CN102643832A - Soybean growth period E1 gene and encoding protein of soybean growth period E1 gene - Google Patents
Soybean growth period E1 gene and encoding protein of soybean growth period E1 gene Download PDFInfo
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Abstract
The invention discloses a soybean growth period E1 gene and encoding protein of the soybean growth period E1 gene and relates to the soybean growth period E1 gene and the encoding protein of the soybean growth period E1 gene. The invention provides the soybean growth period E1 gene and the encoding protein of the soybean growth period E1 gene. The soybean growth period E1 gene sequence is shown as a sequence table sequence identification number (Seq ID No):1. The amino acid sequence of the encoding protein of the soybean growth period E1 gene is shown as the Seq ID No:2. The E1 gene is cloned by a map-based cloning method, and the physiological function of the E1 gene for intensively inhibiting the soybean flowering is verified through transgenosis and screening of a plurality of ethyl methane sulfonate (EMS) mutant libraries. The invention relates to the field of soybean gene.
Description
Technical field
The present invention relates to a kind of soybean growth period E1 gene and proteins encoded thereof.
Background technology
Soybean provides important plant protein and oil content for the mankind.Worldwide; Northern Europe Sweden of north to high latitude and North America Canada; Reaching in the south in the extensive region such as Brazil and Argentina all has soybean culture, but the latitude span of single kind or the general suitable planting of germ plasm resource is less, and gene breeding time of this regional flexibility and soybean is closely related.Found that so far 9 mainly influence soybean bloom and ripening stage (breeding time) gene locus E1-E8 and J.As far back as the twenties in last century, American scholar Owen just finds the main gene of the ripe property of pair of control soybean, and names and be E and e.Thereafter, Bernard (1971) infers that E and e are the not isoallele in same site, names to be E1/e1.The E1 site to the contribution rate in flowering period and ripening stage up to 50%～70%.Soybean growth period gene E1 is not only to the having the greatest impact of soybean bloom and ripening stage, but also closely related with photoperiodic reaction.Modern soybean gene group information shows that the E1 gene is positioned near the kinetochore of the 6th karyomit(e) (Gm06), and its Recombination Fraction is extremely low, thereby clone's difficulty is very big.Though people have successively cloned E4, E3, Dt1 and raq gene with mode crop homologous gene method or meticulous collection of illustrative plates localization method, do not see the relevant report of successfully cloning the E1 gene so far as yet.
Summary of the invention
The purpose of this invention is to provide a kind of soybean growth period E1 gene and proteins encoded thereof.
A kind of soybean growth period E1 gene order of the present invention is shown in sequence table Seq ID No:1.
The aminoacid sequence of the proteins encoded of soybean growth period E1 gene of the present invention is shown in sequence table Seq ID No:2.
The present invention contains the soybean growth period E1 gene order of promotor section shown in sequence table Seq ID No:3.
The present invention contains the soybean growth period E1 gene of promotor section, and (containing E1 gene transcription section is full-length cDNA shown in sequence table Seq ID No:3; Underscore is represented); Wherein, The sequence of 1～2668bp and 3194～4332bp is respectively 5 ' and 3 ' end promoter sequence of E1 gene, and the sequence of 2669bp～3193bp is a soybean growth period E1 gene order, and 2512～2668bp and 3194～3349bp are respectively 5 ' and 3 ' the end non-translational region of coding soybean growth period E1 full length gene cDNA; Wherein, Promotor section (1～2668bp and 3194～4332bp sequence shown in sequence table Seq ID No:3) derives from the sequence A P011814 of BAC clone MiB42G1; Relevant BAC clone's screening and order-checking be with reference to Xia Z, Sato H, Watanabe S; Kawasaki S, Harada K (2005) Construction and characterization of a BAC library of soybean.Euphytica141:129-137.
Beneficial effect of the present invention:
The present invention successfully clones gene E1 breeding time first on molecular level.
The genetic group that the present invention utilizes different near isogenic line Harosoy-E1 of two of soybean varieties Harosoy and Harosoy-e1 intermolecular hybrid to be obtained; Through the map based cloning method; Successfully with near the 17.4Kb zone of the soybean growth period E1 assignment of genes gene mapping kinetochore, this zone is arranged in BAC clone MiB42G1 sequence A P011814, and passes through GenScan; GeneMark, FGENESH and RiceGAAS determine the position of the sequence of soybean growth period E1 gene.The E1 gene coding region full length sequence (Seq ID No:1) that the present invention obtains is corresponding with the Glyma06g23040.1 (gene of soybean gene group Glyma1.0 version is annotated) of announcement among the phytozome (http://www.phytozome.net); But its sequence is also inconsistent, and the protein of the E1 gene translation that the present invention obtains has 174aa (Seq ID No:2).Through transgenic, verified that the E1 gene has the physiological function of strongly inhibited soybean blossoming.
