CN101487048B - Molecular marker method for identifying pepper anti-epidemic disease character - Google Patents
Molecular marker method for identifying pepper anti-epidemic disease character Download PDFInfo
- Publication number
- CN101487048B CN101487048B CN2009100209641A CN200910020964A CN101487048B CN 101487048 B CN101487048 B CN 101487048B CN 2009100209641 A CN2009100209641 A CN 2009100209641A CN 200910020964 A CN200910020964 A CN 200910020964A CN 101487048 B CN101487048 B CN 101487048B
- Authority
- CN
- China
- Prior art keywords
- disease
- capsicum
- primer
- pepper
- pcr amplification
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a molecular marking method that is used for differentiating pepper disease resisting characteristics, which comprises extraction of pepper genomic DNA, PCR argumentation and polymorphism analysis of a specific enzyme incision product, and is characterized in that a pair of nucleotide molecules with a specified sequence that consists of a certain number of basic groups are artificially synthesized according to a plant disease resistance gene sequence and used as a primer, PCR reaction is realized to obtain a molecular marker, and pepper disease resistant materials and pepper disease susceptible materials can be identified through the size of the specific enzyme incision product in the marker. The molecular marking method can be used for disease resistance filtering in pepper seedling or adult phases and can greatly accelerate the process of disease-resistant pepper breeding.
Description
Technical field
The invention belongs to the pepper breeding field, be specifically related to a kind of plant molecular marker technology, particularly a kind of molecule marking method of differentiating pepper anti-epidemic disease character, this method is applicable to anti-eqpidemic disease molecular mark.
Background technology
Capsicum epidemic disease is one of main disease of harm capsicum production; All there is generation in many in the world countries, and big area generation, popular report are all arranged in capsicum main producing regions such as China Qinghai, Xinjiang, Gansu, Shaanxi, Beijing, Liaoning, Shanghai, Zhejiang, Hunan, Sichuan, Yunnan.Along with the development of capsicum industry, capsicum epidemic disease has the trend that increases the weight of year by year, has caused serious economy loss.The pathogenic bacteria of this disease is (Phytophthora capsici Leon), and the seedling stage of capsicum, one-tenth strain phase all can be injured, and stem, leaf and fruit can both be fallen ill.Velocity of propagation is fast, and the morbidity field often has no harvest, and causes serious economy loss.Producing at present and going up the control capsicum epidemic disease is main with chemical prevention, and control effect is poor, and contaminate environment; In addition, phytophthora blight of pepper is prone to morph, and often causes the generation of anti-(resisting) medicine bacterial strain.Though adopt soil disinfection that preventive effect is preferably arranged, production is gone up big area and is used then comparatively difficulty.
The application of anti-eqpidemic disease kind is control or alleviates the effective way of capsicum epidemic disease harm.At present many in the world capsicum main products country all the anti-eqpidemic disease kind of seed selection as one of major objective of pepper breeding work.But at present eqpidemic disease is shown the capsicum resource of high resistance, their drawback is that fruit shape is little, inferior quality, transformation cycle are long, is difficult to satisfy the capsicum industrialization to the update requirement of acceleration of kind.Compare with traditional breeding method, molecular marker breeding sooner, is more saved manpower, is saved time, raises the efficiency.Molecular marking technique mainly plays complementary effect in plant breeding.It is mainly sought and the closely linked molecule marker of target gene, and whether the detection molecules mark exists from hybrid generation then.Detected the plant of molecule marker, shown and contain goal gene, can further optimize and screen.
Utilizing the disease-resistant gene product to have the characteristic of conserved domain, through the similar degenerated primer of conserved sequence of synthetic and known disease-resistant gene product, is that template is carried out pcr amplification with the total DNA of plant, just can obtain the disease resistant gene homologous sequence of this kind of plant.Many researchs prove that some disease resistant gene homologous sequences and disease-resistant gene are chain closely, even are exactly the part of disease-resistant gene.In addition, disease resistant gene homologous sequence also has advantages such as easy acquisition, stable amplification result and cost of use are cheap.Therefore, separate and the disease resistant gene homologous sequence of clone plant becomes one of strategy of mark, location and clone plant disease-resistant gene that current people relatively admit.
