CN109371157A - A kind of labeling method and application of cucumber sterile gene correlation CAPS - Google Patents
A kind of labeling method and application of cucumber sterile gene correlation CAPS Download PDFInfo
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- CN109371157A CN109371157A CN201811491502.3A CN201811491502A CN109371157A CN 109371157 A CN109371157 A CN 109371157A CN 201811491502 A CN201811491502 A CN 201811491502A CN 109371157 A CN109371157 A CN 109371157A
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- 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
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- 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
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Abstract
The invention discloses a kind of identification cucumber male sterility gene Csa3M006660 wild types, Heterozygous mutants, the specific C APS of Mutants homozygous is marked, it is characterized by comprising the label JD-CsaMS-CAPS2.0 for detecting wild type site, and the nucleotide sequence of the label JD-CsaMS-CAPS2.0 is as shown in SEQ ID NO:1；Wild type gene group DNA is as shown in SEQ ID NO:2, and mutant gene group is as shown in SEQ ID NO:3.The method that it is combined by design primer amplification, digestion and agarose gel electrophoresis can carry out seedling stage high frequency zones to two kinds of characters of male sterility in cucumber " YL-5 " strain and male-fertile.Method of the invention has the characteristics that the period is short, at low cost, high-efficient, can be identified in crossbreeding, backcrossing segregating population etc., accurately distinguish heterozygosis, homozygous genotype, improve breeding efficiency.
Description
Technical field
The invention belongs to field of biotechnology, more particularly to a kind of CAPS molecule labelling method for identifying cucumber fertility and
Using.
Background technique
Spring in 2012, we are in one, Tianjin Wuqing test site height for having found cucumber male-sterile mutation in self-mating system
Body, by the method for hybridization, selfing and backcrossing, identifying sterile character is the nuclear male sterility by Dominant gene, and
Sterile character is recessiveness relative to fertile character.Pond is mixed further through building F2, using weight sequencing technologies and constructs genetic map
Method carries out finely positioning to sterile gene, and final result is determined as Csa3M006660(Han, Y., et al. 2017.).
Existing patent CN107058552A is the identification method based on sequencing level, by extracting mixed pond genomic DNA, knot
Weight sequencing technologies are closed, sequence comparison is carried out by analysis of biological information software, determines the genetic background of fertility.
Since existing patent CN107058552A is not appropriate in laboratory the support of sequencing technologies to carry out seedling to material
The identification of phase fertility background, and it is at high cost, generation data volume is big, analyzes time and effort consuming, exists simultaneously certain error, is badly in need of
A kind of completely new, more easy accurate method is developed to complete the mirror to cucumber male sterile line " YL-5 " fertility inheritting background
It is fixed.
Summary of the invention
The present invention overcomes high-flux sequence is relied on to the defect of cucumber fertility background detection technology, provide it is a kind of it is low at
This, when low consumption, efficient cucumber fertility background detection method.Present invention aim to address asking for cucumber fertility background detection
Topic.
To achieve the above object, the invention discloses following technology contents:
A kind of identification cucumber male sterility gene Csa3M006660 wild type, Heterozygous mutants, the specificity of Mutants homozygous
CAPS label, it is characterised in that: including the label JD-CsaMS-CAPS2.0, the label JD- for detecting wild type site
The nucleotide sequence of CsaMS-CAPS2.0 is as shown in SEQ ID NO:1；Wild type gene group DNA as shown in SEQ ID NO:2,
Mutant gene group is as shown in SEQ ID NO:3.
For identifying cucumber male sterility gene Csa3M006660 wild type, Heterozygous mutants, Mutants homozygous it is special
Property CAPS label primer, it is characterised in that the primer are as follows:
Upstream primer JD-CAPS2.0-F:5 '-AGGAAGAGAAGTGAAGCAA-3 ' is as shown in SEQ ID NO:4
Downstream primer JD-CAPS2.0-R:5 '-CCGTAACGTAATATGTGTCT-3 ' as shown in SEQ ID NO:5
The present invention further discloses for identifying cucumber male sterility gene Csa3M006660 wild type, Heterozygous mutants are pure
Close application of the primer of the specific C APS label of mutant in cucumber male sterility breeding.
