CN117051149A - Molecular marker for identifying navel orange M28 easy to peel and application thereof - Google Patents
Molecular marker for identifying navel orange M28 easy to peel and application thereof Download PDFInfo
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- 240000002319 Citrus sinensis Species 0.000 title claims abstract description 109
- 235000005976 Citrus sinensis Nutrition 0.000 title claims abstract description 109
- 239000003147 molecular marker Substances 0.000 title claims abstract description 45
- 210000000349 chromosome Anatomy 0.000 claims abstract description 29
- 238000000034 method Methods 0.000 claims abstract description 14
- 238000003780 insertion Methods 0.000 claims abstract description 7
- 230000037431 insertion Effects 0.000 claims abstract description 7
- 239000002773 nucleotide Substances 0.000 claims abstract description 3
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 3
- 238000001962 electrophoresis Methods 0.000 claims description 6
- 230000003321 amplification Effects 0.000 claims description 5
- 238000010586 diagram Methods 0.000 claims description 5
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 5
- 238000012408 PCR amplification Methods 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 3
- 238000009395 breeding Methods 0.000 claims description 2
- 230000001488 breeding effect Effects 0.000 claims description 2
- 108020004414 DNA Proteins 0.000 description 16
- 239000000463 material Substances 0.000 description 8
- 238000012163 sequencing technique Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 4
- 230000036425 denaturation Effects 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 238000001502 gel electrophoresis Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 3
- 241000207199 Citrus Species 0.000 description 3
- 235000020971 citrus fruits Nutrition 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 2
- 108091081062 Repeated sequence (DNA) Proteins 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000000366 juvenile effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 235000013618 yogurt Nutrition 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000003976 plant breeding Methods 0.000 description 1
- 239000002893 slag Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
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Abstract
The invention discloses a molecular marker for identifying easily peeled navel orange M28 and application thereof, wherein the molecular marker is named Cschr8-M28, and a nucleotide sequence is shown as SEQ ID NO.1 and is positioned on a chromosome of navel orange Chr 8; wherein the 32509814 th site of the Chr8 chromosome is followed by insertion of a DNA transposon flanked by terminal repeats of the 32509807-32509814 th sequence of the Chr8 chromosome. The method is convenient for quick identification of the navel orange germplasm M28 seedlings easy to peel.
Description
Technical Field
The invention relates to a molecular marker for identifying easily peeled navel orange M28 and application thereof, belonging to the technical field of plant breeding.
Background
Citrus is the fruit tree with the widest cultivation area and important economic status in the south of China. The navel orange fruit has special flavor and rich nutrition and medicinal value, so the navel orange fruit is popular with people. However, the white cortex between the navel orange peel and the flesh is compact, and can not be easily pulled out, so that the purchase desire of people on the navel orange is affected to a certain extent, and the sales of the navel orange is affected.
In order to solve the problem, the navel orange easy to peel is bred, wherein the navel orange M28 easy to peel is one navel orange germplasm easy to peel, which is bud-changed and bred from a main cultivated navel orange variety 'Neoher'. The navel orange M28 easy to peel has moderate tree vigor, good yield, orange flesh, tender slag, sweet flavor, obvious easy peeling property of germplasm fruits and similar peeling characteristics to wide-peel oranges. However, due to the long juvenile period of citrus, there is difficulty in identifying varieties by morphology in the juvenile period.
Disclosure of Invention
The invention aims to overcome the defects in the prior art, and provides a molecular marker for identifying navel oranges and application thereof, which are convenient for quick identification of M28 seedlings of navel oranges easy to peel.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides a molecular marker for identifying easily peeled navel orange M28, wherein the molecular marker is named CsChr8-M28, and the nucleotide sequence is shown in SEQ ID NO.1 and is positioned on a chromosome of navel orange Chr 8;
the 32509814 th site of the chromosome of the Chur 8 is followed by insertion of a DNA transposon, which is flanked by terminal repeats formed by the 32509807-32509814 th sequence of the chromosome of the Chur 8.