The gene that utilizes disease resistant gene homologous sequence to obtain, one section lacks the dna sequence dna that 5 ' end and 3 ' is held often, obtains full-length gene order and can adopt unknown flanking sequence amplification---DNA Walking walking method, is called for short walking method.Though the selected primer of this method is complementary with known dna sequence, two primers, 3 ' end is reverse.The chromosome walking technology be mainly used in gene clone, step discover and seize in the new species gene non-conservative region, identify that T-DNA or the insertion site of transposon, the space in the karyomit(e) examining order fill up, thereby obtain complete aspects such as genome sequence.
Summary of the invention
To defective that exists in the prior art or deficiency; The objective of the invention is to; A kind of molecule marking method of differentiating pepper anti-epidemic disease character is provided; This method adopts and the closely linked disease resistant gene homologous sequence molecule marker of capsicum blight-resistant gene, fast, effectively selects anti-eqpidemic disease material in the breeding process thereby be implemented in, and quickens the capsicum blight-resistant breeding process.
To achieve these goals, the present invention adopts following technical solution:
A kind of molecule marking method of differentiating pepper anti-epidemic disease character is characterized in that, may further comprise the steps:
1) the synthetic a pair of nucleotide sequence of artificial design is as primer, and this primer is the oligomer by the particular sequence of based composition, wherein:
Forward primer: 5 '-AGGTGGTCTTCAATGATCAGAGA-3 '
Reverse primer: 5 '-CCTGAGGCAAATTCCAAATCTC-3 '
2) extract the genomic DNA of capsicum, with primer the capsicum genomic dna is carried out pcr amplification, anti-eqpidemic disease material all can amplify the fragment that length is 590bp with sense eqpidemic disease material;
3) pcr amplification product under 37 ℃ of conditions after Bcg I enzyme is cut 16h, the restriction enzyme site of Bcg I enzyme is: anti-eqpidemic disease kind cloned genes sequence 324bp place; Enzyme is cut product electrophoresis, ethidium bromide staining on 2% sepharose, observes and Taking Pictures recording with gel imaging system, and anti-eqpidemic disease plant enzyme is cut product has 325bp and two bands of a spectrum of 265bp, and sense eqpidemic disease plant has only bands of a spectrum of 590bp.
The molecule marking method of discriminating pepper anti-epidemic disease character of the present invention; Step simple and fast, reliable and stable; Identification result and accurate consistent, the good reproducibility of field resistance qualification result; Be specially adapted to anti-eqpidemic disease molecular mark, can improve efficiency of selection well, quicken breeding process.
Description of drawings
Fig. 1 is the analysis of 7 capsicum self-mating system pcr amplification product agarose gel electrophoresis.M：DNAMarker；1：A5；2：A11；3：B17；4：B23；5：B2；6：B32；7：B19
Fig. 2 is that 7 capsicum self-mating system pcr amplification product enzymes are cut the analysis of product agarose gel electrophoresis.M：DNA?Marker；1：A5；2：A11；3：B17；4：B23；5：B2；6：B32；7：B?19
Fig. 3 is the analysis of 7 capsicum self-mating system pcr amplification product agarose gel electrophoresis.M：DNAMarker；1：B3；2：P11；3：B4；4：P30；5：B12；6：P97；7：P45
Fig. 4 is that 7 capsicum self-mating system pcr amplification product enzymes are cut the analysis of product agarose gel electrophoresis.M：DNAMarker；1：B3；2：P11；3：B4；4：P30；5：B12；6：P97；7：P45。
Below in conjunction with accompanying drawing and embodiment the present invention is done more detailed explanation.
Embodiment
Following relevant references is able in the present invention to be used:
The clone and the analysis of capsicum annuum l. RR class disease resistant gene homologous sequence, Ma Wei etc., Xibei Univ. of Agricultural & Forest Science & Technology's journal (natural science edition), the 1st phase, 2008.
A?PCR-based?approach?for?isolating?pathogen?resistance?gene?from?potatowith?potential?for?application?in?plants.Leister?D，Ballvora?A，Salamini?F，Gebhardt?C.Nature?Genet，14，1996。
With the minim DNA process for extracting of PCR evaluation transfer-gen plant, Gong Zhenhui etc., Northwest Agricultural University's journal, the 1st phase, 1997.
The comparison of three kinds of inoculation methods of capsicum epidemic disease, Yi Tuyong etc., China's Vegetable, the 2nd phase, 2003.
