> 4
acgattcctg gacctgcctc atc 23
<210> 5
<211> 20
<212> DNA
<213>Synthesis
<400> 5
tgctccccag aatcatcaaa 20
<210> 6
<211> 22
<212> DNA
<213>Synthesis
<400> 6
aaactcgtcg tctcgtaacc at 22
<210> 7
<211> 22
<212> DNA
<213>Synthesis
<400> 7
ttttacgacc aaggagctga ag 22
<210> 8
<211> 21
<212> DNA
<213>Synthesis
<400> 8
cggtggcatg ttgactctgt a 21
<210> 9
<211> 21
<212> DNA
<213>Synthesis
<400> 9
gcttccatca tcacccttat c 21
<210> 10
<211> 19
<212> DNA
<213>Synthesis
<400> 10
ttggcttctc gcacaactt 19
<210> 11
<211> 20
<212> DNA
<213>Synthesis
<400> 11
ggttgggtaa acgccattct 20
<210> 12
<211> 20
<212> DNA
<213>Synthesis
<400> 12
cattgttccg acgctccatt 20
<210> 13
<211> 22
<212> DNA
<213>Synthesis
<400> 13
ggattgaacc catcatactc ag 22
<210> 14
<211> 22
<212> DNA
<213>Synthesis
<400> 14
cacgagactc ttcctcacct tt 22
Claims (5)
1. application of a kind of arabidopsis SSCD1 mutators under the conditions of short-day in regulation and control plant in jasmonic synthesis,
The SSCD1 mutant gene sequences are as shown in SEQ ID No.2.
2. application as claimed in claim 1, wherein, the application refers to the table for inducing jasmonic biosynthesis gene in plant
Reach.
3. application as claimed in claim 1, wherein, the application refers to the content for lifting jasmonic in plant.
4. application as claimed in claim 1, wherein, the application refers to jasmonic defense pathway downstream responses in induction plant
The expression of gene.
5. application of a kind of arabidopsis SSCD1 mutators under the conditions of short-day in jasmine acid content plant high is cultivated, should
SSCD1 mutant gene sequences are as shown in SEQ ID No.2.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111436297A (en) * | 2020-05-20 | 2020-07-24 | 西南大学 | Method for inhibiting synthesis and response of jasmonic acid, hormone in plant body |
| CN114051856A (en) * | 2021-10-26 | 2022-02-18 | 中国农业科学院茶叶研究所 | Application of plant jasmonic acid pathway inhibitor |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6218169B1 (en) * | 1998-07-31 | 2001-04-17 | E. I. Du Pont De Nemours And Company | Aromatic amino acid catabolism enzymes |
| CN105441466A (en) * | 2016-01-12 | 2016-03-30 | 湖南农业大学 | Application of arabidopsis SSCD1 gene in plant salt tolerance |
-
2016
- 2016-11-28 CN CN201611060797.XA patent/CN106754953B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6218169B1 (en) * | 1998-07-31 | 2001-04-17 | E. I. Du Pont De Nemours And Company | Aromatic amino acid catabolism enzymes |
| CN105441466A (en) * | 2016-01-12 | 2016-03-30 | 湖南农业大学 | Application of arabidopsis SSCD1 gene in plant salt tolerance |
Non-Patent Citations (3)
| Title |
|---|
| CHENGYUN HAN,等: "Disruption of fumarylacetoacetate hydrolase causes spontaneous cell death under short-day conditions in arabidopsis", 《PLANT PHYSIOLOGY》 * |
| 宋云，等: "茉莉酸信号传导在植物抗逆性方面研究进展", 《中国农业科技导报》 * |
| 黄弈，等: "拟南芥sscd1-1点突变对其表达的影响", 《湖南农业大学学报（自然科学版）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111436297A (en) * | 2020-05-20 | 2020-07-24 | 西南大学 | Method for inhibiting synthesis and response of jasmonic acid, hormone in plant body |
| CN114051856A (en) * | 2021-10-26 | 2022-02-18 | 中国农业科学院茶叶研究所 | Application of plant jasmonic acid pathway inhibitor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106754953B (en) | 2018-01-12 |
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