CN102690831A - El-b3a gene in soybean growth period and coding protein thereof - Google Patents
El-b3a gene in soybean growth period and coding protein thereof Download PDFInfo
- Publication number
- CN102690831A CN102690831A CN2012101621768A CN201210162176A CN102690831A CN 102690831 A CN102690831 A CN 102690831A CN 2012101621768 A CN2012101621768 A CN 2012101621768A CN 201210162176 A CN201210162176 A CN 201210162176A CN 102690831 A CN102690831 A CN 102690831A
- Authority
- CN
- China
- Prior art keywords
- gene
- soybean
- growth period
- soybean growth
- sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to an el-b3a gene in a soybean growth period and a coding protein thereof. The gene sequence of the el-b3a gene in the soybean growth period is shown as a sequence list Seq ID No:1, and the amino acid sequence of the coding protein of the el-b3a gene in the soybean growth period is shown as a sequence list Seq ID No:2. By means of relevant heredity and molecular functional verification of varieties, the el-b3a gene has a flowering promoting function as compared with El genes.
Description
Technical field
The present invention relates to soybean growth period e1-b3a gene and proteins encoded thereof.
Background technology
The E1 gene is positioned near the kinetochore, and since Bernard (1971) year reported, people never cloned its functional gene.Because this gene is positioned at nearly kinetochore (pericentromeric region); Recombination Fraction is lower; Be unfavorable for adopting the map based cloning method to clone this gene, in the patent of invention title: a kind of soybean growth period E1 gene and proteins encoded thereof, number of patent application is: in 201210112662.9; After announcing out E1 gene order and corresponding proteins sequence thereof, prove that soybean growth period E1 gene has the physiological function of strongly inhibited soybean blossoming.Though its protein sequence can not identify the B3 structural domain, has the relation of edge far away with the B3 domain gene.The B3 domain gene is one type of comparatively conservative plant specific transcription factor, generally has only 100-120 amino acid, and normal and other structural domains concur.Contain the special site that combines with DNA in the B3 structural domain, major groove general and dna molecular works.Play an important role aspect many in growth that contains the protein antagonism adverse effect of B3 structural domain, plant and growth etc.Simultaneously, the E1 gene has a nuclear localization signal, it is characterized by the whole protein that can guarantee that karyophilic protein generally all contains and can be transported to endonuclear special small peptide aminoacid sequence through nuclear pore complex.In the soybean culture kind, how the sudden change of its E1 gene affects soybean growth period (flowering period and ripening stage) is a very important problem in science, does not also have the correlative study report on molecular level at present.
Summary of the invention
Soybean growth period e1-b3a gene and proteins encoded thereof have been the purpose of this invention is to provide.
The gene order of soybean growth period e1-b3a gene 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-b3a gene of the present invention is shown in sequence table Seq ID No:2.
Soybean growth period e1-b3a gene of the present invention is the (application for a patent for invention number: mutated genes 201210112662.9) of soybean growth period E1 gene.
Beneficial effect of the present invention:
The present invention is cloning the (application for a patent for invention number: on the basis 201210112662.9), further the E1 gene order is goed deep into systematic research of soybean growth period E1 gene.Owing in soybean, have typical B 3 gene structure domain genes, Ben Jiyin represent one type of novel gene family, and we have confirmed that the E1 gene can represent the function in E1 site, promptly have the function of inhibition soybean blossoming.The present invention studies E1 gene order in different varieties and the near isogenic line, and has confirmed the e1-b3a gene, and it gives birth to bloom to soybean has promoter action.
Soybean growth period E1 gene exist in aminoacid sequence 55 ~ 174 zones one with the similar zone of B3 structural domain edge far away; The nucleotides sequence of soybean growth period e1-b3a gene is listed in that 3 base mutations take place at 334 ~ 336 places and 2 base deletions is arranged at 337 ~ 338 places; Do not hold the generation frameshit at proteinic C; 1 ~ 111 place is identical with soybean growth period gene E1 in aminoacid sequence, and 112 ~ 147 places are for the dislocation aminoacid sequence, because of frameshit (frameshift) has formed terminator (seeing Seq ID No:2); And produced terminator codon at 148; Thereby soybean growth period e1-b3a gene of the present invention changes into Isoleucine in 65 in amino acid from Threonine, changed soybean growth period E1 gene (application for a patent for invention number: the integrity of B3 structural domain 201210112662.9), thereby make soybean blossoming in advance; The change that is similar to the structural domain of B3 in the E1 gene is described, has been caused the change of soybean E1 gene function.Kind cigarette with this gene shows as the early blossoming characteristic yellow No. 3.