The molecule marking method of discriminating pepper anti-epidemic disease character of the present invention, anti-with capsicum first, sense eqpidemic disease material is the examination material, according to resistant gene in plant conservative region design primer (Ma Wei etc.; The clone of capsicum annuum l. RR class disease resistant gene homologous sequence and analysis [J]. Xibei Univ. of Agricultural & Forest Science & Technology's journal (natural science edition), 2008,36 (1): 143～148); With the capsicum genomic dna is template amplification; Obtain anti-eqpidemic disease dna homolog sequence fragment, utilize DNA Walking walking method (Leister D, Ballvora A again; Salamini F; Gebhardt C.A PCR-based approach for isolating pathogen resistancegene from potato with potential for application in plants [J] .Nature Genet, 1996,14:421～429.) obtained capsicum epidemic disease resistant gene CanRB.Through CanRB gene order restriction enzyme site is analyzed; The restriction enzyme site that screening is suitable; Cut the product polymorphism analysis in conjunction with pcr amplification and enzyme; Create and optimize the molecule marker system of capsicum epidemic disease resistant gene, develop the molecule marker with capsicum blight-resistant gene linkage, in the hope of accelerating capsicum blight-resistant molecular breeding process.
Specifically comprise the steps:
1, a pair of nucleotide sequence is as primer, and this primer is the oligomer by the particular sequence of based composition, wherein:
Forward primer: 5 '-AGGTGGTCTTCAATGATCAGAGA-3 '
Reverse primer: 5 '-CCTGAGGCAAATTCCAAATCTC-3 '
2, the extraction of capsicum genomic dna: employing improvement SDS method extraction capsicum genomic dna (Gong Zhenhui etc. are with the minim DNA process for extracting [J] of PCR evaluation transfer-gen plant. Northwest Agricultural University's journal, 1997,25 (1): 45-47).
3, be primer with the said nucleic acid molecule of step 1, step 2 capsicum genomic dna is carried out pcr amplification, anti-eqpidemic disease material all can amplify the fragment that length is 590bp with sense eqpidemic disease material.The pcr amplification system is following:
(1) the reaction solution volume is 25 μ L, and the composition of used article and content are:
10 times of PCR buffer 2.5 μ L, 25mmolL
-1MgCl
22.0 μ L, 5U μ L
-1Taq archaeal dna polymerase 0.2 μ L, 5mmolL
-1DNTPs 1.0 μ L, 20 μ molL
-1Forward primer 1.0 μ L, 20 μ molL
-1Downstream primer 1.0 μ L, genomic dna 1.0 μ L (50ng μ L
-1), ddH
2O 16.3 μ L.
(2) the pcr amplification reaction condition is:
Pcr amplification reaction carries out on the PCR appearance, and reaction conditions is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 1min, 58 ℃ of annealing 1min, 72 ℃ are extended 1min, carry out 35 circulations; Last 72 ℃ are extended 10min, subsequent use in 4 ℃ of preservations subsequently.
4, pcr amplification product under 37 ℃ of conditions after Bcg I enzyme is cut 16h, enzyme is cut product electrophoresis, ethidium bromide staining on 2% sepharose, observes and Taking Pictures recording with gel imaging system.Anti-eqpidemic disease plant enzyme is cut product has 325bp and two bands of a spectrum of 265bp, and sense eqpidemic disease plant has only bands of a spectrum of 590bp.
Below be the embodiment that the contriver provides, these embodiment are used to understand the present invention, the invention is not restricted to these embodiment.
Embodiment 1:
1, artificial design synthetic nucleotide sequence is as primer:
Forward primer: 5 '-AGGTGGTCTTCAATGATCAGAGA-3 '
Reverse primer: 5 '-CCTGAGGCAAATTCCAAATCTC-3 '
2, the extraction of capsicum genomic dna: adopt improvement SDS method to extract 7 capsicum self-mating system A5, A11, B17, B23, B2, B32 and B19 genomic dna (Gong Zhenhui etc. respectively; Identify the minim DNA process for extracting [J] of transfer-gen plant with PCR. Northwest Agricultural University's journal; 1997,25 (1): 45-47).
3, be primer with the said nucleic acid molecule of step 1; Respectively above-mentioned 7 capsicum self-mating system genomic dnas are carried out pcr amplification, 7 capsicum self-mating systems all can amplify the fragment that length is 590bp, and (Fig. 1 is the analysis of 7 capsicum self-mating system pcr amplification product agarose gel electrophoresis.M：DNA?Marker；1：A5；2：A11；3：B17；4：B23；5：B2；6：B32；7：B19)。The pcr amplification system is following:
(1) the reaction solution volume is 25 μ L, and the composition of used article and content are:
10 times of PCR buffer 2.5 μ L, 25mmolL
-1MgCl
22.0 μ L, 5U μ L
-1Taq archaeal dna polymerase 0.2 μ L, 5mmolL
-1DNTPs 1.0 μ L, 20 μ molL
-1Forward primer 1.0 μ L, 20 μ molL
-1Downstream primer 1.0 μ L, genomic dna 1.0 μ L (50ng μ L
-1), ddH
2O 16.3 μ L.
