CN107630101A - A kind of method of muskmelon cenospecies " day perfume (or spice) No.1 " Purity Identification based on EST SSR markers - Google Patents
A kind of method of muskmelon cenospecies " day perfume (or spice) No.1 " Purity Identification based on EST SSR markers Download PDFInfo
- Publication number
- CN107630101A CN107630101A CN201610565519.3A CN201610565519A CN107630101A CN 107630101 A CN107630101 A CN 107630101A CN 201610565519 A CN201610565519 A CN 201610565519A CN 107630101 A CN107630101 A CN 107630101A
- Authority
- CN
- China
- Prior art keywords
- cenospecies
- primer
- est
- ssr
- spice
- 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.)
- Pending
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of method of melon variety " day perfume (or spice) No.1 " cenospecies Purity Identification based on EST SSR markers.This method is using the genomic DNA of melon variety " day perfume (or spice) No.1 " cenospecies and its parents as template, by announcing muskmelon est sequence on Gene Bank databases, using 3.0 online design of primers of Primer, designs EST SSR primers.By screening, the complementary banding pattern primer that 1 pair of cenospecies banding pattern is parents is obtained, it is good through field checking test primer stability, and more coincide with field test results, Purity can be carried out to muskmelon Hybrid.The detection method of the present invention can complete Purity Identification work within 3h, have the advantage such as quick, inexpensive, easy to operate.
Description
Technical field
The invention belongs to the vegetable breeding and applied technical field of agricultural, and in particular to muskmelon cenospecies " day perfume (or spice) No.1 " is pure
Authentication method is spent, is a kind of cenospecies seed purity identification method based on EST-SSR molecular marking techniques.
Background technology
" day perfume (or spice) No.1 " is that precocious, high-quality, the white skin red meat thick-skinned melon of the newest culture of Tianjin Ke Run Vegetable Research Institutes is new
Kind.Plant growing way is medium, easily bears fruit, and quality is excellent, aromatic flavour.Fruit ellipse, 35 days puberties, single fruit weight 1.3KG are left
It is right.Pulp is orange, and meat thickness 3.5CM, soluble solid content 18%, fragranced is dense, and flavor is splendid.It is early suitable for spring protecting field
Ripe cultivation.
" day perfume (or spice) No.1 " cultivation management measure：Plantation density is 2000 plants/acre, and mu applies high-quality farm manure 3000KG, compound
Fertile 30KG, potash fertilizer 5KG is as base fertilizer.Single climing training, main stem stay 23-25 piece leaves, and melon is stayed in 13-15 sections position, and side shoot of bearing fruit stays 2 leaves
Pinching, every plant is stayed 1-2 fruit, can also stay regrowth hair fruit.Swollen fruit water is poured during fruit egg size in time, and chases after swollen fruit fertilizer, promotes fruit
Expand in fact.Fruit colour switchs to harvest in time during milky.
The height of muskmelon seedses purity is the direct leading indicator for weighing seed quality quality.Seed purity reduces can be notable
The yield and product quality of crop are reduced, peasant is suffered huge economic loss.Traditional seed purity identification method be with
After planting the field shape of plant is viewed as foundation, and its cycle length, workload are big, and are easily influenceed by environment, seasonal factor, cause
As a result deviation be present.Therefore, fast and accurately seed purity identification method has turned into seed R＆D institution and enterprise is common for exploitation
The problem of concern.
EST-SSR marks are current crop varieties because it has the advantages that codominance, polymorphism are high, experimental arrangement is simple
One of the most frequently used mark in identification and Purity Identification research.In recent years, the continuous improvement with sequencing technologies and molecule
The further investigation of biology, the EST of a large amount of melon crops are sequenced, and so far, international Curcurbitaceae genome plan research is small
Group includes watermelon ESTs7856 bars, muskmelon 126940, cucumber 359105 altogether.