More detailed description of the present invention is as follows:
(1) the present invention solves the technical problem of: propose it is a kind of it is economical, efficiently, it is accurately male for seedling stage assay cucumber
Property infertility wild type, Heterozygous mutants, specific C APS label, the primer and application thereof of Mutants homozygous.
(2) technical solution of the present invention (one) is: a kind of identification cucumber male sterility gene Csa3M006660 wild type,
Heterozygous mutants, the specific C APS label of Mutants homozygous, it is characterised in that: including the label for detecting wild type site
The nucleotide sequence of JD-CsaMS-CAPS2.0, the label JD-CsaMS-CAPS2.0 are shown in SEQ ID NO:1:
TAATATGGACAGCCGATGGCCGGCGAGACGGTTAGAATACGCAGCGGAGGTGATTGTGAA AGCATTGGAAG
AGAAGAAATCAGATAAATTCAGCCATGGCGGAAATGGAATGACTCGTCA AGATGTTCGAGATGCTGCTCGCCTTC
ACATCGGCGACACTGGATTGCTCGATTACGTTCT AAAATCACTGAACAACGTGATCGTAGGTAACCAAATAGTTC
GCCGTGCAGTGAATCCTAA AACACGAATTTTAGAGTACACGATTCATGAACTTAGAAATGGCATTCAATTAACAG
AAGAGCAAGAATCAACAGAAAATTCAGAACCAACCGTAACTCCTGGCAAAGACATTTACAACGA CGTGCTATGTA
TATACAGAAGCATTTTCCTTGACTATCCAGAATCAGAAATGGTAGAATT AGCAACCCAGGGAGTTCTCGATAGTA
AACATTTTGCTAAAGAATGGCCTCTTCAAGATGA AGAAGAGCATCTATTGACGTTCATTATCAAATTGATGCCGA
GGCTAACTTTTACACATAC GGATTTAGAGTTGAAGAGTGATTTCATGCCATCCGGCGAGGTAGTGGTTCTTCCAC
TACACACAACAATCGGAGAAGT
SEQ ID NO:2
TAATATGGACAGCCGATGGCCGGCGAGACGGTTAGAATACGCAGCGGAGGTGATTGTGAA AGCATTGGAA
GAGAAGAAATCAGATAAATTCAGCCATGGCGGAAATGGAATGACTCGTCA AGATGTTCGAGATGCTGCTCGCCTT
CACATCGGCGACACTGGATTGCTCGATTACGTTCT AAAATCACTGAACAACGTGATCGTAGGTAACCAAATAGTT
CGCCGTGCAGTGAATCCTAA AACACGAATTTTAGAGTACACGATTCATGAACTTAGAAATGGCATTCAATTAACA
GAAGA GCAAGAATCAACAGAAAATTCAGAACCAACCGTAACTCCTGGCAAAGACATTTACAACGA CGTGCTATG
TATATACAGAAGCATTTTCCTTGACTATCCAGAATCAGAAATGGTAGAATT AGCAACCCAGGGAGTTCTCGATAG
TAAACATTTTGCTAAAGAATGGCCTCTTCAAGATGA AGAAGAGCATCTATTGACGTTCATTATCAAATTGATGCC
GAGGCTAACTTTTACACATAC GGATTTAGAGTTGAAGAGTGATTTCATGCCATCCGGCGAGGTAGTGGTTCTTCC
ACTACA CACAACAATCGGAGAAGT
SEQ ID NO:3
TAATATGGACAGCCGATGGCCGGCGAGACGGTTAGAATACGCAGCGGAGGTGATTGTGAA AGCATTGGAAG
AGAAGAAATCAGATAAATTCAGCCATGGCGGAAATGGAATGACTCGTCA AGATGTTCGAGATGCTGCTCGCCTTC
ACATCGGCGACACTGGATTGCTCGATTACGTTCT AAAATCACTGAACAACGTGATCGTAGGTAACCAAATAGTTC
GCCGTGCAGTGAATCCTAA AACACGAATTTTAGATTACACGATTCATGAACTTAGAAATGGCATTCAATTAACAG
AAGA GCAAGAATCAACAGAAAATTCAGAACCAACCGTAACTCCTGGCAAAGACATTTACAACGA CGTGCTATGT
ATATACAGAAGCATTTTCCTTGACTATCCAGAATCAGAAATGGTAGAATT AGCAACCCAGGGAGTTCTCGATAGT
AAACATTTTGCTAAAGAATGGCCTCTTCAAGATGA AGAAGAGCATCTATTGACGTTCATTATCAAATTGATGCCG
AGGCTAACTTTTACACATAC GGATTTAGAGTTGAAGAGTGATTTCATGCCATCCGGCGAGGTAGTGGTTCTTCCA
CTACA CACAACAATCGGAGAAGT
(3) technical solution of the present invention (two) is: one kind is used to identify cucumber male sterility gene Csa3M006660 wild type,
Heterozygous mutants, the primer of the specific C APS label of Mutants homozygous, it is characterised in that the primer are as follows:
Upstream primer JD-CAPS2.0-F:5 '-AGGAAGAGAAGTGAAGCAA-3 '
Downstream primer JD-CAPS2.0-R:5 '-CCGTAACGTAATATGTGTCT-3 '