(2) the pcr amplification reaction condition is:
Pcr amplification reaction carries out on the PCR appearance, and reaction conditions is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 1min, 58 ℃ of annealing 1min, 72 ℃ are extended 1min, carry out 35 circulations; Last 72 ℃ are extended 10min.Subsequent use in 4 ℃ of preservations subsequently.
4, pcr amplification product under 37 ℃ of conditions after Bcg I enzyme is cut 16h; Enzyme is cut product electrophoresis, ethidium bromide staining on 2% sepharose, with gel imaging system observe and Taking Pictures recording (Fig. 2 is that 7 capsicum self-mating system pcr amplification product enzymes are cut the analysis of product agarose gel electrophoresis.M：DNAMarker；1：A5；2：A11；3：B17；4：B23；5：B2；6：B32；7：B19)。The result shows that capsicum self-mating system A5, A11, B17, B23, B32 and B19 enzyme are cut product all has 325bp and two bands of a spectrum of 265bp, differentiates to be anti-eqpidemic disease material; And capsicum self-mating system B2 has only bands of a spectrum of 590bp, differentiates to be sense eqpidemic disease material.
Adopt and irritate root inoculation method (Yi Tuyong etc.; The comparison [J] of three kinds of inoculation methods of capsicum epidemic disease. China's Vegetable; 2003 (2): 16～18) the capsicum self-mating system is shown eqpidemic disease disease resistance qualification result: capsicum self-mating system A5, A11, B17, B23, B32 and B19 are anti-eqpidemic disease self-mating system, and capsicum self-mating system B2 is sense eqpidemic disease self-mating system.Therefore, utilize the molecule marker and the true resistance of plant of the discriminating capsicum blight-resistant characteristic that the present invention formulates consistent, be applicable to anti-eqpidemic disease molecular mark.
Embodiment 2:
1, artificial design synthetic nucleotide sequence is as primer:
Forward primer: 5 '-AGGTGGTCTTCAATGATCAGAGA-3 '
Reverse primer: 5 '-CCTGAGGCAAATTCCAAATCTC-3 '
2, the extraction of capsicum genomic dna: adopt improvement SDS method to extract 7 capsicum self-mating system B3, P11, B4, P30, B12, P97 and P45 genomic dnas respectively.
3, be primer with the said nucleic acid molecule of step 1; Respectively above-mentioned 7 capsicum self-mating system genomic dnas are carried out pcr amplification, 7 capsicum self-mating systems all can amplify the fragment that length is 590bp, and (Fig. 3 is the analysis of 7 capsicum self-mating system pcr amplification product agarose gel electrophoresis.M：DNA?Marker；1：B3；2：P11；3：B4；4：P30；5：B12；6：P97；7：P45)。The pcr amplification system is following:
(1) volume of reaction solution is 25 μ L, and the composition of used article and content are:
10 times of PCR buffer 2.5 μ L, 25mmolL
-1MgCl
22.0 μ L, 5U μ L
-1Taq archaeal dna polymerase 0.2 μ L, 5mmolL
-1DNTPs 1.0 μ L, 20 μ molL
-1Forward primer 1.0 μ L, 20 μ molL
-1Downstream primer 1.0 μ L, genomic dna 1.0 μ L (50ng μ L
-1), ddH
2O 16.3 μ L.
(2) the pcr amplification reaction condition is:
Pcr amplification reaction carries out on the PCR appearance, and reaction conditions is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 1min, 58 ℃ of annealing 1min, 72 ℃ are extended 1min, carry out 35 circulations; Last 72 ℃ are extended 10min.Subsequent use in 4 ℃ of preservations subsequently.
4, pcr amplification product under 37 ℃ of conditions after Bcg I enzyme is cut 16h; Enzyme is cut product electrophoresis, ethidium bromide staining on 2% sepharose, with gel imaging system observe and Taking Pictures recording (Fig. 4 is that 7 capsicum self-mating system pcr amplification product enzymes are cut the analysis of product agarose gel electrophoresis.M：DNAMarker；1：B3；2：P11；3：B4；4：P30；5：B12；6：P97；7：P45。)。The result shows that capsicum self-mating system P11, P30, P97 and P45 enzyme are cut product all has 325bp and two bands of a spectrum of 265bp, differentiates to be anti-eqpidemic disease material; And capsicum self-mating system B3, B4 and B12 have only bands of a spectrum of 590bp, differentiate to be sense eqpidemic disease material.