The content of the invention
It is an object of the invention to disclose a kind of muskmelon cenospecies " day perfume (or spice) one based on EST-SSR molecular marking techniques
Number " seed purity identification method, to achieve the above object, the present invention provides following technical scheme：
This method by analyzing carrying out DNA extractions, PCR amplifications and EST-SSR for examination muskmelon cenospecies " day perfume (or spice) No.1 ",
Confirm the codominance complementation banding pattern of cenospecies and its parents' stabilization, it is pure so as to be carried out to muskmelon cenospecies " day perfume (or spice) No.1 " seed
Degree identification.For identifying the SSR primers of muskmelon cenospecies " day perfume (or spice) No.1 " seed purity, including primer forward direction sequence：5’-
TCGAAGTCCAGGAAGAGAG-3 ', reverse primer sequences：5’-TAAAGAATGTGGAGAGCGG-3’.
The method of Rapid identification muskmelon cenospecies seed purity of the present invention, this method is with greenhouse melon kind " day perfume
No.1 " is template for the genomic DNA for planting experimentally son, to known kind " day perfume (or spice) No.1 " parent's DNA otherness fragments analyze.
It is for the sampling of examination muskmelon cenospecies and DNA extraction method：To greenhouse melon grab sample, 50 plants altogether, take close to the tender of root
50~100mg of leaf portion point, liquid nitrogen frozen grinding (Retsch MM400 biologies crusher), adds CTAB lysis buffers (20g/L
CTAB, 1.4M NaCl, 0.1M Tris-HCl, 20mM Na2EDTA) 500 μ L, 65 DEG C of water bath with thermostatic control (Neslab) cracking
30min, follow-up DNA extraction purifications are carried out by resin type genomic DNA purification kit (SBS Genetech) operation.It is dense after DNA extractions
Degree is uniformly diluted to 50 ± 1ng/ μ L, Nanodrop ND1000, Thermol).DNA extraction method can also be by commercial reagent box
Carry out.
It is for examination muskmelon cenospecies PCR method：
(1) SSR upstream primer sequences：5 '-TCGAAGTCCAGGAAGAGAG-3 ', SSR downstream primer sequence：5’-
TAAAGAATGTGGAGAGCGG-3’。
(2) performing PCR amplification, pcr amplification reaction therein are entered using above-mentioned SSR primer pairs experimental cultivar sample template DNA
Cumulative volume be 10 μ L, amplification reaction system is：Master Mix 5 μ L, the μ of upstream and downstream primer 10
Each 0.5 μ L of mol/L, for trying muskmelon cenospecies genomic DNA template (50ng/ μ L) 1 μ L, distilled water supplies 10 μ L；PCR reaction intervals
Sequence is 95 DEG C of pre-degeneration 2min, 32 amplification cycles (94 DEG C of 45sec, 55 DEG C of 45sec, 72 DEG C of 45sec)；72 DEG C extension 7min, 4
DEG C preserve；
(3) gel electrophoresis is carried out for examination muskmelon cenospecies pcr amplification product, non-denaturing polyacrylamide gel concentration is
8%, gel component is：The μ L of deionized water 21mL, 10 × TBE 3mL, 40%PAG 6mL, TMED 30,10% excessively stream acid amides 300
μL；Voltage 250V, electrophoresis 1h are set, power supply is closed, is dyed；Dyeing course：8~10min of silver staining, rapid washing, NaOH
Several minutes of colour developing, until band is clear, wash again；
(4) for examination muskmelon cenospecies EST-SSR interpretations of result, cenospecies and its key band of parents are compared, to for trying
Seed sample carries out codominance complementation band analysis, and gross sample quantity is accounted for by calculating cenospecies quantity in experimental cultivar sample
Ratio-dependent seed purity.
For the detection method of the present invention in the case where not calculating DNA extraction process and taking, PCR takes 90min, EST-SSR
60-90min is analyzed, so the detection method of the present invention can complete Purity Identification work within 3h, is had quick, low
The advantage such as cost, easy to operate.
In order to more clearly illustrate the assay method of the present invention, the test method of the present invention is done with detailed below
Explanation.