Simultaneously, through the homology comparison, find that the E1 gene does not have homologous gene in Arabidopis thaliana and paddy rice; It is the distinctive gene of leguminous plants; And still not about the report of homogenic clone of E1 and functional verification in E1 and the leguminous crop, information biology shows that E1 gene of the present invention contains a nuclear localization signal (putative bipartite nuclear localization signal), has KKRK and RRR diadactic structure territory at present; The signal for locating function can be played in this diadactic structure territory; Can make the E1 gene protein be positioned (being transported to) nucleus, in nucleus, work, lay respectively at 13-16 and 29-31 position among the sequence table Seq ID No:2.In addition, the E1 gene also contain with Arabidopis thaliana (Arabidopsis thaliana) and paddy rice in the similar B3 structural domain of japonica rice (Oryza sativa), and on amino acid levels, have 21%～27% similarity.
Soybean growth period E1 gene of the present invention is 525bp (Seq ID No:1); Soybean growth period E1 expression of gene receives photoperiodic adjusting, and to have received obvious suppression shown in figure 13 for soybean growth period E1 expression of gene amount under short day (12 hours illumination/12 hour dark) condition.
Description of drawings
Fig. 1 does not merge the transient expression carrier structure synoptic diagram of E1 gene for GFP;
Fig. 2 merges the transient expression carrier structure synoptic diagram of E1 gene for GFP;
Fig. 3 does not merge the Subcellular Localization striograph under the laser confocal microscope GFP passage of E1 gene for GFP;
Fig. 4 does not merge the Subcellular Localization striograph under the laser confocal microscope chlorophyll passage of E1 gene for GFP;
Fig. 5 does not merge the ubcellular fixing bitmap under the laser confocal microscope bright field of E1 gene for GFP;
Subcellular Localization striograph after Fig. 6 does not merge the E1 gene for GFP laser confocal microscope GFP passage, chlorophyll passage and the bright field fusion;
Fig. 7 is the Subcellular Localization striograph under the laser confocal microscope GFP passage of GFP fusion E1 gene;
Fig. 8 is the Subcellular Localization striograph under the laser confocal microscope chlorophyll passage of GFP fusion E1 gene;
Fig. 9 is the Subcellular Localization striograph under the laser confocal microscope bright field of GFP fusion E1 gene;
Subcellular Localization striograph after Figure 10 merges the E1 gene for GFP laser confocal microscope GFP passage, chlorophyll passage and the bright field fusion;
Figure 11 is the soybean blossoming state graph after changing the E1 gene over to and not changing the E1 gene over to;
Figure 12 shows the expression level graphic representation of E1 gene in the long day for the Realtime quantitative fluorescent PCR;
Figure 13 shows the expression level graphic representation of E1 gene at short day for the Realtime quantitative fluorescent PCR.
Embodiment
Embodiment one: a kind of soybean growth period E1 gene order of this embodiment is shown in sequence table Seq ID No:1.
Embodiment two: the aminoacid sequence of the proteins encoded of the soybean growth period E1 gene of this embodiment is shown in sequence table Seq ID No:2.
