CN104745702B - EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence - Google Patents
EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence Download PDFInfo
- Publication number
- CN104745702B CN104745702B CN201510150401.XA CN201510150401A CN104745702B CN 104745702 B CN104745702 B CN 104745702B CN 201510150401 A CN201510150401 A CN 201510150401A CN 104745702 B CN104745702 B CN 104745702B
- Authority
- CN
- China
- Prior art keywords
- cotton
- est
- ssr
- mark
- primer
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Botany (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
EST SSR label primers and application the present invention relates to be based on the exploitation of upland cotton transcript profile sequence, EST SSR markers are SCRC1022.The invention further relates to application of the EST SSR label primers in terms of cotton analysis of genetic diversity and dense genetic map structure.The EST SSR primers developed based on upland cotton stem apex transcript profile sequence of the present invention have the advantages that amplification stabilization, electrophoretic band be clear, rich polymorphism, can be effectively used for the research fields such as germplasm resource for cotton analysis of genetic diversity, the structure of high density collection of illustrative plates, variety and authenticity identification, molecular mark.
Description
This case is the divisional application of application number 201410284580.1
Technical field
EST-SSR labeled primers and application, more particularly to base the present invention relates to be based on the exploitation of upland cotton transcript profile sequence
EST-SSR primers and its application in the exploitation of upland cotton Coker312 stem apex transcript profiles sequence, belong to molecular marking technique field.
Background technology
Cotton is one of important industrial crops, is also most important textile fabric crop.Its cultigen has 4, including 2
Individual dliploid cotton seed, i.e. cotton (Gossypium.herbaceum) and Asiatic cotton (G.arboreum) and 2 tetraploid cottons
Plant, i.e. upland cotton (G.hirsutum) and sea island cotton (G.barbadense).Because the yield of upland cotton is high, the spy of adaptability
Point, makes the important kind in cultigen.But the Upland Cotton diversity level reduction being bred as at present, hereditary basis is narrow
It is narrow, cause to be difficult to differentiate between in production and recognized, so as to govern the further raising of yield and quality.
With the development of genomics, molecular labeling turns into the prefered method for improving this problem.Simple repeated sequence
(SSR) it is a kind of highly polymorphic and codominant marker, thus is widely used in molecular labeling genetic analysis.With high-flux sequence
The development of technology, has developed substantial amounts of transcript profile data, development and application of these data in new mark in many species
It is significant in journey.Radish (Raphanus sativus L.), capsicum (Capsicum annuum L.), sesame
The excavation of transcript profile data in crops such as (Sesamum indicum L.), is that it recognizes and develop that new EST-SSR marks are carried
For reliability.New molecular labeling is developed currently with transcript profile sequence to have been applied in cotton, carry out moral and bravely etc. utilize land
670 new EST-SSR marks of the related transcript profile Data Mining of cotton development；Chen Haodong etc. is from Darwinian cotton transcript profile
Develop 780 pairs of EST-SSR primers in sequence, and the labor feature in EST-SSR sites, the polymorphism of primer and can turn
Shifting property；Lv Yuan great etc. is based on Island Cotton Fiber development est sequence and develops 380 pairs of SSR primers；Jena etc. is in cotton transcript profile
In 99780 Unigene, 12471 SSR sites are found out, be distributed in 8900 EST.
At present, in cotton develop EST-SSR mainly with Fibre Development gene-correlation, built up to cotton early forms related
Gene SSR marker exploitation there is not been reported.Organize and plant phyllopodium, the generation of bud former base and plant tip growth at stem end
Advantage is closely related, and developing EST-SSR related to this can increase the quantity of cotton EST-SSR marks, can deeply probe into again
The function of EST-SSR marker gene.
The content of the invention
The present invention is in view of the shortcomings of the prior art, there is provided a kind of EST-SSR marks based on the exploitation of upland cotton transcript profile sequence
Note primer and application.Amplification stabilization, electrophoretic band are had based on the EST-SSR primers that upland cotton stem apex transcript profile sequence is developed
Clearly, the advantages of rich polymorphism, can be effectively used for germplasm resource for cotton analysis of genetic diversity, the structure of high density collection of illustrative plates,
The research field such as variety and authenticity identification, molecular mark.