1st, principle
This method is using a kind of its principle of new method for nucleic acid analysis：EST-SSR is using 1~6 base as repetition
Unit, it is the SSR being present in EST.EST (Expressed sequence tag, EST) is in body by mRNA
Outer reverse transcription is into cDNA and after being cloned into plasmid or phage vector construction cDNA library, extensive random picking cDNA clone,
The Express Sequence Tags for being about 150~500bp obtained after its 5 ' end or 3 ' ends are sequenced.Est sequence comes from laboratory
The cDNA library of structure is downloaded from American National Bioinformatics Institute (NCBI) Genbank.Synthesized using related software
Primer, performing PCR amplification and gel electrophoresis are finally entered to research material, is analyzed by specific spectruming belt, so as to calculate cenospecies
Purity.
2nd, design of primers
By the 214 muskmelon est sequences announced on Gene Bank databases, set using 3.0 online primers of Primer
Meter, primer are synthesized by Shanghai life work, and obtaining 1 pair of EST-SSR primer through screening can be used for muskmelon cenospecies Purity Identification.
Primer is synthesized by Shanghai bio-engineering corporation.
The primer sequence table of table 1 is as follows：
| Primer | Sequence (5'to3') |
| TJTX1- forward primers | TCGAAGTCCAGGAAGAGAG |
| TJTX1- reverse primers | TAAAGAATGTGGAGAGCGG |
3rd, sampling and DNA extractions
Grab sample is carried out to greenhouse melon, takes 50~100mg of tender leaf part close to root, liquid nitrogen frozen grinding
(Retsch MM400 biologies crusher), add CTAB lysis buffers (20g/L CTAB, 1.4MNaCl, 0.1M Tris-
HCl, 20mM Na2EDTA) 500 μ L, 65 DEG C of waters bath with thermostatic control (Neslab) crack 30min, and follow-up DNA extraction purifications press resin type
Genomic DNA purification kit (SBS Genetech) operation is carried out.Concentration is uniformly diluted to 50 ± 1ng/ μ L after DNA extractions,
Nanodrop ND1000, Thermol).DNA extraction method can also be carried out by commercial reagent box.
4th, PCR reacts
Using SSR upstream primer sequences：5 '-TCGAAGTCCAGGAAGAGAG-3 ', SSR downstream primer sequence：5’-
TAAAGAATGTGGAGAGCGG-3’。
SSR primer pair experimental cultivar sample templates DNA enters performing PCR amplification (instrument is ABI Veriti PCR), therein
The cumulative volume of pcr amplification reaction is 10 μ L, and amplification reaction system is：The μ L of Master Mix 5, upstream is with
Primer each 0.5 μ L of 10 μm of ol/L are swum, for trying muskmelon cenospecies genomic DNA template (50ng/ μ L) 1 μ L, distilled water supplies 10 μ L.
PCR response procedures are 95 DEG C of pre-degeneration 2min, 32 amplification cycles (94 DEG C of 45sec, 55 DEG C of 45sec, 72 DEG C of 45sec)；72℃
Extend 7min, 4 DEG C of preservations.
5th, native polyacrylamide gel electrophoresis and silver staining
For examination muskmelon cenospecies pcr amplification product carry out gel electrophoresis (instrument is BIO-RAD electrophoresis apparatuses), non denatured poly- third
Acrylamide gel strength is 8%, and gel component is：The μ of deionized water 21mL, 10 × TBE 3mL, 40%PAG 6mL, TMED 30
L, the μ L of 10% excessively stream acid amides 300；The μ L of loading 1~2, voltage 250V, electrophoresis 1h are set, close power supply, dyed；
Dyeing course (instrument is ORBITAL decolorization swinging tables)：The silver nitrate that 6mL10% is added in 600mL ultra-pure waters is molten
Liquid, 8~10min of silver staining；Washed rapidly in 600mL ultra-pure waters；9g sodium hydroxide powders are added in 600mL ultra-pure waters, it is molten
3mL formaldehyde is added after solution, is developed the color several minutes, until band is clear；Washed again in 600mL ultra-pure waters.