The acquisition and the authentication method of soybean growth period E1 gene are following:
(1) acquisition of soybean growth period E1 gene
One, in 2004, with two near-isogenic line Harosoy-E1 (L68-694, the E1e2E3E4e5 of soybean; PI 547707) and Harosoy-e1 (L58-266; E1e2E3E4e5, PI 547676) hybridize, through the map based cloning method E1 gene is carried out examination then; Two, 117 F2 after hybridization filter out the E1 gene locus in the soybean population between Satt365 and Satt489 through the map based cloning method; Three, further 1442 F2:3 are carried out examination for soybean population; Filter out 7 at the genetic recombination body between positioning area; Wherein, the E1 gene locus of 7 genetic recombination bodies that filter out is between Satt365 and Satt557, and through molecule marker A and molecule marker E5; 7 genetic recombination bodies that obtain are further carried out self progeny's phenotypic evaluation, the E1 gene locus is positioned in the 289kb scope of molecule marker A and E5; Four, through the map based cloning method 13761 F2:5 are carried out examination for soybean population; Filter out 10 recombinant chous; And adopt molecule marker 33 and 10 recombinant chous of 12 pairs of acquisitions of molecule marker to carry out self progeny's phenotypic evaluation, the E1 gene locus is positioned in the 17372bp interval of molecule marker 33 and molecule marker 12; Five, use GenScan; GeneMark; FGENESH (http://linux1.softberry.com/berry.phtml) and RiceGAAS (http://ricegaas.dna.affrc.go.jp) program are further identified the dna sequence dna of the localized 17372bp of step 4, have established unique candidate gene and have named the gene into E1; Six, the blade with soybean Harosoy-E1 strain is a material, extracts the blade genome as template, through K1 (Seq ID No:19) and K2 (Seq ID No:20); Adopt and buy the gene from the high-fidelity enzyme primerSTARHS of Takara company amplification E1, the PCR reaction conditions is following: 94 ℃ of preparatory sex change 10min, 94 ℃ of sex change 30s; 60 ℃ of annealing 15s, 72 ℃ are extended 1min, totally 30 circulations; 72 ℃ are extended 7min again, and the sequence of PCR product is checked order on ABI3130 sequenator (ABI company); Sequencing result shows that soybean growth period E1 gene has the nucleotide sequence of Seq ID NO:1 in the sequence table, and the Seq ID No:1 in the sequence table is by 525 based compositions, and coding has the protein of the aminoacid sequence of SEQ ID NO:2 in the sequence table.
Wherein, employed molecule marker A is that to adopt A1 (Seq ID No:11) and molecule A2 (Seq ID No:12) be primer in the map based cloning method, is template with the genomic dna of above-mentioned F2:5 soybean population; Adopt to buy and go out molecule marker A from the ExTaq of Takara enzymatic amplification, the PCR reaction conditions: 94 ℃ of preparatory sex change 10min, 94 ℃ of sex change 30s; 60 ℃ of annealing 30s; 72 ℃ are extended 1min, totally 35 circulations, and 72 ℃ are extended 10min again;
Molecule marker E5 is that to adopt B1 (Seq ID No:13) and B2 (Seq ID No:14) be primer, and (be template, the employing purchase goes out molecule marker E5 from the ExTaq of Takara enzymatic amplification with the genomic dna of above-mentioned F2:5 soybean population; PCR reaction conditions: 94 ℃ of preparatory sex change 10min; 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ are extended 1min; Totally 35 circulations, 72 ℃ are extended 10min again;
Molecule marker 33 is that to adopt C1 (Seq ID No:15) and C2 (Seq ID No:16) be primer, is template with the genomic dna of above-mentioned F2:5 soybean population, adopts purchase to go out molecule marker 33 from the ExTaq of Takara enzymatic amplification; PCR reaction conditions: 94 ℃ of preparatory sex change 10min; 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ are extended 1min; Totally 35 circulations, 72 ℃ are extended 10min again;
Molecule marker 12 is to adopt D1 (Seq ID No:17) and D2 (Seq ID No:18), is template with the genomic dna of above-mentioned F2:5 soybean population, adopts to buy to go out molecule marker 12 from the ExTaq of Takara enzymatic amplification; PCR reaction conditions: 94 ℃ of preparatory sex change 10min; 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ are extended 1min; Totally 35 circulations, 72 ℃ are extended 10min again;
Wherein, described Harosoy-E1 of step 1 and Harosoy-e1 derive from the unified warehouse-in numbering in USDA gene germplasm storehouse (the USDA-ARS National Plant Germplasm System), are respectively PI 547707 and PI547676; Harosoy-E1 and Harosoy-e1 buy from Japanese Biological resources institute gene pool.
(2) functional verification of soybean growth period E1 gene
1, full length cDNA sequence is measured
One, the blade with soybean Harosoy-E1 strain is a material, extracts the total RNA of blade with the operational manual of buying from the TRIzol of Invitrogen company test kit; Two, the total RNA that adopts DNase I treatment step one to extract, the total RNA after the processing adopts NanoDrop to detect its dna content and quality, after dna content and quality reach standard, carries out next step operation; Three, the total RNA that gets after 1 μ g step 2 is handled is used for the synthetic of cDNA; The synthetic operation of cDNA carries out according to the service manual of buying from the BD SMARTTM RACE of BD Biosciences Clontech company cDNA Amplification Kit test kit, obtains cDNA; Four, be primer with E1 gene F1 (Seq ID No:9) with F2 (Seq ID No:10) respectively; The cDNA that step 3 is obtained carries out 5 ' and 3 ' amplification through Race-PCR; 5 ' and 3 ' the dna segment that obtains is checked order on ABI3130 sequenator (ABI company); The dna segment of 5 ' and 3 ' after will checking order then is spliced into full-length cDNA, shown in sequence table Seq ID No:4; Wherein, DNase I buys from Toyobo company, and NanoDrop buys from Thermo Scientific company; Harosoy-E1 buys from Japanese Biological resources institute gene pool.