With reference to the first aspect, further, the primer pair for amplifying the molecular marker CsChr8-M28 is CsChr8-M28-F and CsChr8-M28-R, wherein one primer is located on the chromosome of navel orange Chr8, and the other primer is located on the inserted DNA transposon.
Further, the sequence of Cschr8-M28-F is shown in SEQ ID NO.2, and the sequence of Cschr8-M28-R is shown in SEQ ID NO. 3.
In a second aspect, the invention provides a kit comprising a primer pair for amplification of the molecular marker Cschr8-M28 as described in any one of the above.
In a third aspect, the invention provides a method for identifying navel orange M28 easy to peel, comprising the step of amplifying genomic DNA of navel orange by detecting primer pair of molecular marker Cschr8-M28.
With reference to the third aspect, further, the method includes the steps of:
extracting genome DNA from navel orange products;
respectively carrying out PCR amplification on the extracted genome DNA by using a primer pair of a molecular marker Cschr8-M28 to obtain an amplification product;
respectively carrying out electrophoresis on the amplified products to obtain an electrophoresis gel diagram;
and screening out the target navel orange according to the pattern of the electrophoresis gel.
Furthermore, the molecular marker, the method or the kit can be applied to breeding of navel orange germplasm easy to peel, and is convenient and rapid.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a transposon insertion site positioned on a chromosome of navel orange M28 germplasm Chr8 easy to peel, and provides a dominant molecular marker CsChr8-M28 for detecting the quality of the navel orange M28 easy to peel based on the site, which is used for identifying whether navel orange seedlings or other propagation materials are the navel orange germplasm M28 easy to peel, and has obvious and rapid effect;
the primer pair of the molecular marker Cschr8-M28 for detecting the easy-to-peel navel orange M28 is provided, so that the quick identification of seedlings of the easy-to-peel navel orange M28 is facilitated.
Drawings
FIG. 1 is a schematic diagram showing the difference between M28 germplasm of a navel orange easy to peel and the chromosome sequence of the Chr8 of the navel orange 'New Yoghurt' provided by the embodiment of the invention;
FIG. 2 is a gel electrophoresis chart of molecular markers Cschr8-M28 for detecting M28 germplasm of navel orange easy to peel and 'Neoher' navel orange according to the embodiment of the invention;
fig. 3 is a gel electrophoresis chart of molecular markers Cschr8-M28 for detecting M28 germplasm of navel orange easy to peel and other navel orange varieties cultivated in the Gannan region according to the embodiment of the invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings. The following examples are only for more clearly illustrating the technical aspects of the present invention, and are not intended to limit the scope of the present invention.
The invention provides a difference site between a Chur 8 chromosome on M28 germplasm of a navel orange which is easy to peel and a 'Newhall' navel orange which is a bud source of the Chur 8 chromosome, and provides a molecular marker CsChur 8-M28 for detecting M28 germplasm of the navel orange which is easy to peel and a primer pair of the CsChur 8-M28.
The invention adopts the technical scheme that: the difference site between M28 germplasm of the navel orange easy to peel and the navel orange of which the bud mutation source is 'New HEL' is positioned on the chromosome of the Chr8, and the reference genome sequence (the chromosome sequence of the Chr8 of the sweet orange) is found in NCBI database (GenBank: CM 030747.1). A transposon sequence is inserted at a position behind 32509814 locus of a Chr8 chromosome of M28 germplasm of the navel orange which is easy to peel, and terminal repeated sequences formed by 32509807-32509814 (9 bp) sequences of the Chr8 chromosome are arranged at two sides of the inserted sequence. Based on the identified chromosomal structural variations, the molecular marker Cschr8-M28 was developed in close linkage with the site of variation.
The molecular marker CsChr8-M28 is located on the chromosome of Chr8, and the sequence is shown as SEQ ID NO. 1.
The invention provides a primer pair corresponding to a molecular marker CsChr8-M28 for identifying navel orange M28 easy to peel, wherein the upstream primer of CsChr8-M28 is CsChr8-M28-F, the sequence is shown as SEQ ID NO.2, and the downstream primer is CsChr8-M28, and the sequence is shown as SEQ ID NO. 3.
