CN110283241A - PtTST1.1 and PtTST2.1 promotes the application in plant growth substance in preparation - Google Patents
PtTST1.1 and PtTST2.1 promotes the application in plant growth substance in preparation Download PDFInfo
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Abstract
The invention discloses a kind of PtTST1.1 and PtTST2.1 to promote the application in plant growth substance in preparation.The method of the present invention is easy；According to transgenic poplar and transgenic arabidopsis as a result, PtTST1.1 and PtTST2.1 can remarkably promote the growth of arabidopsis and poplar, the new variety of plant for cultivating fast growing and high yield has extensive gene value.
Description
Technical field
The present invention relates to the research methods of a kind of purposes of gene and mechanism of action.
Background technique
Xylem is the Transportation Organization of vascular plant, is responsible for up transporting moisture and minerals that root absorbs, in addition also
Have the function of supporting plant.Preferably with sugar by source (mature leaf) to library (seed, fruit, root tuber, stem tuber and timber
Deng) transport mechanism, the especially effect during transportation such as saccharide transporter, for by regulation plant carbon source distribution,
The yield and the speed of growth for improveing plant are of great significance.
Poplar is important bioenergy tree species and environmental protection tree, cultivates and grows the rapid and increased poplar of biomass
Setting new varieties has huge economic value and ecological benefits.
Due to the tree growth period is long, the Genetic Mechanisms of high, many character of genetic heterozygosity are unknown and inter-species geographic isolation,
The factors limitation such as reproduction isolation, it is serious hinder by traditional forest genetics means directive breeding forest fast growing and high yield new varieties into
Journey.The continuous development of Forest-tree Gene Engineering technology and transgenic technology, it is long to compensate for traditional forest genetics period, orderly improvement mistake
Journey is complicated, it is difficult to which the deficiencies of controlling makes significant contribution for Forest Tree Genetic Breeding.High yield, the excavation of fast-growing genetic resource and function
It can parse, cultivate new fast growing and high yield kind, be that raising poplar yield and the speed of growth are most basic, most economical, most effective
Method.
Therefore, using modern molecular biology technique means, it is high that forest fast-growing is improved especially by genetic engineering means
Production capacity power opened up a new way to provide excellent fast growing and high yield Forest Tree New Varieties.
Summary of the invention
The purpose of the present invention is to provide a kind of PtTST1.1 and PtTST2.1 to promote in plant growth substance in preparation
Using the research method with mechanism of action.
The technical solution of the invention is as follows:
A kind of PtTST1.1 and PtTST2.1 promotes the application in plant growth substance in preparation.
A method of research PtTST1.1 and PtTST2.1 is promoting the mechanism of action in plant growth, it is characterized in that:
Include the following steps；
(1) selection of gene and phylogenetic analysis
5 and the TST genetic homology in arabidopsis and beet are found in comospore poplar genome by the method for BLAST
Higher gene order, according to phylogenetic analysis as a result, by this 5 genes be respectively designated as PtTST1.1, PtTST1.2,
PtTST2.1, PtTST2.2 and PtTST3；2 and the ST DNA homolog in arabidopsis are cloned followed by the method for RT-PCR
The highest gene of property, is PtTST1.1 and PtTST2.1 respectively；Wherein, PtTST1.1 includes 2217 pairs of bases, encodes 738 ammonia
Base acid, molecular weight 79.18KD；PtTST2.1 includes 2223 bases, encodes 740 amino acid, molecular weight 79.15KD；
With TMHMM softwarehttp://www.cbs.dtu.dk/services/TMHMM/To two TST albumen of poplar into
Row hydrophobicity analysis, discovery PtTST1.1 have 10 possible transmembrane regions, and PtTST2.1 has 11 possible transmembrane regions, and two
TST albumen has 10 common transmembrane domains, and be also about made of 320 amino acid by one between transmembrane domain 5 and 6
The big ring in hydrophily center, this is consistent with the TSTs protein structure in arabidopsis；
(2) promoter Analysis of PtTST1.1, PtTST2.1 gene
PtTST1.1 and PtTST2.1 gene promoter element is analyzed, it is found that it is red 3 TST genes of poplar have
The binding site E-box of mycin response element GARE-motif and brassinosteroid, and the E-box number of three genes compares
It is more；Meanwhile there are also gibberellin response element P-box, PtTST1.1 the relevant cis regulatory member of 1 auxin by PtTST2.1
Part AuXRR-core, PtTST3 have 1 abscisic acid response element ABRE, thus illustrate, PtTST1.1, PtTST2.1 and PtTST3
Gene may have response to gibberellin, auxin, brassinosteroid and abscisic acid, while also will receive the regulation of these hormones,
The especially regulation of gibberellin and brassinosteroid；
Table: the cis-acting elements relevant to hormone response of the gene promoter area poplar TST distribution；
PtTST1.1 and PtTST2.1 promoter region is analyzed with Plant CARE software；
(3) relevant carriers construct
According to the expression pattern analysis of PtTST1.1, PtTST2.1 and PtTST3 in wild type Turpinia arguta, opened with 2 × 35S
The pCAMBIA2301 carrier of mover driving constructs over-express vector；Target protein and YFP are constructed with pA7-YFP carrier
Fusion protein is used to instantaneous conversion poplar mesophyll protoplast；Target start is constructed with pCAMBIA1300+pBI101
The carrier that son is merged with+gus gene is for converting Turpinia arguta and detect the expression pattern of poplar TST albumen by GUS dyeing；
(4) screening of genetic transformation and transgenic line
It is new that the pCambia2301s2-PtTST1.1/PtTST2.1 over-express vector built is converted into wild type mountain respectively
Poplar has obtained transgenic poplar；By PCR detection, GUS dyeing detection and quantitative fluorescent PCR to the transgenosis young plant of overexpression
It is identified, identifies 15 overexpression strains of PtTST1.1 gene, identify 10 overexpression strains of PtTST2.1 gene
System；PCAMBIA1300+pBI101-PtTST1.1/PtTST2.1 promoter-GUS the carrier built conversion wild type mountain is new
Poplar obtains 10 strains of proPtTST1.1, identifies 11 strains of proPtTST2.1；
To the side for the arabidopsis attst1-2::tDNA mutant three-wheel PCR that H.Ekkehard professor Neuhaus provides