Technical scheme is as follows：
A kind of EST-SSR of upland cotton Coker312 marks the primer of SCRC056, and the primer is a pair, nucleotide sequence
Respectively SEQ ID NO.1 and SEQ ID NO.2.
The primer of above-mentioned EST-SSR marks SCRC056 is in terms of analysis of genetic diversity and dense genetic map structure
Application, mark SCRC056 be positioned on chr.16 (D7) at 20.4cM, the mark of 15.4cM on chr.16 (D7)
On BNL2734 and chr.16 (D7) between the mark CGR6280 of 24.4cM, with mark BNL2734 at a distance of 5cM, with mark
CGR6280 is at a distance of 4cM.
A kind of EST-SSR of upland cotton Coker312 marks the primer of SCRC371, and the primer is a pair, nucleotide sequence
Respectively SEQ ID NO.3 and SEQ ID NO.4.
The primer of above-mentioned EST-SSR marks SCRC371 builds in cotton analysis of genetic diversity and dense genetic map
The application of aspect, mark SCRC371 is positioned on chr.18 (D13) at 63.2cM, the mark of 52.7cM on chr.18 (D13)
On note SHIN1403 and chr.18 (D13) between the mark CGR5021 of 78.4cM, with mark SHIN1403 at a distance of 10.5cM.
A kind of EST-SSR of upland cotton Coker312 marks the primer of SCRC878, and the primer is a pair, nucleotide sequence
Respectively SEQ ID NO.5 and SEQ ID NO.6.
The primer of above-mentioned EST-SSR marks SCRC878 builds in cotton analysis of genetic diversity and dense genetic map
The application of aspect, mark SCRC878 is positioned on chr.2 (A2) at 42.5cM, positioned at linkage group end, and positioned at chr.2
(A2) the mark HAU880 of 35.5cM is closest on, at a distance of 7.0cM.
A kind of EST-SSR of upland cotton Coker312 marks the primer of SCRC1022, and the primer is a pair, nucleotide sequence
Respectively SEQ ID NO.7 and SEQ ID NO.8.
The primer of above-mentioned EST-SSR marks SCRC1022 builds in cotton analysis of genetic diversity and dense genetic map
The application of aspect, mark SCRC1022 is positioned on chr.23 (D9) at 19.5cM, the mark of 16.6cM on chr.23 (D9)
On note CGR5392 and chr.23 (D9) between the mark HAU2017 of 24.4cM, with mark CGR5392 at a distance of 2.9cM, with mark
HAU2017 is at a distance of 4.9cM.
The method that the primer of above-mentioned EST-SSR marks carries out cotton analysis of genetic diversity, step is as follows：
(1) DNA is extracted
Testing sample STb gene is extracted using the CTAB methods of improvement
(2) primer amplification and electrophoresis detection
Testing sample with step (1) extraction enters performing PCR and expands as template, using above-mentioned primer；Amplified production uses 8%
Native polyacrylamide gel electrophoresis detects that silver staining develops the color.
PCR system (10 μ L) includes that 1.0 μ 10 × Taq of L Buffer (contain Mg2+), 0.2 μ L dNTP, 0.1 μ L (2.5U)
Taq enzyme, each 0.6 μ L of upstream and downstream primer, 2.0 μ L (20~50ng) DNA profilings, plus sterile distilled water are to 10 μ L.
PCR programs are：94 DEG C of predegeneration 3min；94 DEG C of 45s, (Tm) DEG C 45s, 72 DEG C of 45s, totally 32 circulations；72 DEG C are prolonged
Stretch 7min；25 DEG C of preservations.
PCR primer adds point sample after 2.0 μ L bromophenol blues, is detected using 8% native polyacrylamide gel electrophoresis.Electrophoresis
Carried out in Electrophoresis Power Supply-EPS301 (Made in Sweden) nucleic acid electrophoresis system.Take 1.2
μ L samples, are DNA molecular amount standard, 300W invariable power electrophoresis 40min, silver with Takara 20bp DNA Ladder Marker
Dye colour developing.