The full-length cDNA of the E1 gene that obtains is 838bp, shown in sequence table Seq ID No:4.The upper reaches 5 ' the UTR and the downstream 3 ' UTR of E1 gene are respectively 157bp and 156bp.
2, soybean growth period E1 gene is appraised and decided position research
E1 gene cDNA to obtain in the step 1 is a template, through sequences Design G1 (Seq ID No:19) and G2 (the Seq ID No:20) primer of Seq ID No:3 in the sequence table, adopts and buys the gene from the high-fidelity enzyme primerSTARHS of Takara company amplification E1; The PCR reaction conditions is following: 94 ℃ of preparatory sex change 10min, 94 ℃ of sex change 30s, 60 ℃ of annealing 15s; 72 ℃ are extended 1min; Totally 30 circulations, 72 ℃ are extended 7min again, obtain E1 gene coding region product (Seq ID No:1); Two, the E1 gene coding region product that step 1 is obtained adopts ClaI enzyme and SpeI enzyme to carry out enzyme and cuts; Then endonuclease bamhi is cloned in the pBSK plasmid, forms E1 and eGFP syzygy plasmid (being CaMV 35S:E1:eGFP) that a CaMV 35S promoter drives; Three, preparation Arabidopis thaliana protoplastis (Arabidopsis protoplasts); Step 2 is obtained CaMV 35S:E1:eGFP plasmid through calcium chloride polyoxyethylene glycol method transfection Arabidopis thaliana protoplastis (Arabidopsis protoplasts); Simultaneously; With the pBSK that does not contain E1 (CaMV 35S:eGFP) plasmid as control group, through calcium chloride polyoxyethylene glycol method transfection Arabidopis thaliana protoplastis (Arabidopsis protoplasts); Place then under the laser confocal microscope and observe, wherein, the pBSK plasmid is bought from Stratagene company;
Wherein, the preparation method is following for Arabidopis thaliana protoplastis (Arabidopsis protoplasts): one, preparation cellulase Digestive system, and mix the back and under 55 ℃ of temperature, heat 10min, add the CaCl that final concentration is 10mM/L again
2Solution, the beta-mercaptoethanol of 5mM/L and 0.1% (w/v) BSA; Two, get three to around age the Arabidopis thaliana lotus throne blade of bolting not, be cut into the shape of about 0.5mm * 0.5mm, be immersed in then in the cellulase Digestive system that step 1 makes; Place 3h in dark, filter with 200 eye mesh screens again, collect filtered solution and place centrifuge tube with the centrifugal 2min of the speed of 100g/min; Remove supernatant, with the resuspended cleaning of solution of the W5 of precooling, once more with the speed of 100g/min centrifugal after; Collecting precipitation is resuspended and place 30min on ice with W5, and is centrifugal with the speed of 100g/min once more, collecting precipitation; MMg solution with 1mL is resuspended, promptly gets Arabidopis thaliana protoplastis (Arabidopsis protoplasts); Wherein, the cellulase Digestive system the ratio of each component is: the cellulase R10 of 1.25% (w/v), the macerozyme R10 of 0.3% (w/v), the N.F,USP MANNITOL of 0.4M/L, the KCl of 20mM/L, the MES of 20mM/L (pH=5.7); W5 the ratio of each component is: the NaCl of 154mM/L, the CaCl of 125mM/L
2, the MES damping fluid (pH=5.7) of the KCl of 5mM/L and 2mM/L; The proportioning of MMg is: the N.F,USP MANNITOL of 0.4M/L, 15mM/MgCl
2The MES of solution and 4mM/L.