Method is identified, and pCambia2301s2-PtTST1.1/PtTST2.1 over-express vector is converted the mutant, is obtained
The strain of the PtTST1.1/PtTST2.1-attst1-2::tDNA complementation of T3 generation homozygosis；
(5) expression pattern analysis
Using three months big wild type Turpinia argutas as material, expression pattern analysis has been carried out to two TST genes of poplar,
Result of study finds that two TST genes are generally expressed in multiple histoorgans, wherein PtTST2.1 expression quantity in climax leaves
Highest, also there is a higher expression in xylem, bast and spire, PtTST1.1 expression quantity highest in xylem,
There is higher expression in climax leaves；With the protoplast of Turpinia arguta mesophyll cell to PtTST1.1 and PtTST2.1 albumen
Subcellular localization is carried out, research finds that the two TST albumen of poplar are both positioned on tonoplast；
The wild type Turpinia arguta tissue-cultured seedling of same state is handled one month with the BR of various concentration, is found at 10nM BR
After reason, PtTST1.1 and PtTST2.1 expression quantity is lowered；After the BR processing of 50nM and 100nM, PtTST1.1 and PtTST2.1's
Expression quantity increases；2 months big wild type Turpinia arguta stems are handled with the BR of 100nM, and when 0h, 0.5h, 1h, 2h, 4h samples, and studies
It was found that PtTST1.1 and PtTST2.1 gene expression quantity when BR handles 0.5h and 1h significantly increases, expression quantity is lowered when 2h, 4h
When expression quantity without significant changes, illustrate that TST gene may take part in the response of BR；
GUS dyeing, alcohol decoloration are carried out to the proPtTST1.1 and proPtTST2.1 that are grown on MS root media 20d
After take pictures, the study found that PtTST1.1 and PtTST2.1 has expression at multiple positions of poplar；
(6) functional analysis of transgenic arabidopsis
By the strain of the PtTST1.1/PtTST2.1-attst1-2::tDNA complementation of T3 generation homozygosis, wild type col and intend
Southern mustard mutant attst1-2::tDNA sterilizes 10min with 10% liquor natrii hypochloritis, then with the deionized water punching of sterilizing
It washes 5 times, water is added to be placed on 4 DEG C of dark vernalization 3d；After spring China, seed is sowed on the MS culture medium of not added with antibiotic, 10d
Seedling is transferred to inside soil afterwards: fertile soil: vermiculite=1:2；
Daily fixed point observation growth of seedling situation, record is taken pictures in time；When seedling stem is evacuated to 1cm, it is positioned as the plant
Strain bolting, lotus throne leaf and stem leaf number, flowering time when recording bolting time, bolting in time；Arabidopsis grows to 48d
When, young plant growing state is photographed to record, seed amount in single plant pod number and single pod is counted；
The study found that 18d after seed sprouting, compared with wild type, the growth of seedling of mutant is smaller, the seedling of complementary plant
Sub- growing way is larger；26 days after seed sprouting, the phenotype of wild type and mutant is not significantly different, but the growing way of complementary plant
It is better than wild type and mutant, thus speculates that PtTST1.1 and PtTST2.1 can significantly promote the growth of Arabidopsis thaliana Seedlings phase；
Research finds that PtTST1.1 and PtTST2.1 can significantly promote blooming for arabidopsis, compared with wild type, mutant
The bolting time and flowering time be not significantly different, the flowering time of complementary plant and bolting time significantly shift to an earlier date；
(7) functional analysis of transgenic poplar
It is raw to the MS containing 200mM Ticarcillin/Clavulanate Acid that pCambia2301s2-PtTST1.1/PtTST2.1 is overexpressed poplar subculture
It in root culture medium, cultivates 4 weeks or so, measures the plant height of transgenic plant, the number of blade, internode length, fresh weight compared with wild type, turn
The plant height of gene poplar, average internode length are significantly higher than wild type, and the number of blade is not significantly different.
The method of the present invention is easy；According to transgenic poplar and transgenic arabidopsis as a result, PtTST1.1 and PtTST2.1
The growth that arabidopsis and poplar can be remarkably promoted, the new variety of plant for cultivating fast growing and high yield are worth with extensive gene.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples.
Fig. 1 is the amino acid alignment of the TST albumen of different plant origins and the location drawing of transmembrane domain and hydrophilic loop.
Homologous sequence is compared to be completed with DNAMAN analysis software.
Fig. 2 is the phylogenetic analysis schematic diagram of PtTST1.1, PtTST2.1 DNA encoding the protein.
Chadogram is constructed using MEGA5.1 software ortho position phase connection, bootstrap value is represented in the number of each node and shows
It is intended to.
Fig. 3 is PtTST1.1 and PtTST2.1 protein tertiary structure.
The transmembrane domain of poplar (a) PtTST1.1 He (b) PtTST2.1 are analyzed respectively with TMHMM software.
Fig. 4 is relevant carriers building structure chart.
(a) the vector construction structure chart of PtTST1.1 and PtTST2.1 over-express vector；(b) PtTST1.1 and PtTST2.1
The vector construction structure chart that promoter is merged with gus gene；(c) carrier of PtTST1.1 and PtTST2.1 and YFP fusion protein
Construct structure chart.
Fig. 5 is the identification schematic diagram of poplar PtTST1.1 gene overexpression transgenic line.
Fig. 6 is the identification schematic diagram of poplar PtTST2.1 gene overexpression transgenic line.
Fig. 7 is the tissue specific expression analysis schematic diagram of PtTST1.1 and PtTST2.1.
Transcriptional expression using quantitative fluorescent PCR analysis ' Turpinia arguta ' Cao target gene in each tissue Cao is horizontal.Poplar EF1
β is reference gene.
Fig. 8 is the subcellular localization schematic diagram of PtTST1.1 and PtTST2.1 albumen Yu YFP fusion protein.
Fig. 9 is the response mode schematic diagram of PtTST1.1 and PtTST2.1 to gibberellin.
Figure 10 is the response mode of PtTST1.1 and PtTST2.1 to brassinosteroid.
(a) the brassinosteroid BL of wild type Turpinia arguta tissue-cultured seedling various concentration is handled 1 month, utilizes fluorescent quantitation
PCR analyzes the gene expression amount variation of PtTST1.1, PtTST2.1 and PtTST3；(b) 2 months big wild type Turpinia arguta stems are used
100nM BL handles 0h, 0.5h, 1h, 2h and 4h, utilizes the gene expression of quantitative fluorescent PCR analysis PtTST1.1 and PtTST2.1
Amount variation.
