CN102154285A - Molecular marker SIsv0737 closely linked with millet leaf color gene - Google Patents
Molecular marker SIsv0737 closely linked with millet leaf color gene Download PDFInfo
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Abstract
The invention belongs to the field of molecular biology, and relates to a molecular marker, in particular to a molecular marker SIsv0737 closely linked with millet leaf color gene. The molecular marker comprises the sequence as shown in Seq ID No.1. The invention further relates to a primer which is used for amplifying the molecular marker, applications of the molecular marker and the primer in the millet leaf color gene mapping or the millet inheritance breeding, and a millet breeding method. The molecular marker is used for linking the genome DNA sequence with the millet leaf color gene to be good for building an auxiliary breeding system of the millet molecular marker; and the inheritance close linkage distance between the molecular marker and the millet leaf color gene is 2.1cM. The molecular marker and the primer used for amplifying the molecular marker can be conveniently and fast applied to the millet breeding practice and the resource and breed verification with high throughput.
Description
Technical field
The invention belongs to biology field, relate to a kind of molecule marker, particularly relate to a kind of and the closely linked molecule marker of millet leaf color gene.The invention still further relates to primer, this molecule marker and the primer purposes in the assignment of genes gene mapping of millet leaf color or millet genetic breeding of this molecule marker of amplification.
Background technology
China is the country of origin of millet (Setaria italica L.Beauv.), is the concentrated plantation state of millet in the world, and millet occupies an important position in the national economy of China and social production, and is significant to the dry farming ecological agriculture construction.Therefore, the breeding process of acceleration millet is particularly important.Because millet is regional importance crop, therefore current research means about millet is relative with method more backward, and how the research means of application of advanced science is done the problem that the millet breeding is a sternness well.Along with development of molecular biology, the appearance of molecular marking technique is for new thinking and means have been opened up in genetic research and the breeding of millet.Develop the molecule marker of important character gene and carry out assisted selection research, will play promoter action improving China's millet breeding level with China's characteristic.
The leaf look is the general performance of various pigments in the chloroplast(id), and chlorophyll is preponderated in the normal blade, is usually expressed as green.Leaf variegation is the higher and mutant character that is easy to identify of mutation frequency in the higher plant.Up to now, in nearly all higher plants such as paddy rice, wheat, barley, corn, soybean, cotton, tobacco, corn, Sunflower Receptacle, tomato, cucumber, potato, mulberry tree, all found leaf look mutant.Its mutator gene directly or indirectly influences photosynthetic pigments, and particularly chlorophyllous synthetic and degraded causes the content of various pigments and ratio to change, thereby causes the leaf chromatic variation.In recent years, the utility value of leaf look mutant more and more receives publicity, now become the exotic materials of research photosynthesis of plant mechanism, chlorophyll biosynthetic pathway, chloroplast development and Genetic Control mechanism, leaf look mutant also has important use value in the utilization of the breeding of high photosynthetic efficiency and mark property simultaneously.Some leaf variegations are used as mark property and are used for stock breeding and cross-fertilize seed production.Along with finishing of Arabidopis thaliana, the order-checking of paddy rice isotype Plant Genome, the researching value of leaf look mutant on functional genomics receives increasing concern, and they have become the ideal material of research photosynthetical system structure, gene function and regulatory mechanism thereof.According to incompletely statistics, the genes involved of the paddy rice leaf look of report sudden change at present surpasses 80, and these mutant are broadly divided into 5 big classes such as albinism, yellow type, stripe, green changing type, zebra leaf.But there is not bibliographical information substantially for millet Ye Sejiyin and closely linked with it Study on Molecular Marker.
Summary of the invention
The purpose of this invention is to provide a kind of and the closely linked molecule marker of millet leaf color gene.
It is right that another object of the present invention provides a kind of primer that can be used for pcr amplification and the closely linked molecule marker of millet leaf color gene, and the molecule marker that amplification is obtained by this primer.
A further object of the present invention provide above-mentioned molecule marker in the assignment of genes gene mapping of millet leaf color, detection and millet assistant breeding purposes and the detection method of above-mentioned molecule marker.