Calcium chloride polyoxyethylene glycol method concrete steps are following: the plasmid (CaMV 35S:E1:eGFP and the CaMV35S:eGFP that one, get 10 μ l respectively; Plasmid concentration is 1 μ g/ μ L) add in the Arabidopis thaliana protoplastis for preparing (Arabidopsis protoplasts) of 100 μ l, add the PEG-CaCl of 110 μ l again
2Solution, the pressure-vaccum mixing transforms 20min; Two, transformed after, add the W5 solution mixing of 440 μ l after, then with the centrifugal 3min of the speed of 100g/min, remove to add behind the supernatant W5 solution incubated overnight 16h of 1mL, promptly accomplish calcium chloride polyoxyethylene glycol method; Wherein, PEG-CaCl
2Be PEG4000 by 4g, the H of 3mL
2O, the 0.8M/L N.F,USP MANNITOL of 2.5mL, the 1M/L CaCl of 1mL
2Solution is processed; The NaCl solution that consists of 154mM/L of W5, the CaCl of 125mM/L
2Solution, the MES damping fluid (pH=5.7) of the KCl solution of 5mM/L and 2mM/L.
The distribution of soybean growth period E1 gene in the Arabidopis thaliana protoplastis Different Organs under laser confocal microscope after the observation transfection, the definite kernel location confirms that tentatively the soybean growth period E1 assignment of genes gene mapping is in nucleus.
Wherein, Fig. 1 and Fig. 2 appraise and decide the structural representation that used carrier is studied in the position for the E1 gene, and Fig. 1 is a control vector, and promptly eGFP (green fluorescent protein) does not merge E1; E1 and eGFP fusion rotein carrier that Fig. 2 makes up for this test.
Under fluorescent microscope, observe soybean growth period E1 gene protein in the distribution situation in the Different Organs shown in Fig. 3 to 10.Can know that by Fig. 3 to Figure 10 E1 albumen (Seq ID No:2) mainly is positioned in the nucleus, also exists a small amount of E1 albumen in tenuigenin.
3, transgenic is carried out functional verification
Concrete steps are: one, the blade with soybean Harosoy-E1 strain is a material, extracts full genome as template, is primer with H1 (Seq ID No:6) and H2 (Seq ID No:5); Adopt to buy and buy from the ExTaq of Takara enzymatic amplification from the high-fidelity enzyme primerSTARHS of Takara company; PCR reaction conditions: 94 ℃ of preparatory sex change 10min, 94 ℃ of sex change 30s, 60 ℃ of annealing 30s; 72 ℃ are extended 5min; Totally 35 circulations, 72 ℃ are extended 10min again, obtain to contain the soybean growth period E1 gene (Seq ID No:3) of promotor; The soybean growth period E1 gene that contains promotor (Seq ID No:3) that two, will obtain is cloned in the pMDC123-GFP plasmid, and the pMDC123-GFP plasmid is transferred in the soybean Kariyutaka kind; Other establishes the pMDC123-GFP empty plasmid as control group, and the pMDC123-GFP plasmid that does not promptly contain the E1 gene compares test, relatively the influence of the bloom of E1 gene pairs plant; Wherein, Related plasmid is transformed and transgenic method carries out with reference to the method for " Sato H; Yamada T; Kita Y, Ishimoto M, Kitamura K (2007) Production of transgenic plants and their early seed set in Japanese soybean variety; Kariyutaka.Plant Biotechnol 24:533-536 " and " Curtis MD, Grossniklaus U (2003) A gateway cloning vector set for high-throughput functional analysis of genes in planta.Plant Physiololgy 133 (2): 462-9 "; Wherein, Soybean Harosoy-E1 and Kariyutaka strain are buied from Japanese Biological resources institute gene pool; The pMDC123-GFP plasmid is through transforming available from the PMDC123 (CD3-747) of The Arabidopsis Biological Resource Center (ABRC) (http://www.arabidopsis.org), and the method for transformation is with reference to Sato H, Yamada T; Kita Y; Ishimoto M, Kitamura K (2007) Production of transgenic plants and their early seed set in Japanese soybean variety, Kariyutaka.Plant Biotechnol 24:533-536 carries out.
The soybeans they grow and the bloom of this test are shown in figure 11, can be known by Figure 11, and 1 is the E1 gene high expression; Demonstrate flower-shaped attitude in tangible evening, and 2 be the empty plasmid contrast, obviously early blossoming; And born pods, explain that the function of E1 gene has the function that suppresses soybean blossoming.