Figure 11 is proTST1.1 (A) and proTST2.1 (B) promoter GUS staining analysis schematic diagram.
Figure 12 is the phenotype schematic diagram of PtTST1.1-attst1-2::tDNA complementation.
A is the phenotype of 18d after germination, and B is the phenotype of 26d after germination.C is the expression that real-time detects PtTST1.1
Amount, AtTUB is reference gene.D is PCR qualification result.
Figure 13 is the phenotype schematic diagram of PtTST2.1-attst1-2::tDNA complementation.
A is the phenotype of 18d after germination, and B is the phenotype of 26d after germination.C is the expression that real-time detects PtTST1.1
Amount, AtTUB is reference gene.D is PCR qualification result.
Figure 14 is the phenotype schematic diagram of PtTST1.1-attst1-2::tDNA complementation.
A is the phenotype of 32d after germination, and B is that (lotus throne number of sheets mesh, blade amt mesh, is bloomed at stem leaf number for data statistics
Time, bolting time).
Figure 15 is the phenotype schematic diagram of PtTST2.1-attst1-2::tDNA complementation.
A is the phenotype of 32d after germination, and B is that (lotus throne number of sheets mesh, blade amt mesh, is bloomed at stem leaf number for data statistics
Time, bolting time).
Figure 16 is the phenotype schematic diagram that poplar PtTST1.1 is overexpressed.A is the phenotype of tissue-cultured seedling 30d.B is plant height, blade
The statistics of number, average internode length, rootage duration.
Figure 17 is the phenotype schematic diagram that poplar PtTST2.1 is overexpressed.
A is the phenotype of tissue-cultured seedling 30d.B is plant height, the number of blade, the statistics of average internode length, rootage duration.
Specific embodiment
A kind of PtTST1.1 and PtTST2.1 promotes the application in plant growth substance in preparation.
A method of research PtTST1.1 and PtTST2.1 is promoting the mechanism of action in plant growth, it is characterized in that:
Include the following steps；
(1) selection of gene and phylogenetic analysis
5 and the TST genetic homology in arabidopsis and beet are found in comospore poplar genome by the method for BLAST
Higher gene order, according to phylogenetic analysis as a result, by this 5 genes be respectively designated as PtTST1.1, PtTST1.2,
PtTST2.1, PtTST2.2 and PtTST3；2 and the ST DNA homolog in arabidopsis are cloned followed by the method for RT-PCR
The highest gene of property, is PtTST1.1 and PtTST2.1 respectively；Wherein, PtTST1.1 includes 2217 pairs of bases, encodes 738 ammonia
Base acid, molecular weight 79.18KD；PtTST2.1 includes 2223 bases, encodes 740 amino acid, molecular weight 79.15KD；
With TMHMM softwarehttp://www.cbs.dtu.dk/services/TMHMM/To two TST albumen of poplar into
Row hydrophobicity analysis, discovery PtTST1.1 have 10 possible transmembrane regions, and PtTST2.1 has 11 possible transmembrane regions, and two
TST albumen has 10 common transmembrane domains, and be also about made of 320 amino acid by one between transmembrane domain 5 and 6
The big ring in hydrophily center, this is consistent with the TSTs protein structure in arabidopsis；
(2) promoter Analysis of PtTST1.1, PtTST2.1 gene
PtTST1.1 and PtTST2.1 gene promoter element is analyzed, it is found that it is red 3 TST genes of poplar have
The binding site E-box of mycin response element GARE-motif and brassinosteroid, and the E-box number of three genes compares
It is more；Meanwhile there are also gibberellin response element P-box, PtTST1.1 the relevant cis regulatory member of 1 auxin by PtTST2.1
Part AuXRR-core, PtTST3 have 1 abscisic acid response element ABRE, thus illustrate, PtTST1.1, PtTST2.1 and PtTST3
Gene may have response to gibberellin, auxin, brassinosteroid and abscisic acid, while also will receive the regulation of these hormones,