Purpose of the present invention also comprises provides a kind of recombinant vectors that comprises above-mentioned molecule marker, and the reconstitution cell that contains this recombinant vectors; A kind of millet auxiliary breeding means that comprises the localization method of the millet leaf color gene that uses above-mentioned molecule marker and use described molecule marker is provided.
To achieve these goals, the present invention has adopted following technical scheme:
The invention discloses a kind of and the closely linked molecule marker of millet leaf color gene, described molecule marker contains sequence shown in the Seq ID No.1; Preferred described molecule marker has sequence shown in the Seq ID No.1.
It is right with the primer of the closely linked molecule marker of millet leaf color gene to the invention also discloses a kind of amplification, and the right primer 1 of described primer contains sequence shown in the Seq ID No.2, and primer 2 contains sequence shown in the Seq ID No.3; Preferred described primer 1 has sequence shown in the Seq ID No.2, and primer 2 has sequence shown in the Seq ID No.3;
Seq?ID?No.2：5’-GGTGGTTGTTTACCATTTTCG-3’；
Seq?ID?No.3：5’-AACTACATTCCAATGTCCAGTT-3’。
The invention also discloses a kind of and the closely linked molecule marker of millet leaf color gene, described molecule marker be by above-mentioned primer to blade partially the millet genomic dna of green be that template obtains through pcr amplification.
The preferred molecule marker that amplification is obtained by above-mentioned primer contains sequence shown in the Seq ID No.1.
In one embodiment of the invention, described molecule marker (dna fragmentation that contains nucleotide sequence shown in the Seq ID No.1) is the dna fragmentation of nucleotide sequence shown in the Seq ID No.1 in the millet genome, 5 ' the end of the Seq ID No.1 that is promptly comprised and/or the nucleotide sequence beyond the 3 ' end also are the sequences in the millet genome, preferably, be the 5 ' end of Seq ID No.1 in the millet genome and/or the upstream and downstream sequence of 3 ' end.It will be understood by those skilled in the art that as long as amplification or detect blade this molecule marker in the millet genomic dna of green partially, must detect or increase and obtain containing the sequence shown in the Seq ID No.1.The length of the 5 ' end of Seq ID No.1 and/or the upstream and downstream sequence of 3 ' end is suitable length, be not particularly limited, for example, the length that satisfies molecule marker is less than 10,000bp, less than 5,000bp, less than 2,000bp, less than 1,500bp, less than 1,200bp, less than 1,000bp or less than 800bp.
In one embodiment of the invention, 5 ' the end and/or 3 ' of the Seq ID No.1 that described molecule marker (dna fragmentation that contains nucleotide sequence shown in the Seq ID No.1) is comprised is held be operably connected artificial sequence and/or control sequence, for example promotor, enhanser, terminator, restriction enzyme site, primer sequence or the like.Wherein, term " operationally " is defined as a kind of following conformation in the present invention, in this conformation, control sequence for example promotor is suitably placed on the position of Seq ID No.1, so that this control sequence instructs the generation of Seq ID No.1 encoded polypeptides.
The invention also discloses a kind of recombinant vectors, it contains molecule marker of the present invention.Described recombinant vectors can be expression vector or the cloning vector that is inserted with molecule marker of the present invention.After obtaining above-mentioned recombinant vectors, it will be understood by those skilled in the art that recombinant vectors to be transformed in the suitable cell, obtain containing the reconstitution cell of this recombinant vectors according to different needs.Therefore, the invention also discloses a kind of reconstitution cell that contains described recombinant vectors.
The invention also discloses the preparation method of molecule marker of the present invention, comprise the steps: to use leaf color partially the genomic dna of green millet as template, to carrying out pcr amplification, the 920bp amplified production that obtains promptly contains described molecule marker with above-mentioned primer; Preferably, also comprise the step of pcr amplification product being carried out purifying.
To those skilled in the art, be appreciated that also can the DNA chemosynthesis method obtain molecule marker of the present invention.