(3) data statistics.Using two kinds of methods of data statistics：1. pleomorphism site is recorded.According to DNA molecular amount standard
With primer amplification statistics.There is clear band and is recorded as " 1 " in same equipotential point, is recorded as " 0 " without band；2. difference
Property bar tape recording.As standard, the banding pattern of parent 1 is recorded as " 1 " band amplified in parent with primer, the banding pattern of parent 2
" 2 " are recorded as, different the first banding pattern of parent's banding pattern are come across and is recorded as " 3 ", by that analogy.
The analysis of genetic diversity is：Using 1. data of method statistic in above-mentioned (3), calculated using PopGene32
The polymorphism information content (PIC) of primer.
The structure of the dense genetic map is to use Jion- using the data 2. recorded in above-mentioned steps (3)
Map4.0 software buildings Genetic Linkage Map composes (LOD value=6.5), with Kosambi function constructions；Using MapChart2.2
Genetic map is drawn by mapping software.
Beneficial effect
EST-SSR primers of the present invention are all from upland cotton stem apex transcript profile sequence, with conventional upland cotton fiber
There is function difference in the SSR primers developed in related transcript profile, described primer is distributed in upland cotton stem apex transcript profile
Uniformly, rich polymorphism, amplification stabilization, amplified band are easy to identification, and success is chain onto dense genetic map, enriches
Upland cotton is based on the quantity that transcript profile excavates SSR primers, can be effectively used for germplasm resource for cotton analysis of genetic diversity, highly dense
Spend structure, variety and the authenticity identification of collection of illustrative plates.
Brief description of the drawings
Fig. 1 is SSR Locator software search result schematic diagrams；
Fig. 2 is the electrophoresis result figure of the polymorphism that primer SCRC056 is detected in 13 parts of test materials；
In figure：M, Marker (20bp DNA Ladder TAKARA), 1, Tm-1,2, Coker 312,3, AcalasJ-5,
4th, Zhongmiansuo 12,5, CCRI 10,6, Shandong cotton grind No. 22,7, Shandong original 343,8,3-79,9, sea 7124,10, Ah must not Buddhist nun,
11st, Lei Mengdeshi cottons, 12, A Feilijia cottons, 13, stone system it is sub- No. 1；
Fig. 3 is the electrophoresis result figure of the polymorphism that primer SCRC371 is detected in 13 parts of test materials；
In figure：M, Marker (20bp DNA Ladder TAKARA), 1, Tm-1,2, Coker 312,3, AcalasJ-5,
4th, Zhongmiansuo 12,5, CCRI 10,6, Shandong cotton grind No. 22,7, Shandong original 343,8,3-79,9, sea 7124,10, Ah must not Buddhist nun,
11st, Lei Mengdeshi cottons, 12, A Feilijia cottons, 13, stone system it is sub- No. 1；
Fig. 4 is the electrophoresis result figure of the polymorphism that primer SCRC878 is detected in 13 parts of test materials；
In figure：M, Marker (20bp DNA Ladder TAKARA), 1, Tm-1,2, Coker 312,3, AcalasJ-5,
4th, Zhongmiansuo 12,5, CCRI 10,6, Shandong cotton grind No. 22,7, Shandong original 343,8,3-79,9, sea 7124,10, Ah must not Buddhist nun,
11st, Lei Mengdeshi cottons, 12, A Feilijia cottons, 13, stone system it is sub- No. 1；
Fig. 5 is the electrophoresis result figure of the polymorphism that primer SCRC1022 is detected in 13 parts of test materials；
In figure：M, Marker (20bp DNA Ladder TAKARA), 1, Tm-1,2, Coker 312,3, AcalasJ-5,
4th, Zhongmiansuo 12,5, CCRI 10,6, Shandong cotton grind No. 22,7, Shandong original 343,8,3-79,9, sea 7124,10, Ah must not Buddhist nun,
11st, Lei Mengdeshi cottons, 12, A Feilijia cottons, 13, stone system it is sub- No. 1；
Fig. 6 is the electrophoresis result figure after 5 pairs of primers are expanded in RIL parent；
In figure：M, Marker (20bp DNA Ladder TAKARA), 5 pairs of primers：SCRC056(Ⅰ)、SCRC371(Ⅱ)、
SCRC584(Ⅲ)、SCRC878(Ⅳ)、SCR1022C(Ⅴ)；1st, parent 1,2, parent 2；
Fig. 7 is positioning schematic diagram of 4 pairs of primers on genetic map.