Description of drawings
Fig. 1 is the e1-b3a electrophorogram, wherein, be for No. 1 middle yellow 13, No. 2 be Jack for closing rich 55, No. 3, No. 4 is yellow No. 3 of cigarette, No. 5 for cultivate rich 18, No. 6 for closing rich 50;
The soybean varieties cigarette of Fig. 2 tool e1-b3a gene yellow No. 3 under 16 hours long day illumination/8 hour dark condition, the back 45 days photos of emerging, wherein, arrow refers to the soybean flower of having opened;
The soybean varieties Harosoy-E1 of Fig. 3 tool soybean growth period E1 gene under 16 hours long day illumination/8 hour dark condition, the back 45 days photo of emerging.
Embodiment
Embodiment one: the mutator gene sequence of the soybean growth period e1-b3a gene of this embodiment is shown in sequence table Seq IDNo:1.
This embodiment is to clone (the application for a patent for invention number: on the basis 201210112662.9), further the E1 gene order is goed deep into systematic research of soybean growth period E1 gene.Owing in soybean, have typical B 3 gene structure domain genes, Ben Jiyin represent one type of novel gene family, and we have confirmed that the E1 gene can represent the function in E1 site, promptly have the function of inhibition soybean blossoming.This embodiment is studied E1 gene order in different varieties and the near isogenic line, and has confirmed the e1-b3a gene, and it gives birth to bloom to soybean has promoter action.
Soybean growth period E1 gene exist in aminoacid sequence 55 ~ 174 zones one with the similar zone of B3 structural domain edge far away; The nucleotides sequence of soybean growth period e1-b3a gene is listed in that 3 base mutations take place at 334 ~ 336 places and 2 base deletions is arranged at 337 ~ 338 places; Do not hold the generation frameshit at proteinic C; 1 ~ 111 place is identical with soybean growth period gene E1 in aminoacid sequence, and 112 ~ 147 places are for the dislocation aminoacid sequence, because of frameshit (frameshift) has formed terminator (seeing Seq ID No:2); And produced terminator codon at 148; Thereby 65 in the amino acid of the soybean growth period e1-b3a gene of this embodiment changes into Isoleucine from Threonine, changed soybean growth period E1 gene (application for a patent for invention number: the integrity of B3 structural domain 201210112662.9), thereby make soybean blossoming in advance; The change that is similar to the structural domain of B3 in the E1 gene is described, has been caused the change of soybean E1 gene function.Kind cigarette with this gene shows as the early blossoming characteristic yellow No. 3.
Embodiment two: the aminoacid sequence of the proteins encoded of the soybean growth period e1-b3a gene of this embodiment is shown in sequence table Seq ID No:2.
This embodiment is to clone (the application for a patent for invention number: on the basis 201210112662.9), further the E1 gene order is goed deep into systematic research of soybean growth period E1 gene.Owing in soybean, have typical B 3 gene structure domain genes, Ben Jiyin represent one type of novel gene family, and we have confirmed that the E1 gene can represent the function in E1 site, promptly have the function of inhibition soybean blossoming.This embodiment is studied E1 gene order in different varieties and the near isogenic line, and has confirmed the e1-b3a gene, and it gives birth to bloom to soybean has promoter action.
Soybean growth period E1 gene exist in aminoacid sequence 55 ~ 174 zones one with the similar zone of B3 structural domain edge far away; The nucleotides sequence of soybean growth period e1-b3a gene is listed in that 3 base mutations take place at 334 ~ 336 places and 2 base deletions is arranged at 337 ~ 338 places; Do not hold the generation frameshit at proteinic C; 1 ~ 111 place is identical with soybean growth period gene E1 in aminoacid sequence, and 112 ~ 147 places are for the dislocation aminoacid sequence, because of frameshit (frameshift) has formed terminator (seeing Seq ID No:2); And produced terminator codon at 148; Thereby 65 in the amino acid of the soybean growth period e1-b3a gene of this embodiment changes into Isoleucine from Threonine, changed soybean growth period E1 gene (application for a patent for invention number: the integrity of B3 structural domain 201210112662.9), thereby make soybean blossoming in advance; The change that is similar to the structural domain of B3 in the E1 gene is described, has been caused the change of soybean E1 gene function.Kind cigarette with this gene shows as the early blossoming characteristic yellow No. 3.