6th, EST-SSR spectral band analysis
By analyzing EST-SSR results, compare the cenospecies of test sample and its key band of parents, to for examination hybridization
Kind carries out Purity.The SSR primers obtained with screening, female parent show as 1 key band, and male parent shows as being different from female parent
1 key band, homozygous cenospecies is to show as the set of this 2 key bands, that is, codominant complementary banding pattern；
It is non-it is homozygous can not show as codominant complementary banding pattern, maternal key band banding pattern may be shown as, that is, occur it is maternal from
Hand over.
7th, Purity
By the ratio-dependent seed purity for calculating cenospecies in experimental cultivar sample:
Seed purity=cenospecies quantity/test sample quantity * 100%.
Brief description of the drawings:
Fig. 1 is that 1 pair of SSR primer that screening obtains is used for muskmelon cenospecies " day perfume (or spice) No.1 " and its parents' amplified production
Electrophoretic analysis collection of illustrative plates.Being followed successively by 1~6 maternal banding pattern, 7~12 cenospecies banding patterns, 13~18 male parent banding patterns from left to right, (figure is
The TJTX1SSR primers that screening obtains are used for the electrophoretic analysis collection of illustrative plates of muskmelon cenospecies and its parents' amplified production.From left to right
It is followed successively by female parent 1~2, cenospecies 3~6, male parent 7~8.)
Embodiment
In order to more clearly illustrate the method for the present invention, the test method of the present invention is made with specifically below
Bright, need to be illustrated herein is：Primer sequence of the present invention is shown in Table 1.
Embodiment 1
(1) reagent：The production of Promega companiesMaster Mix solution；The EST- of Shanghai life work synthesis
SSR primers；
(2) EST-SSR primer sequences table is as follows：
| Primer | Sequence (5'to3') |
| TJTX1- forward primers | TCGAAGTCCAGGAAGAGAG |
| TJTX1- reverse primers | TAAAGAATGTGGAGAGCGG |
(3) sampling and DNA extractions
90 plants of experimental cultivar seedling, liquid nitrogen frozen grinding (Retsch MM400 biologies crusher), it is slow to add CTAB cracking
Fliud flushing (20g/L CTAB, 1.4M NaCl, 0.1M Tris-HCl, 20mM Na2EDTA) 500 μ L, 65 DEG C of waters bath with thermostatic control
(Neslab) 30min is cracked, follow-up DNA extraction purifications are carried out by resin type genomic DNA purification kit (SBS Genetech) operation.
Concentration is uniformly diluted to 50 ± 1ng/ μ L, Nanodrop ND1000, Thermol after DNA extractions).DNA extraction method can also
Carried out by commercial reagent box.
(4) PCR reacts
(instrument is to entering performing PCR amplification for examination seedling DNA profiling and its parent DNA for pcr amplification primer thing as described in upper table
AB Veriti PCR), the cumulative volume of pcr amplification reaction therein is 10 μ L, and amplification reaction system is：
Master Mix 5 each 0.5 μ L of μ L, 10 μm of ol/L of upstream and downstream primer, experimental cultivar female parent gene group DNA profiling (50ng/ μ
L) 1 μ L, distilled water supply 10 μ L.PCR response procedures are 95 DEG C of pre-degeneration 2min, and 32 amplification cycles (94 DEG C of 45sec, 55 DEG C
45sec, 72 DEG C of 45sec)；72 DEG C of extension 7min, 4 DEG C of preservations.
(5) gel electrophoresis and silver staining
The μ L of experimental cultivar female parent PCR primer loading 1~2, voltage 250V, electrophoresis 1h are set, close power supply, dyed；
Dyeing course：8~10min of silver staining, rapid washing, NaOH develop the color several minutes, until band is clear, washed again；
(6) EST-SSR interpretations of result：90 plants for having that 3 plants of electrophoresis banding pattern is consistent with maternal banding pattern in examination seedling, be non-miscellaneous
Hand over kind.
(7) Purity
By the ratio-dependent seed purity for calculating cenospecies in experimental cultivar sample.
Seed purity=87/90*100%=96.7%.