Specific embodiment
Technical scheme is further elaborated with reference to embodiment, but institute's protection domain of the present invention is not limited to
This.
Embodiment
Applications of the upland cotton Coker312 transcript profiles EST-SSR in Gossypium inter-species and Interspecific polymorphism analysis
1. the extraction of cotton genomic dna
(1) material
5 cotton seeds totally 13 parts of materials (being shown in Table 1) are chosen from Cotton Germplasms material, for the evaluation of primer newly developed, bag
Include expanding effect and polymorphism analysis.
It is used for the material information of primer polymorphism analysis in the present invention of table 1
(2) CTAB methods extract genomic DNA
Cotton DNA is extracted using the CTAB methods of improvement, is comprised the following steps that：
1) the fresh blade 0.2g of cotton material to be tested is taken, is cleaned, dried, be put into after tearing up in 2mL A centrifuge tubes, added
Liquid nitrogen, is ground using Retsch (MM400) tissue grinder instrument.
2) to 700 μ L Extraction buffers (4 DEG C of preservations, composition is as shown in table 2) are added in centrifuge tube, it is vortexed uniform, ice bath
10min, 7200 turns/min are centrifuged 10min, abandoning supernatant.
The Extraction buffer formula (500mL) of table 2
3) to 700 μ L lysis buffers (65 DEG C of preheatings, composition is as shown in table 3) are added in precipitation, gently stirred evenly with pipette tips,
65 DEG C of water-bath 40min, during gently overturn several times.
The lysis buffer formula (500mL) of table 3
4) after water-bath, chloroform is added：Isoamyl alcohol (volume ratio 24:1) 700 μ L, upset more than 30 times, 15 DEG C 10000 turns/
Min is centrifuged 10min.
5) supernatant is transferred in 1.5mL centrifuge tubes, adds 0.6 times of isopropanol of supernatant volume (- 20 DEG C of precoolings), turned over
Turn, until there is flocculent deposit, to stand 10min, 10000 turns/min centrifugation 8min, abandoning supernatant.
6) ethanol of 700 μ L 70% is added to wash (this step can be overnight) in precipitation, 10000 turns/min centrifugation 5min are abandoned
Supernatant, untill putting the alcohol-free smell of ventilation drying.
7) 700 μ L TE buffer solutions, 65 DEG C of water-bath 30min are added.
8) after DNA dissolvings, 700 μ L chloroforms are added：Isoamyl alcohol (volume ratio 24:1), slow upset more than 30 times, stand
10000 turns/min centrifugations 10min after 5min.
9) take supernatant to be transferred in new 1.5mL centrifuge tubes, add in 0.1 times of sodium acetate of supernatant volume, addition etc.
(- 20 DEG C) of the isopropanol of supernatant volume overturns more than 30 times, stands 30min, 10000 turns/min centrifugation 5min, abandoning supernatant.
10) the small groups of 700 μ L70% ethanol washing DNA are added, 10000 turns/min centrifugation 5min, abandoning supernatant is dried
The small groups of DNA.
11) 200 μ L TE buffer solutions, 4 DEG C of preservations are added.
12) using DNA concentration detector detection DNA concentration, the agarose gel electrophoresis of 1% (quality specific volume) and
Eppendorf Biophotometer DNA concentrations instrument detects DNA mass.
2. the exploitation of upland cotton Coker312 stem apexs transcript profile EST-SSR marks
(1) lookup in upland cotton stem apex transcript profile sequence SSR sites
Based on 73518 unigene sequences that the sequencing of upland cotton stem apex transcript profile is obtained, using software MISA and SSR
Locator searches the SSR sites of 2~10bp, search criterion difference >=6,5,4,4,4,3,3,3.