Through following verification experimental verification effect of the present invention:
(1) e1-b3a gene of the present invention obtains through following method
Get the blade that grows to yellow No. 3 soybean varieties of three leaf leaf period cigarettes, adopt the CTAB method to extract the blade genomic dna; DNA to extract is that template is passed through K1 and K2 primer, adopts and buys the gene from the high-fidelity enzyme primerSTAR of Takara company HS amplification e1-b3a, and 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 PCR product is checked order on ABI3130 sequenator (ABI company).
Sequencing result shows that soybean growth period e1-b3a 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 522 based compositions, and coding has the protein of the aminoacid sequence of Seq ID No:2 in the sequence table.
(2) e1-b3a gene identification in the soybean E1 gene:
One, gets the blade that grows to three leaf leaf periods kind to be measured, adopt the CTAB method to extract the blade genomic dna; Two, the genomic dna that extracts with step 1 is a template, and F1 and F2 are primer, adopts to buy and increases from the rTaq of Takara company enzyme, and the PCR reaction conditions is following: 95 ℃ of preparatory sex change 3min; 95 ℃ of sex change 20s, 58 ℃ of annealing 30s, 72 ℃ are extended 40s, totally 32 circulations, 72 ℃ are extended 7min again; Three, learn from else's experience 8 μ l (its concentration is about 300ng/ μ l) behind the step 2 PCR, dilute 1 times after, through BfuI restriction enzyme (purchase) from Takara company, under 37 ° of C temperature, digestion 3h; Four, the product after then the step 3 enzyme being cut adopts 13% non-sex change PAGE gel electrophoresis to detect; Wherein, yellow 13 in the described kind to be measured of step 1, close rich 55, Jack, yellow No. 3 of cigarette, cultivate rich 18 with close rich 50.
Electrophoresis detection result is as shown in Figure 1, can be known by Fig. 1, and No. 42 bands (shown in BC) appear in yellow No. 3 kinds of cigarette; Be the e1-b3a genotype; And 1,2,3,5 and No. 6 had only a band (shown in A), promptly do not have and can not be digested by BfuI, and then be other genotype of E1.
(3) functional verification of e1-b3a gene
One, soybean varieties has the phenotype analytical of e1-b3a gene
For further verifying the genotypic function of e1-b3a; To contain the cigarette of e1-b3a gene yellow No. 3 with contain (the application for a patent for invention number: Harosoy-E1 kind 201210112662.9) of soybean growth period E1 gene; Be planted in that (the MC duration of day: the long day is 16 hours illumination/8 hour dark in the artificial culture case; In isometric photograph be 13.5 hours/9.5 hours; The short day condition is 12 hours illumination/12 hour dark), light source is luminescent lamp and incandescent light, its light intensity is 235 ~ 270 μ mols
-1M
-2, simultaneously,, observe and to contain yellow No. 3 kinds of e1-b3a gene cigarette and to contain (the application for a patent for invention number: the fate of Harosoy-E1 kind from emerging to blooming 201210112662.9) of soybean growth period E1 gene at the field planting of natural lighting.
The result is as shown in table 1; From table 1, can find out; Soybean is a short day crop; Under short day (12 hours illumination/12 hour dark) condition, the Harosoy-E1 kind that contains soybean growth period E1 gene is bloomed fate (blooming from being seeded into) quite, about about 30 days with yellow No. 3 kinds of the cigarette that contains the e1-b3a gene; And under long day condition (16 hours illumination/8 hour dark); The fate of blooming of Harosoy-E1 kind that contains soybean growth period E1 gene was above 70 days; And yellow No. 3 kinds of the cigarette that contains the e1-b3a gene are about 43 days; Under medium sunshine condition, yellow No. 3 kinds of cigarette that contain the e1-b3a gene were bloomed (Fig. 2) in the time of 45 days, and the Harosoy-E1 kind that contains soybean growth period E1 gene then is in the situation of nourishing and growing (Fig. 3).