The invention also provides a method for detecting the easy-to-peel navel orange M28, which comprises the following steps:
the primer pair of the molecular marker Cschr8-M28 is used for amplifying the genome DNA of the navel orange, and after 1% agarose gel electrophoresis of the amplified product, if the amplified fragment corresponding to the variation in M28 germplasm is obtained, the amplified fragment is indicated to be the M28 germplasm of the navel orange which is easy to peel. If the amplified fragment corresponding to the variation in the easy-to-peel navel orange M28 is not obtained, the specific variation position contained in the easy-to-peel navel orange M28 germplasm does not exist, and the seedling is predicted to be not the easy-to-peel navel orange M28 germplasm easy to peel.
Specifically, the molecular marker Cschr8-M28 for detecting the M28 species of the navel orange easy to peel disclosed by the invention is selected by the following steps:
(1) In the process of continuously bud-changing seed selection of navel orange for many years, a strain of easy-to-peel bud-changing navel orange is found in 'New Yoghurt' navel orange, and M28 germplasm of the easy-to-peel navel orange is obtained.
(2) Analysis by re-sequencing techniques identified the sequence differences of 'newhol' and its budding germplasm easy-to-peel navel orange M28, transposon sequence insertion was identified after the 32509814 th site on the Chr8 chromosome of easy-to-peel navel orange M28, and terminal repeats formed by sequences flanking the insertion sequence with respect to the 32509807-32509814 (9 bp) th sequence of the Chr8 chromosome.
(3) Based on the sequence difference found above, a molecular marker CsChr8-M28 for detecting M28 species of navel orange easy to peel is developed, one amplification primer of the marker is positioned on a chromosome of the Chr8, and the other primer is positioned on an inserted transposon sequence, so that the specificity of the sequence is ensured.
Embodiment one:
in this example, the identification of the differential sites and the development of the molecular markers were performed as follows.
(1) Identification of the differential sites
Samples of M28 germplasm and 'New HEL' navel orange materials which are identified to be easy to peel are taken from a national navel orange engineering research center nursery, and three biological repeats are selected from different sample materials. And (3) extracting M28 germplasm of the navel orange easy to peel and DNA of the navel orange leaves of 'Newhol' by using a CTAB method, then constructing a library, and carrying out second generation sequencing with the depth of 30 multiplied.
According to the invention, the sweet orange genome is taken as a reference (http:// citrus. Hzau. Edu. Cn), sequencing data are analyzed in a Linux server, M28 in FIG. 1 represents a chromosome sequence diagram of M28 germplasm of navel orange easy to peel, and NHE represents a chromosome sequence diagram of 'Newhol' navel orange Chr 8. By comparison, as shown in the box-selected position of FIG. 1, sequencing sequences which cannot be completely mapped exist before and after 32509814 locus of one Chr8 chromosome of M28 germplasm of the navel orange easy to peel, and a plurality of repeats exist. Indicating that an insertion/deletion site was identified after the Chr8 chromosome 32509814 site.
(2) Differential site sequence clone analysis
In order to further define the sequence variation characteristics, primers are designed according to the reads of the variation site obtained by sequencing and the sequences at the two sides of the 32509814 site of the reference genome, M28 germplasm of the navel orange which is easy to peel and the DNA of the navel orange 'Neohol' are taken as templates, the sequence at the variation position is cloned, and the terminal repeated sequence formed by the 29470204-29470212 (9 bp) sequence of the Chr8 chromosome is found at the two wings of the inserted sequence, so that the inserted sequence is a DNA transposon.
(3) Molecular marker design
According to the characteristics of the inserted sequence, the invention designs a molecular marker CsChr8-M28, wherein one primer pair of the molecular marker CsChr8-M28 is located on a chromosome of navel orange Chr8, and the other primer pair is located on an inserted transposon sequence, as shown in figure 1. The primer pair is Cschr8-M28-F and Cschr8-M28-R, wherein the sequence of Cschr8-M28-F is shown as SEQ ID NO.2, and the sequence of Cschr8-M28-R is shown as SEQ ID NO. 3.
