CN102154284B - Molecular marker SIsv0408 capable of being closely linked to Setaria italica pollen color gene - Google Patents
Molecular marker SIsv0408 capable of being closely linked to Setaria italica pollen color gene Download PDFInfo
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Abstract
The invention belongs to the field of molecular biology and relates to a molecular marker, particularly relating to a molecular marker capable of being closely linked to a Setaria italica pollen color gene. The molecular marker has a sequence as shown in Seq ID No.1. The invention also relates to primers for amplifying the molecular marker, the applications of the molecular marker and the primers in pollen color gene localization of Setaria italica or genetic breeding of Setaria italica, and a Setaria italica breeding method. The molecular marker links the genome DNA sequence with the Setaria italica pollen color gene and helps the establishment of Setaria italica molecular marker-assisted breeding systems. The distance of genetic close linkage between the molecular marker and the Setaria italica pollen color gene is 0.3 cM. The molecular marker and the molecular marker amplification primers provided in the invention are designed for the convenient, rapid and high-throughout application in the breeding practices and the resource and variety identification of Setaria italica.
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 pollen color gene.The invention still further relates to primer, this molecule marker and the primer purposes in millet pollen color gene location 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 and method about millet is more backward relatively, 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.
Grain color color and luster and male sterile after millet pollen color and the millet maturation all have certain being associated.But the research to control millet pollen color genes involved does not have bibliographical information basically at present.
Summary of the invention
The purpose of this invention is to provide a kind of and the closely linked molecule marker of millet pollen 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 pollen color gene, and the molecule marker that amplification is obtained by this primer.
A purpose more of the present invention provides purposes and the detection method of above-mentioned molecule marker of above-mentioned molecule marker in millet pollen color gene location, detection and millet assistant breeding.
The object of the invention also comprises provides a kind of carrier that comprises above-mentioned molecule marker, and the reconstitution cell that contains this carrier; A kind of millet auxiliary breeding means that comprises the localization method of the millet pollen color gene that uses above-mentioned molecule marker and use said 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 pollen color gene, said molecule marker contains sequence shown in the Seq ID No.1; Preferred said 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 pollen color gene to the invention also discloses a kind of amplification, and the right primer 1 of said primer contains sequence shown in the Seq ID No.2, and primer 2 contains sequence shown in the Seq ID No.3; Preferred said 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’-CCATTTGACCCATGAAGCTC-3’；
Seq?ID?No.3：5’-GCTACCTTCCCCAAGATTCG-3’。
The invention also discloses a kind of and the closely linked molecule marker of millet pollen color gene, said molecule marker is to being that template obtains through pcr amplification with pollen yellow-white or orange millet genomic dna by above-mentioned primer.
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; Said 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 ' end and/or the nucleotide sequence beyond the 3 ' end of the Seq ID No.1 that is promptly comprised 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 needing only amplification perhaps detects this molecule marker in pollen yellow-white or the orange millet genomic dna, must detect or increase to 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, is 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 said 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.Said 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 said recombinant vectors.
The invention also discloses the preparation method of molecule marker of the present invention; The genomic dna that comprises the steps: to use pollen color yellow-white or orange millet is as template; To carrying out pcr amplification, the 672bp amplified production that obtains promptly contains said 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 said molecule marker, comprise step: the nucleotide sequence design primer according to above-mentioned molecule marker is that template increases with millet genomic dna to be detected, and judges whether there is this molecule marker in the amplified production.Preferably, said 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 millet to be detected template, (Seq ID No.2 and Seq ID No.3) carries out pcr amplification with above-mentioned primer, obtains amplified production.Can the amplified production that obtain be checked order or gel electrophoresis.
The invention also discloses the purposes of described molecule marker in millet pollen color gene location or detection.
The invention also discloses the localized method of a kind of millet pollen color gene, said 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, said 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 the molecular mark from now on; It will be understood by those skilled in the art that such as through detect whether exist molecule marker of the present invention to screen millet pollen whether to contain control pollen color be that yellow-white or orange color gene (for example, can references; 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).Said detection can be the method that PCR detects, and particularly, can use the primer of above-mentioned molecule marker of the present invention right.Said detection can also be carried out through sequence measurement.This millet auxiliary breeding means has easy, quick, high-throughout advantage.
