CN104789562A - Molecular marker Indel-T-47 for close linkage with cucumber parthenocarpic main-effect QTL (quantitative trait loci) - Google Patents
Molecular marker Indel-T-47 for close linkage with cucumber parthenocarpic main-effect QTL (quantitative trait loci) Download PDFInfo
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- CN104789562A CN104789562A CN201510235861.2A CN201510235861A CN104789562A CN 104789562 A CN104789562 A CN 104789562A CN 201510235861 A CN201510235861 A CN 201510235861A CN 104789562 A CN104789562 A CN 104789562A
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Abstract
The invention discloses a molecular marker Indel-T-47 for close linkage with cucumber parthenocarpic main-effect QTL (quantitative trait loci), and belongs to the technical field of biology. The invention firstly discloses a molecular marker Indel-T-47 for close linkage with QTL Parth2.1, and further discloses a primer Indexl-T-47F/Indel-T-47R of the marker. In a segregating population containing Parth2.1, the marker is significantly related to parthenocarpy; in an all-female selfing line material containing the Parth2.1, a 155bp product can be amplified from a strong parthenocarpy strain by the marker primer Index-T-47F/Index-T-47R; the marker primer Index-T-47F/Index-T-47R is a peak marker of the Parth2.1, and is closely linked with the Parth2.1. The molecular marker for close linkage with the Parth2.1 disclosed by the invention can be applied to assistant breeding of an all-female cucumber parthenocarpy molecular marker.
Description
Technical field
The invention discloses the molecule marker of Parthenocarpy in Cucumber main effect QTL, belong to vegetables molecular genetic breeding technical field, can be used for the qualification of parthenocarpy ability and the molecular mark of the complete female kind of matter of cucumber.
Background technology
Cucumber is one of vegetables of China's commerial growing, has high social value and economic benefit.Parthenocarpy refers to that ovary stimulates without Pollination Fertilization or other and develops into the phenomenon of fruit.The cucumber variety that parthenocarpy ability is strong, has significant yield potential, generally can improve output more than 20%, and the fruit of parthenocarpy is because of without seed, and melon pulp is few, and pulp is thick, and quality is improved, and commodity is good.Therefore, parthenocarpy is and cucumber yield and the closely-related important economical trait of quality, is one of objective trait of cucumber variety seed selection, in breed cucumber, has utility value.
At present, the seed selection of Parthenocarpy in Cucumber material or kind mainly relies on field bundle flower isolation to carry out phenotypic evaluation, the method is length consuming time not only, need a large amount of man power and materials, and parthenocarpy is easily by the impact of ambient conditions, Phenotypic Selection efficiency is not high, and seed selection difficulty is large, it is unstable to there is parthenocarpy in the cucumber variety be bred as, and is subject to the drawbacks such as planting environment impact.In recent years, along with the development of molecular marking technique, many investigators utilize the molecule marker chain with objective trait QTL to carry out assisted Selection to kind of matter, this method is not subject to the impact of genetic expression and environmental factors, can carry out selecting in early generation and simple to operate, greatly enhance speed and the accuracy of selection.Wherein InDel (Insertion/Deletion) molecule marker be based on full-length genome resurvey sequence exploitation mark, because of its distribute in genome extensively, density is large, stability and polymorphic rate high, detect easy and there is codominant inheritance, reproducible, to advantages such as DNA specification of quality are lower, can, efficiently and accurately for genetic map construction, the molecule marker chain with objective trait be utilized to carry out assistant breeding.
Based on above-mentioned purpose, the present invention has carried out Parthenocarpy in Cucumber genetic mechanism and parthenocarpy QTL Position Research.Be maternal with strong parthenocarpy self-mating system ' EC1 ', non-parthenocarpy self-mating system ' 8419s-1 ' is male parent, constructs F
2:3colony and F
4:5residual heterozygous line colony (RHL, Residual heterozygous line), utilizes F
2:3colony positions Parthenocarpy in Cucumber main effect QTL site, utilizes F
4:5residual heterozygous line colony verifies this main effect QTL, weighs the analysis of sequencing result in conjunction with parents simultaneously, develops and screen to mark with the closely linked InDel in main effect QTL site, and utilizes F
3:4colony and 7 parts of complete female cucumber germplasm materials verify the Selection effect of this mark to parthenocarpy.This mark can be the breeding of Parthenocarpy in Cucumber molecular marker assisted selection and provides novel, efficient, practical molecule marker.
