CN104152446A - SSR molecular markers cosegregated from cucumber powdery mildew-resistant major QTL - Google Patents
SSR molecular markers cosegregated from cucumber powdery mildew-resistant major QTL Download PDFInfo
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Abstract
The invention discloses three SSR molecular markers cosegregated from a cucumber powdery mildew-resistant major QTL. The three SSR molecular markers are respectively named as SSR-N1, SSR-N2 and SSR-N3, wherein the nucleotide sequence of the SSR-N1 is shown in the SEQ ID No.1; the nucleotide sequence of the SSR-N2 is shown in the SEQ ID No.2; the nucleotide sequence of the SSR-N3 is shown in the SEQ ID No.3. The three SSR molecular markers disclosed by the invention all have high stability, can be simply and rapidly applied to auxiliary screening of powdery mildew-resistant individual plants at the cucumis sativus seedling stage, and lay a foundation for assistant breeding of powdery mildew-resistant molecular markers, so that the process of breeding cucumber powdery mildew-resistant molecules is accelerated greatly. Moreover, the cosegregated molecular markers lay a foundation for cloning of the cucumber powdery mildew-resistant major QTL.
Description
Technical field
The present invention relates to genetic engineering technique, be specifically related to cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker.
Background technology
Cucumber (Cucumis sativus L.) is Curcurbitaceae (Cucurbitaceae) the Cucumis annual herb climber that overgrows.Cucumber is as one of large important vegetable crops in the world ten, and one of Ye Shi China main cultivation vegetable crop accounts for 10% left and right of national vegetables area.Cucumber not only always enjoys the attention of breeding man as important vegetable crop, and cucumber hypoploidy 2n=2x=14, genome is less, in 2009, the successful order-checking of Cucumber germplasm provided very big facility for cucumber carries out molecular biology research as model plant cucurbitaceous especially.
In cucumber production, face the harm of numerous disease, wherein Powdery Mildew is one of the most serious disease.Powdery mildew of cucumber is the fungal disease being caused by obligate parasite (Podosphaera xanthii), can be in morbidity breeding time of whole cucumber, main harm blade, when serious, stem can be endangered climing, particularly serious in the middle and late growth stage morbidity, plant is uprooted plants after their edible portions have been harvested ahead of time, cause serious production loss.Field rotating medicine, causes pesticide residue, affects fruit quality, jeopardizes food safety, and pollutes the environment.Long-term dispenser also can promote Physiological Races of Powdery Mildew to produce resistance, thereby increases difficulty of prevention and cure, increases plantation family production cost.Cultivating disease-resistant cucumber variety is the best method that solves Powdery Mildew harm.Conventional breeding for disease resistance length consuming time, need to be through too much generation hybridization and backcross process; Disease occurs closely related with envrionment conditions, needs special sick garden to carry out inoculated identification; These have all increased the difficulty of cultivating cucumber disease-resistant variety.
Molecular breeding can be accelerated breeding process greatly, significantly shortens breeding cycle.The fast development of modern biotechnology, has opened up new approach for breeding for disease resistance, and utilizing biotechnology to cultivate disease-resistant variety has become current focus.The prerequisite of molecular breeding is functional gene or the molecule marker closely linked with it that obtains correlated character.Utilize molecular marker analysis system, in genetic group, identify disease-resistant genetic development, binding molecule mark genetic linkage maps simultaneously, location and clone's powdery mildew resistance gene, study the molecular regulation mechanism of its function and resistance, can provide theoretical foundation for the molecular mark of cucumber resistant gene and Molecular design breeding.Utilize and the closely linked molecule marker of disease-resistant gene, carry out molecular marker assisted selection breeding, multiple disease-resistant genes can be incorporated into a kind, significantly improve breeding efficiency, shorten breeding time, and having increased substantially disease-resistant dynamics and persistence, for cultivating, to meet the cucumber disease-resistant varieties of grower's demand significant for this.