The invention also discloses the detection method of described molecule marker, comprise step: the nucleotide sequence design primer according to above-mentioned molecule marker is that template increases with detected millet genomic dna, and judges whether there is this molecule marker in the amplified production.Preferably, described primer is that the above-mentioned primer that contains Seq ID No.2 and Seq ID No.3 respectively is right.
For example, can with the genomic dna of detected millet template, (Seq ID No.2 and Seq ID No.3) carries out pcr amplification with above-mentioned primer, obtains amplified production.The amplified production that obtains can be checked order or gel electrophoresis.
The invention also discloses the purposes of described molecule marker in the assignment of genes gene mapping of millet leaf color or detection.
The invention also discloses the method for a kind of millet leaf color assignment of genes gene mapping, described method comprises the step of using molecule marker of the present invention.
The invention also discloses the purposes of described molecule marker in the millet assistant breeding.
The invention also discloses a kind of millet auxiliary breeding means, described method comprises detection molecule marker of the present invention or the right step of molecule marker primer.
Molecule marker of the present invention can be used in from now on the molecular mark, it will be appreciated by those skilled in the art that, such as by detect whether exist molecule marker of the present invention screen the millet blade whether contain the control leaf color for the color gene of green partially (for example, can reference, the purposes of dna molecular marker in wheat breeding for disease resistance, east, Gansu Province institute's journal (natural science edition), the 16th the 1st phase of volume of April in 2006, P65-69).Described detection can be the method that PCR detects, and particularly, can use the primer of above-mentioned molecule marker of the present invention right.Described detection can also be undertaken by sequence measurement.This millet auxiliary breeding means has easy, quick, high-throughout advantage.
In the present invention, particularly, described millet can be for opening paddy No. 1, millet A2 sterile line, No. 3, a confused flour beetle or opening the F2 generation that No. 3 selfings of confused flour beetle produce.Wherein, open paddy No. 1, confused flour beetle No. 3 or open the part F2 that No. 3 selfings of confused flour beetle produce green partially for leaf color; The part F2 that millet A2 sterile line, No. 3 selfing of confused flour beetle produce is yellow partially for leaf color.
Because adopt above technical scheme, the beneficial effect that the present invention is possessed is:
The invention provides and the closely linked molecule marker of millet leaf color gene, this molecule marker connects genomic dna sequence and millet leaf color gene, helps the foundation of millet molecular mark system; The hereditary close linkage distance of described molecule marker and millet leaf color gene is 2.1cM.Molecule marker of the present invention and molecule marker amplimer can be applied to millet breeding practice and resource and cultivar identification easy, quick, high-throughput.
Description of drawings
Fig. 1: molecule marker primer (Seq ID No.2 and Seq ID No.3) is to the partial results of F2 generation 480 individual plant amplifications.Wherein:
Swimming lane 1-12 is the 12 strain millet leaf colors pcr amplification product of the individual plant of green partially in F2 generation; Swimming lane 13-24 is the pcr amplification product of F2 inclined to one side xanchromatic individual plant of 12 strain leaf colors in generation.Swimming lane M is marker, and it is 100bp DNALadder; Its molecular weight comprises:, 1500bp, 1000bp, 900bp, 800bp, 700bp, 600bp, 500bp, 400bp, 300bp, 200bp and 100bp.
Embodiment
The invention discloses a kind of primer to and with the closely linked molecule marker SIsv0737 of millet leaf color gene.Utilizing primer of the present invention right, is that template is carried out PCR with the millet genomic dna, can obtain and the closely linked molecule marker of millet leaf color gene, and this molecule marker is called after molecule marker SIsv0737 in the present invention.It is pointed out that to it will be understood by those skilled in the art that, can also obtain molecule marker of the present invention by chemosynthesis except obtaining the molecule marker of the present invention by above-mentioned pcr amplification.