In the present invention, particularly, said 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 No. 1 pollen color of paddy yellow-white; Millet A2 sterile line pollen color is brown; It is orange to open No. 3 pollen colors of confused flour beetle; Open F2 seville orange flower powder color part yellow-white that No. 3 selfings of confused flour beetle produce, part is orange, part is brown.
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 pollen color gene, this molecule marker connects genomic dna sequence and millet pollen color gene, helps the foundation of millet molecular mark system; The hereditary close linkage distance of said molecule marker and millet pollen color gene is 0.3cM.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 pcr amplification product of 12 strain millet pollen color yellow-whites or orange individual plant in F2 generation; Swimming lane 13-24 is the pcr amplification product of the F2 individual plant that 12 strain pollen colors are brown in generation.Swimming lane M is marker, and it is 100bp DNA Ladder; 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 SIsv0408 of millet pollen 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 pollen color gene, and this molecule marker is called after molecule marker SIsv0408 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 through chemosynthesis except obtaining the molecule marker of the present invention through 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’-CCATTTGACCCATGAAGCTC-3’；
Seq?ID?No.3：5’-GCTACCTTCCCCAAGATTCG-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 confirm 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 paternal pollen yellow-white and the brown homozygote hybridization of maternal pollen, 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 from 1105 pairs of primers, filters out 616 pairs of primers with polymorphum and validity.616 pairs of primers that employing is developed; 480 individuals of F2 colony are carried out PCR detect, the line data statistical study of going forward side by side is carried out genetic map with Map Maker3.0 software and is drawn; Obtain of the present invention and the closely linked molecule marker of pollen color gene, and amplimer.The male parent sequence that the primer that employing filters out obtains amplification, i.e. molecule marker SIsv0408 among the present invention.The F2 individuality is carried out character analysis, and millet pollen color gene is positioned on the genetic map according to gene character data and phenotypic character data.
Below through specific embodiment and combine accompanying drawing that the present invention is done further explain.Following examples are only further explained 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, plant type is high, and boot leaf is long and narrow, and bristle is red, and clever shell is red, can educate, 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, and bristle is green, and clever shell is green, partial sterility, the leaf colour cast is yellow, and 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 pollen color is orange), and the F1 selfing obtains F2.Wherein F1 is No. 3 seeds of a confused flour beetle.Altogether F2 for individual plant 480 strains, wherein, the pollen color is 120 strains that have of yellow-white, orange have 240 strains, brown have 120 strains.
No. 1 seed of above-mentioned paddy, millet A2 male-sterile seed and open No. 3 seeds of confused flour beetle can be referring to one Chinese patent application " with the closely linked molecule marker SIsv0372 of millet anti-herbicide gene ", publication number CN101974521A, date of publication February 16 in 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 following:
(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 (1L) as follows 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 absolute ethyl 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 deposition 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 ℃ subsequent use.
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 following:
The PCR response procedures is following:
94 ℃ of preparatory 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 through 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, then based on the sequence of difference, with the increase primer of diversity sequence of primer premier software design; 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 1105 pairs of random primers of table 1
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 preparatory sex change 5min; Get into 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 polymorphum: be meant in this validity whether amplified production is arranged, polymorphum 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 polymorphum with 616 couple of exploitation carries out PCR to 480 individuals of F2 colony and detects, the genomic dna of template used 480 individuals for the F2 colony that makes.
The PCR product is carried out agarose gel electrophoresis, obtain the result of molecule marker primer the amplification of 480 individuals.
Whole electrophoresis result are carried out data statistic analysis; Concrete grammar is following: 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, and 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 F2 colony 480 individuals.Such as, the data of 480 individuals that obtain with first pair of primer are a, b, and h ,-; B ... totally 480 data, the data of using second pair of primer to obtain 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/EXP3.0, S Lincoln, M Daly; E Lander-Cambridge; MA:Whitehead Institute, 1992) carry out the genetic linkage maps drafting, obtain genetic linkage map.Can confirm from this genetic linkage map that obtains 616 pairs of primers the position and with the genetic distance of millet pollen color gene.