The especially regulation of gibberellin and brassinosteroid；
Table: the cis-acting elements relevant to hormone response of the gene promoter area poplar TST distribution；
PtTST1.1 and PtTST2.1 promoter region is analyzed with Plant CARE software；
(3) relevant carriers construct
According to the expression pattern analysis of PtTST1.1, PtTST2.1 and PtTST3 in wild type Turpinia arguta, opened with 2 × 35S
The pCAMBIA2301 carrier of mover driving constructs over-express vector；Target protein and YFP are constructed with pA7-YFP carrier
Fusion protein is used to instantaneous conversion poplar mesophyll protoplast；Target start is constructed with pCAMBIA1300+pBI101
The carrier that son is merged with+gus gene is for converting Turpinia arguta and detect the expression pattern of poplar TST albumen by GUS dyeing；
(4) screening of genetic transformation and transgenic line
It is new that the pCambia2301s2-PtTST1.1/PtTST2.1 over-express vector built is converted into wild type mountain respectively
Poplar has obtained transgenic poplar；By PCR detection, GUS dyeing detection and quantitative fluorescent PCR to the transgenosis young plant of overexpression
It is identified, identifies 15 overexpression strains of PtTST1.1 gene, identify 10 overexpression strains of PtTST2.1 gene
System；PCAMBIA1300+pBI101-PtTST1.1/PtTST2.1 promoter-GUS the carrier built conversion wild type mountain is new
Poplar obtains 10 strains of proPtTST1.1, identifies 11 strains of proPtTST2.1；
To the side for the arabidopsis attst1-2::tDNA mutant three-wheel PCR that H.Ekkehard professor Neuhaus provides
Method is identified, and pCambia2301s2-PtTST1.1/PtTST2.1 over-express vector is converted the mutant, is obtained
The strain of the PtTST1.1/PtTST2.1-attst1-2::tDNA complementation of T3 generation homozygosis；
(5) expression pattern analysis
Using three months big wild type Turpinia argutas as material, expression pattern analysis has been carried out to two TST genes of poplar,
Result of study finds that two TST genes are generally expressed in multiple histoorgans, wherein PtTST2.1 expression quantity in climax leaves
Highest, also there is a higher expression in xylem, bast and spire, PtTST1.1 expression quantity highest in xylem,
There is higher expression in climax leaves；With the protoplast of Turpinia arguta mesophyll cell to PtTST1.1 and PtTST2.1 albumen
Subcellular localization is carried out, research finds that the two TST albumen of poplar are both positioned on tonoplast；
The wild type Turpinia arguta tissue-cultured seedling of same state is handled one month with the BR of various concentration, is found at 10nM BR
After reason, PtTST1.1 and PtTST2.1 expression quantity is lowered；After the BR processing of 50nM and 100nM, PtTST1.1 and PtTST2.1's
Expression quantity increases；2 months big wild type Turpinia arguta stems are handled with the BR of 100nM, and when 0h, 0.5h, 1h, 2h, 4h samples, and studies
It was found that PtTST1.1 and PtTST2.1 gene expression quantity when BR handles 0.5h and 1h significantly increases, expression quantity is lowered when 2h, 4h
When expression quantity without significant changes, illustrate that TST gene may take part in the response of BR；
GUS dyeing, alcohol decoloration are carried out to the proPtTST1.1 and proPtTST2.1 that are grown on MS root media 20d
After take pictures, the study found that PtTST1.1 and PtTST2.1 has expression at multiple positions of poplar；
(6) functional analysis of transgenic arabidopsis
By the strain of the PtTST1.1/PtTST2.1-attst1-2::tDNA complementation of T3 generation homozygosis, wild type col and intend
Southern mustard mutant attst1-2::tDNA sterilizes 10min with 10% liquor natrii hypochloritis, then with the deionized water punching of sterilizing