Primer of the present invention is to containing sequence shown in ordered list Seq ID No.2 and the Seq ID No.3 respectively,
Seq?ID?No.2：5’-GGTGGTTGTTTACCATTTTCG-3’；
Seq?ID?No.3：5’-AACTACATTCCAATGTCCAGTT-3’。
Those skilled in the art know, in sequence shown in above-mentioned Seq ID No.2 and the Seq ID No.3, can increase by 1～10 base respectively at its 5 ' end or 3 ' end, the base type that is increased can according on the millet genomic dna with Seq ID No.2 and Seq ID No.3 be complementary the zone the base type and determine the primer that obtains thus pair and the amplified production of Seq ID No.2 and Seq ID No.3 basic identical (dna sequence dna between the upstream and downstream primer is identical) according to basepairing rule.Therefore, above-mentioned at Seq ID No.2 and Seq ID No.3 5 ' end or 3 ' end increases by 1～10 base respectively and the primer that obtains basic identical dna fragmentation of increasing is right, include primer centering of the present invention.In the concrete embodiment of the present invention, primer of the present invention is to being preferably sequence shown in Seq ID No.2 and the Seq ID No.3.The present invention obtains F1 generation (opening confused flour beetle No. 3) with inclined to one side green of male parent blade and the inclined to one side xanchromatic homozygote hybridization of maternal blade, produces F2 for colony with F1 for selfing again, totally 480 individual plants.The preferred SV molecular markers development method that adopts is at first carried out de novo order-checking (60X contig N50:22K, scaffold N50:320K to male parent; Total size:400Mb), the female parent preface (10X) of resurveying; Then according to this sequencing data of father and mother, the SOAP software that utilizes the big independent development of China is the sequence difference of father and mother between this relatively, hold about 50bp position, the outside at the 5 ' end and 3 ' of male parent diversity sequence respectively, the primer of the Design of length diversity sequence amplification about picked at random 20bp has designed 1105 pairs of primers according to different diversity sequences.DNA with father and mother's basis and F1 is that template increases, and filters out 616 pairs of primers with polymorphism and validity from 1105 pairs of primers.616 pairs of primers that employing is developed, 480 individualities to F2 colony carry out the PCR detection, and the line data statistical study of going forward side by side is carried out genetic map with Map Maker3.0 software and drawn, obtain of the present invention and the closely linked molecule marker of leaf color gene, and amplimer.The male parent sequence that the primer that employing filters out obtains amplification, i.e. molecule marker SIsv0737 among the present invention.The F2 individuality is carried out character analysis, and according to gene character data and phenotypic character data with the assignment of genes gene mapping of millet leaf color on genetic map.
Also in conjunction with the accompanying drawings the present invention is described in further detail below by specific embodiment.Following examples only are further detailed the present invention, should not be construed as limitation of the present invention.
Embodiment 1: millet F2 is for the structure of segregating population
Male parent: anti-Sethoxydin, the plant type height, boot leaf is long and narrow, the bristle redness, clever shell redness can be educated, and the leaf colour cast is green, the pollen yellow-white, be late period heading stage.Male parent is for opening No. 1 seed of paddy.
Maternal: not anti-Sethoxydin, plant type is short, and boot leaf is short and wide, the bristle green, clever shell green, partial sterility, leaf colour cast Huang, pollen is brown, and be early stage heading stage.Female parent is a millet A2 male-sterile seed.
F2 colony makes up: male parent and hybridization of female parent obtain F1 generation (F1 leaf look is for green partially), and the F1 selfing obtains F2.Wherein F1 is No. 3 seeds of a confused flour beetle.Altogether F2 for individual plant 480 strains, No. 1 seed of above-mentioned paddy, millet A2 male-sterile seed and No. 3 seed of confused flour beetle can be referring to Chinese patent application " with the closely linked molecule marker SIsv0372 of millet anti-herbicide gene ", publication number CN101974521A, date of publication on February 16th, 2011.
Embodiment 2: in father and mother this and F1 generation, F2, are for the extraction of genes of individuals group DNA
With the CTAB method extract respectively among the embodiment 1 father and mother this, F1 generation and 480 F2 be for the genomic dna of individuality, concrete grammar is as follows:
(1) takes by weighing the fresh blade of 1.0g, shred and put into mortar,, grind to form homogenate and change in the centrifuge tube of 15mL, add 1mL 1.5 * CTAB then in the mortar and wash and change in the centrifuge tube again with adding 3mL 1.5 * CTAB after the liquid nitrogen grinding.Behind the mixing in 65 ℃ of water-bath 30min, during slowly shake up frequently.