Summary of the invention
The object of this invention is to provide molecule marker InDel-T-47 closely linked with Parthenocarpy in Cucumber main effect QTL Parth2.1, and apply the qualification that this mark carries out Parthenocarpy in Cucumber germplasm materials.
The present invention is achieved by the following technical programs:
The closely linked molecule marker InDel-T-47 of parthenocarpy main effect QTL Parth2.1 in cucumber self-mating system ' EC1 ', its upstream primer sequence is TCAAACGAAAGGTAAGGAGGAATG, and downstream primer sequence is AGGCCTGAAGAGAGCTTCAAAGTA.
The invention also discloses the method for this primer qualification Parthenocarpy in Cucumber material, namely treat expert evidence with this labeled primer and increase through performing PCR, amplify the material of 155bp specific band, all there is stronger parthenocarpy ability.
Above-mentioned primer for the identification of the concrete grammar of Cucumber Germplasm material unisexuality ability is:
1., using the DNA of material to be identified as template, carry out pcr amplification with InDel-T-47 molecule marker primer;
2.PCR amplification reaction system consists of: 10 × buffer is (containing Mg
2+) 2.0 μ L, dNTP2.0 μ L, each 1 μ L of forward and reverse primer, Taq enzyme (5U μ L
-1) 0.25 μ L, DNA (10ng) 1.0 μ L, ddH
2o, 12.75 μ L, reaction cumulative volume is 20 μ L.Response procedures is 94 DEG C of denaturation 5min, each circulation 95 DEG C of denaturation 30s, 60 DEG C of annealing 30S, and 72 DEG C extend 1min 20s, 35 circulations; 72 DEG C extend 5min, 10 DEG C of preservations;
3.PCR product detects: reaction product is electrophoresis on the non-denaturing polyacrylamide gel of 7%, uses cma staining.If the plant that electrophoresis result can amplify 155bp specific band is the plant containing Parth2.1.
Beneficial effect:
1. in ' EC1 ' disclosed by the invention, closely linked molecule marker primer (InDel-T-47F/InDel-T-47R) designs according to the resurvey result of sequence of parents' genome with parthenocarpy main effect QTL Parth2.1 site, whether be used for qualification plant containing Parth2.1, compared with marking with the ALFP etc. announced in existing document, method is simple, convenient, amplification is stable.
2. the product that labeled primer InDel-T-47F/InDel-T-47R increases is the peak markers of Parth2.1, compared with molecule marker disclosed in existing document, the genetic distance of molecule marker of the present invention and Parth2.1 is nearer, utilizes the accuracy of molecular marker assisted selection Parth2.1 higher.
Accompanying drawing explanation
Fig. 1: utilize F
2:3colony carries out Parth2.1 location, and white box and twill frame represent the detected result in spring in 2013 and autumn respectively.
Fig. 2: utilize F
4:5the Parth2.1 section that RHL colony shortens and the mark position of Indel-T-47 on No. 2 karyomit(e)s
Note: reticulate pattern frame table shows the Parth2.1 section of shortening, and solid black round dot represents the peak of Parth2.1, LOD value is 9.1.
Fig. 3: with parents, strong parthenocarpy, non-parthenocarpy self-mating system DNA as template, pcr amplification is carried out with the labeled primer Indel-T-47F/Indel-T-47R in the present invention, result is presented at the single slice amplifying 155bp in EC1 (swimming lane 1) and strong parthenocarpy self-mating system (swimming lane 4-8), and does not all amplify corresponding band in non-parthenocarpy parent (swimming lane 2) and self-mating system (swimming lane 9-10).
Note: swimming lane 1,12:Marker; Swimming lane 2:.EC1; Swimming lane 3:8419s-1; Swimming lane 4:F1; Swimming lane 5-9: strong parthenocarpy self-mating system; Swimming lane 10-11: non-parthenocarpy self-mating system
Embodiment
At the F that spring in 2013, season in autumn two build " EC1 × 8419s-1 "
2:3colony has carried out parthenocarpy phenotypic evaluation.Select 1335 pairs of SSR primers and 173 pairs of InDel primers to carry out polymorphic primer screening between ' EC1 ' and ' 8419s-1 ', then by 232 to polymorphic primer at 16 F
2individual plant continues screening, and final 133 SSR primers and 9 InDel primers are at 145 F
2carry out pcr amplification in individual plant, build linkage map with JoinMap4.0, and to be correlated with QTL with Windows QTL Cartographer v2.5 software location Parthenocarpy in Cucumber.Result shows, and the main effect QTL Parth2.1 be arranged on No. 2 karyomit(e) all can be detected at two environment, and explainable phenotypic variation rate is respectively 17.4% and 10.2%.From F
3:4in colony, screening contains the residue heterozygosis individual plant of Parth2.1 and builds F
4:5colony.Utilize this colony to carry out mark encryption to main effect interval, and in conjunction with the phenotypic evaluation of individual plant parthenocarpy in this colony, shortened further by Parth2.1 section, Indel-T-47 is the peak markers of this section, with Parth2.1 close linkage.