(4) Molecular marker identification of M28 germplasm of navel orange easy to peel and 'Newhall' navel orange material
The CTAB method is used for extracting the blade DNA of the navel orange material which is easy to peel, and the blade DNA is used for molecular marker identification of the navel orange material which is easy to peel, and the navel orange M28.
The extracted leaf DNA was amplified, and the PCR reaction system (10. Mu.l) contained 0.5. Mu.l of the DNA template, 0.25. Mu.l of each of the upstream and downstream primers (1 mmol/L), 5. Mu.l of Mix, and 4. Mu.l of ddH2O.
PCR reaction procedure: denaturation at 95℃for 5min; followed by 45 cycles of denaturation at 95℃for 30s, annealing at Tm (58.5 ℃) for 30s and elongation at 72℃for 30s; then extending for 5min at 72 ℃; finally, the mixture is preserved at 10 ℃.
The PCR amplification product of the present invention was electrophoresed on 1% agarose. Film was scanned and analyzed in a ChemiScope imaging system.
(5) Results and analysis
The molecular marker Cschr8-M28 can distinguish the M28 germplasm of the navel orange which is easy to peel from the navel orange of which the bud becomes the source 'Newhall'. In the identified gel plots, cschr8-M28 was linearly labeled with two bands, 1069bp in size and no band, respectively. The navel orange M28 with 1069bp band is easy to peel, and the navel orange with no band is 'Newhall'.
As shown in FIG. 2, the molecular marker Cschr8-M28 provided by the invention is used for detecting the M28 germplasm of the navel orange easy to peel and the gel electrophoresis chart of the 'Newhol' navel orange; wherein M is DL-2000marker,1-3 channels are Cschr8-M28 to detect M28 germplasm band type of easy-peeling navel orange, and 4-6 channels are Cschr8-M28 to detect 'Neoher' navel orange band type;
embodiment two:
this example carried out molecular markers Cschr8-M28 to identify `M 28` and other navel orange germplasm.
(1) Molecular marker detection is carried out on M28 germplasm of navel orange easy to peel and other navel orange germplasm.
The CTAB method is used for extracting the DNA of the navel orange M28 germplasm easy to peel and other domestic navel orange material leaves, and is used for molecular marker identification of the navel orange M28 easy to peel and other domestic navel orange materials.
The extracted DNA was subjected to PCR amplification by a specific PCR reaction system (10. Mu.l) containing 0.5. Mu.l of the DNA template, 0.25. Mu.l of each of the upstream and downstream primers (1 mmol/L), 5. Mu.l of Mix, and 4. Mu.l of ddH2O.
PCR reaction procedure: denaturation at 95℃for 5min; followed by 45 cycles of denaturation at 95℃for 30s, annealing at Tm (58.5 ℃) for 30s and elongation at 72℃for 30s; then extending for 5min at 72 ℃; finally, the mixture is preserved at 10 ℃.
The PCR amplified products were electrophoresed on 1% agarose. Film was scanned and analyzed in a ChemiScope imaging system.
(2) Results and analysis.
The molecular marker Cschr8-M28 can distinguish the M28 germplasm of the navel orange which is easy to peel from other germplasm of the domestic main cultivated navel orange. In the identified gel plots, cschr8-M28 was linearly labeled with two bands, 1069bp in size and no band, respectively. The M28 germplasm of the navel orange which has a 1069bp band is easy to peel, and other navel orange germplasm are all without bands.
As shown in FIG. 3, the molecular marker Cschr8-M28 provided by the invention is used for detecting the M28 germplasm of the navel orange easy to peel and the gel electrophoresis chart of other navel orange varieties in the Gannan region; wherein, 1-2 channels are Cschr8-M28 to detect the band type of the easy-peeling navel orange M28, and 3-24 channels are Cschr8-M28 to detect the band type of other navel orange varieties.