(4) technical solution of the present invention (three) is: it is above-mentioned for identifying cucumber male sterility gene Csa3M006660 wild type,
Heterozygous mutants, application of the primer of the specific C APS label of Mutants homozygous in cucumber male sterility breeding technique.It is real
Test as the result is shown: CAPS that the present invention obtains label can precise Identification cucumber male sterility gene Csa3M006660 wild type,
Heterozygous mutants, Mutants homozygous can be applied to cucumber male sterility assistant breeding, realize seedling stage to cucumber male sterility material
Expect the identification of genetic background.
(5) present invention is further, and aforementioned applications comprise the steps of:
I, cucumber sample gene group DNA is extracted;
Ii, pass through primer JD-CAPS2.0-F and JD-CAPS2.0-R, PCR is carried out to Cucumber germplasm DNA obtained in step i
Amplification, obtains single goal segment;
Iii, PCR product obtained in step ii is subjected to agarose through restriction enzyme CviQI digestion, and to digestion products
Gel electrophoresis analysis, if gel electrophoresis shows the characteristic bands of 908bp, 362bp, 256bp, the sample is heterozygous mutant
Body；If gel electrophoresis shows the characteristic bands of a 908bp, the sample is Mutants homozygous；
If gel electrophoresis shows the characteristic bands of 362bp, 256bp, the sample is wild type.
(6) further, in step ii, the PCR amplification system and program are as follows:
PCR system (20ul):
Template 1ul
JD-CAPS-F 1ul
JD-CAPS-R 1ul
Mix 10ul
ddH2O 7ul
PCR program:
94℃ 3min
94℃ 15s
51.5℃ 15s
72℃ 30s
72℃ 5min
4 DEG C of preservations
In step iii, the endonuclease reaction system are as follows:
Digestion system (10ul)
Restriction endonuclease 1ul
DNA 1ug
10XNEBuffer 1ul
Total Rxn Volume 10ul
25 DEG C of temperature
Time 2h
The present invention has mainly investigated the Molecular Identification for carrying out precise and high efficiency to cucumber male sterile material how in seedling stage, and emphasis is
Solve the problems, such as it is high for seedling stage cucumber male sterility sample gene background appraisal cost at present, the difficult point of invention be improve seedling
The efficiency of phase cucumber male sterility sample gene background identification, the innovation of the invention consists in that first identification cucumber male sterility base
Because the CAPS of background is marked.
Test effect of the present invention is as follows:
Seedling stage can be completed to the mirror of cucumber male sterile line fertility inheritting background by simple PCR and endonuclease reaction in the present invention
It is fixed.
Conclusion (of pressure testing)
Agarose gel electrophoresis is carried out to digestion products:
If gel electrophoresis shows the characteristic bands of 618bp, 362bp, 256bp, the sample is Heterozygous mutants.If solidifying
Gel electrophoresis shows the characteristic bands of a 618bp, and the sample is Mutants homozygous.If gel electrophoresis show 362bp,
The characteristic bands of 256bp, the sample are wild type.
