CN108384873A - SSR marker and method for the green phoenix hybrid seed purity identification of watermelon - Google Patents
SSR marker and method for the green phoenix hybrid seed purity identification of watermelon Download PDFInfo
- Publication number
- CN108384873A CN108384873A CN201810160859.7A CN201810160859A CN108384873A CN 108384873 A CN108384873 A CN 108384873A CN 201810160859 A CN201810160859 A CN 201810160859A CN 108384873 A CN108384873 A CN 108384873A
- Authority
- CN
- China
- Prior art keywords
- watermelon
- green phoenix
- phoenix
- green
- dna
- 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
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
The present invention provides a kind of SSR markers and method for the green phoenix hybrid seed purity identification of watermelon.The specific steps are with the CTAB methods of improvement extract respectively green phoenix male parent, female parent and F1 seedling leaves to be measured genomic DNA；Agarose gel electrophoresis and spectrophotometer detect the quality and concentration of put forward DNA；Using the DNA of said extracted as template, PCR amplification is carried out with SSR marker ClSSR01623；Polyacrylamide gel electrophoresis is carried out to the product of amplification；Electrophoresis result is analyzed.The heterozygosis band 119/110bp containing Parent banding pattern simultaneously is amplified in the green phoenix of Hybrid, the purity for the identification green phoenix of watermelon hybrid kind that can be apparent from.The method of the present invention establishes a kind of method for quickly, efficiently identifying the green phoenix Purity Identification of watermelon hybrid kind, greatly shortens qualification time, saves appraisal cost, improves identification accuracy, has stronger commercial application value.
Description
Technical field
The present invention relates to a kind of SSR markers and its method for the green phoenix hybrid seed purity identification of variety of watermelon, belong to
In biotechnology.
Background technology
Watermelon（Citrullus lanatus(Thunb.) Matsum. & Nakai）Belong to Curcurbitaceae
(Cucurbitaceae) Citrullus（Citrullus）, it is cross-pollinatd plant, fruit is tender and crisp, sweet succulence, and nutrition is rich
Richness is rich in several mineral materials and vitamin, extensively in various regions cultivating and growing.China is the maximum producing country of watermelon and country of consumption
（Ren et al., 2014）, since the 80's of 20th century, watermelon hybrid kind is promoted in China's rapid proliferation, cultivates face
Product expands year by year, it has also become one of the important crops increased farmers' income and promote rural economic development（Liu Ziji etc., 2016;
Zhou Xianlin etc., 2011）.
Seed purity is the important indicator of seed quality, and seed purity belongs to seed inferior below 95%, can be substantially reduced
The yield and quality of crop causes heavy losses to production（Zhou Xianlin etc., 2011;Li Li etc., 2015）.Therefore, watermelon
Cenospecies seed must be by purity detecting before packing and selling, this is to ensureing seed quality and improving productivity effect with important
Meaning.Currently watermelon Purity main method is traditional field plot field plot test method, and the entire qualification process period is long, takes
When take a lot of work, floor space it is larger, and it is easily affected by environment cause qualification result inaccurate, current year's production needs (side cannot be met
Refined osmanthus etc., 2009;When Luan Yu etc., 1998).
In recent years, with the rapid development of sequencing technologies so that detecting purity of hybrid from genomic level becomes
It may.Molecular marking technique can detect the fine difference between filial generation and parent from DNA levels, not limited by space-time, can be with
Greatly shorten detection time（Tan Meilian etc., 2012）.In eukaryotic gene group, there is SSR labels quantity to enrich, be distributed
Uniformly, easy to operate, codominant inheritance, stability are good etc. advantage, is the ideal mark type for carrying out Hybrid seed purity test.
Watermelon genome has been completed to be sequenced, and has the SSR marker much developed.All prominent personages etc.（2012）With 3 watermelon hybrid generation product
Kind and its parent are test material, and Rapid identification has been carried out to hybrid water melon purity using EST-SSR labels.Liu Ziji etc.
Purity is done to small watermelon kind " the U.S. moon " with SSR marker, qualification result is consistent with field test.Molecular marking technique
Efficient, the accurate detection variety of watermelon purity of developing into technical support is provided.