1.5 * CTAB prescription following (1L) wherein:
Add deionized water and be settled to 1L, adding final concentration before using is the mercaptoethanol of 0.2% (2ml).
(2) to be cooled to room temperature, add equal-volume chloroform/primary isoamyl alcohol (24: 1), mixing gently becomes deep green to subnatant.
(3) the centrifugal 10min of 4200rpm moves on to new 15mL centrifuge tube mutually with upper water, adds the dehydrated alcohol of 2 times of volume precoolings, mixes static 5min.Place 30min deposit D NA in-20 ℃.
(4) the centrifugal 10min of 4200rpm discards supernatant, adds 1mL 75% washing with alcohol precipitation 1 time, is inverted the centrifuge tube dry DNA, adds 200 μ L TE dissolving DNAs.
(5) detect genomic dna with 0.8% sepharose.
(6) with the father and mother that obtain this and F1 generation, F2 for the genomic dna of individuality be stored in-20 ℃ standby.
Embodiment 3: the preparation of molecule marker
Genomic dna with the male parent extracted among the embodiment 2, F1 generation or F2 generation is a template, with the molecule marker amplimer (Seq ID No.2 and Seq ID No.3) is carried out pcr amplification.
The PCR reaction system is as follows:
The PCR response procedures is as follows:
94 ℃ of pre-sex change 5 minutes; 94 ℃ of sex change 30 seconds, 60 ℃ of annealing 30 seconds, 72 ℃ were extended 40 seconds, and moved 35 circulations; Last 72 ℃ were extended 3 minutes.Pcr amplification product can be 4 ℃ of preservations.
Obtain molecule marker through above-mentioned amplification procedure, after the preferred amplification amplified production is carried out purification process.Check order behind the purifying, the result is shown in Seq ID No.1.
To those skilled in the art, be appreciated that also and can obtain this molecule marker by the method for DNA chemosynthesis.
Embodiment 4:SV molecular markers development
Male parent: de novo order-checking, 60X contig N50:22K, scaffold N50:320K; Total size:400Mb; Maternal: the preface of resurveying 10X.
According to this sequencing data of father and mother, utilize the SOAP software (SOAP2.20 for example of the big independent development of China, can download from http://soap.genomics.org.cn/, also can use other sequence alignment software) sequence difference of father and mother between this relatively, based on the sequence of difference, use the primer of primer premier software design amplification diversity sequence then; Based on different diversity sequences, 1105 pairs of primers have been designed altogether.In the following table 1 part primer sequence wherein (Seq ID No.2-Seq ID No.41) has been shown
Part primer in the table 11105 pair random primer
Genomic dna with the father and mother that extract this and F1 generation is a template respectively, carries out pcr amplification with 1105 pairs of primers of design.
PCR reaction system (25 μ L):
PCR response procedures: 94 ℃ of pre-sex change 5min; Enter 35 circulations then: 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ are extended 40s; 72 ℃ are extended 3min after the loop ends; 4 ℃ of preservations.
PCR product electrophoresis detection: 1.2% sepharose 120v electrophoresis 25min, the EB 10min that dyes, according to glue and record.
The validity of primer and polymorphism: be meant in this validity whether amplified production is arranged, polymorphism is meant that the clip size of this amplified production of father and mother is variant.
Carry out the screening of primer according to following screening criteria: father and mother's basis and F1 all have amplified production, and this amplified production of father and mother all has only a distinct banding pattern and big or small variant, F1 shows as the heterozygosis banding pattern of this banding pattern of father and mother, and two bands of male parent and maternal banding pattern are promptly arranged.
The selection result:, from 1105 pairs of primers of design, filter out 616 pairs of primers according to above-mentioned screening criteria.
Embodiment 5: the genetic map construction and the assignment of genes gene mapping
(1) genetic map construction
The molecule marker that has a polymorphism with 616 couple of exploitation carries out PCR to 480 individualities of F2 colony and detects, the genomic dna of template used 480 individualities for the F2 colony that makes.