Although the generation of ground family crop powdery mildew of cucumber is commonplace and serious, but the research about its powdery mildew resistance gene location is relatively weak, not yet find with powdery mildew resistance gene/QTL be divided into from mark, say nothing of the clone of gene and the research of Resistance mechnism, rest at present the stage of major gene/QTL location.Because many reports show that cucumber powdery mildew resistance is by multiple recessive gene control, investigator has carried out the analysis of QTL to it.2006, Sakata etc. utilized the recombinant inbred lines of the anti-sense combination of 97 strain cucumber, had located first the QTL of anti-cucumber powdery mildew proterties; In 4 linkage groups, detect 6 with the QTL of temperature correlation, wherein a main effect QTL on LGII all shows resistance at 20 DEG C and 26 DEG C.The F that Liu etc. (2008a) utilize cucumber high sense Powdery Mildew self-mating system S94 and high mildew-resistance self-mating system S06 to build
2: 3family has been carried out QTL location, 5 powder mildew resistance QTL detected altogether, is distributed in linkage group 1,2,5, and the contribution rate of single QTL is between 3.4%～45%; The RIL also building by this two parent detects 4 powder mildew resistance QTL altogether, lays respectively in linkage group 1,2,4,6, and the contribution rate of single QTL is (Liu et al., 2008b) between 5.2%～21.0%.Shen Liping (2009) application ISSR (inter-simple sequence repeats) and SRAP (sequence-related amplified polymorphism) labeling technique, with the F of height sense Powdery Mildew cucumber variety D8 and high powdery mildew resisting green cucumber variety JIN5
2crowd surveillance, to 2 QTL that control cucumber powdery mildew resistance, is all positioned in the 3rd linkage group, and contribution rate is 7.6% and 13.5%.Open the F of holy equality (2011) with K8 (disease-resistant) × K18 (susceptible) combination
2and F
2: 3family is research object, the QTL of 4 powder mildew resistances detected altogether.Recently, Fukino etc. (2013) utilize RIL 9 QTL to be detected, lay respectively on karyomit(e) 1,3,4,5,6, the contribution rate of single QTL is between 5%～44%, wherein the effect in 4 sites has obtained confirmation by residue heterozygote (residual heterozygous lines, RHLs).He etc. (2013) utilize F
2: 3family has been carried out qtl analysis to the white powder resistance of cucumber hypocotyl, cotyledon and true leaf simultaneously, and result detects 6 QTL on 1,3,4, No. 5 karyomit(e), and the contribution rate of single QTL is between 6.1%～74.5%; Wherein 2 main effect QTLs are positioned at the interval of No. 5 chromosomal 40 cM, explain the contribution rate of 21.0-74.5%, and hypocotyl Resistance QTL has played most important effect to cucumber white powder resistance.
Above-mentioned most research shows, cucumber is to the resistance of Powdery Mildew by multiple gene actings in conjunction, and resistant gene shows as recessive effect, and these factors have increased disease-resistant gene Fine Mapping and the difficulty separating.Because powdery mildew of cucumber is very large on cucumber production impact, research to cucumber powdery mildew resistance gene is a lot, but not yet there is the report of this cloned resistance gene and Study on Molecular Mechanism at present, acquired linked marker genetic distance is far away, be unfavorable for carrying out of molecular mark, hindered the process of disease-resistant variety molecular breeding.Therefore, find with cucumber powdery mildew resistance gene/QTL close linkage, be divided into from molecule marker, it is carried out Fine Mapping, is separated and clone, and this not only can be for its disease-resistant molecular breeding provides good technical support, also for the molecular mechanism of opening cucumber powdery mildew resistance lays the foundation.
Summary of the invention
Object of the present invention, is to overcome the problem that QTL positioning difficulty is large, provide three with cucumber powdery mildew resistance main effect QTL pm5.1 be divided into from codominance SSR molecule marker.The present invention utilizes BSA (Bulked Segregant Analysis) and QTL localization method, finds a main effect QTL of controlling powder mildew resistance.For this main effect QTL of Fine Mapping, we have built chromosome segment substitution line (Chromosome Segment Substitution Lines, CSSL) and the segregating population that backcrosses thereof containing main effect QTL.By the exploitation of Fine Mapping and mark, obtain three with cucumber powdery mildew resistance main effect QTL be divided into from SSR mark so that the foundation of molecular mark system.Molecule marker of the present invention can be applied to breeding practice easy, quick, high-throughput.
The present invention is achieved by the following technical solutions:
One with cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, called after SSR-N1, its nucleotide sequence is as shown in SEQ ID NO.1.