1. design with Parth2.1 compact linkage molecule labeled primer
In the research that above-mentioned molecular marker linkage maps builds and Parthenocarpy in Cucumber main effect QTL Parth2.1 excavates, molecule marker Indel-T-47 is the peak markers (Fig. 2) of Parth2.1.This labeled primer to be resurveyed sequence by parents' full-length genome, and the small segment utilizing SAMTOOLS software detection length to be less than 50bp inserts and deletion segment, and according to the sequence of this site upstream and downstream 200bp, form by Premier 5.0 software design.
Mark Indel-T-47 primer sequence is: Indel-T-47F:5 '-TCAAACGAAAGGTAAGGAGGAATG-3 ', Indel-T-47R:5 '-AGGCCTGAAGAGAGCTTCAAAGTA-3 '.
2. labeled primer Indel-T-47F/Indel-T-47R is at F
3:4molecular Detection in colony
In order to verify the accuracy of this molecule marker, this laboratory utilizes Indel-T-47F/Indel-T-47R primer at F
3:4carry out pcr amplification detection in colony's individual plant, the phenotypic evaluation in conjunction with individual plant carries out Chi-square test, result display χ
2=20.14 > χ
2 0.01, 8=18.48, show Indel-T-47 mark and parthenocarpy pole significant correlation.The mean value that amplified production contains the plant parthenocarpy rate of 155bp specific band is extremely significantly high, the plant (table 1) of this band of Yu Buhan.
The significance analysis of table 1 Indel-T-47F/Indel-T-47R primer different genotype plant parthenocarpy rate
Note: a, b, c represent that P < 0.05, A, B, C represents P < 0.01, and parthenocarpy rate has carried out inverse sine conversion
Pcr amplification reaction system consists of: 10 × buffer is (containing Mg
2+) 2.0 μ L, dNTP 2.0 μ L, each 1 μ L of forward and reverse primer, Taq enzyme (5U μ L
-1) 0.25 μ L, DNA (10ng) 1.0 μ L, ddH
2o, 12.75 μ L, reaction cumulative volume is 20 μ L.Response procedures is 94 DEG C of denaturation 5min, each circulation 95 DEG C of denaturation 30s, 60 DEG C of annealing 30S, and 72 DEG C extend 1min 20s, 35 circulations; 72 DEG C extend 5min, 10 DEG C of preservations.PCR primer detects: reaction product is electrophoresis on the non-denaturing polyacrylamide gel of 7%, uses cma staining.
3. the Molecular Detection of labeled primer Indel-T-47F/Indel-T-47R in the complete female germplasm materials of cucumber
Utilize labeled primer Indel-T-47F/Indel-T-47R in parents, 5 parts of strong parthenocarpies and 2 parts of complete female germplasm materials (table 2) of non-parthenocarpy, carry out pcr amplification detection (condition and method the same).Result shows: the single slice that can amplify 155bp in ' EC1 ' parent and strong parthenocarpy germplasm materials, and does not all amplify this band (Fig. 3) in ' 8419s-1 ' parent and non-parthenocarpy material.
The complete female cucumber material of the different parthenocarpy ability of 7 parts, table 2
Claims (5)
1. the molecule marker Indel-T-47 of Parthenocarpy in Cucumber main effect QTL Parth2.1, is characterized in that: the primer sequence of this molecule marker is:
Indel-T-47F：5’-TCAAACGAAAGGTAAGGAGGAATG-3’
Indel-T-47R：5’-AGGCCTGAAGAGAGCTTCAAAGTA-3’
And Indel-T-47 is the peak markers of Parth2.1.
2. the molecule marking method of Parthenocarpy in Cucumber main effect QTL Parth2.1, its feature comprises: using the DNA of material to be identified as template, carries out pcr amplification with the primer pair of molecule marker Indel-T-47 according to claim 1; Polyacrylamide gel electrophoresis compartment analysis is carried out to amplified production; The plant that can amplify 155bp specific band is the plant containing Parthenocarpy in Cucumber main effect QTL Parth2.1.