Adopt and irritate root inoculation method (Yi Tuyong etc.; The comparison [J] of three kinds of inoculation methods of capsicum epidemic disease. China's Vegetable; 2003 (2): 16～18) the capsicum self-mating system is shown eqpidemic disease disease resistance qualification result: capsicum self-mating system P11, P30, P97 and P45 are anti-eqpidemic disease self-mating system, and capsicum self-mating system B3, B4 and B12 are sense eqpidemic disease self-mating system.Therefore, utilize the molecule marker and the true resistance of plant of the discriminating capsicum blight-resistant characteristic that the present invention formulates consistent, be applicable to anti-eqpidemic disease molecular mark.
Claims (2)
1. a molecule marking method of differentiating pepper anti-epidemic disease character is characterized in that, carries out according to the following steps:
1) the synthetic a pair of nucleotide sequence of artificial design is as primer, and this primer is the oligomer by the particular sequence of based composition, wherein:
Forward primer: 5 '-AGGTGGTCTTCAATGATCAGAGA-3 '
Reverse primer: 5 '-CCTGAGGCAAATTCCAAATCTC-3 '
2) extract the genomic DNA of capsicum, with primer the capsicum genomic dna is carried out pcr amplification, anti-eqpidemic disease material all can amplify the fragment that length is 590bp with sense eqpidemic disease material;
3) pcr amplification product under 37 ℃ of conditions after Bcg I enzyme is cut 16h, the restriction enzyme site of Bcg I enzyme is: anti-eqpidemic disease kind cloned genes sequence 324bp place; Enzyme is cut product electrophoresis, ethidium bromide staining on 2% sepharose, observes and Taking Pictures recording with gel imaging system, and anti-eqpidemic disease plant enzyme is cut product has 325bp and two bands of a spectrum of 265bp, and sense eqpidemic disease plant has only bands of a spectrum of 590bp.
2. the method for claim 1 is characterized in that, the pcr amplification reaction of said step (2) is:
(1) 25 μ L PCR reaction system is by 10 times of PCR buffer, 2.5 μ L, 25mmo1L
-1MgCl
22.0 μ L, 5U μ L
-1Taq archaeal dna polymerase 0.2 μ L, 5mmolL
-1DNTPs 1.0 μ L, 20 μ molL
-1Forward primer 1.0 μ L, 20 μ molL
-1Reverse primer 1.0 μ L, genomic dna 1.0 μ L, ddH
2O 16.3 μ L form;
(2) pcr amplification reaction carries out on the PCR appearance, and reaction conditions is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 1min, 58 ℃ of annealing 1min, 72 ℃ are extended 1min, carry out 35 circulations; Last 72 ℃ are extended 10min, subsequent use in 4 ℃ of preservations subsequently.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100209641A CN101487048B (en) | 2009-01-19 | 2009-01-19 | Molecular marker method for identifying pepper anti-epidemic disease character |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100209641A CN101487048B (en) | 2009-01-19 | 2009-01-19 | Molecular marker method for identifying pepper anti-epidemic disease character |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101487048A CN101487048A (en) | 2009-07-22 |
| CN101487048B true CN101487048B (en) | 2012-05-23 |
Family
ID=40890108
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100209641A Expired - Fee Related CN101487048B (en) | 2009-01-19 | 2009-01-19 | Molecular marker method for identifying pepper anti-epidemic disease character |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101487048B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102812836B (en) * | 2012-08-20 | 2013-09-11 | 西北农林科技大学 | Method for rapidly identifying functions of related disease resisting genes of hot pepper |
| CN104878093B (en) * | 2015-04-30 | 2017-06-16 | 中国农业大学 | A kind of molecular labeling and its application with capsicum blight-resistant gene close linkage |
| CN115927699B (en) * | 2022-07-18 | 2024-01-26 | 江西省农业科学院蔬菜花卉研究所 | Molecular marker for identifying germplasm of pepper with epidemic disease resistance and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1970790A (en) * | 2006-11-29 | 2007-05-30 | 广东省农业科学院蔬菜研究所 | Molecular marker-assisted selection method for capsicum blight-resistant breeding |