Above-mentioned SSR-N1 is obtained by upstream primer SSR-N1-F and downstream primer SSR-N1-R pcr amplification, the sequence of described upstream primer SSR-N1-F is 5 '-CCACAACAGCAGAAGGCTAACA-3 ', and the sequence of described downstream primer SSR-N1-R is 5 '-CCAATGGGTTGATAGAGGGAGA-3 '.
One with cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, called after SSR-N2, its nucleotide sequence is as shown in SEQ ID NO.2.
Above-mentioned SSR-N2 is obtained by upstream primer SSR-N2-F and downstream primer SSR-N2-R pcr amplification, the sequence of described upstream primer SSR-N2-F is 5 '-CTTCATTGTTGATTTCCAGGC-3 ', and the sequence of described downstream primer SSR-N2-R is 5 '-TGTTACGACCTATAACCACAAAAT-3 '.
One with cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, called after SSR-N3, its nucleotide sequence is as shown in SEQ IDNO.3.
Above-mentioned SSR-N3 is obtained by upstream primer SSR-N3-F and downstream primer SSR-N3-R pcr amplification, the sequence of described upstream primer SSR-N3-F is 5 '-GAAGATGCATCGAATTGAAACA-3 ', and the sequence of described downstream primer SSR-N3-R is 5 '-ATGATGTCCCAACTTATCCAAA-3 '.
BSA and QTL localization method for the present invention, utilize F
2colony finds a main effect QTL of controlling powder mildew resistance.For this main effect QTL of Fine Mapping, utilize the method constantly backcrossing to build chromosome segment substitution line (Chromosome Segment Substitution Lines, CSSL) and the segregating population that backcrosses thereof containing main effect QTL.By to the segregating population BC that backcrosses
3f
1, BC
2f
2resistance Identification analysis, powder mildew resistance has become single Mendelian factor heredity, by Dominant gene, resistance is recessive inheritance.By Fine Mapping, resistant gene navigates between mark UW065021 and UW065094, physical distance 170kb.By Cucumber germplasm sequence, developing SSR mark therebetween, finally obtain three with cucumber powdery mildew resistance main effect QTL be divided into from SSR mark, be named as respectively SSR-N1, SSR-N2, SSR-N3.3 of this patent invention are divided into from SSR molecule marker to be codominant marker, can to distinguish homozygote and heterozygote, by being conducive to the foundation of molecule marker assistant system of powder mildew resistance breeding, can be applied to breeding practice easy, quick, high-throughput.
Compared with prior art, the present invention has following beneficial effect: conventional traditional breeding for disease resistance length consuming time, and need to be through too much generation hybridization and backcross process; Disease occurs closely related with envrionment conditions, needs special sick garden to carry out inoculated identification; These have all increased the difficulty of cultivating cucumber disease-resistant variety.Because Powdery Mildew is obligate parasite, Resistance Identification needs inoculation, and after morbidity, plant easily dies; And powdery mildew resistance gene shows as recessive effect more, ordinary method back cross breeding needs a selfing generation again after first backcross generation to confirm the infiltration of resistant gene, and these factors have all increased cycle and the difficulty of powder mildew resistance breeding.It is of the present invention that to be divided into from SSR molecule marker be codominance, can identify plant at cucumber at seedling stage, and can distinguish homozygote and heterozygote, in the infiltration process that backcrosses, save the step of per generation selfing, follow the tracks of resistant gene with molecule marker, both saved time also accurate, so can be used for the molecular mark of cucumber powdery mildew resistance, greatly accelerated the process of cucumber powdery mildew resistance breeding.Separation marking also will promote the clone of powder mildew resistance QTL/ gene altogether simultaneously, thereby the molecular mechanism forming for announcement powder mildew resistance lays the foundation.
Brief description of the drawings
Fig. 1 is the polyacrylamide gel electrophoresis effect of molecule marker SSR-N1;
Fig. 2 is the polyacrylamide gel electrophoresis effect of molecule marker SSR-N2;
Fig. 3 is the polyacrylamide gel electrophoresis effect of molecule marker SSR-N3.
Shown in figure, S1001 is disease-resistant parent; S05 is Susceptible parent; F
1represent two parent filial generations; R and S represent respectively BC
2f
2the detected result of the disease-resistant and disease plant of random choose in segregating population backcrosses.