Embodiment 2
(1) reagent：The production of Promega companiesMaster Mix solution；The EST- of Shanghai life work synthesis
SSR primers；
(2) EST-SSR primer sequences table is as follows：
| Primer | Sequence (5'to3') |
| TJTX1- forward primers | TCGAAGTCCAGGAAGAGAG |
| TJTX1- reverse primers | TAAAGAATGTGGAGAGCGG |
(3) sampling and DNA profiling extraction
93 plants of experimental cultivar seedling, liquid nitrogen frozen grinding (Retsch MM400 biologies crusher), it is slow to add CTAB cracking
Fliud flushing (20g/L CTAB, 1.4M NaCl, 0.1M Tris-HCl, 20mM Na2EDTA) 500 μ L, 65 DEG C of waters bath with thermostatic control
(Neslab) 30min is cracked, follow-up DNA extraction purifications are carried out by resin type genomic DNA purification kit (SBS Genetech) operation.
Concentration is uniformly diluted to 50 ± 1ng/ μ L, Nanodrop ND1000, Thermol after DNA extractions).DNA extraction method can also
Carried out by commercial reagent box.
(4) PCR reacts
(instrument is to entering performing PCR amplification for examination seedling DNA profiling and its parent DNA for pcr amplification primer thing as described in upper table
AB Veriti PCR), the cumulative volume of pcr amplification reaction therein is 10 μ L, and amplification reaction system is：
Master Mix 5 each 0.5 μ L of μ L, 10 μm of ol/L of upstream and downstream primer, experimental cultivar male parent gene group DNA profiling (50ng/ μ
L) 1 μ L, distilled water supply 10 μ L.PCR response procedures are 95 DEG C of pre-degeneration 2min, and 32 amplification cycles (94 DEG C of 45sec, 55 DEG C
45sec, 72 DEG C of 45sec)；72 DEG C of extension 7min, 4 DEG C of preservations.
(5) gel electrophoresis and silver staining
The μ L of experimental cultivar male parent PCR primer loading 1~2, voltage 250V, electrophoresis 1h are set, close power supply, dyed；
Dyeing course：8~10min of silver staining, rapid washing, NaOH develop the color several minutes, until band is clear, washed again；
(6) EST-SSR interpretations of result：98 plants for having that 1 plant of electrophoresis banding pattern is consistent with maternal banding pattern in examination seedling, be non-miscellaneous
Hand over kind.
(7) Purity
By the ratio-dependent seed purity for calculating cenospecies in experimental cultivar sample.
Seed purity=97/98*100%=99.0%.
Embodiment 3
(1) reagent：The production of Promega companiesMaster Mix solution；The EST- of Shanghai life work synthesis
SSR primers；
(2) EST-SSR primer sequences table is as follows：
| Primer | Sequence (5'to3') |
| TJTX1- forward primers | TCGAAGTCCAGGAAGAGAG |
| TJTX1- reverse primers | TAAAGAATGTGGAGAGCGG |
(3) sampling and DNA extractions
Experimental cultivar takes cenospecies individual plant 120, liquid nitrogen frozen grinding (Retsch MM400 biologies crusher), adds CTAB
Lysis buffer (20g/L CTAB, 1.4MNaCl, 0.1M Tris-HCl, 20mM Na2EDTA) 500 μ L, 65 DEG C of waters bath with thermostatic control
(Neslab) 30min is cracked, follow-up DNA extraction purifications are carried out by resin type genomic DNA purification kit (SBS Genetech) operation.
Concentration is uniformly diluted to 50 ± 1ng/ μ L, Nanodrop ND1000, Thermol after DNA extractions).DNA extraction method can also
Carried out by commercial reagent box.
(4) PCR reacts
Pcr amplification primer thing as described in upper table enters performing PCR amplification to experimental cultivar cenospecies individual plant template DNA, and (instrument is
AB Veriti PCR), the cumulative volume of pcr amplification reaction therein is 10 μ L, and amplification reaction system is：
Each 0.5 μ L of μ L, 10 μm of ol/L of upstream and downstream primer of Master Mix 5, experimental cultivar cenospecies genomic DNA template
(50ng/ μ L) 1 μ L, distilled water supply 10 μ L.PCR response procedures are 95 DEG C of pre-degeneration 2min, 32 (94 DEG C of amplification cycles
45sec, 55 DEG C of 45sec, 72 DEG C of 45sec)；72 DEG C of extension 7min, 4 DEG C of preservations.