4, the Real-time quantitative fluorescent PCR carries out functional verification
The concrete operations step is: one, the blade with soybean Harosoy-E1 strain is a material, extracts the total RNA of blade with the operational manual of buying from the TRIzol of Invitrogen company test kit; Two, the total RNA that adopts DNase treatment step one to extract, the total RNA after the processing adopt the NanoDrop that buys from Toyobo company to detect its dna content and quality; Three, the step 2 NanoDrop that learns from else's experience detects the total RNA of 1 μ g after the standard up to standard, adopts the test kit of buying from the ReverTraAce of TOYOBO company, and the dT20 primer worn of test box, according to the synthetic cDNA of the operational manual of ReverTra Ace test kit; Four, get cDNA after 3 times of the 2 μ l step 3 dilutions as the RT-PCR template; With J1 (Seq ID No:7) and J2 (Seq ID No:8) is primer; Operation steps is undertaken by buying the operation steps of in the SYBR of BioRad company Green Supermix kit test kit, being introduced, and the total reaction volume of each sample is 20 μ L; Five, adopt purchase to detect from the iCycle of BioRad company iQ PCR in real time detection system; The expression of gene level is to represent that with respect to the relative value of reference gene GmTubulin the numerical value of each point is the mensuration result of three individual vanes.
The E1 gene is shown in figure 12 at the expression level graphic representation of long day, and the E1 gene is shown in figure 13 at the expression level graphic representation of short day, and wherein TUB is reference gene GmTubulin; Dotted line is represented the E1 gene in the Harosoy-E1 kind among Figure 12, and solid line is represented the E1 gene in the Harosoy-E1 kind among Figure 13, and wherein TUB is reference gene GmTubulin.
Can know that by Figure 12 and 13 E1 gene of the present invention is expressed and is bordering on zero under the short day condition, explain that the effect that E1 suppresses to bloom is bordering on zero; The soybean varieties prematurity; And the article difference between species disappears, side light the function of E1 gene, after this test repetitive operation once; Obtained analog result, the repetitive operation evidence low this rule of short day expression amount.Promotor is an integral part of gene (gene) on the broad sense, and controlling gene is expressed the time of origin of (transcribing) and the degree of expression.Promotor (Promoters) determines the activity of gene just as " switch ".E1 promotor section (1～2694bp shown in sequence table Seq IDNo:3 and 3436～4332bp sequence) is certainly existing the cis element of controlling E1 genetic expression in test; Cause the E1 gene expression amount height under the long day condition, the low characteristic of expression amount under the short day condition to occur.
Embodiment three: the soybean growth period E1 gene order that contains the promotor section of this embodiment is shown in sequence table Seq ID No:3.
Claims (2)
1. a soybean growth period E1 gene is characterized in that soybean growth period E1 gene order is shown in sequence table Seq ID No:1.
2. the proteins encoded of a kind of soybean growth period E1 gene as claimed in claim 1, the aminoacid sequence of proteins encoded that it is characterized in that soybean growth period E1 gene is shown in sequence table Seq ID No:2.
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| CN102994516A (en) * | 2012-12-12 | 2013-03-27 | 中国科学院东北地理与农业生态研究所 | Soybean flowering gene ft2a-1 and protein coded by the gene |
| CN102994516B (en) * | 2012-12-12 | 2014-04-02 | 中国科学院东北地理与农业生态研究所 | Soybean flowering gene ft2a-1 and protein coded by the gene |
| CN109055393A (en) * | 2018-08-20 | 2018-12-21 | 中国科学院东北地理与农业生态研究所 | Soybean growth period QNE1 gene and its coding albumen and application |
| CN113308478A (en) * | 2021-05-28 | 2021-08-27 | 中国科学院东北地理与农业生态研究所 | Application of soybean E1 gene in regulating pod bearing habit |
| WO2023168691A1 (en) * | 2022-03-11 | 2023-09-14 | Syngenta Crop Protection Ag | Methods and compositions for modifying flowering time genes in plants |
| WO2023173003A3 (en) * | 2022-03-11 | 2023-10-26 | Syngenta Crop Protection Ag | Methods and compositions for modifying flowering time genes in plants |
| CN118440960A (en) * | 2024-06-28 | 2024-08-06 | 吉林省农业科学院(中国农业科技东北创新中心) | Soybean genotype and functional molecular marker InDel-E1 for increasing yield of semi-dwarf and application thereof |
| CN118440960B (en) * | 2024-06-28 | 2024-09-10 | 吉林省农业科学院(中国农业科技东北创新中心) | Soybean genotype and functional molecular marker InDel-E1 for increasing yield of semi-dwarf and application thereof |
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| CN102643832B (en) | 2013-05-08 |
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