(2) design of upland cotton stem apex transcript profile SSR primers
Centered on SSR sequences according to obtained in step (1), each 200bp sequences of SSR upstream and downstream are extracted, if upstream
Or downstream sequence then extracts the full sequence design primer in upstream or downstream less than 200bp.With newly-designed SSR primers to draw
Thing, with CMD (http：//www.cottonmarker.org/) est sequence for template run ePCR, obtain without repeat
New EST-SSR primers, are named as SCRCXXX (ShanDong Cotton Research Center).The major parameter of design
For：18~21bp of primer length, optimal length 20bp, PCR primer length 100~395；Most suitable 58 DEG C of Tm values.Design of primers into
Synthesized by Shanghai bioengineering Co., Ltd after work(.
(3) amplification and screening of upland cotton stem apex transcript profile SSR primers
4 parts of cotton materials of selection, the amplification situation for detecting upland cotton transcript profile EST-SSR marks.
Using the primer of synthesis in step (2), expanded by template of 4 parts of DNA of material, screened according to expanding effect
Amplification, banding pattern clearly primer 650 pairs can be stablized.
The primer of above-mentioned 650 pairs of amplifications stabilization, is expanded, in 13 parts of cotton materials used in the present invention wherein having
83 pairs of primers can expand polymorphic bandses in 13 parts of materials, and wherein 23 pairs bands of selection are clear, the abundant bar of polymorphism information
Band, polymorphism analysis are carried out in different inter-species and in planting.
Table 4 is based on 23 pairs of EST-SSR primer information of upland cotton stem apex transcript profile sequence exploitation
PCR system (10 μ L) includes：
1.0 μ 10 × Taq of L Buffer (contain Mg2+), 0.2 μ L dNTP, 0.1 μ L (2.5U) Taq enzyme, upstream and downstream primer is each
0.6 μ L, 2.0 μ l (20~50ng) DNA profilings, plus sterile distilled water are to 10 μ L.
PCR programs are：94 DEG C of predegeneration 3min；94 DEG C of denaturation 45s, (Tm) DEG C annealing 45s, 72 DEG C extend 45s, totally 32
Circulation；72 DEG C of extension 7min；25 DEG C of preservations.
PCR primer adds point sample after 2.0 μ L bromophenol blues, is detected using 8% native polyacrylamide gel electrophoresis.Electrophoresis
Carried out in Electrophoresis Power Supply-EPS301 (Made in Sweden) nucleic acid electrophoresis system.Take 1.2
μ L samples, are DNA molecular amount standard, 300W invariable power electrophoresis 40min, silver with Takara 20bp DNA Ladder Marker
Dye colour developing.
(4) data analysis
1) pleomorphism site is recorded.According to DNA molecular amount standard and primer amplification statistics.Same equipotential is pointed out
Now clear band is recorded as " 1 ", is recorded as " 0 " without band；Set up material to be tested SSR polymorphism information databases.
Calculated using PopGene32 softwares, material to be tested interior and interspecies variation information (PIC) not of the same race.In 13 parts of confessions
In examination material, the PIC luffings between total material：0.121~0.648, average PIC are 0.422, wherein (Tm- between five cotton seeds
1,3-79, A Feilijia cotton, Lei Mengdeshi cottons, sub- No. 1 of stone system) average PIC=0.424, hence it is evident that higher than upland cotton and island
Between cotton seedDetails are shown in Table 5.
Polymorphism analysis of the 5 23 pairs of upland cotton EST-SSR primers of table in Gossypium inter-species and kind
The primer (see Fig. 6) of the variant band in the RIL parent of this laboratory is selected in above-mentioned primer,
Respectively：SCRC056, SCRC371, SCRC584, SCRC878, SCRC1022.Using this 5 pairs of primers in RIL 359
Memory amplification in individual colony.
Being configured to of above-mentioned this laboratory RIL grinds the hybridization of No. 22 (male parents) and Shandong original 343 (female parent) using Shandong cotton
Afterwards, in continuous selfing to F8 generations, obtain since F2 generations, and operating method is this area conventional meanses.
2) otherness bar tape recording.As standard, the banding pattern of parent 1 is recorded as the band amplified in parent with primer
" 1 ", the banding pattern of parent 2 is recorded as " 2 ", comes across different the first banding pattern of parent's banding pattern and is recorded as " 3 ", by that analogy.