It washes 5 times, water is added to be placed on 4 DEG C of dark vernalization 3d；After spring China, seed is sowed on the MS culture medium of not added with antibiotic, 10d
Seedling is transferred to inside soil afterwards: fertile soil: vermiculite=1:2；
Daily fixed point observation growth of seedling situation, record is taken pictures in time；When seedling stem is evacuated to 1cm, it is positioned as the plant
Strain bolting, lotus throne leaf and stem leaf number, flowering time when recording bolting time, bolting in time；Arabidopsis grows to 48d
When, young plant growing state is photographed to record, seed amount in single plant pod number and single pod is counted；
The study found that 18d after seed sprouting, compared with wild type, the growth of seedling of mutant is smaller, the seedling of complementary plant
Sub- growing way is larger；26 days after seed sprouting, the phenotype of wild type and mutant is not significantly different, but the growing way of complementary plant
It is better than wild type and mutant, thus speculates that PtTST1.1 and PtTST2.1 can significantly promote the growth of Arabidopsis thaliana Seedlings phase；
Research finds that PtTST1.1 and PtTST2.1 can significantly promote blooming for arabidopsis, compared with wild type, mutant
The bolting time and flowering time be not significantly different, the flowering time of complementary plant and bolting time significantly shift to an earlier date；
(7) functional analysis of transgenic poplar
It is raw to the MS containing 200mM Ticarcillin/Clavulanate Acid that pCambia2301s2-PtTST1.1/PtTST2.1 is overexpressed poplar subculture
It in root culture medium, cultivates 4 weeks or so, measures the plant height of transgenic plant, the number of blade, internode length, fresh weight compared with wild type, turn
The plant height of gene poplar, average internode length are significantly higher than wild type, and the number of blade is not significantly different.
(8) it summarizes
According to transgenic poplar and transgenic arabidopsis as a result, PtTST1.1 and PtTST2.1 can remarkably promote quasi- south
The growth of mustard and poplar, the new variety of plant for cultivating fast growing and high yield are worth with extensive gene.
Sequence relevant information:
PtTST1.1-CDS sequence, protein sequence
>PtTST1.1-2217bp
ATGAAGGGAGCATCTCTAGTGGCTATTGCTGCTTGCGTTGGTAACTTCTTGCAAGGATGGGATAATGCT
ACCATTGCTGGCGCTGTCATTTACGTCAAGAAAGACCTCAAGTTGCAATCGAGTGTGGAAGGTCTTGTTGTGGCCAT
GTCACTTATTGGAGCCGCTGCTATCACAACATGCTCAGGACCCATATCAGATTGGATTGGTCGGCGTCCAATGCTAA
TAAGCTCATCAATTCTCTACTTTGTCAGTGGTTTGGTAATGTTTTGGTCACCCAATGTTTATGTCTTGTGCATAGGA
AGACTGTTGGATGGATTTGGCGTTGGTTTAGCAGTTACTCTTATTCCACTCTATATTTCTGAGACCGCCCCATCAGA
TATAAGGGGAATGTTAAATACTCTACCTCAGTTTGCCGGTTCAGGTGGCATGTTTCTGTCGTACTGTATGGTTTTTG
GGATGTCATTGACAACTTCACCTAGTTGGAGGATGATGCTTGGAATTCTTTCCATTCCTTCTTTACTATATTTTGTA
CTCACAGTGTTTTACTTGCCTGAATCTCCTCGATGGCTTGTAAGTAAAGGAAAGATGCTTGAGGCAAAGCAGGTTCT
CCAGAGATTGCGTGGCAGGGAAGATGTTTCAGGCGAGATGGCTTTACTGGCTGAAGGTCTTGGTATCGGGGGTGAAA
CATCCATAGAAGAATACATAATAGGGCCTGCTGATGAAGTCGCTGATGGTCAAGAACCCATTGTTGATAAAGACAAA
ATCAAGTTATATGGACCTGAAGAAGGCCTTTCCTGGGTTGCTAAACCCGTAACTGGACAGAGTTCTCTTGCTCTTGT
ATCGCGCCAAGGAAGCATGGTGAACCAAGGCGTGCCTCTTATGGACCCTCTTGTGACTCTTTTTGGTAGTGTTCATG
AAAAGCTCCCTGAGACAGGAAGCATGCGGAGCATGCTTTTCCCTAATTTTGGCAGCATGTTTAGTACAGCAGAACCT
CACTTTAGGACTGAGCAGTGGGATGAAGAGAGTGTACAAAGAGAAGGTGAGGGCTACACATCAGAGGCTGGTGGTGA
GGATTCTGATGACAATTTGCACAGTCCACTAATATCACGCCAGACGACAAGCATGGAAAAGGATATGGCCCACCCAA
CTTCCCATGGCAGTGCTCTGAGCATGAGACGGCATAGCAGTCTATTGCAAGGAGCTGGGGAGGCAGTTGACGGTACT
GGCATTGGTGGGGGTTGGCAGTTGGCATGGAAATGGTCCGAGAGAGAAGGTGAGGATGGAAAGAAGGAAGGGGGGTT
TAAAAGGATTTATTTGCACCAAGAGGGAGTTCCTGGATCCCGACGCGGGTCTGTTGTTTCACTTCCTGGTGGTGATG
TTCCTGTTGAAGGTGAGTATATCCAGGCTGCTGCTCTGGTAAGCCAGCCAGCTCTTTATTCAAAGGAGCTTATGGAT
CAGCATCCAGTTGGACCCGCAATGGTTCACCCATCTCAAACAGCTACAAAAGCTCCGATATGGGCCGCTCTGCTTGA
