CN102994516A - Soybean flowering gene ft2a-1 and protein coded by the gene - Google Patents
Soybean flowering gene ft2a-1 and protein coded by the gene Download PDFInfo
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- CN102994516A CN102994516A CN2012105349734A CN201210534973A CN102994516A CN 102994516 A CN102994516 A CN 102994516A CN 2012105349734 A CN2012105349734 A CN 2012105349734A CN 201210534973 A CN201210534973 A CN 201210534973A CN 102994516 A CN102994516 A CN 102994516A
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Abstract
A Soybean flowering gene ft2a-1 and a protein coded by the gene. The invention relates to the soybean flowering gene ft2a-1 and protein coded by the gene. The invention aims to provide the soybean flowering gene ft2a-1 and the protein coded by the gene. The soybean flowering gene ft2a-1 provided by the invention has a gene sequence shown as a Seq ID No:1 in the sequence table. An amino acid sequence of the protein coded by the soybean flowering gene ft2a-1 is shown as a Seq ID No:2 in the sequence table. According to Arabidopsis genetic transformation verification, it illustrates that the ft2a-1 gene has obviously decreased function of promoting flowering compared with an FT2a gene. The invention can be applied to soybean gene engineering.
Description
Technical field
The present invention relates to soybean blossoming gene ft2a-1 and proteins encoded thereof.
Background technology
Soybean provides important plant protein and oil content for the mankind.Worldwide, north is to Northern Europe Sweden and the North America Canada of high latitude, reach in the south in the extensive region such as Brazil and Argentina soybean culture is all arranged, but the latitude span of single kind or the general suitable planting of germ plasm resource is less, this Regional suitability and soybean photoperiod and breeding time gene or quantitative trait locus (Quantitative Trait Locus, QTL) closely related.The wide good soybean varieties of seed selection ecological suitability is the fundamental way that realizes high yield, high-quality, efficient Soybean Industry.But conventional breeding is made slow progress in the Wide-adaptive breed breeding, utilizes the molecular breeding means to be expected to orientation adjustment soybean photoperiod and breeding time, accelerates the seed selection of Wide-adaptive kind.The result of study of model plant Arabidopis thaliana and paddy rice shows, the element of blooming (Florigen, FT albumen) is transported to stem apex from blade and induces colored formation (Corbesier etc., 2007; Tamaki etc., 2007).We have cloned 10 FT genes from soybean, being replicated on the soybean linkage group of this 10 pair of genes.The result of gene expression analysis shows, be under the short day condition in the induction of flowering condition, there are two FT genes (GmFT2a and GmFT5a) to form the phase (about 25～30 days) at bud, great expression in ternately compound leaf, and other gene expression amounts are extremely low or do not express.And these two genes present the genetic expression type of physiological clock rule, begin 4 hours expression intensities in illumination and weaken gradually after the highest.Under non-long day condition of inducing, to only have a FT gene (GmFT5a) relative expression, thereby caused with this understanding soybean bloom evening, prolong breeding time.Utilize double-mutant (Harosoy-e3e4) research of soybean phytochrome A gene then to find, under the long day condition, the flowering period of Harosoy-e3e4 is consistent the flowering period under the short day condition with its wild-type plant.And, in double-mutant, forming the phase at bud, the expression of GmFT2a and GmFT5a is induced in a large number and is presented genetic expression with the identical physiological clock rule of short day.These results of study have tentatively disclosed the molecule mechanism of soybean bloom and breeding time from the molecular level, namely (1) is under the short day condition of inducing, having two FT genes to participate in expressing causes the florescence very short, yet under non-long day condition of inducing, cause flowering period elongated thereby only have a FT gene to participate in expressing; (2) soybean adapts in the existence of high latitude zone phytochrome A gene inactivation through its own evolution, and this evolutionary process is finally to realize by regulating and control FT genetic expression main in two soybean; (3) under the long day condition, the expression of the gene induced FT gene of phytochrome A of long day plant Arabidopis thaliana, and the phytochrome A gene of short day plant soybean suppresses the expression of FT gene, illustrates that short day plant contains the photoperiodic reaction regulatory mechanism different from long day plant (Kong etc. 2010).In addition, by the Genetic Transformation of Soybean technology, the investigator has verified that further GmFT2a has soybean blossoming promotion functions (Sun etc. 2011).But how the sudden change of GmFT2a gene affects soybean growth period (flowering period and ripening stage) is a very important problem in science, does not also have at present the correlative study report on molecular level.
Summary of the invention
The purpose of this invention is to provide soybean blossoming gene ft2a-1 and proteins encoded thereof.
The gene order of soybean blossoming gene ft2a-1 of the present invention is shown in sequence table Seq ID No:1.
The aminoacid sequence of the proteins encoded of soybean blossoming gene ft2a-1 of the present invention is shown in sequence table Seq ID No:2.