(5) gel electrophoresis and silver staining
The μ L of experimental cultivar cenospecies PCR primer loading 1~2, voltage 250V, electrophoresis 1h are set, close power supply, contaminated
Color；Dyeing course：8~10min of silver staining, rapid washing, NaOH develop the color several minutes, until band is clear, washed again；
(6) EST-SSR interpretations of result：Experimental cultivar cenospecies banding pattern is different from 1 of female parent for maternal 1 with male parent,
The set of this 2 key bands, that is, codominant complementary banding pattern.
(7) Purity
By the ratio-dependent seed purity for calculating cenospecies in experimental cultivar sample.
Seed purity=111/120*100%=92.5%.
Claims (2)
1. 1 pair of SSR primer for identifying muskmelon cenospecies " day perfume (or spice) No.1 " seed purity, it is characterised in that draw including upstream
Thing sequence：5 '-TCGAAGTCCAGGAAGAGAG-3 ', downstream primer sequence：5’-TAAAGAATGTGGAGAGCGG-3’.
2. one kind uses primer described in claim 1, muskmelon cenospecies " day perfume (or spice) No.1 " seed purity based on EST-SSR marks
The method of identification, it is characterised in that comprise the following steps：
(1) performing PCR amplification is entered using the SSR primer pair experimental cultivar sample templates DNA described in claim 1, PCR therein expands
The cumulative volume for increasing reaction is 10 μ L, and amplification reaction system is：The μ L of Master Mix 5, upstream and downstream draw
Each 0.5 μ L of 10 μm of ol/L of thing, for trying muskmelon cenospecies genomic DNA template (50ng/ μ L) 1 μ L, distilled water supplies 10 μ L；PCR
Response procedures are 95 DEG C of pre-degeneration 2min, 32 amplification cycles (94 DEG C of 45sec, 55 DEG C of 45sec, 72 DEG C of 45sec)；72 DEG C of extensions
7min, 4 DEG C of preservations；
(2) gel electrophoresis is carried out to pcr amplification product, non-denaturing polyacrylamide gel concentration is 8%, and gel component is：Go
Ionized water 21mL, 10 × TBE 3mL, 40%PAG 6mL, TMED 30 μ L, the μ L of 10% excessively stream acid amides 300；Voltage 250V is set,
Electrophoresis 30min, power supply is closed, is dyed；Dyeing course：8~10min of silver staining, rapid washing, NaOH develop the color several minutes, directly
It is clear to band, wash again；
(3) by analyzing EST-SSR results, cenospecies and its key band of parents are compared, to being total to for planting experimentally subsample
Dominant complementary band analysis, by calculate cenospecies quantity in experimental cultivar sample account for gross sample quantity ratio-dependent seed it is pure
Degree.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610565519.3A CN107630101A (en) | 2016-07-15 | 2016-07-15 | A kind of method of muskmelon cenospecies " day perfume (or spice) No.1 " Purity Identification based on EST SSR markers |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610565519.3A CN107630101A (en) | 2016-07-15 | 2016-07-15 | A kind of method of muskmelon cenospecies " day perfume (or spice) No.1 " Purity Identification based on EST SSR markers |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107630101A true CN107630101A (en) | 2018-01-26 |
Family
ID=61113073
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610565519.3A Pending CN107630101A (en) | 2016-07-15 | 2016-07-15 | A kind of method of muskmelon cenospecies " day perfume (or spice) No.1 " Purity Identification based on EST SSR markers |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107630101A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140039866A (en) * | 2012-09-25 | 2014-04-02 | 한국생명공학연구원 | Ssr primer sets for discrimination of oriental melon line or cultivar and uses thereof |
| CN105755168A (en) * | 2016-05-25 | 2016-07-13 | 天津科润农业科技股份有限公司 | Method for identifying purity of muskmelon 'bud' hybrid seeds based on EST-SSR marks |
-
2016