Embodiment
One, the qualification of cucumber powdery mildew resistance main effect QTL/gene
1. colony builds and Resistance Identification
That preliminary Resistance QTL is located use is F
2colony, disease-resistant parent is S1003, and Susceptible parent is S1001, and they all belong to the cucumber self-mating system of North China type.Two parents are hybridized the F of generation
1produce F for selfing
2for colony.Powder mildew resistance identifies that the bacterial classification using is to separate and obtain from the cucumber plant of Shanghai Communications University's greenhouse morbidity.The random 148 strain F that select
2for the individual Resistance Identification that carries out in seedling stage.Gather the Powdery Mildew of purifying from susceptible seedling, make spore suspension with sterilized water, being adjusted to concentration is 1 × 10
5individual mL
-1.In the time that the 3rd true leaf of cucumber just launched, deployed spore suspension is evenly sprayed onto on true leaf, not become drops to be as the criterion.Inoculation culture 12d " Invest, Then Investigate " incidence.The incidence of plant is divided into 5 grades according to (2003) such as Morishita.0 grade and 1 grade is considered as disease-resistantly, and 2 grades are considered as susceptible above.
According to incidence survey, S1003 is high resistance, and S1001 is high sense, F
1for susceptible, be partial to Susceptible parent; F
2colony's disease resistance presents bimodal distribution, and incidence trend is partial to susceptiblely, and intermediate type is less, and the disease resistance that has recessive major gene control white powder is described.
2.BSA method and qtl analysis are determined the chromosome position of major gene
Therefore we take the chromosomal region at the first antagonism major gene of BSA method place to analyze.From F
2in segregating population, choose at random respectively high resistance and individual each 10 strains of high sense, set up anti-, sense gene pool.Screen with two parents of 780 pairs of SSR primer pairs that are evenly distributed on karyomit(e) and two gene pools.
SSR reaction system is genomic dna 30ng, primer 0.2 μ mol/L, and 200 μ mol/L dNTPs, 2mmol/L MgCl2,1 μ l 10 × PCR reacfions buffer, 0.5U TaqDNA polysaccharase, total reaction system is 10 μ l.Pcr amplification program is: 94 DEG C of 5min; 35 circulations: 94 DEG C of 30s, 55-60 DEG C of 30s, 72 DEG C of 30s; 72 DEG C of 5min.Amplified production separates with 6% denaturing polyacrylamide gel electrophoresis, and electrophoretic buffer is 1 × TBE, the permanent power of 45W, electrophoresis 1.5h-2h.
After electrophoresis, carrying out silver dyes.Silver staining method is: the sheet glass with glue is put into stationary liquid, shake gently to indicator color fade, wherein consisting of of stationary liquid on shaking table: the volume ratio of Glacial acetic acid, dehydrated alcohol, distilled water is 1: 10: 100; With ultrapure washing 1min-3min; Offset plate after rinsing is put into staining fluid and shake half an hour, wherein the component of staining fluid is 2g/L Silver Nitrate; After offset plate after dyeing is put into ultrapure water rinsing 5s, put into the plastics casing that developing solution is housed, it is clear to shake gently to band, puts into tap water and rinses 3min; Dry under room temperature, take pictures after dry, wherein developing solution adds 15g NaOH and 3ml formaldehyde to mix to obtain in 1L distilled water.
Screen by BSA, be marked between pond and show as polymorphism at continuous 7 SSR of the long-armed end of the 5th karyomit(e), therefore powdery mildew resistance main effect gene is positioned at this region.In order further to determine the position of resistance main effect gene, we have carried out qtl analysis.From 7 karyomit(e)s of cucumber average selection 73 polymorphism SSR marks to 148 strain F
2carry out electrophoretic analysis for individuality.Use JoinMap 3.0 to build linkage map, wherein LOD >=5.0, adopt Kosambi function that recombination fraction is converted into genetic distance.By F
2the average of three blade grades of colony's individual plant is carried out QTL mapping as the disease-resistant disease index of plant.Qtl analysis uses QTL Cartographer 2.5, adopts composite interval mapping method (CIM) to carry out QTL mapping.By the result of qtl analysis, find a main effect QTL pm5.1 at the 5th chromosomal long-armed end, the position that this and BSA method draw coincide, and therefore we determine that this position exists a main effect QTL of controlling powder mildew resistance.