-
2009
- 2009-01-19 CN CN2009100209641A patent/CN101487048B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1970790A (en) * | 2006-11-29 | 2007-05-30 | 广东省农业科学院蔬菜研究所 | Molecular marker-assisted selection method for capsicum blight-resistant breeding |
Non-Patent Citations (4)
| Title |
|---|
| A. Palloix et al..Breeding transgressive lines of pepper for resistance to Phytophthora capsici in a recurrent selection system.《Euphytica》.1990,第51卷(第2期),141-150. * |
| A. Thabuis · V. Lefebvre · G. Bernard ·A. M. Daubèze · T. Phaly.Phenotypic and molecular evaluation of a recurrent selection program for a polygenic resistance to Phytophthora capsici in pepper.《Theoretical And Applied Genetics》.2004,第109卷(第2期),342-351. * |
| 易图永等.几个抗疫病性不同的辣椒材料抗病基因同源序列的分离与比较.《园艺学报》.2003,第30卷(第5期),540-544. * |
| 杜晓华等.辣椒抗疫病的遗传与育种.《西北农业学报》.2005,第14卷(第1期),30-36. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101487048A (en) | 2009-07-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Alvarez et al. | Characterization of a phytoplasma associated with frogskin disease in cassava | |
| KR101516190B1 (en) | SSR primer sets for discrimination of oriental melon line or cultivar and uses thereof | |
| CN101942520B (en) | Marker of avirulence gene PsAvr3b of phytophthora sojae | |
| Tohidi et al. | Association of a 16SrIX-C phytoplasma with eggplant phyllody in Iran | |
| CN101487048B (en) | Molecular marker method for identifying pepper anti-epidemic disease character | |
| KR101047322B1 (en) | DNA marker for the identification of Anguilla eel species | |
| Karimi et al. | Phytoplasma detection and identification in declining pomegranate in Iran | |
| CN102304587A (en) | Method for rapidly identifying erect panicle of rice | |
| CN103866038B (en) | For detecting tobacco to the N gene-specific primer of TMV resistance to, detection method and test kit | |
| Sbissi et al. | Diversity of the desert truffle Terfezia boudieri Chatin. in southern Tunisia | |
| Nowicka et al. | Precise karyotyping of carrot mitotic chromosomes using multicolour-FISH with repetitive DNA | |
| KR101716021B1 (en) | Primer and method for diagnosting sex of ginkgo | |
| CN110468229A (en) | Rice wide spectrum high resistance to hoja blanca gene Xa45's (t) isolates molecular labeling Hxjy-1 | |
| KR101299557B1 (en) | SNP MAKER FOR DISCRIMINATION OF Schizandrae fructus, SPECIFIC-IDENTIFICATION METHODS OF Schizandrae fructus SPECIES USING SNP MAKER, AND SPECIFIC-IDENTIFICATION KITS OF Schizandrae fructus SPECIES USING SNP MAKER KIT | |
| Liu et al. | Development of expressed sequence tag-simple sequence repeat markers for Chrysanthemum morifolium and closely related species | |
| Jamali et al. | Nested-PCR for detecting Terfezia claveryi in roots of Helianthemum species in field and greenhouse conditions | |
| KR101719719B1 (en) | Kit for detection of Pseudomonas syringae pv. actinidiae of Psa1 group | |
| KR100998571B1 (en) | DNA marker for discrimination of Angelica decursiva Franch. et Savatier=Peucedanum decursivum Maxim., Peucedanum praeruptorum Dunn. and Anthricus sylvestris L. Hoffman | |
| KR101855984B1 (en) | composition comprising SNP markers for a differentiation of Cudrania tricuspidata Bureau lines, and method for a differentiation of Cudrania tricuspidata Bureau lines and hybrid using the same | |
| CN110016515B (en) | Method for detecting kiwi fruit rot germs by using PCR primers | |
| Chung et al. | Occurrence of stolbur phytoplasma disease in spreading type Petunia hybrida cultivars in Korea | |
| JP5849317B2 (en) | Variety identification marker of vegetative propagation crop | |
| CN107130019B (en) | Molecular marker primer of rice local variety thin rice panicle blast resistance gene and application thereof | |
| CN105936939B (en) | SNP (Single nucleotide polymorphism) marker related to lignin content of larch and application thereof | |
| KR101133104B1 (en) | Method for distingusishment of Zanthoxylum schinifolium subspecies |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120523 Termination date: 20130119 |