ACCCGGAGTTAAGCATGCTTTGTTTGTTGGGATGGGAATTCAATTGCTTCAGCAGTTTGCTGGTATAAATGGAGTTC
TTTACTACACGCCTCAAATTCTTGAAGACGCAGGTGTTTCGGTTCTTCTTGCAAACTTGGGCCTCAGCACAAACTCT
GCATCATTCCTTATAAGTGCATTTACAAACCTCCTTATGCTTCCATGTATAGGAGTAGCGATGAAGCTTATGGATAT
CTCAGGGAGAAGGACGCTCCTACTTACCACAATTCCTGTGCTGATACTTTCCCTCGTCGTCTTAATTATTTTTGAAC
TAGTGACTGTGAGCGCAATCGTCAGTGCTGCAATCTTAACTGCCTGTGTTATCATCTTCATCTGCTGTTTTGTGTCG
GCTTATGGACCAATTCCTAATATCCTCTGCTCGGAGATCTTCCCGACAAGAGTCCGAGGCCTCTGCATTGCCATTTG
TGCCATGGTTTACTGGATTGGAGACATCATTGTCACCTACACACTGCCTGTGATGCTAACTTCCATCGGCCTAGTTG
GTATCTTCAGCATTTACGCCGCTGTGTGCGTCATCTCTTGGATCTTTGTTTTCTTGAAGGTCCCAGAGACCAAAGGA
ATGCCTCTTGAAGTCATTACTGAGTTCTTTGCTGTGGGCGCAAGACAAGCTGCTGCTGCCAAGAATTAA
>PtTST1.1
MKGASLVAIAACVGNFLQGWDNATIAGAVIYVKKDLKLQSSVEGLVVAMSLIGAAAITTCSGPISDWIG
RRPMLISSSILYFVSGLVMFWSPNVYVLCIGRLLDGFGVGLAVTLIPLYISETAPSDIRGMLNTLPQFAGSGGMFLS
YCMVFGMSLTTSPSWRMMLGILSIPSLLYFVLTVFYLPESPRWLVSKGKMLEAKQVLQRLRGREDVSGEMALLAEGL
GIGGETSIEEYIIGPADEVADGQEPIVDKDKIKLYGPEEGLSWVAKPVTGQSSLALVSRQGSMVNQGVPLMDPLVTL
FGSVHEKLPETGSMRSMLFPNFGSMFSTAEPHFRTEQWDEESVQREGEGYTSEAGGEDSDDNLHSPLISRQTTSMEK
DMAHPTSHGSALSMRRHSSLLQGAGEAVDGTGIGGGWQLAWKWSEREGEDGKKEGGFKRIYLHQEGVPGSRRGSVVS
LPGGDVPVEGEYIQAAALVSQPALYSKELMDQHPVGPAMVHPSQTATKAPIWAALLEPGVKHALFVGMGIQLLQQFA
GINGVLYYTPQILEDAGVSVLLANLGLSTNSASFLISAFTNLLMLPCIGVAMKLMDISGRRTLLLTTIPVLILSLVV
LIIFELVTVSAIVSAAILTACVIIFICCFVSAYGPIPNILCSEIFPTRVRGLCIAICAMVYWIGDIIVTYTLPVMLT
SIGLVGIFSIYAAVCVISWIFVFLKVPETKGMPLEVITEFFAVGARQAAAAKN
PtTST2.1-CDS sequence, protein sequence
>PtTST2.1-2223bp
ATGAATGGAGCTGTGCTTGTAGCTGTTGCTGCTGCTATTGGCAACTTATTGCAAGGATGGGATAATGCA
ACCATCGCAGGGGCTGTTTTATACATAAAAAGGGAATTTCATTTGGAAAGTGAACCTACTATTGAAGGATTAATCGT
GGCTACATCACTTGTTGGAGCCACTTTAATTACTACGTGTTCTGGACCCATATCTGATTGTCTAGGCCGCCGTCCTT
TGTTGATAATCTCATCAATACTTTATTTTGTTAGTGGTCTTGTAATGTTATGGTCTCCCAATGTTTACGTTCTGCTT
TTGGCAAGGCTTTTGGATGGATTTGGCATTGGTTTGGCAGTAACTCTTGTTCCAGTTTATATATCTGAGACAGCACC
ACCTGAAATAAGGGGGTTGTTGAATACCCTTCCTCAGTTCACTGGATCTGGTGGAATGTTTCTGTCATATTGCATGG
TGTTCGGAATGTCCTTGATGGAGGCTCCAAGTTGGAGAGTGATGCTTGGAGTTCTTTTTATTCCCTCAATCATCTAT
TTTCTATTGACTGTATTTTTCTTGCCTGAGTCTCCAAGGTGGCTTGTAAGTAAAGGACGGATGCTTGAGGCCAAGAA
GGTTCTGCAGAGGCTCCGTGGCAGAGAAGATGTTTCTGGTGAGCTGGCTTTACTGGTTGAGGGACTTGGAGTTGGGA
CTGACATATCAATAGAGGAGTACATAATTGGCCCTGCCAACGATTTCACCGATGACCATGATATAGCTGCCGACAAA
GATCATATCAAGTTATATGGGCCTGAACAAGGTCACTCCTGGGTTGCCAGACCTGTCAGTGGGCAGAGTGCTATTGG
TCTTGCGTCTAGGCATGGAAGCATGGCAAACCAGAGTCTAGCTCTCATGGATCCTCTTGTCACCCTCTTTGGTAGCG
TCCATGAGAAGCTCCCTGAACAAGGAAGCATGAGAAGCATGCTTTTCCCTCACTTTGGAAGCATGTTCAGTGTAGGA
GGGAATCATCCTAGAAATGAAGATTGGGATGAGGAGAGCCAAGCCAGAGATGGTGAGGATTATGCATCTGATGGTGC
TGCTGGTGATTCTGATGACAATTTGCAGAGTCCATTGATCTCACGTCAGGCAACAAGCATGGACAAGGACATGGTCC
CGCCTGCCCATGGAAGCATGTCAAGCATGAGACATGGGAGTCTGATTACAGGAAATGCTGGAGATCCAGTTGGTAAC
ACAGGGATTGGTGGTGGTTGGCAGCTGGCATGGAAATGGTCCGAGAGAGAAGGTCAAGATGGAAAGAAGGAAGGGGG
CTTTAAGAGAATTTATTTGCATCAAGAGGGAGCCCCTGGTTCTCGCCGTGGATCTCTGGTTTCTCTGACTGGTGCTG
ATGCCCATGCAGACAGTGAATACATCCAGGCTGCTGCTCTGGTGAGTCAATCAGCTCTTTATCCCAAGGAGCTTGTG
AATGAGAATCCAGCTGGACCAGCTATGGTCCACCCATCTGAAACTGTAGCTAAAGGACCAAGCTGGAGAGATCTTTT
TGAACCAGGAGTCAAGCATGCCTTGGCTGTTGGTGTGGGAATTCAAATACTTCAGCAGTTCGCTGGCATAAATGGGG
TTCTCTACTATACTCCTCAAATTCTTGAGCAGGCAGGAGTTGGAGTTCTTCTTTCAAACTTGGGCCTCAGTTCAGCT
TCTACATCTCTGCTTATCAGCGCCCTTACAACATTGTTGATGCTCCCTTGTATAGCTGTTGCCATGAGGCTCATGGA
TATCTCTGGGAGAAGGACTTTGCTGCTCACCACCATCCCCGTGTTGATCATATCCCTCATTTTGTTAGTCCTTGGAA
GCTTGGTGGATATGGGCAGTGTTGTTAATGCATCAATCTCAACTGTTAGCGTTGTGCTCTACTTCTGTTTTTTCGTC
ATGGGTTTTGGGCCAATTCCCAACATATTATGTGCAGAGATCTTCCCTACTCGTGTTCGTGGCCTTTGCATAGCCAT
ATGCGCCCTTACTTTCTGGATTTGCGACATCATTGTGACGTATACGCTCCCAGTTATGCTTAAATCTATCGGCCTTG
CTGGTGTTTTTGGCTTATATGCAATCGTGTGCGTCATATCATTTGTGTTTGTCTACTTGAAAGTTCCAGAGACCAAG
GGCATGCCTCTGGAAGTGATTTCCGAGTTCTTTGCCGTTGGTGCAAAGCAGGCTGCAGCTGCTAAGGAAAACTGA
>PtTST2.1