Detailed description of the invention
Fig. 1 is cucumber male sterility sample through above-mentioned steps i, as a result, in figure on duct after step ii, step iii processing
Lower correspondence, from left to right successively are as follows: Marker indicates D2000 Marker；A indicates fertile 643a；B indicates fertile 643b；C table
Show fertile 643c；D indicates infertility 641d；E indicates infertility 642e；F indicates infertility 642；G indicates fertile g；H indicates fertile h；I table
Show fertile i；J indicates infertility j；K indicates infertility k；L indicates infertility l；M indicates positive control；N indicates negative control.If gel
Electrophoresis showed has the characteristic bands of 618bp, 362bp, 256bp, and the sample is Heterozygous mutants.If gel electrophoresis is shown
The characteristic bands of one 618bp, the sample are Mutants homozygous.If gel electrophoresis shows the feature of 362bp, 256bp
Band, the sample are wild type.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.Cucumber " YL-5 " male sterile line sample source is in (Han, Y., et al. 2017.).This
Raw materials used and reagent is invented to be commercially available.
Embodiment 1
The present embodiment shooting identification cucumber male sterility gene Csa3M006660 wild type, Heterozygous mutants, Mutants homozygous
The foundation of specific C APS label
(1) design of primers:
Essence by Tianjin Cucumber Inst. about cucumber (Cucumis sativus L.) mutant male sterility gene ms-3
Fine positioning (Han, Y., et al. (2017) " Fine mapping of a male sterility gene ms-3 in
A novel cucumber (Cucumis sativus L.) mutant. " Theor Appl Genet.) it studies it is found that such as
The 1258 bit base of fruit Csa3M006660 gene are T, then the plant is sterile plant.Utilize Primer premier 6.0
Two sides design of the software in the sudden change region 1bp includes the upstream and downstream primer JD-CAPS2.0 in the site.
Upstream primer JD-CAPS2.0-F:5 '-AGGAAGAGAAGTGAAGCAA-3 '
Downstream primer JD-CAPS2.0-R:5 '-CCGTAACGTAATATGTGTCT-3 '
(2) amplification and endonuclease bamhi theory analysis:
Wild type gene group DNA can amplify the segment of a 618bp (column are shown in SEQ ID NO:2);Mutant gene group DNA energy
Amplify segment (column are shown in SEQ ID NO:3) of a 618bp
Amplified production is through restriction enzyme CviQI digestion and carries out 2% agarose gel electrophoresis:
If gel electrophoresis shows the characteristic bands of 618bp, 362bp, 256bp, the sample is Heterozygous mutants；
If gel electrophoresis shows the characteristic bands of a 618bp, the sample is Mutants homozygous；
If gel electrophoresis shows the characteristic bands of 362bp, 256bp, the sample is wild type.
(3) verifying of molecular labeling
Material to be tested: A, fertile 643a；B, fertile 643b；C, fertile 643c；D, infertility 641d；E, infertility 642e；F, infertility
642；G, fertile g；H, fertile h；I, fertile i；J, infertility j；K, infertility k；L, infertility l；Above-mentioned material carries out stringent phenotype
It is sampled after identification.