The e1-b3a gene is in yellow No. 3 of soybean varieties cigarette that China oneself cultivates, to find, explain that the e1-b3a gene representing extremely important one type of variation, plays a decisive role for the mechanism that fullys understand the E1 gene.
Table a kind has the e1-b3a gene and has the genotypic comparison of E1
Annotate: n.d. (not done) does not test accordingly; L=Light (illumination length); D=Dark (dark length)
Two, edge far away is similar to the influence of the sudden change of B3 structure to flowering period in the E1 gene
In kind good fortune rich (Fukuyutaka) (available from Japanese agriculture Biological resources institute); Handle through ethane methylsulphonic acid (EMS); Obtained series mutation body storehouse; Screening and the (application for a patent for invention number: related mutants 201210112662.9) of soybean growth period E1 gene in its mutant library; We have obtained at similar B3 structural domain (65 places of amino acid; Two mutants from the Threonine of E1 gene to Isoleucine is named and is e1-m3, and its foundation about mutant library, mutant choice process are with reference to (2000) Purification such as (2000) Targeting induced local lesions IN genomes (TILLING) for plant functional genomics.Plant Physiol 123:439 – 442 such as McCallum and Yang; Cloning, and characterization ofthe CEL I nuclease.Biochemistry 39:3533 – 3541] carry out.
The result is as shown in table 2; Can know by table 2; During its heterozygote offspring separates; The mutant of isozygotying (e1-m3/e1-m3) be its flowering period 37.33 than band isozygoty the genotypic strain of E1/E1 be flowering period 46.75 and wild-type have the E1 gene flowering period MV 48.57 days in advance, reach level of signification difference.The result is illustrated in except that soybean growth period E1 gene under the identical situation of all genotype, and the structural domain that edge far away is similar to the B3 gene in the E1 gene is undergone mutation, and causes blooming in advance of plant.Equally, contain in yellow No. 3 kinds of cigarette of e1-as gene, the structural domain that edge far away is similar to the B3 gene also takes place in sudden change, impacts the function of some effects E1 gene at least, causes to bloom in advance.
Separation in the two mutants heterozygous self progeny's of B3 structural domain flowering period in the E1 gene in the table 2 soybean varieties good fortune rich (Fukuyutaka)
Annotate: the E1 genotype of each strain plant is that the sequencing sequencing result according to its E1 gene confirms
Significance test is to carry out with the ANOVA among the SigmaPlot (V12.1).
Harosoy-E1 in the above-mentioned test 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, Harosoy-e1, Williams 82, Bay, Enrei, slope are originally early given birth to (Sakamotowase) and Jack etc. and from Japanese Biological resources institute gene pool, are bought and obtain.Yellow 13 in other kinds, close richly 55, cultivating rich 18, bluish yellow beans (Aokimame), Beijing Semen glycines sojae (Peking) is that " present in soybean national project research centre with closing rich 50 by soybean research center, Jilin Academy of Agricultural Science.Yellow buy from crop investigations institute of Chinese Academy of Agricultural Sciences germ plasm resource research department for No. 3 of cigarette obtains.
Claims (2)
1. soybean growth period e1-b3a gene, the gene order that it is characterized in that soybean growth period e1-b3a gene is shown in sequence table SeqID No:1.