MNGAVLVAVAAAIGNLLQGWDNATIAGAVLYIKREFHLESEPTIEGLIVATSLVGATLITTCSGPISDC
LGRRPLLIISSILYFVSGLVMLWSPNVYVLLLARLLDGFGIGLAVTLVPVYISETAPPEIRGLLNTLPQFTGSGGMF
LSYCMVFGMSLMEAPSWRVMLGVLFIPSIIYFLLTVFFLPESPRWLVSKGRMLEAKKVLQRLRGREDVSGELALLVE
GLGVGTDISIEEYIIGPANDFTDDHDIAADKDHIKLYGPEQGHSWVARPVSGQSAIGLASRHGSMANQSLALMDPLV
TLFGSVHEKLPEQGSMRSMLFPHFGSMFSVGGNHPRNEDWDEESQARDGEDYASDGAAGDSDDNLQSPLISRQATSM
DKDMVPPAHGSMSSMRHGSLITGNAGDPVGNTGIGGGWQLAWKWSEREGQDGKKEGGFKRIYLHQEGAPGSRRGSLV
SLTGADAHADSEYIQAAALVSQSALYPKELVNENPAGPAMVHPSETVAKGPSWRDLFEPGVKHALAVGVGIQILQQF
AGINGVLYYTPQILEQAGVGVLLSNLGLSSASTSLLISALTTLLMLPCIAVAMRLMDISGRRTLLLTTIPVLIISLI
LLVLGSLVDMGSVVNASISTVSVVLYFCFFVMGFGPIPNILCAEIFPTRVRGLCIAICALTFWICDIIVTYTLPVML
KSIGLAGVFGLYAIVCVISFVFVYLKVPETKGMPLEVISEFFAVGAKQAAAAKEN
PtTST3-CDS sequence, protein sequence
>PtTST3-2160bp
ATGAGGGGAGCTGTTCTTGTAGCACTTGCTGCTACGGTGGGGAATCTGTTACAAGGATGGGATAATTC
AACCATTGCAGGATCCATTCCTTACATCAAGGAGGAGTTAAATTTGCAGTCTCAACCAGCAGTAGAAGGGCTGATT
GTAGCCATGTCAATTATCGGTGGCACTACTATCACAACATTTTCTGGAACAGTATCAGATATCTTCGGAAGACGTC
CGATGCTGATAATGTCGTCAATTCTTTATTTTCTAAGCAGCATAATTATACTATGGGCTCCCAATGTTTATGTCCT
ACTTTTGGCGAGACTACTTGATGGTTTTGGAGTAGGTCTTGCTGTCACTCTTGTTCCTTTATATATATCTGAAACG
GCTCCATCTGAGATGAGGGGGCAATTAAATACACTTCCACAGTTTATGGGTTCAGGAGGAATGTTTTTGTCATATT
GCATGGTGTTTTTCATGTCAATGATGGATTCACCAAGCTGGCGGCTGATGCTTGGGACTCTGTCAATCCCCGCTGT
CATTTATCTAGCATTGACGCTTTTTTTCTTGCCTGAATCTCCAAGGTGGCTTGTGAGTAAAGGAAAGATGATCGAA
GCTAAACAGGTTTTGCAGAGACTTCGTGGTAGGGAAGACGTTTCAGGTGAGCTGGCTTTGCTACTTGAAGGTCTTG
GTGTTGGGACCGAAACAACAATAGAAGAGTACATTATTGGCCCAGCCAATGAGATCACTGGTGAAACTGATGCGAA
AGAACATGTTAAATTATATGGGCCCGAGGAAGGTGTCTCGTGGATTGCCAAACCCGTCACCGCTGGATTCAGTAGT
TTAGGGATGTTGTCCCGTAACGGCAGTTTGGTAAACCAAACCGTGCCTTTGATGGATCCACTTGTCACTCTATTTG
GAAGTGTCCATGAGAACATGCCTACAACGGGAAGCACACGCAGCTTGCTTTTTCCAAACACTGCAAGCATGGTGAG
TGTTGGAGAAAATCAAGGTAGAAATGAACAGTGGGACGAAGAGGGCGACAAGGATGGCGAGGACTCTTATCCAGAA
GCTTCTAGAGCTGATTCTGATGACAATTTGCGAAGCCCGTTACTATCACACCAACATTCTAGCATGGAAAAGGGCA
TCAGTCATTGGCGCAATAGCAGCCTCGTAAATTCTGGGGAAGAAGGTGCGATGGGTATTGGTGGCGGATGGCAGCT
GGCATATAAATGGTCTGAGAAGATAGGTAAAGATGGTAGTAAGGAAGGAGGGCTTCAGAGGATATATTTGCACCAG
GAAGGTACGATTGGTTCTCAGAAGCATTCAGTTACTTCCTCTGCTGGAATTGACATTCCTGAAGATGAGTTTGTAC
AAGCTGCTGCTCTGGTTAGCCAACCTGCTGTTTGTTCTAAGGACATACTGGGTCAAGCATCAGAAGGTCTAGCAGC
TATTCACCCATCAGAAATTGCTGCTAAAGGTCCAAGCTGCGGTGACCTTTTTGAACCAGGAGTTAAGCGTGCATTG
ATTGTTGGAGTAGGGCTTCAAATACTCCAGCAGGTTGCTGGCATAAATGGTGTTCTCTACTACACACCCCAGATTC
TTGAGCAAGCAGGGGTGGTAGTTCTTCTATCAAGTCTGGGCTTGAGTTCAGCTTCTGCATCTTACTTGATGAGTAT
TCTCACTACATTCTTGATGCTTCCTTGCATATTTCTTGCCATGAGACTAATGGATGTCTCTGGCAGAAGATCTATT
TTGCTGTACACCATTCCTATCTTGGTAGCATCGCTAGTTGCTTTTGTCCTTGGCAGTATTGTCAACATGGACTCGT
CTTTGAAAGCAGTGATCTCTACTGGCAGTGTTATGATCTATTTGAGTTGTTTCGTCATGGGCTTCGGAGTAATTCC
AAACATCCTCTGTGCTGAAATTTTCCCGACTCGTGTTCGTGGCATTTGCATCACAATATGTTCTCTGACATATTGG
ATTGGAAACATCACGATCACGTACTCGCTTCCTGTTATGCTAAACTTCTTTGGCCTTTCTGGTGTCTTCACAATCT
ATGCCATTGGATGCGCGGTGTCATGGATTTTTGTTTTCTTGAAGGTTCCTGAGACAAAGGGCATGCCCCTTGAAGT
CATCACCGAGTTCTTTGCTGTGGGTTCCAAGAATGATTGA>PtTST3
MRGAVLVALAATVGNLLQGWDNSTIAGSIPYIKEELNLQSQPAVEGLIVAMSIIGGTTITTFSGTVSD
IFGRRPMLIMSSILYLLSSIIILWAPNVYVLLLARLLDGFGVGLAVTLVPLYISETAPSEMRGQLNTLPQFMGSGG
MFLSYCMVFFMSMMDSPSWRLMLGTLSIPAVIYLALTLFFLPESPRWLVSKGKMIEAKQVLQRLRGREDVSGELAL
LLEGLGVGTETTIEEYIIGPANEITGETDAKEHVKLYGPEEGVSWIAKPVTAGFSSLGMLSRNGSLVNQTVPLMDP
LVTLFGSVHENMPTTGSTRSLLFPNTASMVSVGENQGRNEQWDEEGDKDGEDSYPEASRADSDDNLRSPLLSHQHS
SMEKGISHWRNSSLVNSGEEGAMGIGGGWQLAYKWSEKIGKDGSKEGGLQRIYLHQEGTIGSQKHSVTSSAGIDIP
EDEFVQAAALVSQPAVCSKDILGQASEGLAAIHPSEIAAKGPSCGDLFEPGVKRALIVGVGLQILQQVAGINGVLY
YTPQILEQAGVVVLLSSLGLSSASASYLMSILTTFLMLPCIFLAMRLMDVSGRRSILLYTIPILVASLVAFVLGSI
VNMDSSLKAVISTGSVMIYLSCFVMGFGVIPNILCAEIFPTRVRGICITICSLTYWIGNITITYSLPVMLNFFGLS
GVFTIYAIGCAVSWIFVFLKVPETKGMPLEVITEFFAVGSKND。
Claims (2)
1. a kind of PtTST1.1 and PtTST2.1 promotes the application in plant growth substance in preparation.