Cucumber sample gene group DNA is extracted according to CTAB method, utilizes identification primer JD-CAPS2.0-F and JD-CAPS2.0-
R expands genomic DNA, and reaction system and method are as follows:
PCR system (20ul):
Template 1ul
JD-CAPS-F 1ul
JD-CAPS-R 1ul
Mix 10ul
ddH2O 7ul
PCR program:
94℃ 3min
94℃ 15s
51.5℃ 15s
72℃ 30s
72℃ 5min
4 DEG C of preservations
Amplified production passes through restriction enzyme CviQI digestion, and digestion system is as follows:
Digestion system (10ul)
Restriction endonuclease 1ul
DNA 1ug
10XNEBuffer 1ul
Total Rxn Volume 10ul
25 DEG C of temperature
Time 2h
As a result as shown in Figure 1, duct is corresponding up and down in figure, from left to right successively are as follows: Marker indicates D2000 Marker；A table
Show fertile 643a；B indicates fertile 643b；C indicates fertile 643c；D indicates infertility 641d；E indicates infertility 642e；F indicates infertility
642；G indicates fertile g；H indicates fertile h；I indicates fertile i；J indicates infertility j；K indicates infertility k；L indicates infertility l；M indicates sun
Property control；N indicates negative control.If gel electrophoresis shows the characteristic bands of 618bp, 362bp, 256bp, the sample is
Heterozygous mutants.If gel electrophoresis shows the characteristic bands of a 618bp, the sample is Mutants homozygous.If solidifying
Gel electrophoresis shows the characteristic bands of 362bp, 256bp, and the sample is wild type.Digestion interpretation result and phenotypic evaluation result
It is completely the same, it was demonstrated that CAPS that the present invention develops label can precise Identification cucumber male sterility gene Csa3M006660 it is wild
Type, Heterozygous mutants, Mutants homozygous can be applied to cucumber male sterility assistant breeding, realize seedling stage to cucumber male not
Educate the identification of material genetic background.
After the preferred embodiment of detailed description, it is familiar with this skilled worker and is clearly understood that, it is above-mentioned not departing from
Various change and modification can be carried out under claim and spirit, it is all according to the technical essence of the invention to above embodiments institute
Make any simple modification, equivalent change and modification, belongs to the range of technical solution of the present invention.And the present invention is not also by specification
The limitation of middle example embodiment.
SEQUENCE LISTING
<110>Tianjin Kerun Agricultural Science & Technology Co., Ltd.
<120>a kind of labeling method and application of cucumber sterile gene correlation CAPS
<160> 5
<170> PatentIn version 3.5
<210> 1
<211> 618
<212> DNA
<213>artificial sequence
<400> 1
taatatggac agccgatggc cggcgagacg gttagaatac gcagcggagg tgattgtgaa 60
agcattggaa gagaagaaat cagataaatt cagccatggc ggaaatggaa tgactcgtca 120
agatgttcga gatgctgctc gccttcacat cggcgacact ggattgctcg attacgttct 180
aaaatcactg aacaacgtga tcgtaggtaa ccaaatagtt cgccgtgcag tgaatcctaa 240
aacacgaatt ttagagtaca cgattcatga acttagaaat ggcattcaat taacagaaga 300
gcaagaatca acagaaaatt cagaaccaac cgtaactcct ggcaaagaca tttacaacga 360
cgtgctatgt atatacagaa gcattttcct tgactatcca gaatcagaaa tggtagaatt 420
agcaacccag ggagttctcg atagtaaaca ttttgctaaa gaatggcctc ttcaagatga 480
agaagagcat ctattgacgt tcattatcaa attgatgccg aggctaactt ttacacatac 540
ggatttagag ttgaagagtg atttcatgcc atccggcgag gtagtggttc ttccactaca 600
cacaacaatc ggagaagt 618
<210> 2
<211> 618
<212> DNA
<213>artificial sequence
<400> 2
taatatggac agccgatggc cggcgagacg gttagaatac gcagcggagg tgattgtgaa 60
agcattggaa gagaagaaat cagataaatt cagccatggc ggaaatggaa tgactcgtca 120
agatgttcga gatgctgctc gccttcacat cggcgacact ggattgctcg attacgttct 180
aaaatcactg aacaacgtga tcgtaggtaa ccaaatagtt cgccgtgcag tgaatcctaa 240
aacacgaatt ttagagtaca cgattcatga acttagaaat ggcattcaat taacagaaga 300