2. the proteins encoded of soybean growth period e1-b3a gene as claimed in claim 1, the aminoacid sequence that it is characterized in that soybean growth period e1-b3a gene coded protein is shown in sequence table Seq ID No:2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210162176 CN102690831B (en) | 2012-05-23 | 2012-05-23 | El-b3a gene in soybean growth period and coding protein thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210162176 CN102690831B (en) | 2012-05-23 | 2012-05-23 | El-b3a gene in soybean growth period and coding protein thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102690831A true CN102690831A (en) | 2012-09-26 |
| CN102690831B CN102690831B (en) | 2013-05-08 |
Family
ID=46856568
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201210162176 Expired - Fee Related CN102690831B (en) | 2012-05-23 | 2012-05-23 | El-b3a gene in soybean growth period and coding protein thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102690831B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN114231558A (en) * | 2021-12-14 | 2022-03-25 | 东北农业大学 | Application of GmREM16 gene in plant stress resistance |
| WO2023168691A1 (en) * | 2022-03-11 | 2023-09-14 | Syngenta Crop Protection Ag | Methods and compositions for modifying flowering time genes in plants |
-
2012
- 2012-05-23 CN CN 201210162176 patent/CN102690831B/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| XIA Z ET AL: "Genbank:AB552966.1", 《GENBANK》 * |
| XIA Z ET AL: "Genbank：AB552968.1", 《GENBANK》 * |
| XIA Z ET AL: "Positional cloning and characterization the molecular basis for soybean maturity locus E1 that regulates photoperiodic flowering", 《PNAS》 * |
| 宁慧霞等: "大豆生育期研究进展", 《2005年全国作物遗传育种学研讨会暨中国作物学分子育种分会成立大会论文集》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN114231558A (en) * | 2021-12-14 | 2022-03-25 | 东北农业大学 | Application of GmREM16 gene in plant stress resistance |
| WO2023168691A1 (en) * | 2022-03-11 | 2023-09-14 | Syngenta Crop Protection Ag | Methods and compositions for modifying flowering time genes in plants |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102690831B (en) | 2013-05-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106011167B (en) | The method of the application and rice fertility restorer of male sterility gene OsDPW2 | |
| EP3805391B1 (en) | Use of yr4ds gene of aegilops tauschii in stripe rust resistance breeding of triticeae plants | |
| CN110213961A (en) | Crop based on genome editor is engineered and produces plant of short stem | |
| CN102586277B (en) | Set of DTH2 genes for controlling heading stage of paddy rice and haplotypes and application thereof | |
| JP2014517687A (en) | Plants with useful characteristics and related methods | |
| Shan et al. | Breeding of an elite cultivar Haibao No. 1 of Undaria pinnatifida (Phaeophyceae) through gametophyte clone crossing and consecutive selection | |
| CN107012243A (en) | A kind of molecular labeling for identifying the true hybrid of lichee and its application | |
| CN103583470A (en) | Molecular-assisted-selection cultivation method for green-shin recessive-white-feather chicken line | |
| CN105950651A (en) | Application of male-sterility gene OsGEN and method for restoring fertility | |
| CN104823919A (en) | Cultivation method and application of recessive white feather new strain of Xiaoxiang chicken in southeast of Guizhou province | |
| CN111492057A (en) | CMV resistance allele | |
| CN102690831B (en) | El-b3a gene in soybean growth period and coding protein thereof | |
| WO2014115680A1 (en) | Method for breeding brassica rapa plant having self-compatibility | |
| CN105441456B (en) | A kind of cabbage type rape genic male sterile gene Bnms4bAnd preparation method and application | |
| CN107418956A (en) | Rice photaesthesia genic male sterile gene pms1 separation clone and application | |
| CN109971763A (en) | Florescence control gene C MP1 and relevant carrier and its application | |
| CN102676543A (en) | Soybean growth period e1-as gene and encoded protein thereof | |
| CN109089875B (en) | Method for creating broad-spectrum and lasting rice blast-resistant excellent germplasm of japonica rice in lower Yangtze river | |
| CN113943732B (en) | SNP (Single nucleotide polymorphism) marker, primer set, kit and application related to heat resistance of cucumber in adult stage | |
| CN115369120B (en) | Temperature regulation gene for fertility transfer starting point of temperature-sensitive dual-purpose sterile line of rice and application thereof | |
| CN102732529A (en) | Soybean growth period e1-fs gene and protein encoded by the same | |
| CN106797883A (en) | A kind of breeding method of paddy rice | |
| Kawamura et al. | The identification of the Rosa S-locus provides new insights into the breeding and wild origins of continuous-flowering roses | |
| CN106317214B (en) | The heat-resisting GAP-associated protein GAP TaXPD of wheat and its encoding gene and application | |
| CN107022628A (en) | A kind of molecular labeling for identifying the early evening true hybrid of flower lichee filial generation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130508 Termination date: 20140523 |