Beneficial effect of the present invention:
The present invention is on the basis that clones soybean blossoming gene GmFT2a (Kong etc. 2010), further the GmFT2a gene order is goed deep into systematic research.We have confirmed that the GmFT2a gene has the function that promotes soybean blossoming.The present invention is studied GmFT2a gene order in the different varieties, and has determined the ft2a-1 gene, and it is in close relations to soybean growth period.
The ft2a-1 gene is compared with soybean blossoming gene GmFT2a (Kong etc. 2010), both sequences only have a Nucleotide (No. 506) difference dna level, but cause 169 missense mutation from the glycine to the aspartic acid of aminoacid sequence, thereby changed the function of original GmFT2a gene.The genetic transformation of Arabidopis thaliana is tested and is shown, ft2a-1 gene of the present invention is compared with soybean blossoming gene GmFT2a, and the function of its Accelerate bloom obviously weakens.
Description of drawings
Fig. 1 is pMDC100IG-ft2a-1 plasmid expression vector structural representation;
Fig. 2 is the flowering time figure that changes the ft2a-1 gene over to and change GmFT2a gene Arabidopis thaliana and wild-type Arabidopis thaliana over to; Wherein, Col-0 is the wild-type Arabidopis thaliana, and GmFT2a-OX is the Arabidopis thaliana that changes over to behind the GmFT2a gene, and ft2a-1-OX is the Arabidopis thaliana that changes over to behind the ft2a-1 gene.
Embodiment
Embodiment one: the gene order of the soybean blossoming gene ft2a-1 of present embodiment is shown in sequence table Seq ID No:1.
Present embodiment is studied GmFT2a gene order in the different varieties, and has determined the ft2a-1 gene, and it is in close relations to soybean growth period.
Present embodiment is expressed GmFT2a gene and ft2a-1 gene by Arabidopis thaliana genetic transformation means in Arabidopis thaliana, verified that ft2a-1 gene of the present invention compares with soybean blossoming gene GmFT2a, and the function of its Accelerate bloom obviously weakens.
Embodiment two: the aminoacid sequence of the proteins encoded of the soybean blossoming gene ft2a-1 of present embodiment is shown in sequence table Seq ID No:2.
By following verification experimental verification effect of the present invention:
(1) acquisition of gene ft2a-1 sequence
One, take the blade of soybean varieties K3 as 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; Three, the total RNA that gets after 1 μ g step 2 is processed 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 of BD BiosciencesClontech company RACE cDNA Amplification Kit test kit, obtains cDNA; Four, take the cDNA that obtains as template by F1 and R1 primer amplification ft2a-1 gene, the PCR reaction conditions is as follows: 94 ℃ of denaturation 5min, 94 ℃ of sex change 30s, 56 ℃ of annealing 30s, 72 ℃ are extended 1min, totally 35 circulations, 72 ℃ are extended 10min again, and the PCR product is checked order at ABI3130 sequenator (ABI company);
Wherein, the sequence of primers F 1 is 5 ' ccatgcctagtggaagtagg; The sequence of primer R1 is gagtgtgggagattgccaat-3 '.
Sequencing result shows that soybean blossoming gene ft2a-1 has the nucleotide sequence of Seq ID No:1 in the sequence table, and the Seq ID No:1 in the sequence table is comprised of 622 Nucleotide, and coding has the protein of the aminoacid sequence of Seq ID No:2 in the sequence table.
(2) functional verification of soybean blossoming gene ft2a-1
One, take the blade of soybean varieties K3 as 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; Three, the total RNA that gets after 1 μ g step 2 is processed 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 of BD BiosciencesClontech company RACE cDNA Amplification Kit test kit, obtains cDNA; Four, respectively take ft2a-1 gene F2 and R2 as primer, the cDNA that step 3 is obtained carries out 5 ' and 3 ' amplification by PCR, PCR reaction conditions: 94 ℃ of denaturation 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 blossoming gene ft2a-1 of the two enzyme restriction enzyme sites of XbaI-SacI; The soybean blossoming gene ft2a-1 that contains restriction enzyme site that five, will obtain is cloned in the pMDC100IG plasmid, obtain the pMDC100IG-ft2a-1 plasmid, take Arabidopis thaliana (Col-0) as material, utilize agrobacterium-mediated transformation to carry out genetic transformation, acquisition turns ft2a-1 gene Arabidopis thaliana, to turn ft2a-1 gene Arabidopis thaliana, turn GmFT2a gene Arabidopis thaliana and wild-type Arabidopis thaliana (Col-0) illumination in 16 hours and planted under the dark condition in 8 hours, observe their bloom; Wherein, pMDC100IG plasmid and turn GmFT2a gene Arabidopis thaliana discloses in the article that is called " Two Coordinately Regulated Homologs of FLOWERING LOCUS T AreInvolved in the Control of Photoperiodic Flowering in Soybean " that we publishs in 2010 " Plant Physiology "; Soybean varieties K3 belongs to a Cultivar of Thailand, publication is so kind as to give by professor Abe of Hokkaido University in 2006 " Kasetsart J " " Molecular marker analysis of days to flowering invegetable soybean (Glycme max) ".