(2) assignment of genes gene mapping
According to the pollen color phenotype of 480 individuals, similar with the male parent type proterties a (pollen yellow-white) that is designated as, similar with the maternal type proterties b (pollen is brown) that is designated as, proterties occupy and is designated as h (pollen is orange) between male parent and the female parent.Obtain the phenotypic data of 480 individuals; The phenotypic data of the 480 individuals genotype data with 480 individuals that obtain is before compared; Similar Gao Ze represents this mark and pollen color proterties close linkage, and the pollen color gene is positioned on the genetic linkage maps.
Embodiment 6: with the checking of the closely linked molecule marker of millet pollen color gene
1. on the basis of the genetic linkage maps that embodiment 5 makes; According to the genetic linkage distance of millet pollen color gene; With the hereditary close linkage distance of millet pollen color gene for having confirmed molecule marker primer (Seq ID No.2 and Seq ID No.3) in the position of 0.3cM; And find corresponding male parent sequence location, and the 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’-CCATTTGACCCATGAAGCTC-3’；
Seq?ID?No.3：5’-GCTACCTTCCCCAAGATTCG-3’。
2. in addition; In the electrophoresis in embodiment 5 to the pcr amplification product of 480 individuals in F2 generation; Amplification for molecule marker primer (Seq ID No.2 and Seq ID No.3) is: pollen color yellow-white or orange plant amount to about 360 strains (wherein about 120 strains of the plant of pollen color yellow-white; About 240 strains of the orange plant of pollen color); Their amplified production all has the band of 672bp size, and the pcr amplification product of the plant that about 120 strain pollen colors are brown does not all have the band (the part amplification is as shown in Figure 1) of 672bp.And the fragments sequence through this 672bp of order-checking proof is identical with Seq ID No.1.
It is thus clear that molecule marker of the present invention (Seq ID No.1) is and the closely linked molecule marker of millet pollen color gene.
Embodiment 7: molecular marker clone
The fragment cloning of the 672bp 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.From reconstitution cell, extract plasmid, said 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 is to combine concrete embodiment to the further explain that the present invention did, and can not assert that practical implementation of the present invention is confined to these explanations.For the those of ordinary skill of technical field under the present invention, under the prerequisite that does not break away from the present invention's design, can also make some simple deduction or replace, all should be regarded as belonging to protection scope of the present invention.
Claims (11)
- One kind with the closely linked molecule marker of millet pollen color gene, it is characterized in that: said 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 pollen color gene is right, it is characterized in that: the right primer 1 of said primer is 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’-CCATTTGACCCATGAAGCTC-3’；Seq?ID?No.3：5’-GCTACCTTCCCCAAGATTCG-3’。
- One kind with the closely linked molecule marker of millet pollen color gene, it is characterized in that: said molecule marker by the described primer of claim 2 to being that template obtains through pcr amplification with pollen yellow-white or orange millet genomic dna.
- 4. molecule marker according to claim 3 is characterized in that: said molecule marker is a sequence shown in the Seq IDNo.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 millet genomic dna to be detected is that template increases, and judges whether there is this molecule marker in the amplified production.
- 8. detection method according to claim 7 is characterized in that: said 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 millet pollen color gene location or the purposes in detecting.
- 10. localized method of millet pollen color gene is characterized in that: said method comprises the right step of molecule marker primer of using each described molecule marker in the claim 1,3,4 or claim 2.
- 11. a millet auxiliary breeding means is characterized in that: said 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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| CN106916880A (en) * | 2015-12-28 | 2017-07-04 | 深圳华大农业与循环经济科技有限公司 | With the molecular labeling SVhd1 of millet breeding time gene close linkage |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101142894A (en) * | 2007-09-26 | 2008-03-19 | 云南省农业科学院生物技术与种质资源研究所 | Wild-rice distant hybridization high-efficient cultivating superior progeny method |
| CN101760539A (en) * | 2008-12-19 | 2010-06-30 | 李祥 | Development and class of genetic marker |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101142894A (en) * | 2007-09-26 | 2008-03-19 | 云南省农业科学院生物技术与种质资源研究所 | Wild-rice distant hybridization high-efficient cultivating superior progeny method |
| CN101760539A (en) * | 2008-12-19 | 2010-06-30 | 李祥 | Development and class of genetic marker |
Non-Patent Citations (1)
| Title |
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| 李志江等.谷子分子标记与功能基因组研究进展.《中国农业科技导报》.2009,第11卷(第4期),16-22. * |
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