Two, the backcross Fine Mapping of structure and main effect QTL of segregating population
Because parent S1003 and S1001 are North China type cucumber, sibship is near, and polymorphism mark is few, therefore builds while backcrossing segregating population and has adopted the European greenhouse cucumber self-mating system S05 of high sense Powdery Mildew as donor parents.Selfing after backcrossing by many generations in conjunction with MAS, has built the only backcross population BC of main effect QTL pm5.1 region disconnecting
3f
1, BC
2f
2.Respectively to 2 BC
3f
1colony's (114 strains, 480 strains) and BC
2f
2the Resistance Identification of colony's (483 strain), anti-, sense ratio meets 1: 1 and 1: 3 through chi-square analysis, therefore, Resistance QTL has been converted into single Mendelian factor, i.e. Dominant gene, and disease-resistant be recessive inheritance.The molecule marker of having developed by main effect QTL region carries out linkage analysis to these 1074 individual plants, and result arrives disease-resistant gene Fine Mapping between SSR mark UW065021 and UW065094, physical distance 170kb.By Cucumber germplasm sequence, develop therebetween 15 pairs of SSR marks, result only has 3 pairs between parent, to have polymorphism, i.e. three molecule marker SSR-N1 of the present invention, SSR-N2, SSR-N3.By colony's linkage analysis, finally draw these three SSR molecule markers and cucumber powdery mildew resistance main effect QTL be divided into from, the disease-resistant plant banding pattern of colony is all consistent with disease-resistant parent's banding pattern, and disease plant banding pattern is all Susceptible parent or F
1banding pattern.
Fig. 1 is the polyacrylamide gel electrophoresis effect of molecule marker SSR-N1, and Fig. 2 is the polyacrylamide gel electrophoresis effect of molecule marker SSR-N2, and Fig. 3 is the polyacrylamide gel electrophoresis effect of molecule marker SSR-N3.
Claims (6)
- One with cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, called after SSR-N1, its nucleotide sequence is as shown in SEQ ID NO.1.
- According to claim 1 and cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, it is characterized in that, described SSR-N1 is obtained by upstream primer SSR-N1-F and downstream primer SSR-N1-R pcr amplification, the sequence of described upstream primer SSR-N1-F is 5 '-CCACAACAGCAGAAGGCTAACA-3 ', and the sequence of described downstream primer SSR-N1-R is 5 '-CCAATGGGTTGATAGAGGGAGA-3 '.
- One with cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, called after SSR-N2, its nucleotide sequence is as shown in SEQ IDNO.2.
- According to claim 3 and cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, it is characterized in that, described SSR-N2 is obtained by upstream primer SSR-N2-F and downstream primer SSR-N2-R pcr amplification, the sequence of described upstream primer SSR-N2-F is 5 '-CTTCATTGTTGATTTCCAGGC-3 ', and the sequence of described downstream primer SSR-N2-R is 5 '-TGTTACGACCTATAACCACAAAAT-3 '.
- One with cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, called after SSR-N3, its nucleotide sequence is as shown in SEQ ID NO.3.
- According to claim 5 and cucumber powdery mildew resistance main effect QTL be divided into from SSR molecule marker, it is characterized in that, described SSR-N3 is obtained by upstream primer SSR-N3-F and downstream primer SSR-N3-R pcr amplification, the sequence of described upstream primer SSR-N3-F is 5 '-GAAGATGCATCGAATTGAAACA-3 ', and the sequence of described downstream primer SSR-N3-R is 5 '-ATGATGTCCCAACTTATCCAAA-3 '.