Green phoenix is our company（Jiangsu Lv Gang modern agricultural developments Co., Ltd）The first cross kind of independent research, possesses
It is female parent that the new varieties of entirely autonomous intellectual property, which are with " L55 ", and " F92 " is the yellow flesh gift type watermelon of precocity that male parent is bred as
Kind.The kind field growing gesture is stronger, easily bears fruit, in resistance, disease resistance, suitable for Jiangsu Province and similar ecological condition area
Cultivation.In recent years, green phoenix gradually becomes gift watermelon main breed, and cultivated area expands year by year.Therefore, developing can be fast
Speed efficiently identifies that the SSR molecular marker technology of green phoenix seed purity will ensure the maximization of economic benefit of the improved seeds, has
There is higher commercial application prospect.
Invention content
In order to overcome the qualification cycle of hybrid water melon purity it is long, it is time-consuming, take up a large area, it is easily affected by environment etc.
The deficiency of field plot field plot test method, the purpose of the present invention is to provide for the green phoenix hybrid seed purity mirror of variety of watermelon
Fixed SSR primers, and establish a kind of side quickly, efficiently, accurately identifying the green phoenix hybrid seed purity identification of variety of watermelon
Method.
The present invention expands parent DNA using 32 pairs of watermelon SSR markers of selection, electrophoresis, and screening has polymorphism
Primer is successfully chosen and arrives SSR marker ClSSR01623, forward primer sequence：
5'- AGAGGAAACCTTGCAGCTTG -3', reverse primer sequences：5'- TCCTTCCTGTGGATGTAGGC -3',
The banding pattern of green phoenix Parent can be used for clearly distinguishing（Fig. 1）, and the pcr amplification product of Parent is sequenced, it is sequenced
As a result（Fig. 2）, find the sequence that 110bp is amplified in female parent（SEQ ID N0.3）, the sequence of 119bp is amplified in male parent
（SEQ ID N0.4）.
Provided by the present invention for the method for the green phoenix Purity Identification of watermelon hybrid kind, it is as follows：
（1）Extract respectively green phoenix male parent, female parent and F1 seedling leaves to be measured genomic DNA；
（2）The quality and concentration of put forward DNA are detected with agarose gel electrophoresis and spectrophotometer
（3）Using the DNA of said extracted as template, SSR primers ClSSR01623 is selected to carry out PCR amplification；
（4）Polyacrylamide gel electrophoresis is carried out to the product of amplification；
（5）Electrophoresis result is analyzed：
The SSR standard diagrams of green phoenix are while tool is there are two the collection of illustrative plates of parent's specific band, and 119bp is amplified in male parent
Specific band, the specific band of 110bp is amplified in maternal.The electrophoresis pattern of detected seed and green phoenix standard
When collection of illustrative plates is consistent, it is accredited as green phoenix.
In the present invention, PCR amplification uses 25 μ l reaction systems, including 12.5 μ 2 × Taq of l MasterMix, μ l DNA to carry
Take object, 10 μM of forward and reverse primer each 1 μ l, 9.5 μ l ddH2O.The program of PCR amplification is：94 DEG C of pre-degeneration 2min, [94 DEG C of changes
Property 30s, 55 DEG C annealing 30s, 72 DEG C extension 30s], 35 cycle, 72 DEG C extension 2min, then reaction be maintained at 16 DEG C.
In the present invention, the detection method of the electrophoresis is：3 μ l loading Buffer are added in 25 μ l PCR products, take
1 μ l are splined on 8% native polyacrylamide gel electrophoresis, silver staining detection.
Beneficial effects of the present invention：The SSR marker ClSSR01623 that the present invention filters out, amplifies 119bp in male parent
Specific band, amplify the specific band of 110bp in maternal, and amplify in the green phoenix of Hybrid and contain simultaneously
The heterozygosis band 119/110bp of Parent banding pattern, and band is clear, genetic stability, can by the green phoenix of watermelon hybrid kind with
Its Parent seed, other accessory seeds distinguish, and quickly detect the purity of the green phoenix of watermelon hybrid kind.By this hair
It is bright, water melon leaf genomic DNA need to be simply only extracted, then carries out PCR amplification, polyacrylamide gel electrophoresis, you can effectively
Identification the green phoenix of watermelon hybrid kind purity.The invention avoids traditional field plot qualification cycle it is long, it is time-consuming, account for
The big equal deficiency of ground area, the method that can substitute conventional hybridization Purity Identification greatly shorten qualification time, save identification
Cost improves identification accuracy, has stronger commercial application value.
Description of the drawings
Fig. 1 is the electrophoretogram of green phoenix parent and Hybrid that SSR marker amplification is carried out using ClSSR01623 primers.
Fig. 2 is green phoenix Parent sequencing result.
Wherein：M indicates the Marker of 500bp in Fig. 1.In Fig. 2 green phoenix Parent respectively sequencing twice, be expressed as 1,
2；Wherein preceding 20 bases are preceding primer sequence, and rear 20 bases are rear primer reverse sequence.
Specific implementation mode
With reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto,
Protection scope of the present invention cannot be limited with following embodiments.
Experimental method used in following embodiments is conventional method unless otherwise specified；Used material,
Reagent etc., is commercially available unless otherwise specified.
（One）Materials and methods
1. vegetable material
This research is experiment material using the green phoenix of watermelon hybrid kind and its maternal, male parent seed, existing according to Jiangsu Lv Gang
For agricultural development company（Referred to as green port）Experimental method is sowed and is managed.
2. the extraction and detection of DNA
Using the CTAB methods of improvement, watermelon genomic DNA is extracted from 21 days or so fresh plant blades.The CTAB of the improvement
Method includes the following steps：
1）It takes fresh blade 30mg to be put into the centrifuge tube of 2ml, the steel ball of a diameter 4mm is added, covers tightly lid, puts it into
Then 2min in liquid nitrogen utilizes tissue grinder instrument grind away；
2）600 μ l CTAB, 55 DEG C of water-bath 15min are added in the sample of milled；
3）12000rpm centrifuges 5min, draws 500 μ l supernatants in the centrifuge tube of clean 1.5ml, 250 μ l chloroforms and different are added
Amyl alcohol mixture mixes well；The volume ratio of chloroform and isoamyl alcohol is 24 in chloroform and iso pentane alcohol mixture：1；
4）13000rpm centrifuges 90s, takes 350 μ l supernatants in another new 1.5ml centrifuge tubes, and addition carries under the conditions of -20 DEG C
600 μ l of absolute ethyl alcohol of preceding precooling, 60 μ l ammonium acetates are added, mixing puts -20 DEG C of refrigerator 1h；
5）13000rpm centrifuges 5min, outwells supernatant, is placed at room temperature for；
6）When alcohol-free taste in centrifuge tube, 100 μ l ddH are added2O dissolving DNAs.
The quality and concentration of agarose gel electrophoresis and spectrophotometer detection DNA.
3. PCR amplification and detection
PCR amplification carries out on Chinese Hua Sheng companies LY96G ThermocyclerTM amplification instruments, and reaction system is 25 μ L, including
12.5 μ 2 × Taq of l MasterMix, 1 μ l DNA extracts, 10 μM of forward and reverse primer each 1 μ l, 9.5 μ l ddH2O.PCR expands
The program of increasing is：94 DEG C of pre-degeneration 2min, [94 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 30s], 35 cycles, 72 DEG C
Extend 2min, then reaction is maintained at 16 DEG C.The segment that PCR amplification is detached on 8% polyacrylamide gel electrophoresis, carries out silver
Dye, and show under white light.
4. label screening and Purity with polymorphism
Choose 32 pairs of watermelon SSR markers（Zhu et al., 2016）, there is polymorphism primer using parental line selection.Pass through label
The amplification of parent DNA is carried out, polyacrylamide gel electrophoresis is carried out, filters out the label that can obviously distinguish Parent, as
There is the label of polymorphism.
With the label with polymorphism screened, the amplification of the green phoenix of watermelon hybrid kind is carried out, its purity mirror is carried out
It is fixed.
（Two）As a result with analysis
1. the detection and analysis of the genomic DNA of selected materials
Use ultramicron ultraviolet-uisible spectrophotometer（DeNovix DS-11）To 96 of the extraction of this research institute（Wherein parent
This each 1, F1 groups 94）Water melon leaf genomic DNA carries out Concentration Testing, it is found that the DNA concentration of extraction is above 100ng
/ μ l, and OD260/OD280 is substantially between 1.8 ~ 2.0.Various concentration DNA is taken to be detected with 2% agarose electrophoresis, electricity
Swimming figure is clear bright, without apparent hangover, illustrates that the DNA mass of extraction is preferable.So the high quality water melon leaf gene extracted
Group DNA is suitable for PCR amplification, the biological test that SSR equimoleculars mark.
2. PCR amplification and Purity
Parent DNA is expanded using 32 pairs of watermelon SSR markers of selection, electrophoresis, screening has the primer of polymorphism.Success is selected
SSR marker ClSSR01623 is got, can be used for clearly distinguishing the banding pattern of green phoenix Parent（Fig. 1）, and to the PCR of Parent
Amplified production is sequenced, and sequencing result is shown in Fig. 2, finds the sequence that 110bp is amplified in female parent（SEQ ID N0.3）, father
The sequence of 119bp is amplified in this（SEQ ID N0.4）.
The sequence of above-mentioned SSR marker ClSSR01623 is：Forward primer sequence：5'- AGAGGAAACCTTGCAGCTTG -
3'（SEQ ID N0.1）, reverse primer sequences：5'- TCCTTCCTGTGGATGTAGGC - 3'（SEQ ID N0.2）
Green phoenix germline SEQ ID N0.3：
agaggaaaccttgcagcttgaagaaatcaaagaagaagaagaagaaga
agaagaactaagacagtgttttacgtggttcggctaagatcagcctacatccacaggaagga
Green phoenix male parent sequence SEQ ID N0.4：
agaggaaaccttgcagcttgaagaaatcaaagaagaagaagaagaaga
agaagaagaagaagaactaagacagtgttttacgtggttcggctaagatcagcctacatccacaggaagga
The green phoenix group of watermelon hybrid kind is expanded using ClSSR01623 labels and carries out Purity, the result is shown in Figure 1, in figure
It can clearly find out, the specific band of 119bp is amplified in male parent, the specific band of 110bp is amplified in maternal,
The electrophoresis pattern for being detected seed is consistent with green phoenix standard diagram, is while having the band 119/ there are two parent's specificity
110bp（Fig. 1）, show that the green phoenix purity of watermelon hybrid kind has reached 100%, SSR marker ClSSR01623 used can be efficient
The purity of the stable identification green phoenix of watermelon hybrid kind.
<110>Jiangsu Lv Gang modern agricultural developments Co., Ltd
<120>SSR marker and method for the green phoenix hybrid seed purity identification of watermelon
<140>201810160859.7
<160> 4
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence
<400> 1
agaggaaacc ttgcagcttg 20
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence
<400> 2
tccttcctgt ggatgtaggc 20
<210> 3
<211> 110
<212> DNA
<213>Artificial sequence
<400> 3
agaggaaacc ttgcagcttg aagaaatcaa agaagaagaa 40
gaagaagaag aagaactaag acagtgtttt acgtggttcg 80
gctaagatca gcctacatcc acaggaagga 110
<210> 4
<211> 119
<212> DNA
<213>Artificial sequence
<400> 4
agaggaaacc ttgcagcttg aagaaatcaa agaagaagaa 40
gaagaagaag aagaagaaga agaactaaga cagtgtttta 80
cgtggttcgg ctaagatcag cctacatcca caggaagga 119
Claims (5)
1. the SSR marker for the green phoenix hybrid seed purity identification of watermelon, which is characterized in that including forward primer sequence：5'-
AGAGGAAACCTTGCAGCTTG -3', reverse primer sequences：5'-
TCCTTCCTGTGGATGTAGGC - 3'。
2. a kind of being used for the SSR marker method of the green phoenix hybrid seed purity identification of watermelon using primer described in claim 1,
It is characterized in that, being carried out by following step：
（1）With the CTAB methods of improvement extract green phoenix male parent, female parent and F1 seedling leaves to be measured genomic DNA；
（2）The quality and concentration of put forward DNA are detected with agarose gel electrophoresis and spectrophotometer；
（3）Using the DNA of said extracted as template, SSR primers ClSSR01623 is selected to carry out PCR amplification；
（4）Polyacrylamide gel electrophoresis is carried out to the product of amplification；
（5）Electrophoresis result is analyzed.
3. according to the SSR marker method for the green phoenix hybrid seed purity identification of watermelon described in claim 2, feature exists
In the step（3）The reaction system of middle PCR amplification is 25 μ L, including 12.5 μ 2 × Taq of l MasterMix, 1 μ l DNA are carried
Take object, 10 μM of forward and reverse primer each 1 μ l, 9.5 μ l ddH2O；The program of PCR amplification is：94 DEG C of pre-degeneration 2min, [94 DEG C of changes
Property 30s, 55 DEG C annealing 30s, 72 DEG C extension 30s], 35 cycle, 72 DEG C extension 2min, then reaction be maintained at 16 DEG C.
4. special according to the SSR marker method for the green phoenix hybrid seed purity identification of watermelon described in claim 2 or 3
Sign is, polyacrylamide gel electrophoresis, non-denaturing polyacrylamide gel a concentration of 8% are carried out to pcr amplification product；In 25 μ
3 μ l loading Buffer are added in l PCR products, and 1 μ l is taken to be splined on 8% native polyacrylamide gel electrophoresis, silver staining inspection
It surveys.
5. a kind of SSR marker for the green phoenix hybrid seed purity identification of watermelon utilized as described in claim 2~4 is any
Method, which is characterized in that the SSR standard diagrams of green phoenix are while having the figure there are two parent's specific band 110/119bp
Spectrum amplifies the specific band of 119bp in male parent, the specific band of 110bp is amplified in maternal, is detected seed
When electrophoresis pattern is consistent with green phoenix standard diagram, it is accredited as green phoenix.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810160859.7A CN108384873A (en) | 2018-02-27 | 2018-02-27 | SSR marker and method for the green phoenix hybrid seed purity identification of watermelon |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810160859.7A CN108384873A (en) | 2018-02-27 | 2018-02-27 | SSR marker and method for the green phoenix hybrid seed purity identification of watermelon |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108384873A true CN108384873A (en) | 2018-08-10 |
Family
ID=63069854
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810160859.7A Pending CN108384873A (en) | 2018-02-27 | 2018-02-27 | SSR marker and method for the green phoenix hybrid seed purity identification of watermelon |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108384873A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109234437A (en) * | 2018-10-31 | 2019-01-18 | 宁夏泰金种业股份有限公司 | A kind of watermelon hybrid seed Purity method based on SSR marker method |
| CN111549172A (en) * | 2020-06-12 | 2020-08-18 | 中国农业科学院郑州果树研究所 | Watermelon leaf posterior green gene linkage site and CAPS marker |
| CN114853856A (en) * | 2021-02-03 | 2022-08-05 | 北京市农林科学院 | Application of ClZISO gene in preparation of yellow-flesh watermelons and application of ClZISO gene in identification of yellow-flesh watermelons |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104711361A (en) * | 2015-03-26 | 2015-06-17 | 浙江省农业科学院 | Method for quickly identifying purity of new watermelon species namely red peace hybrid seeds as well as primer and kit adopted by method |
| CN105154550A (en) * | 2015-09-11 | 2015-12-16 | 山东省华盛农业股份有限公司 | Method for quickly identifying purity of watermelon variety Zhentian 1217 by using EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker |
-
2018
- 2018-02-27 CN CN201810160859.7A patent/CN108384873A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104711361A (en) * | 2015-03-26 | 2015-06-17 | 浙江省农业科学院 | Method for quickly identifying purity of new watermelon species namely red peace hybrid seeds as well as primer and kit adopted by method |
| CN105154550A (en) * | 2015-09-11 | 2015-12-16 | 山东省华盛农业股份有限公司 | Method for quickly identifying purity of watermelon variety Zhentian 1217 by using EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker |
Non-Patent Citations (2)
| Title |
|---|
| HUAYU ZHU 等: "Genome wide characterization of simple sequence repeats in watermelon genome and their application in comparative mapping and genetic diversity analysis", 《BMC GENOMICS》 * |
| 李超汉 等: "基于SSR标记的5个西瓜新品种纯度鉴定及特异性分析的研究 ", 《中国农学通报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109234437A (en) * | 2018-10-31 | 2019-01-18 | 宁夏泰金种业股份有限公司 | A kind of watermelon hybrid seed Purity method based on SSR marker method |
| CN111549172A (en) * | 2020-06-12 | 2020-08-18 | 中国农业科学院郑州果树研究所 | Watermelon leaf posterior green gene linkage site and CAPS marker |
| CN111549172B (en) * | 2020-06-12 | 2023-02-28 | 中国农业科学院郑州果树研究所 | Watermelon leaf posterior green gene linkage site and CAPS marker |
| CN114853856A (en) * | 2021-02-03 | 2022-08-05 | 北京市农林科学院 | Application of ClZISO gene in preparation of yellow-flesh watermelons and application of ClZISO gene in identification of yellow-flesh watermelons |
| CN114853856B (en) * | 2021-02-03 | 2023-07-07 | 北京市农林科学院 | Application of ClZISO gene in preparation of yellow pulp watermelons and application of ClZISO gene in identification of yellow pulp watermelons |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106755481B (en) | SSR molecular marker VI for identifying progeny plants of Gala apples and application thereof | |
| CN104711361B (en) | The method of the red peaceful hybrid seed purity of Rapid identification new water melon breed and the primer and kit of use | |
| CN105154550B (en) | A kind of method using 1217 purity of EST-SSR molecular labeling Rapid identification variety of watermelon treasure sweet tea | |
| CN109762920B (en) | SNP marker tightly linked with cucumber fruit thorny gene ns and application thereof | |
| CN108342465A (en) | A method of crop rhizosphere prokaryotic micro-organisms is detected based on high-flux sequence | |
| CN108546774A (en) | A kind of SSR primers and identification method for purple chili Hybrid Purity | |
| CN108384873A (en) | SSR marker and method for the green phoenix hybrid seed purity identification of watermelon | |
| CN105543222B (en) | The molecular labeling InDeL_33 of soybean 100-grain weight main effect QTL and its application | |
| CN106521021B (en) | Gene marker for identifying haplotype of rice grain width and grain weight GS5 gene and application | |
| CN104928396A (en) | Method for rapidly identifying hot pepper species and golden pepper purity degree by using EST-SSR molecular markers | |
| CN108384879A (en) | A kind of SSR primers and method for watermelon hybrid object innovation | |
| CN113151567B (en) | SSR molecular marker and method for identifying Lepista sordida N006# strain | |
| CN105821154A (en) | SSR primers and method for purity identification of luffa hybrid seeds | |
| CN108796119A (en) | Specific molecular marker PCMI-M001 for Cathay poplar male seedling Rapid identification | |
| CN102226189B (en) | Seed number per pod character major gene site of rape and application thereof | |
| CN109880932B (en) | SSR primer pair for identifying cotton 1279 in hybrid cotton variety, product thereof and identification method | |
| CN107058494A (en) | Simplify the method for Jian Kuo Foreign Banks' Entries Purities using SCoT molecular labelings | |
| CN108165652A (en) | For the specific molecular marker TGMI001 of Chinese torreya seedling stage sex identification | |
| CN103014151A (en) | Rapid detection method for purity of cucumber seeds | |
| CN108531636A (en) | A kind of molecular marked compound TJcM01 and its application for identifying muskmelon unisexual flower | |
| CN108754015A (en) | Specific molecular marker PCMI-F001 for Cathay poplar female seedling Rapid identification | |
| CN113943732A (en) | SNP (Single nucleotide polymorphism) marker related to heat resistance of cucumber in adult stage, primer group, kit and application | |
| CN105420235B (en) | Cucumber fruits paliform epidermal cell gene Pe Indel marks and its application | |
| CN106244683B (en) | Primer for detecting " Qiong Li " small watermelon purity of hybrid combines and its methods and applications | |
| CN110229929A (en) | Identify specific molecular marker and its application of the alfalfa seed true and false |
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 | ||
| CB02 | Change of applicant information |
Address after: Jiangsu green port agricultural park, No.1 Buzhang Road, Nancai Township, Sucheng District, Suqian City, Jiangsu Province 223800 Applicant after: JIANGSU GREENPORT MODERN AGRICULTURAL DEVELOPMENT Co.,Ltd. Address before: Jiangsu green port agricultural park, No.1 Buzhang Road, Nancai Township, Sucheng District, Suqian City, Jiangsu Province 223800 Applicant before: JIANGSU GREENPORT MODERN AGRICULTURAL DEVELOPMENT Co.,Ltd. |
|
| CB02 | Change of applicant information | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180810 |
|
| WD01 | Invention patent application deemed withdrawn after publication |