2. a kind of research PtTST1.1 and PtTST2.1 is in the method for promoting the mechanism of action in plant growth, it is characterized in that: packet
Include the following steps；
(1) selection of gene and phylogenetic analysis
It is found in comospore poplar genome by the method for BLAST 5 higher with the TST genetic homology in arabidopsis and beet
Gene order, according to phylogenetic analysis as a result, by this 5 genes be respectively designated as PtTST1.1, PtTST1.2,
PtTST2.1, PtTST2.2 and PtTST3；2 and the ST DNA homolog in arabidopsis are cloned followed by the method for RT-PCR
The highest gene of property, is PtTST1.1 and PtTST2.1 respectively；Wherein, PtTST1.1 includes 2217 pairs of bases, encodes 738 ammonia
Base acid, molecular weight 79.18KD；PtTST2.1 includes 2223 bases, encodes 740 amino acid, molecular weight 79.15KD；
With TMHMM softwarehttp://www.cbs.dtu.dk/services/TMHMM/Two TST albumen of poplar are dredged
Water quality analysis, discovery PtTST1.1 have 10 possible transmembrane regions, and PtTST2.1 has 11 possible transmembrane regions, two TST eggs
It is white to have 10 common transmembrane domains, and be also about made of 320 amino acid by one between transmembrane domain 5 and 6 hydrophilic
Property center big ring, this is consistent with the TSTs protein structure in arabidopsis；
(2) promoter Analysis of PtTST1.1, PtTST2.1 gene
PtTST1.1 and PtTST2.1 gene promoter element is analyzed, it is found that 3 TST genes of poplar have gibberellin
The binding site E-box of response element GARE-motif and brassinosteroid, and the E-box number of three genes compares
It is more；Meanwhile there are also gibberellin response element P-box, PtTST1.1 the relevant cis-regulating element of 1 auxin by PtTST2.1
AuXRR-core, PtTST3 have 1 abscisic acid response element ABRE, thus illustrate, PtTST1.1, PtTST2.1 and PtTST3 base
Because that may have response to gibberellin, auxin, brassinosteroid and abscisic acid, while it also will receive the regulation of these hormones, especially
It is the regulation of gibberellin and brassinosteroid；
Table: the cis-acting elements relevant to hormone response of the gene promoter area poplar TST distribution；
PtTST1.1 and PtTST2.1 promoter region is analyzed with Plant CARE software；
(3) relevant carriers construct
According to the expression pattern analysis of PtTST1.1, PtTST2.1 and PtTST3 in wild type Turpinia arguta, with 2 × 35S promoter
The pCAMBIA2301 carrier of driving constructs over-express vector；Merging for target protein and YFP is constructed with pA7-YFP carrier
Albumen is used to instantaneous conversion poplar mesophyll protoplast；Constructed with pCAMBIA1300+pBI101 target promoter with+
The carrier of gus gene fusion is used to convert Turpinia arguta and detects the expression pattern of poplar TST albumen by GUS dyeing；
(4) screening of genetic transformation and transgenic line
The pCambia2301s2-PtTST1.1/PtTST2.1 over-express vector built is converted into wild type Turpinia arguta respectively,
Transgenic poplar is obtained；The transgenosis young plant of overexpression is carried out by PCR detection, GUS dyeing detection and quantitative fluorescent PCR
Identification, identifies 15 overexpression strains of PtTST1.1 gene, identifies 10 overexpression strains of PtTST2.1 gene；It will
PCAMBIA1300+pBI101-PtTST1.1/PtTST2.1 promoter-GUS carrier conversion wild type the Turpinia arguta built, is obtained
10 strains for obtaining proPtTST1.1 identify 11 strains of proPtTST2.1；
The arabidopsis attst1-2::tDNA mutant that H.Ekkehard professor Neuhaus is provided with the method for three-wheel PCR into
It has gone identification, and pCambia2301s2-PtTST1.1/PtTST2.1 over-express vector is converted into the mutant, obtained T3 generation
The strain of homozygous PtTST1.1/PtTST2.1-attst1-2::tDNA complementation；
(5) expression pattern analysis
Using three months big wild type Turpinia argutas as material, expression pattern analysis has been carried out to two TST genes of poplar, has been studied
As a result, it has been found that two TST genes are generally expressed in multiple histoorgans, wherein PtTST2.1 expression quantity highest in climax leaves,
Also there are higher expression, PtTST1.1 expression quantity highest in xylem, in maturation in xylem, bast and spire
There is higher expression in leaf；Asia is carried out to PtTST1.1 and PtTST2.1 albumen with the protoplast of Turpinia arguta mesophyll cell
Cellular localization, research find that the two TST albumen of poplar are both positioned on tonoplast；
The wild type Turpinia arguta tissue-cultured seedling of same state is handled one month with the BR of various concentration, after discovery 10nM BR processing,
PtTST1.1 and PtTST2.1 expression quantity is lowered；After the BR processing of 50nM and 100nM, the expression quantity of PtTST1.1 and PtTST2.1
It increases；2 months big wild type Turpinia arguta stems are handled with the BR of 100nM, when 0h, 0.5h, 1h, 2h, 4h, samples, the study found that
PtTST1.1 and PtTST2.1 gene expression quantity when BR handles 0.5h and 1h significantly increases, and expression quantity is lowered when 2h, table when 4h
Up to amount without significant changes, illustrate that TST gene may take part in the response of BR；
GUS dyeing is carried out to the proPtTST1.1 and proPtTST2.1 that are grown on MS root media 20d, alcohol decoloration is laggard
Row is taken pictures, the study found that PtTST1.1 and PtTST2.1 has expression at multiple positions of poplar；
(6) functional analysis of transgenic arabidopsis
By strain, wild type col and the arabidopsis of the PtTST1.1/PtTST2.1-attst1-2::tDNA complementation of T3 generation homozygosis
Mutant attst1-2::tDNA sterilizes 10min with 10% liquor natrii hypochloritis, then rinses 5 with the deionized water of sterilizing
It is secondary, add water to be placed on 4 DEG C of dark vernalization 3d；After spring China, seed is sowed on the MS culture medium of not added with antibiotic, after 10d
Seedling is transferred to inside soil: fertile soil: vermiculite=1:2；
Daily fixed point observation growth of seedling situation, record is taken pictures in time；When seedling stem is evacuated to 1cm, the plant has been positioned as it
Through bolting, lotus throne leaf and stem leaf number, flowering time when recording bolting time, bolting in time；When arabidopsis grows to 48d,
Young plant growing state is photographed to record, seed amount in single plant pod number and single pod is counted；
The study found that 18d after seed sprouting, compared with wild type, the growth of seedling of mutant is smaller, and the young plant of complementary plant is long
Gesture is larger；26 days after seed sprouting, the phenotype of wild type and mutant is not significantly different, but the growing way of complementary plant is eager to excel
In wild type and mutant, thus speculate that PtTST1.1 and PtTST2.1 can significantly promote the growth of Arabidopsis thaliana Seedlings phase；
Research finds that PtTST1.1 and PtTST2.1 can significantly promote blooming for arabidopsis, compared with wild type, the pumping of mutant
A kind of sedge time and flowering time are not significantly different, and the flowering time of complementary plant and bolting time significantly shift to an earlier date；
(7) functional analysis of transgenic poplar
PCambia2301s2-PtTST1.1/PtTST2.1 is overexpressed poplar subculture and takes root training to the MS containing 200mM Ticarcillin/Clavulanate Acid
It supports in base, cultivates 4 weeks or so, measure the plant height of transgenic plant, the number of blade, internode length, fresh weight compared with wild type, transgenosis
The plant height of poplar, average internode length are significantly higher than wild type, and the number of blade is not significantly different.
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| CN111187777A (en) * | 2020-02-06 | 2020-05-22 | 东北农业大学 | Application of soybean GmTST2.1 gene in soybean breeding |
| CN111187777B (en) * | 2020-02-06 | 2023-03-24 | 东北农业大学 | Application of soybean GmTST2.1 gene in soybean breeding |
| CN111235125A (en) * | 2020-03-19 | 2020-06-05 | 山东师范大学 | Rhodanese EsSTR4A related to salt tolerance, oxidation resistance and antifungal capacity of plants, and coding gene and application thereof |
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