Wherein, the sequence of primers F 2 is 5 ' gctctagaccatgcctagtggaagtagg; The sequence of primer R2 is gcgagctcgagtgtgggagattgccaat-3 '.
The pMDC100IG-ft2a-1 plasmid over-express vector structural representation that step 5 obtains as shown in Figure 1;
The flowering time figure that changes the ft2a-1 gene over to and change GmFT2a gene and wild-type Arabidopis thaliana over to as shown in Figure 2, wherein, Col-0 is the wild-type Arabidopis thaliana, and GmFT2a-OX is the Arabidopis thaliana that changes over to behind the GmFT2a gene, and ft2a-1-OX is the Arabidopis thaliana that changes over to behind the ft2a-1 gene.As can be seen from Figure 2, the flowering time of Col-0 is 31 days, the flowering time of GmFT2a-OX is 24.8 days, show the Arabidopis thaliana flowering time that changes over to behind the GmFT2a gene than the flowering time of wild-type Arabidopis thaliana early than 6 days, thereby explanation GmFT2a gene has the function of Accelerate bloom; And the flowering time of the flowering time of ft2a-1-OX and wild-type Arabidopis thaliana is basic identical, thereby illustrates that ft2a-1 gene of the present invention compares with soybean blossoming gene GmFT2a, and the function of its Accelerate bloom obviously weakens or afunction.
Claims (2)
1. soybean blossoming gene ft2a-1 is characterized in that the gene order of soybean blossoming gene ft2a-1 is shown in sequence table Seq ID No:1.
2. the proteins encoded of soybean blossoming gene ft2a-1 as claimed in claim 1 is characterized in that the aminoacid sequence of proteins encoded of soybean blossoming gene ft2a-1 is shown in sequence table Seq ID No:2.
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| CN103288942A (en) * | 2013-05-02 | 2013-09-11 | 东北林业大学 | African agapanthus flowering gene ApFT protein and coding gene and probe thereof |
| CN104357442A (en) * | 2014-10-25 | 2015-02-18 | 中国科学院东北地理与农业生态研究所 | QTL mapping region for soybean flowering stage and obtaining method as well as application |
| CN106489492A (en) * | 2016-10-22 | 2017-03-15 | 杨继周 | A kind of method for promoting soybean blossoming |
| CN110438133A (en) * | 2019-08-16 | 2019-11-12 | 安徽省农业科学院作物研究所 | A kind of Mung Bean Blooming gene VrFT2a and its application |
| CN112961842A (en) * | 2021-03-23 | 2021-06-15 | 中国科学院东北地理与农业生态研究所 | Soybean phytochrome chromophore synthesis gene GmHY2 and encoding protein and application thereof |
| US11225671B2 (en) | 2015-04-20 | 2022-01-18 | Monsanto Technology Llc | Compositions and methods for altering flowering and plant architecture to improve yield potential |
| US11555201B2 (en) | 2016-10-19 | 2023-01-17 | Monsanto Technology Llc | Compositions and methods for altering flowering and plant architecture to improve yield potential |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103288942A (en) * | 2013-05-02 | 2013-09-11 | 东北林业大学 | African agapanthus flowering gene ApFT protein and coding gene and probe thereof |
| CN104357442A (en) * | 2014-10-25 | 2015-02-18 | 中国科学院东北地理与农业生态研究所 | QTL mapping region for soybean flowering stage and obtaining method as well as application |
| US11225671B2 (en) | 2015-04-20 | 2022-01-18 | Monsanto Technology Llc | Compositions and methods for altering flowering and plant architecture to improve yield potential |
| US11555201B2 (en) | 2016-10-19 | 2023-01-17 | Monsanto Technology Llc | Compositions and methods for altering flowering and plant architecture to improve yield potential |
| CN106489492A (en) * | 2016-10-22 | 2017-03-15 | 杨继周 | A kind of method for promoting soybean blossoming |
| CN110438133A (en) * | 2019-08-16 | 2019-11-12 | 安徽省农业科学院作物研究所 | A kind of Mung Bean Blooming gene VrFT2a and its application |
| CN110438133B (en) * | 2019-08-16 | 2021-06-04 | 安徽省农业科学院作物研究所 | Application of expression vector containing mung bean flowering gene VrFT2a |
| CN112961842A (en) * | 2021-03-23 | 2021-06-15 | 中国科学院东北地理与农业生态研究所 | Soybean phytochrome chromophore synthesis gene GmHY2 and encoding protein and application thereof |
| CN112961842B (en) * | 2021-03-23 | 2022-11-29 | 中国科学院东北地理与农业生态研究所 | Soybean phytochrome chromophore synthesis gene GmHY2 and encoding protein and application thereof |
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