The PCR product is carried out agarose gel electrophoresis, obtain the result of molecule marker primer 480 individual amplifications.
Whole electrophoresis result are carried out data statistic analysis, concrete grammar is as follows: be a that is designated as of male parent type with F2 colony individual plant amplified band, amplified band is the b that is designated as of maternal type, amplified band contains the h that is designated as of male parent type and maternal type simultaneously, banding pattern blurs or disappearance is designated as-, be equivalent to the data disappearance, finally obtain the genotype data of 616 pairs of primer amplifications of 480 individualities of F2 colony.Such as, the data of 480 individualities that obtain with first pair of primer are a, b, and h ,-, b ... totally 480 data, the data that obtain with second pair of primer are b, a, h, a ,-... totally 480 data, totally 616 pairs of primers are added up respectively, and gained is the genotype data of this F2 colony.
With MapMaker 3.0 softwares (Constructing genetic maps with MAPMAKER/EXP 3.0, S Lincoln, M Daly, E Lander-Cambridge, MA:Whitehead Institute, 1992) carry out the genetic linkage maps drafting, obtain genetic linkage map.Can determine from this genetic linkage map that obtains 616 pairs of primers the position and with the genetic distance of millet leaf color gene.
(2) assignment of genes gene mapping
According to the leaf color phenotype of 480 individualities, similar to the male parent type proterties a (blade is green partially) that is designated as, similar to the maternal type proterties b (blade is yellow partially) that is designated as, proterties occupy the h that is designated as between male parent and the female parent.Obtain the phenotypic data of 480 individualities, the phenotypic data of 480 individualities genotype data with 480 individualities that obtain is before compared, similar Gao Ze represents this mark and leaf color proterties close linkage, and with the leaf color assignment of genes gene mapping on genetic linkage maps.
Embodiment 6: with the checking of the closely linked molecule marker of millet leaf color gene
1. on the basis of the genetic linkage maps that embodiment 5 makes, according to the genetic linkage distance of millet leaf color gene, with the hereditary close linkage distance of millet leaf color gene for having determined molecule marker primer (Seq ID No.2 and Seq ID No.3) in the position of 0.5cM, and find corresponding male parent sequence location, sequence between the upstream and downstream primer is molecule marker, and its nucleotide sequence is shown in Seq ID No.1.
Seq?ID?No.2：5’-GGTGGTTGTTTACCATTTTCG-3’；
Seq?ID?No.3：5’-AACTACATTCCAATGTCCAGTT-3’。
2. in addition, in the electrophoresis in embodiment 5 to the pcr amplification product of 480 individualities in F2 generation, amplification for molecule marker primer (Seq ID No.2 and Seq ID No.3) is: the amplified production of the plant that about 360 strain leaf colors are green partially all has the band of 920bp size, and the pcr amplification product of the inclined to one side xanchromatic plant of about 120 strain leaf colors does not all have the band (the part amplification as shown in Figure 1) of 920bp.And the fragments sequence through this 920bp of order-checking proof is identical with Seq ID No.1.
As seen, molecule marker of the present invention (Seq ID No.1) is and the closely linked molecule marker of millet leaf color gene.
Embodiment 7: molecular marker clone
The fragment cloning of the 920bp that obtains increasing among the embodiment 6 obtains recombinant vectors in the pMD18-T carrier.This recombinant vectors is transformed in the e. coli jm109, chooses mono-clonal, cultivate and obtain reconstitution cell.Extract plasmid from reconstitution cell, described plasmid is a recombinant vectors, adopts M13 universal primer (sequence information is with reference to the TaKaRa goods catalogue) that cloned sequence is checked order, and the result shows, contains molecule marker of the present invention (Seq ID No.1) in the recombinant vectors.Steps such as above-mentioned clone, conversion, cultivation, plasmid extraction are with reference to " the molecular cloning experiment guide third edition ", and Huang Peitang etc. translate, and Science Press publishes in September, 2002.