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| CN201610338869.6A CN105838712B (en) | 2014-08-06 | 2014-08-06 | The SSR molecular marker isolated with cucumber powdery mildew resistance main effect QTL |
| CN201610341936.XA CN105838812B (en) | 2014-08-06 | 2014-08-06 | The SSR molecular marker isolated with cucumber powdery mildew resistance main effect QTL |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498486A (en) * | 2014-12-02 | 2015-04-08 | 中国农业科学院蔬菜花卉研究所 | Indel marker of powdery mildew resistant gene pm-h of cucumber and application of Indel marker |
| CN104498484A (en) * | 2014-12-01 | 2015-04-08 | 北京市农林科学院 | Linked molecular marker for powdery mildew resistant gene pm1 of cucurbita pepo L. and application of linked molecular marker |
| CN104560983A (en) * | 2015-01-30 | 2015-04-29 | 扬州大学 | Two SNP markers in close linkage with cucumber powdery mildew resistance and application thereof |
| CN105734057A (en) * | 2016-04-29 | 2016-07-06 | 新疆农业科学院哈密瓜研究中心 | SSR mark linked with pseudoperonospora cubensis resistance main effect QTL and application of SSR mark |
| CN105969860A (en) * | 2016-05-17 | 2016-09-28 | 青岛市农业科学研究院 | Anti-cucumber-powdery-mildew major QTL (quantitative trait locus) mapping |
| CN108546778A (en) * | 2018-07-19 | 2018-09-18 | 上海市农业科学院 | A kind of SNP marker and its application for detecting anti-cucumber powdery mildew character |
Families Citing this family (1)
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| CN109234429B (en) * | 2018-08-01 | 2021-07-20 | 扬州大学 | Two SNP markers closely linked with powdery mildew resistance of cucumber and application thereof |
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| CN101240342B (en) * | 2008-03-13 | 2010-06-23 | 上海交通大学 | Cucumber powdery mildew resistance main effect QTL compact linkage molecule labeling method and applying method |
-
2014
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Non-Patent Citations (3)
| Title |
|---|
| JINGTAO NIE ET AL.: "Identification and fine mapping of pm5.1: a recessive gene for powdery mildew resistance in cucumber (Cucumis sativus L.)", 《MOL BREEDING》, vol. 35, no. 7, 15 January 2015 (2015-01-15), pages 1 - 13, XP 035448210, DOI: doi:10.1007/s11032-015-0206-8 * |
| 张圣平 等: "黄瓜白粉病抗性基因QTL定位", 《中国农业科学》, vol. 44, no. 17, 1 September 2011 (2011-09-01), pages 3584 - 3593 * |
| 林肖剑 等: "黄瓜抗白粉病染色体片段导入系的SSR鉴定", 《园艺学报》, vol. 39, no. 3, 25 March 2012 (2012-03-25), pages 485 - 492 * |
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| CN104498484A (en) * | 2014-12-01 | 2015-04-08 | 北京市农林科学院 | Linked molecular marker for powdery mildew resistant gene pm1 of cucurbita pepo L. and application of linked molecular marker |
| CN104498484B (en) * | 2014-12-01 | 2018-01-16 | 北京市农林科学院 | The dominant disease-resistant gene pm1 of Pumpkin powdery mildew linkage molecule mark and its application |
| CN104498486A (en) * | 2014-12-02 | 2015-04-08 | 中国农业科学院蔬菜花卉研究所 | Indel marker of powdery mildew resistant gene pm-h of cucumber and application of Indel marker |
| CN104498486B (en) * | 2014-12-02 | 2017-03-01 | 中国农业科学院蔬菜花卉研究所 | The Indel labelling of anti-cucumber powdery mildew gene pm h and its application |
| CN104560983A (en) * | 2015-01-30 | 2015-04-29 | 扬州大学 | Two SNP markers in close linkage with cucumber powdery mildew resistance and application thereof |
| CN104560983B (en) * | 2015-01-30 | 2017-03-08 | 扬州大学 | Two SNP marker and its application with anti-cucumber powdery mildew close linkage |
| CN105734057A (en) * | 2016-04-29 | 2016-07-06 | 新疆农业科学院哈密瓜研究中心 | SSR mark linked with pseudoperonospora cubensis resistance main effect QTL and application of SSR mark |
| CN105734057B (en) * | 2016-04-29 | 2019-05-10 | 新疆农业科学院哈密瓜研究中心 | With the SSR marker and its application of muskmelon downy mildew resistance main effect QTL linkage |
| CN105969860A (en) * | 2016-05-17 | 2016-09-28 | 青岛市农业科学研究院 | Anti-cucumber-powdery-mildew major QTL (quantitative trait locus) mapping |
| CN108546778A (en) * | 2018-07-19 | 2018-09-18 | 上海市农业科学院 | A kind of SNP marker and its application for detecting anti-cucumber powdery mildew character |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105838812B (en) | 2019-09-27 |
| CN105838712A (en) | 2016-08-10 |
| CN105838712B (en) | 2018-11-16 |
| CN104152446B (en) | 2017-03-15 |
| CN105838812A (en) | 2016-08-10 |
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