3. the molecule marking method of Parthenocarpy in Cucumber main effect QTL Parth2.1 according to claim 2, is characterized in that comprising the following steps:
(1) with the DNA of material to be identified for template, carry out pcr amplification with the primer pair of molecule marker Indel-T-47 according to claim 1; Pcr amplification reaction be totally 20 μ l, comprising: 10 × buffer is (containing Mg
2+) 2.0 μ L, dNTP 2.0 μ L, each 1 μ L of forward and reverse primer, Taq enzyme (5U μ L
-1) 0.25 μ L, DNA (10ng) 1.0 μ L, ddH
2o, 12.75 μ L.Response procedures is 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 60 DEG C of annealing 30S, 72 DEG C extend 1min 20s, 35 circulations; 72 DEG C extend 5min, 10 DEG C of preservations.
PCR primer detects: reaction product is electrophoresis on the non-denaturing polyacrylamide gel of 7%, uses cma staining.
(2) labeled primer qualification Parth2.1: the plant that can amplify 155bp specific band is the plant containing Parth2.1.
4. the application of molecule marker according to claim 1 in molecular mark.
5. the application of molecule marker according to claim 1 in the complete female Cucumber Germplasm parthenocarpy ability of qualification.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367505A (en) * | 2016-09-06 | 2017-02-01 | 中国农业科学院郑州果树研究所 | A group of InDel sites closely linked with mountain peach aphid resistance main-effect QTL qGPAR-3-1 and application thereof |
| CN107586882A (en) * | 2017-11-01 | 2018-01-16 | 南京农业大学 | One molecular labeling InDel-RT that character close linkage is pushed up with cucumber circle fruit |
| CN108929916A (en) * | 2018-08-12 | 2018-12-04 | 扬州大学 | With the InDel label of Parthenocarpy in Cucumber gene close linkage and its application and primer |
| CN110195125A (en) * | 2019-07-05 | 2019-09-03 | 南京农业大学 | The linkage molecule of Parthenocarpy in Cucumber main effect QTL Parth2.1 marks and its application |
| CN114381548A (en) * | 2022-01-25 | 2022-04-22 | 南京农业大学 | Molecular marker assisted selective breeding method for cucumber strong parthenocarpy character multi-site selection |
-
2015
- 2015-05-06 CN CN201510235861.2A patent/CN104789562A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| ZHE WU ET AL.: "Identification of a stable major-effect QTL (Parth 2.1) controlling parthenocarpy in cucumber and associated candidate gene analysis via whole genome re-sequencing", 《BMC PLANT BIOLOGY》 * |
| ZHE WU ET AL.: "QTL Mapping of Parthenocarpy in Gynoecious Cucumber and Evaluation of a Marker Linked to a Major QTL", 《PLANT AND ANIMAL GENOME XXIII CONFERENCE》 * |
| 武喆 等: "黄瓜单性结实QTL定位分析", 《中国园艺学会黄瓜分会第四届年会论文摘要集》 * |
| 武喆 等: "黄瓜单性结实性状的QTL定位", 《中国农业科学》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367505A (en) * | 2016-09-06 | 2017-02-01 | 中国农业科学院郑州果树研究所 | A group of InDel sites closely linked with mountain peach aphid resistance main-effect QTL qGPAR-3-1 and application thereof |
| CN106367505B (en) * | 2016-09-06 | 2020-10-09 | 中国农业科学院郑州果树研究所 | Group of InDel loci closely linked with aphid-resistant major QTL qGPAR-3-1 of wild peach and application thereof |
| CN107586882A (en) * | 2017-11-01 | 2018-01-16 | 南京农业大学 | One molecular labeling InDel-RT that character close linkage is pushed up with cucumber circle fruit |
| CN107586882B (en) * | 2017-11-01 | 2021-06-11 | 南京农业大学 | Molecular marker InDel-RT closely linked with cucumber round fruit top character |
| CN108929916A (en) * | 2018-08-12 | 2018-12-04 | 扬州大学 | With the InDel label of Parthenocarpy in Cucumber gene close linkage and its application and primer |
| CN110195125A (en) * | 2019-07-05 | 2019-09-03 | 南京农业大学 | The linkage molecule of Parthenocarpy in Cucumber main effect QTL Parth2.1 marks and its application |
| CN114381548A (en) * | 2022-01-25 | 2022-04-22 | 南京农业大学 | Molecular marker assisted selective breeding method for cucumber strong parthenocarpy character multi-site selection |
| CN114381548B (en) * | 2022-01-25 | 2023-09-26 | 南京农业大学 | Molecular marker assisted selective breeding method for multi-site selection of strong parthenocarpy trait of cucumber |
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