Above content be in conjunction with concrete embodiment to further describing that the present invention did, can not assert that concrete enforcement of the present invention is confined to these explanations.For the general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, can also make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (10)
- One kind with the closely linked molecule marker of millet leaf color gene, it is characterized in that: described molecule marker contains sequence shown in the Seq ID No.1; Preferably, described molecule marker is a sequence shown in the Seq ID No.1.
- 2. the primer of amplification and the closely linked molecule marker of millet leaf color gene is right, and it is characterized in that: the right primer 1 of described primer contains sequence shown in the Seq ID No.2, and primer 2 contains sequence shown in the Seq ID No.3; Preferably, primer 1 is a sequence shown in the Seq ID No.2, and primer 2 is a sequence shown in the Seq ID No.3;Seq?ID?No.2：5’-GGTGGTTGTTTACCATTTTCG-3’；Seq?ID?No.3：5’AACTACATTCCAATGTCCAGTT-3’。
- One kind with the closely linked molecule marker of millet leaf color gene, it is characterized in that: described molecule marker be by the described primer of claim 2 to blade partially the green the millet genomic dna be that template obtains through pcr amplification.
- 4. molecule marker according to claim 3 is characterized in that: described molecule marker contains sequence shown in the Seq ID No.1; Preferably, described molecule marker is a sequence shown in the Seq ID No.1.
- 5. a carrier is characterized in that: contain each described molecule marker in the claim 1,3,4.
- 6. a reconstitution cell is characterized in that: contain the described carrier of claim 5.
- 7. the detection method of the described molecule marker of claim 1, comprise step: according to the nucleotide sequence design primer of the molecule marker of claim 1, with detected millet genomic dna is that template increases, and judges whether there is this molecule marker in the amplified production; Preferably, described primer is that the described primer of claim 2 is right.
- In the claim 1,3,4 the molecule marker primer of each described molecule marker or claim 2 in millet assistant breeding or the assignment of genes gene mapping of millet leaf color or the purposes in detecting.
- 9. the method for the millet leaf color assignment of genes gene mapping is characterized in that: described method comprises the right step of molecule marker primer of using each described molecule marker in the claim 1,3,4 or claim 2.
- 10. millet auxiliary breeding means is characterized in that: described method comprises that test right requires each described molecule marker in 1,3,4 or with the step of molecule marker primer to detecting of claim 2.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107385105A (en) * | 2017-09-22 | 2017-11-24 | 山西省农业科学院谷子研究所 | With molecular labeling, primer and the application of millet glume color trait close linkage |
| CN108660234A (en) * | 2017-12-08 | 2018-10-16 | 深圳华大小米产业股份有限公司 | With the molecular labeling SIsv0362 of millet leaf color gene close linkage |
| CN108660235A (en) * | 2017-12-08 | 2018-10-16 | 深圳华大小米产业股份有限公司 | A kind of molecular labeling SIsv0686 with millet leaf color gene close linkage |
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| CN101974520A (en) * | 2010-11-22 | 2011-02-16 | 深圳华大基因科技有限公司 | Molecular maker SIsv1118 closely linked with high gene of millet strain |
| CN101974521A (en) * | 2010-11-22 | 2011-02-16 | 深圳华大基因科技有限公司 | Molecular marker SIsv0372 in close linkage with foxtail millet herbicide resistant gene |
| CN101982545A (en) * | 2010-11-22 | 2011-03-02 | 深圳华大基因科技有限公司 | Molecular marker SIsv0053 closely linked with millet plant height genes |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107385105A (en) * | 2017-09-22 | 2017-11-24 | 山西省农业科学院谷子研究所 | With molecular labeling, primer and the application of millet glume color trait close linkage |
| CN108660234A (en) * | 2017-12-08 | 2018-10-16 | 深圳华大小米产业股份有限公司 | With the molecular labeling SIsv0362 of millet leaf color gene close linkage |
| CN108660235A (en) * | 2017-12-08 | 2018-10-16 | 深圳华大小米产业股份有限公司 | A kind of molecular labeling SIsv0686 with millet leaf color gene close linkage |
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