Build genetic map data storehouse.
(LOD value=6.5) are composed using Jion-Map4.0 software buildings Genetic Linkage Map, is made with Kosambi functions
Figure；Genetic map is drawn using MapChart2.2 mapping softwares.
Have in above-mentioned 5 pairs of primers on 4 pairs of chain genetic maps built to early stage of success (see Fig. 7), respectively
SCRC056、SCRC371、SCRC878、SCRC1022.Wherein, mark SCRC056 is positioned on chr.16 (D7) at 20.4cM,
Between the mark CGR6280 of 24.4cM on mark BNL2734 and chr.16 (D7) of 15.4cM on chr.16 (D7), with mark
BNL2734 is remembered at a distance of 5cM, with mark CGR6280 at a distance of 4cM；Mark SCRC371 is positioned on chr.18 (D13) at 63.2cM,
Between the mark CGR5021 of 78.4cM on mark SHIN1403 and chr.18 (D13) of 52.7cM on chr.18 (D13),
With mark SHIN1403 at a distance of 10.5cM；Mark SCRC878 is positioned on chr.2 (A2) at 42.5cM, positioned at linkage group end,
Mark HAU880 with the 35.5cM on chr.2 (A2) is closest, at a distance of 7.0cM；Mark SCRC1022 is positioned at
On chr.23 (D9) at 19.5cM, the 24.4cM on the mark CGR5392 and chr.23 (D9) of 16.6cM on chr.23 (D9)
Mark HAU2017 between, with mark CGR5392 at a distance of 2.9cM, with mark HAU2017 at a distance of 4.9cM.
Claims (2)
1. a kind of EST-SSR of upland cotton Coker312 based on upland cotton stem apex transcript profile sequence marks SCRC1022's
Primer, it is characterised in that the primer is a pair, and nucleotide sequence is respectively SEQ ID NO.7 and SEQ ID NO.8.
2. the primer of the EST-SSR marks SCRC1022 described in claim 1 is in cotton analysis of genetic diversity and high density
Application in terms of genetic map construction, mark SCRC1022 is positioned on chr.23 (D9) at 19.5cM, positioned at chr.23
(D9) on mark CGR5392 and chr.23 (D9) of 16.6cM between the mark HAU2017 of 24.4cM, with mark
CGR5392 at a distance of 2.9cM, with mark HAU2017 at a distance of 4.9cM.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510150401.XA CN104745702B (en) | 2014-06-23 | 2014-06-23 | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410284580.1A CN104017896B (en) | 2014-06-23 | 2014-06-23 | EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application |
| CN201510150401.XA CN104745702B (en) | 2014-06-23 | 2014-06-23 | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410284580.1A Division CN104017896B (en) | 2014-06-23 | 2014-06-23 | EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104745702A CN104745702A (en) | 2015-07-01 |
| CN104745702B true CN104745702B (en) | 2017-07-04 |
Family
ID=51434890
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510150401.XA Active CN104745702B (en) | 2014-06-23 | 2014-06-23 | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence |
| CN201510150375.0A Active CN104745701B (en) | 2014-06-23 | 2014-06-23 | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence |
| CN201410284580.1A Active CN104017896B (en) | 2014-06-23 | 2014-06-23 | EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application |
Family Applications After (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510150375.0A Active CN104745701B (en) | 2014-06-23 | 2014-06-23 | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence |
| CN201410284580.1A Active CN104017896B (en) | 2014-06-23 | 2014-06-23 | EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (3) | CN104745702B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106987648B (en) * | 2017-06-01 | 2020-07-17 | 中国农业大学 | High-flux SSR molecular marking method related to plant organ development |
| CN108085406A (en) * | 2017-12-13 | 2018-05-29 | 山东棉花研究中心 | A kind of identification method in upland cotton precocity molecular breeding correlation SSR marker site |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1796572A (en) * | 2004-12-22 | 2006-07-05 | 中国农业科学院棉花研究所 | Molecule marker of characteristics of cotton yield or quality of fibre |
| CN101020924A (en) * | 2006-12-30 | 2007-08-22 | 华中农业大学 | Prepn process and application of sea island cotton EST SSR marker |
| CN101880714B (en) * | 2010-06-04 | 2012-08-22 | 华中农业大学 | Method for building cotton fiber transcription genetic linkage map by EST-SSR sign |