gcaagaatca acagaaaatt cagaaccaac cgtaactcct ggcaaagaca tttacaacga 360
cgtgctatgt atatacagaa gcattttcct tgactatcca gaatcagaaa tggtagaatt 420
agcaacccag ggagttctcg atagtaaaca ttttgctaaa gaatggcctc ttcaagatga 480
agaagagcat ctattgacgt tcattatcaa attgatgccg aggctaactt ttacacatac 540
ggatttagag ttgaagagtg atttcatgcc atccggcgag gtagtggttc ttccactaca 600
cacaacaatc ggagaagt 618
<210> 3
<211> 618
<212> DNA
<213>artificial sequence
<400> 3
taatatggac agccgatggc cggcgagacg gttagaatac gcagcggagg tgattgtgaa 60
agcattggaa gagaagaaat cagataaatt cagccatggc ggaaatggaa tgactcgtca 120
agatgttcga gatgctgctc gccttcacat cggcgacact ggattgctcg attacgttct 180
aaaatcactg aacaacgtga tcgtaggtaa ccaaatagtt cgccgtgcag tgaatcctaa 240
aacacgaatt ttagattaca cgattcatga acttagaaat ggcattcaat taacagaaga 300
gcaagaatca acagaaaatt cagaaccaac cgtaactcct ggcaaagaca tttacaacga 360
cgtgctatgt atatacagaa gcattttcct tgactatcca gaatcagaaa tggtagaatt 420
agcaacccag ggagttctcg atagtaaaca ttttgctaaa gaatggcctc ttcaagatga 480
agaagagcat ctattgacgt tcattatcaa attgatgccg aggctaactt ttacacatac 540
ggatttagag ttgaagagtg atttcatgcc atccggcgag gtagtggttc ttccactaca 600
cacaacaatc ggagaagt 618
<210> 4
<211> 19
<212> DNA
<213>artificial sequence
<400> 4
aggaagagaa gtgaagcaa 19
<210> 5
<211> 20
<212> DNA
<213>artificial sequence
<400> 5
ccgtaacgta atatgtgtct 20
Claims (3)
1. a kind of identification cucumber male sterility gene Csa3M006660 wild type, Heterozygous mutants, the specificity of Mutants homozygous
CAPS label, it is characterised in that: including the label JD-CsaMS-CAPS2.0, the label JD- for detecting wild type site
The nucleotide sequence of CsaMS-CAPS2.0 is as shown in SEQ ID NO:1；Wild type gene group DNA as shown in SEQ ID NO:2,
Mutant gene group is as shown in SEQ ID NO:3.
2. one kind is for identifying cucumber male sterility gene Csa3M006660 wild type, Heterozygous mutants, the spy of Mutants homozygous
The primer of anisotropic CAPS label, it is characterised in that the primer are as follows:
Upstream primer JD-CAPS2.0-F:5 '-AGGAAGAGAAGTGAAGCAA-3 ' is as shown in SEQ ID NO:4
Downstream primer JD-CAPS2.0-R:5 '-CCGTAACGTAATATGTGTCT-3 ' is as shown in SEQ ID NO:5.
3. for identifying cucumber male sterility gene Csa3M006660 wild type described in claim 2, Heterozygous mutants are homozygous
Application of the primer of the specific C APS label of mutant in cucumber male sterility breeding.
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| CN201811491502.3A CN109371157A (en) | 2018-12-07 | 2018-12-07 | A kind of labeling method and application of cucumber sterile gene correlation CAPS |
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| CN201811491502.3A CN109371157A (en) | 2018-12-07 | 2018-12-07 | A kind of labeling method and application of cucumber sterile gene correlation CAPS |
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| CN (1) | CN109371157A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113604592A (en) * | 2021-07-20 | 2021-11-05 | 天津市农业科学院 | InDel molecular marker for changing white cauliflower ball flower purple character and primer and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105420408A (en) * | 2016-01-27 | 2016-03-23 | 天津科润农业科技股份有限公司黄瓜研究所 | Cucumber male sterility gene related SNP (Single Nucleotide Polymorphism) marker and application |
| CN107058552A (en) * | 2017-05-05 | 2017-08-18 | 天津科润农业科技股份有限公司黄瓜研究所 | Cucumber male sterility gene, molecular labeling, screening technique and application thereof |
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- 2018-12-07 CN CN201811491502.3A patent/CN109371157A/en active Pending
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| CN113604592B (en) * | 2021-07-20 | 2023-10-27 | 天津市农业科学院 | InDel molecular marker with white cauliflower ball purple character, primer and application thereof |
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