- 2016-07-15 CN CN201610565519.3A patent/CN107630101A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140039866A (en) * | 2012-09-25 | 2014-04-02 | 한국생명공학연구원 | Ssr primer sets for discrimination of oriental melon line or cultivar and uses thereof |
| CN105755168A (en) * | 2016-05-25 | 2016-07-13 | 天津科润农业科技股份有限公司 | Method for identifying purity of muskmelon 'bud' hybrid seeds based on EST-SSR marks |
Non-Patent Citations (3)
| Title |
|---|
| ILKNUR SOLMAZ等: "Genetic Characterization of Turkish Snake Melon (Cucumis melo L. subsp. melo flexuosus Group) Accessions Revealed by SSR Markers", 《BIOCHEM GENET》 * |
| 刘鹏等: "利用黄瓜、甜瓜和西瓜SSR开发葫芦科作物穿梭标记", 《分子植物育种》 * |
| 李丽等: "SSR标记对甜瓜品种纯度和真实性的鉴定", 《分子植物育种》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104059992B (en) | A kind of method of the muskmelon cross-fertilize seed Purity Identification based on EST-SSR mark | |
| CN102443654B (en) | Cauliflower hybrid seed purity identification method based on EST-SSR (expressed sequence tag-simple sequence repeat) marker | |
| CN107164533B (en) | SNP (Single nucleotide polymorphism) marker of watermelon peel stripe, detection method and application | |
| CN109652412B (en) | SNP molecular marker, method for detecting melon flower type and application | |
| CN105154550B (en) | A kind of method using 1217 purity of EST-SSR molecular labeling Rapid identification variety of watermelon treasure sweet tea | |
| CN105779643B (en) | The method of muskmelon cenospecies No. 2 Purity Identifications of bud based on EST-SSR label | |
| CN107217099A (en) | A kind of SNP marker identified available for snakehead genetic sex and supermale fish and its application | |
| CN104419762A (en) | Method for identifying cauliflower varieties by EST-SSR labels | |
| CN103789441A (en) | SSR (Simple Sequence Repeat) molecular marker method for identifying two hippocampus populations | |
| CN105695454B (en) | A kind of molecular labeling and its identification method for identifying sesame genie male sterile line | |
| CN111057784B (en) | SSR molecular marker primer related to walnut black spot and application thereof | |
| CN105755168B (en) | The method of muskmelon cenospecies bud Purity Identification based on EST-SSR label | |
| CN107630100A (en) | A kind of method of muskmelon cenospecies " beautiful exquisite " Purity Identification based on EST SSR markers | |
| CN106755465A (en) | The molecular labeling of QTL QFll.sicau 2D close linkage long with wheat flag leaf and application | |
| CN107630101A (en) | A kind of method of muskmelon cenospecies " day perfume (or spice) No.1 " Purity Identification based on EST SSR markers | |
| CN113462813B (en) | Site linked with pear peel red character, molecular marker and application thereof | |
| CN114032332B (en) | Molecular marker cm17-16 for rapidly detecting pumpkin naked kernel characteristics and application thereof | |
| CN113502334B (en) | Molecular marker C27449 for rapidly identifying genetic sex of Penaeus japonicus and application thereof | |
| CN113481320B (en) | Site linked with pear peel red character, molecular marker and application thereof | |
| CN109868327A (en) | A method of the muskmelon cenospecies sweet tea ninth-seeded Purity based on EST-SSR label | |
| CN105420354B (en) | Conventional rice variety Huaihe River rice No. 5 and Huaihe River rice No. 18 identification methods based on InDel label | |
| CN109868325A (en) | A method of the crisp sweet tea Purity Identification in muskmelon cenospecies east based on EST-SSR label | |
| CN108165652A (en) | For the specific molecular marker TGMI001 of Chinese torreya seedling stage sex identification | |
| CN109868326A (en) | A method of 100 Purity Identification of muskmelon cenospecies saliva sweet tea based on EST-SSR label | |
| CN102586459B (en) | Method for identifying varieties of Chinese cabbage by using composite expressed sequence tag-simple sequence repeat (EST-SSR) marker |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180126 |
|
| WD01 | Invention patent application deemed withdrawn after publication |