| CN102181442B (en) * | 2011-05-10 | 2013-07-10 | 中国农业科学院棉花研究所 | Molecular label relevant with cotton fiber strength and arising from high-quality variety Xinluzao No.24 |
| CN103160506A (en) * | 2013-03-14 | 2013-06-19 | 南通大学 | Salt-tolerance EST-SSR molecular marker of cotton |
| CN103255139B (en) * | 2013-05-07 | 2015-04-15 | 南京农业大学 | Major QTL (Quantitative Trait Locus) of cotton high-strength fiber and molecular marker and application thereof |
-
2014
- 2014-06-23 CN CN201510150401.XA patent/CN104745702B/en active Active
- 2014-06-23 CN CN201510150375.0A patent/CN104745701B/en active Active
- 2014-06-23 CN CN201410284580.1A patent/CN104017896B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| Analysis of ESTs from multiple Gossypium hirsutum tissues and identification of SSRs;Taliercio et al.;《Genome》;20060328;第49卷;第306-319页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104745701B (en) | 2017-07-04 |
| CN104017896B (en) | 2015-06-24 |
| CN104745702A (en) | 2015-07-01 |
| CN104745701A (en) | 2015-07-01 |
| CN104017896A (en) | 2014-09-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110117673B (en) | Molecular marker of brassica napus dwarf trait locus and application thereof | |
| CN101250523B (en) | Molecule marker tightly linked with cucumber pubescence gene G1 | |
| CN103305510A (en) | Rice blast resistance gene Pi9 gene specificity molecular marker Pi9SNP as well as preparation and application thereof | |
| Stalker et al. | Molecular markers of Arachis and marker-assisted selection | |
| CN105907884A (en) | Rice blast resistant gene Pita specific molecular marker of rice and application | |
| CN110295251A (en) | Chain SNP marker and its application with wheat available tillering QTL | |
| CN110066883A (en) | With the molecular labeling R112146 of Rice Bacterial Blight Xa23 close linkage | |
| CN101117645B (en) | Molecule labeling method for non-heading Chinese cabbage late bolting gene | |
| CN108048593A (en) | A kind of molecular labeling that resistance to verticillium wilt can be improved from sea island cotton sea 1 | |
| CN104745702B (en) | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence | |
| CN107937397A (en) | SNP marker and its preparation method and application with tomato male sterility gene close linkage | |
| CN100447254C (en) | Molecular marker-assisted selection method for capsicum blight-resistant breeding | |
| CN106498048A (en) | A kind of QTL related to soybean nodulation number, SNP marker and application | |
| CN105483248B (en) | From sea island cotton molecular labeling related with fibre strength and its application | |
| CN111926104B (en) | SSR molecular marker for identifying authenticity of sugarcane and festuca arundinacea filial generation and method thereof | |
| CN104694661B (en) | EST SSR label primers and application based on the exploitation of upland cotton transcript profile sequence | |
| CN105524998B (en) | A kind of and chain molecular labeling of cotton Island Cotton Fiber intensity | |
| CN104278028B (en) | It is positioned at haynaldia villosa 6VS DNA and penetrates into wheat anti-powdery mildew NIL sequence and application | |
| CN106755413A (en) | Nitrogen in Rice absorbs site qNUE6 and its molecule labelling method | |
| CN109207628A (en) | A kind of molecular labeling and application suitable for detecting purple radish | |
| CN104278025B (en) | Cabbage type rape full-length genome SCAR molecular labelings and preparation method and application | |
| CN104313149A (en) | Molecular marker suitable for detecting radish leaf margin lobe trait and application of molecular marker | |
| CN101565745A (en) | Method for performing variety identification and seed purity check of celery cabbage by using SCAR marker | |
| CN109321670A (en) | The molecular labeling of rice number of grain per ear gene NOG1 and its application | |
| CN107980618A (en) | A kind of molecular breeding method using the mono- QTL fragments substitution line improvement cotton fiber strengths of chr.7 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |