CN103740838A - Rapid purity identification method for indica-japonica hybrid rice seeds - Google Patents
Rapid purity identification method for indica-japonica hybrid rice seeds Download PDFInfo
- Publication number
- CN103740838A CN103740838A CN201410023390.4A CN201410023390A CN103740838A CN 103740838 A CN103740838 A CN 103740838A CN 201410023390 A CN201410023390 A CN 201410023390A CN 103740838 A CN103740838 A CN 103740838A
- Authority
- CN
- China
- Prior art keywords
- seed
- purity
- hybrid rice
- quick
- rice seeds
- 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 invention provides a rapid purity identification method for indica-japonica hybrid rice seeds. The rapid purity identification method comprises the steps of direct extraction of seed DNA; polymerase chain reaction (PCR) amplification reaction, wherein two selections are available for one pair of SSR primers in the PCR reaction system, sequences for one selection are as follows: forward primer 5'-CCAAAGATGAAACCTGGATTG-3' and reverse primer 5'-GCACAAGGTGAGCAGTCC-3', and sequences for the other selection are as follows: forward primer 5'-CAAAAACAGAGCAGATGAC-3' and reverse primer 5'-CTCAAGATGGACGCCAAGA-3'; gel electrophoresis detection; seed purity calculation. The identification method disclosed by the invention has the characteristics of being accurate, simple and rapid.
Description
Technical field
The present invention relates to seed detection technique field, refer to especially a kind of quick Purity method of Xian round-grained rice friendship type hybrid rice seeds.
Background technology
Indica-Japonica Hybrid Rice is to utilize paddy rice indica-japonica hybrid to gather the good strong advantage Hybrid Rice Combinations of subspecies beneficial gene seed selection Comprehensive Traits, it is the strategic emphasis of China's breeding of hybridized rice research, our province grain aggregate will stablized, ensure grain ration safety, promote peasant's increasing both production and income to play a significant role.
Xian round-grained rice is handed in rice paddy seed and is contained much starch (65%-70%), protein (7%-9%) and fat (2%-3%), and the existence of these materials has affected the quality of the DNA directly extracting in seed to a great extent.Now traditional way is that seed is carried out to standard germination, waits seedling to grow to the DNA that extracts seed after a certain size in seedling leaves, and this method has extended the cycle of DNA extraction greatly, thereby has reduced rapidly and efficiently property of indoor molecular markers for identification seed purity.
Seed authenticity and purity are the core index of Seed Quality of Hybrid Rice, it is the main standard of seed quality classification, it is the key foundation of enterprise's purchase and sale seed, improved seeds impurity of seeds, purity drop will hinder giving full play to of the good heritability of kind, cause the obvious reduction of crop yield and quality, not only to having an immense impact on, also will cause heavy losses to country, government, peasant.Along with increasing and breeding material hereditary basis day by day narrow of crop Cultivars number, make interracial hereditary difference more and more less, cause the difficulty of variety authentication and Purity to continue to increase.Therefore, set up a set of easy, quick, economical, accurately variety authentication and Purity technical system be protection breeder intellecture property, hit fake and forged seed, standard Seed Market and promote that China's seed industry takes part in international competition in the urgent need to.DNA molecular marker has reflected the hereditary difference between biont on DNA level, has that quantity is abundant, polymorphism is high, not affected by environment, detects the advantage such as quick and be applied gradually (Li Zhaohua etc., 2006).Wherein, SSR (simple sequence repeat) mark, because polymorphism is good, repeatability is high, easy and simple to handle, is considered to the good DNA fingerprint technology of a kind of development prospect.SSR molecule marker is to equal by Moore a kind of molecule marker (Moore etc. based on specific primer PCR technology that create for 1989,1991), belong to codominant marker, there is rich polymorphism, simple to operate, the advantage such as reproducible, detected result accurately and reliably, reproducible, easy and simple to handle, quick, and not high to DNA quantity and specification of quality, be widely used in the aspects such as genetic map construction, the research of icp gene group, analysis of genetic diversity and Idioplasm identification, be one of at present most popular DNA fingerprinting technology.The application of SSR molecular marking technique in Purity Identification of Hybrid Rice Seeds, is applied among a small circle seed purity detection and puts into practice (Li Zhaohua etc., 2006).Peng Suotang etc. (2003) have carried out SSR evaluation with primer RM17 to Hybrid Rice Combination Shanyou 63 and two line system; Nandakumar etc. (2004) utilize SSR molecule marker successfully to build 4 rice varieties and parent's thereof finger printing, and successfully for the Purity of kind.Xiao little Yu etc. (2006) filter out from 208 pairs of primers that to have the primer 123 of polymorphism right, have set up the DNA fingerprint database of Sichuan Province's Main Parents of Hybrid Rice.This research has been carried out the optimization of technical system from aspects such as DNA extraction, PCR reaction system and program and gel electrophoresises, has set up the technical system of Rapid identification Xian round-grained rice friendship rice seed purity accurate, simple, quick, with low cost.
Summary of the invention
The present invention proposes a kind of quick Purity method of Xian round-grained rice friendship type hybrid rice seeds, solved the problem of seed authenticity and purity in prior art.
Technical scheme of the present invention is achieved in that
A kind of quick Purity method of Xian round-grained rice friendship type hybrid rice seeds, comprising:
(1) the direct extraction of seed DNA (without germinateing);
(2) pcr amplification reaction;
In the system of described PCR reaction, 1 pair of SSR primer has two kinds of selections, and a kind of Selective sequence is forward primer 5 '-CCAAAGATGAAACCTGGATTG-3 ' and reverse primer
5’-GCACAAGGTGAGCAGTCC-3’；
Another kind of Selective sequence is forward primer 5 '-CAAAAACAGAGCAGATGAC-3 ' and reverse primer 5 '-CTCAAGATGGACGCCAAGA-3 ';
(3) detected through gel electrophoresis;
(4) seed purity is calculated;
The seed that has father and mother's feature master tape concurrently is target seed, counting;
Calculation formula is as follows: examined seed lot seed purity (%)=examined and in seed, have Parent feature banding pattern seed number/examined seed sum × 100 concurrently.
As preferred technical scheme, the step of the direct extraction of described step (1) seed DNA is as follows: after handing over hybrid rice seeds to peel off bran shell Xian round-grained rice, put into centrifuge tube; Organizing the 30s that polishes on sample grinding machine; Successively add 400 μ l extracting solutions and 120 μ l5mol/L Potassium ethanoates; After mixing under 4 ℃ of environment standing 10-15min; The centrifugal 10min-15min of 12000rpm at 25 ℃; Get supernatant, successively add dehydrated alcohol 400 μ l and 3mol/L sodium-acetate 15 μ l; At 25 ℃, after the centrifugal 10min-15min of 12000rpm, remove supernatant; Add 400 μ l70% absolute ethanol washings; The centrifugal 4min-6min of 12000rpm at 4 ℃; Remove supernatant, after drying, add 100 μ l distilled waters to dissolve ,-20 ℃ of preservations.
As preferred technical scheme, described extracting solution is by 1mol/LTris-HCl, 5mol/LNaCl, and 10% sodium lauryl sulphate, 0.5mol/L quadrol tetrem disodium salt and distilled water form.
As preferred technical scheme, the concrete steps of described step (2) pcr amplification reaction are as follows: (DNApolymerasemixture, containing Taq enzyme, dNTP and Mg in 96 hole PCR plates, to put into respectively DNA profiling, Mixed mixed solution
2+deng), the 1 pair of SSR primer and distilled water; First 94 ℃ of sex change 3min; 94 ℃ of sex change 30s again, 55 ℃ of annealing 45s, 72 ℃ are extended 1min, 30 circulations; 72 ℃ are extended 5min, 4 ℃ of preservations.
As preferred technical scheme, the concrete steps of described step (3) detected through gel electrophoresis are as follows: mounting glass plate is on electrophoresis chamber; The glue configuring is shaken up, and encapsulating, inserts comb, waits the rear point sample of gelling knot; In electrophoresis chamber, add damping fluid, 110V, the about 2h of electrophoresis; After lower glue, gel is immersed in and in staining fluid, contaminates 10min; With distilled water flushing gel twice, 5min develops in developing solution; Rinse after twice of gel Taking Pictures recording in gel electrophoresis imaging system instrument with tap water.
As preferred technical scheme, in described electrophoresis chamber, damping fluid is 1 × TBE, by Tutofusin tris, boric acid and 0.5mol/L quadrol tetrem disodium salt, is mixed.
As preferred technical scheme, described glue is mixed by 5 × TBE, 40% acrylamide, 10% ammonium persulphate, Tetramethyl Ethylene Diamine, distilled water, and their mixed volume ratio is 8: 6: 0.3: 0.024: 26.
Accompanying drawing explanation
In order to be illustrated more clearly in embodiment of the present invention or technical scheme of the prior art, to the accompanying drawing of required use in embodiment or description of the Prior Art be briefly described below, apparently, accompanying drawing in the following describes is only embodiments more of the present invention, for those of ordinary skills, do not paying under the prerequisite of creative work, can also obtain according to these accompanying drawings other accompanying drawing.
Fig. 1 is indica-japonica hybrid paddy rice spring excellent 84 and parent's thereof key band.Wherein, numbering 1-4 is maternal key band (80bp), and numbering 5-28 is F1 seed characteristics bands of a spectrum (having Parent band 80bp and 120bp concurrently), and numbering 29-32 is male parent key band (120bp).
Embodiment
To the technical scheme in the embodiment of the present invention be clearly and completely described below, obviously, described embodiment is only the present invention's part embodiment, rather than whole embodiment.Based on the embodiment in the present invention, those of ordinary skills, not making the every other embodiment obtaining under creative work prerequisite, belong to the scope of protection of the invention.
Embodiment 1
A kind of quick Purity method of Xian round-grained rice friendship type hybrid rice seeds, comprising:
Step S1: DNA extraction in seed
1, by centrifuge tube numbering, from submitted sample, choose at random clean seed, put into respectively centrifuge tube, be sure not many of pipes or blank pipe;
2, add 400 μ lSDS extracting solutions, extracting solution is by 1MTris-Hcl60 μ l, 5MNacl40 μ l, and 10%SDS80 μ l, 0.5MEDTA80 μ l and distilled water 140 μ l mix;
3, organizing the 30s that polishes on sample grinding machine, add 120 μ l5MKac, shake up the quiet 10-15min that carries on ice bath or under 4 ℃ of environment;
4, the centrifugal 15min of 12000 turn/min at 25 ℃, gets supernatant;
5, add dehydrated alcohol 400 μ l, 3MNaAc15 μ l, the centrifugal 8min of 12000 turn/min at 25 ℃;
6, outwell gently liquid, add 400 μ l70% dehydrated alcohols to rock washing and stick to DNA and the mixtures of impurities on centrifuge tube;
7, the centrifugal 5min of 12000 turn/min at 4 ℃, outwells supernatant, and centrifuge tube is inverted on paper handkerchief and is dried;
8, add 100 μ l distilled water dissolving DNAs after drying, be placed in-20 ℃ of preservations.
Step S2:PCR amplified reaction
1, reaction system (10 μ l): in 96 hole PCR plates, put into respectively 1 μ lDNA template, 5 μ lMixed mixed solutions (DNApolymerasemixture, containing Taq enzyme, dNTP and Mg2+ etc.), 0.25 μ lSSR forward primer, 0.25 μ lSSR reverse primer and 3.5 μ l distilled waters.
2, primer sequence: forward primer 5 '-CCAAAGATGAAACCTGGATTG-3 ' and reverse primer 5 '-GCACAAGGTGAGCAGTCC-3 '; Another kind of Selective sequence is forward primer 5 '-CAAAAACAGAGCAGATGAC-3 ' and reverse primer 5 '-CTCAAGATGGACGCCAAGA-3 '.
3, amplification program: first 94 ℃ of sex change 3min; 94 ℃ of sex change 30s again, 55 ℃ of annealing 45s, 72 ℃ are extended 1min, 30 circulations; 72 ℃ are extended 5min, 4 ℃ of preservations.
Step S3: gel electrophoresis
1, with alcohol, firmly clean sheet glass, after being dried, assemble and be installed on electrophoresis chamber;
2, glue encapsulating: during the preparation of polyacrylamide gel, each piece glue glue is mixed by 4ml5 × TBE, 3ml40% acrylamide, 150 μ l10%APS, 12 μ lTEMED, 13ml distilled water.To after liquid mixing, shake up encapsulating immediately, polymerization time 40min to 1h;
3, in electrophoresis chamber, add 1 × tbe buffer liquid, point sample 1.5-2.5 μ l, 110V electrophoresis, approximately 2h;
4, silver dyes: gel is immersed in and is equipped with 0.2%AgNO
3shaking table in 10min, with distilled water rinse gel twice, then gel is immersed in developing solution (1.5%NaOH, 0.1% formaldehyde) and soaks 5min, with tap water rinse gel twice, Taking Pictures recording.
Step S4: seed lot purity is calculated
Obtain as stated above Xian japonica hybrid rice and parent's thereof feature electrophoretogram, the seed that has Parent feature master tape concurrently is target seed, counting, and calculated purity as follows: examined seed lot seed purity (%)=examined and in seed, have Parent feature banding pattern seed number/examined seed sum × 100 concurrently.
Its technique effect is: the present invention utilizes SDS rapid fractionation method directly from Xian round-grained rice, to hand over hybrid rice seed and extract DNA, then utilize a pair of SSR primer pair Xian round-grained rice to hand over Parent and first-filial generation seed DNA thereof to increase, and separate with polyacrylamide gel electrophoresis the first-filial generation complementary characteristic bands of a spectrum that obtain having concurrently Parent feature band.To thering is the seed number counting of this type of key band, and calculate seed lot purity.Through the Purity Fields of continuous 3 years, identify contrast, this method qualification result and field inspection result, without significant difference, refer to table 1.
Table 1
| Batch number | Purity Field qualification result % | Indoor Purity result % |
| 1 | 98.4 | 97.3 |
| 2 | 98.6 | 98.5 |
| 3 | 97.8 | 97.3 |
| 4 | 98.6 | 98.9 |
| 5 | 98.6 | 96.8 |
| 6 | 99 | 98 |
| 7 | 99 | 100 |
| 8 | 99.6 | 99 |
| 9 | 99.4 | 100 |
| 10 | 99.4 | 98 |
| 11 | 97.8 | 97.6 |
| 12 | 98.8 | 97.9 |
| 13 | 97 | 95.8 |
| 14 | 95.6 | 95.7 |
| 15 | 99 | 99 |
| 16 | 98.8 | 98.5 |
| 17 | 99 | 98.9 |
| 18 | 96.9 | 96 |
| 19 | 99.4 | 98.3 |
| 20 | 99.6 | 99.2 |
| 21 | 100 | 99 |
| On average | 98.6 | 98.1 |
As can be seen from Table 1, the indoor purity of most of batch is all a little less than Purity Field assay, and this may be because some hybrid strain phenotype and true plant are very approaching, artificial erroneous judgement cause.Through statistical study, result shows that indoor purity test result and field result are without significant difference.The consistence of two kinds of method of inspection results is described, this seed DNA rapid extraction and SSR molecule marker method of inspection are applicable to Xian round-grained rice and hand over the Purity of hybrid rice.
The present invention is applicable to Xian round-grained rice and hands over and directly extract DNA in hybrid rice seeds, and apply SSR molecule marker carry out verity and Purity accurately, simply, the method for inspection fast.
The foregoing is only preferred embodiment of the present invention, in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (7)
1. the quick Purity method of Xian round-grained rice friendship type hybrid rice seeds, comprising:
(1) the direct extraction of seed DNA; Wherein seed is without germination;
(2) pcr amplification reaction;
In the system of described PCR reaction, 1 pair of SSR primer has two kinds of selections, and a kind of Selective sequence is forward primer 5 '-CCAAAGATGAAACCTGGATTG-3 ' and reverse primer
5’-GCACAAGGTGAGCAGTCC-3’；
Another kind of Selective sequence is forward primer 5 '-CAAAAACAGAGCAGATGAC-3 ' and reverse primer 5 '-CTCAAGATGGACGCCAAGA-3 ';
(3) detected through gel electrophoresis;
(4) seed purity is calculated;
The seed that has father and mother's feature master tape concurrently is target seed, counting;
Calculation formula is as follows: examined seed lot seed purity (%)=examined and in seed, have Parent feature banding pattern seed number/examined seed sum × 100 concurrently.
2. the quick Purity method of a kind of Xian round-grained rice friendship type hybrid rice seeds according to claim 1, is characterized in that, the step of the direct extraction of described step (1) seed DNA is as follows: after handing over hybrid rice seeds to peel off bran shell Xian round-grained rice, put into centrifuge tube; Organizing the 30s that polishes on sample grinding machine; Successively add 400 μ l extracting solutions and 120 μ l5mol/L Potassium ethanoates; After mixing under 4 ℃ of environment standing 10-15min; The centrifugal 10min-15min of 12000rpm at 25 ℃; Get supernatant, successively add dehydrated alcohol 400 μ l and 3mol/L sodium-acetate 15 μ l; At 25 ℃, after the centrifugal 10min-15min of 12000rpm, remove supernatant; Add 400 μ l70% absolute ethanol washings; The centrifugal 4min-6min of 12000rpm at 4 ℃; Remove supernatant, after drying, add 100 μ l distilled waters to dissolve ,-20 ℃ of preservations.
3. the quick Purity method of a kind of Xian round-grained rice friendship type hybrid rice seeds according to claim 2, it is characterized in that, described extracting solution is by 1mol/LTris-HCl, 5mol/LNaCl, 10% sodium lauryl sulphate, 0.5mol/L quadrol tetrem disodium salt and distilled water form.
4. the quick Purity method of a kind of Xian round-grained rice friendship type hybrid rice seeds according to claim 1, it is characterized in that, the concrete steps of described step (2) pcr amplification reaction are as follows: in 96 hole PCR plates, put into respectively DNA profiling, Mixed mixed solution, the 1 pair of SSR primer and distilled water; First 94 ℃ of sex change 3min; 94 ℃ of sex change 30s again, 55 ℃ of annealing 45s, 72 ℃ are extended 1min, 30 circulations; 72 ℃ are extended 5min, 4 ℃ of preservations.
5. the quick Purity method of a kind of Xian round-grained rice friendship type hybrid rice seeds according to claim 1, is characterized in that, the concrete steps of described step (3) detected through gel electrophoresis are as follows: mounting glass plate is on electrophoresis chamber; The glue configuring is shaken up, and encapsulating, inserts comb, waits the rear point sample of gelling knot; In electrophoresis chamber, add damping fluid, 110V, electrophoresis 2h; After lower glue, gel is immersed in and in staining fluid, contaminates 10min; With distilled water flushing gel twice, 5min develops in developing solution; Rinse after twice of gel Taking Pictures recording in gel electrophoresis imaging system instrument with tap water.
6. the quick Purity method of a kind of Xian round-grained rice friendship type hybrid rice seeds according to claim 1, it is characterized in that, in described electrophoresis chamber, damping fluid is 1 × TBE, by Tutofusin tris, boric acid and 0.5mol/L quadrol tetrem disodium salt, is mixed.
7. the quick Purity method of a kind of Xian round-grained rice friendship type hybrid rice seeds according to claim 6, it is characterized in that, described glue is mixed by 5 × TBE, 40% acrylamide, 10% ammonium persulphate, Tetramethyl Ethylene Diamine, distilled water, and their mixed volume ratio is 8: 6: 0.3: 0.024: 26.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410023390.4A CN103740838A (en) | 2014-01-18 | 2014-01-18 | Rapid purity identification method for indica-japonica hybrid rice seeds |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410023390.4A CN103740838A (en) | 2014-01-18 | 2014-01-18 | Rapid purity identification method for indica-japonica hybrid rice seeds |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103740838A true CN103740838A (en) | 2014-04-23 |
Family
ID=50497903
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410023390.4A Pending CN103740838A (en) | 2014-01-18 | 2014-01-18 | Rapid purity identification method for indica-japonica hybrid rice seeds |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103740838A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402280A (en) * | 2018-08-01 | 2019-03-01 | 中国水稻研究所 | A kind of SSR marker for identifying Xian round-grained rice, screening technique and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1813517A (en) * | 2005-01-31 | 2006-08-09 | 云南农业大学 | Yunnan-type hybridrice restoring line molecular auxiliary selecting method |
| CN101942518A (en) * | 2010-09-26 | 2011-01-12 | 中国农业大学 | Method for authenticating rice indica subspecies and special primer composition thereof |
| CN102031301B (en) * | 2010-09-06 | 2012-09-12 | 南京农业大学 | Molecular marking method for indica-japonica segment substitution of fertility loci of indica-japonica hybrid |
-
2014
- 2014-01-18 CN CN201410023390.4A patent/CN103740838A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1813517A (en) * | 2005-01-31 | 2006-08-09 | 云南农业大学 | Yunnan-type hybridrice restoring line molecular auxiliary selecting method |
| CN102031301B (en) * | 2010-09-06 | 2012-09-12 | 南京农业大学 | Molecular marking method for indica-japonica segment substitution of fertility loci of indica-japonica hybrid |
| CN101942518A (en) * | 2010-09-26 | 2011-01-12 | 中国农业大学 | Method for authenticating rice indica subspecies and special primer composition thereof |
Non-Patent Citations (4)
| Title |
|---|
| 任辉: "SSR分子标记在杂交水稻品种纯度鉴定中的应用研究", 《万方数据库学位论文》 * |
| 陈荣林等: "SSR分子标记检测超级杂交稻桂两优2号种子纯度的研究", 《杂交水稻》 * |
| 隋志伟 等: "转基因水稻种子核酸提取方法的比较研究", 《中国稻米》 * |
| 黄庭旭等: "超级稻宜优673遗传构成与籼粳属性分析", 《中国农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402280A (en) * | 2018-08-01 | 2019-03-01 | 中国水稻研究所 | A kind of SSR marker for identifying Xian round-grained rice, screening technique and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101545004B (en) | Waxy maize inbred line Shen-W22 specific molecular marker and application thereof in offspring variety identification | |
| CN101824484B (en) | DNA fingerprint detection method for authenticity identification of cotton variety | |
| CN105154550B (en) | A kind of method using 1217 purity of EST-SSR molecular labeling Rapid identification variety of watermelon treasure sweet tea | |
| CN101921866A (en) | Method for identifying cotton variety by utilizing SSR core primers | |
| CN103361425B (en) | Primer and method for identifying purity of cucumber 'Yuexiu No.3' hybrid seed | |
| CN109266776B (en) | Kit and method for identifying citrus pomelo hybrid progeny by using InDel marker | |
| CN101560556B (en) | Method for detecting DNA fingerprint purity in cotton crossbreeds | |
| CN102031253A (en) | Molecular marker method for identifying indica type rice and japonica rice by using rice grain | |
| CN101696450B (en) | Method for distinguishing conventional cotton and hybrid cotton in assistant mode and special kit thereof | |
| CN108165647B (en) | Molecular marking method for identifying non-heading Chinese cabbage Suzhou green, dwarf yellow and Wuta-tsai | |
| CN106350584A (en) | Primer set and method for identifying variety authentication and seed purity of tomatoes | |
| CN106967797A (en) | The specific sequence and detection method of muskmelon seedses purity detecting and application | |
| CN104928396A (en) | Method for rapidly identifying hot pepper species and golden pepper purity degree by using EST-SSR molecular markers | |
| CN112626262B (en) | InDel molecular marker linked with radish petal color gene, primer and application thereof | |
| CN105821154A (en) | SSR primers and method for purity identification of luffa hybrid seeds | |
| CN104004845A (en) | Method and special primer set for identifying facticity to-be-tested variety belonging to CCRI 63 | |
| CN107058518B (en) | SSR molecular marker closely linked with sesame stem blight-resistant major gene locus and application thereof | |
| CN110453008B (en) | Molecular marker ZMM6206 closely linked with length and width major gene locus of sesame leaf and application thereof | |
| CN109852717B (en) | Molecular marking method for identifying non-heading Chinese cabbage glossy dwarf yellow, yellow rose and green star | |
| CN112592996A (en) | Molecular marker ZMM1776 closely linked with sesamin content major gene locus of sesame seeds and application thereof | |
| CN103740838A (en) | Rapid purity identification method for indica-japonica hybrid rice seeds | |
| CN102181540B (en) | Method for verifying seed purity of cotton | |
| CN101619358B (en) | Method for identifying breeds of Chinese cabbage and special kit thereof | |
| CN103937873B (en) | The DNA fingerprint detection method of cotton variety ' No. 49, CCRI ' | |
| CN113265481B (en) | Lycoris fluorescent EST-SSR molecular marker primer, method for identifying lycoris interspecific hybrid F1 generation and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Cao Dongdong Inventor after: Yuan Wei Inventor after: Ruan Xiaoli Inventor after: Sun Min Inventor after: Weng Shuangli Inventor before: Cao Dongdong Inventor before: Ruan Guanhai Inventor before: Ruan Xiaoli Inventor before: Zhan Yan Inventor before: Chen Heyun |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140423 |