The molecular marker Cschr8-M28 or the primer pair, the kit and the screening step of the easy-to-peel navel orange M28 can be applied to detection of the easy-to-peel navel orange M28 germplasm.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that modifications and variations could be made by those skilled in the art without departing from the technical principles of the present invention, and such modifications and variations should also be regarded as being within the scope of the invention.
The DNA sequence related to the invention is as follows:
SEQ ID NO.1
CACATCCGTCATGTAAAACAAAAGGGCCTTCCAACTGAACTAAAGCTATTTTTCCTGCAATTATG
ACCCAGCAAACATGAGAGGAACTAGGCAATCACATTTACAATCTTTACTAACTGATGTGATGAA
TAATCGAAATAAACTAGCTTTTGGATAATGCTATCACTCTCTACTCATCTTCAAACATAAAAGAA
CAAATTTCAAACTTAAATCTCTCTCTATCACTATGAACAGTTGCAATTTCACCCCTTCCAAAAGA
AAATCATCCTTGTCCGGTAACACACAAATAAATAATTCACAATAGTCAATGACAAGCAATACTC
ACGAAATTTGCCATTACCTACACAATTTGAGAGCAGCAGAGACGATAGAGAAGAGAGAAAGCA
GACCCACGGTGAGAGACCAACGACGAGAGATCCACGGCCAGAGACAAATGCCAAGCACCACCG
CCGAGAAAGTTTGAGAATAGAGACGACAGGGAAGAGAGACAGCAGACCCACGGCCAGAGACCA
ACGCCGAGCACCCTCAGAAGGTTTGAGAACAGTGAGATTTCTGTGAGAGCAAATGGAGAAAAGT
TGTGCGTCGCGTGAGATGGATTGCGCGTGTGTTGGGATTGCTATGTGTATTACACGTGAGTTGGC
TCATTTTATAGTATGTCCAAAACGCACCGTTTGGTGCTTATTGTGGTGCTTAACGTGGGAAGAAT
CTCATTTTCCTTTTTCCCTCTTCACTCTCCTTTTCTTCTCTCTCTTTCTCTCTCTTTTTGTTTTAACAA
ACAAACAAACAGGATTGTTTTTTTTTAAAAAAAAAATACACACACTTATTGTTATTATTATTATT
ATTATTATTATTATTATTATTATTATTATTATCATCGTTATTTTATTTTATTCAAGTGATTAATTCT
TTATTATTTTTTTATTCATTTGTTGTTGTTATTACGATTATTTATTAGAATATGAACTTTTGCTTAT
AAATTTCAGGGAAAGAAGGCTATTGATGCACGTAATTGTCGCACGTGTTGCGAATGAGACTTTSEQ ID NO.2
GAAATCTAAATCAAGGGCACTG
SEQ ID NO.3
AAAGTCTCATTCGCAACACG
Claims (7)
1. the molecular marker for identifying the easily peeled navel orange M28 is characterized in that the molecular marker is named Cschr8-M28, the nucleotide sequence of the molecular marker is shown as SEQ ID NO.1, and the molecular marker is positioned on a chromosome of the navel orange Chr 8;
the 32509814 th site of the chromosome of the Chur 8 is followed by insertion of a DNA transposon, which is flanked by terminal repeats formed by the 32509807-32509814 th sequence of the chromosome of the Chur 8.
2. The molecular marker of claim 1, wherein the primer pair for amplifying the molecular marker CsChr8-M28 is CsChr8-M28-F and CsChr8-M28-R, wherein one primer is located on the chromosome of navel orange Chr8 and the other primer is located on the inserted DNA transposon.
3. The molecular marker according to claim 2, wherein the sequence of Cschr8-M28-F is shown in SEQ ID NO.2, and the sequence of Cschr8-M28-R is shown in SEQ ID NO. 3.
4. A kit comprising a primer pair for amplification of the molecular marker CsChr8-M28 of any one of claims 2-3.
5. A method for identifying navel orange M28 susceptible to peeling comprising the step of amplifying genomic DNA of navel orange by detecting primer pair of molecular marker CsChr8-M28 according to claim 2.
6. The method according to claim 5, comprising the steps of:
extracting genome DNA from navel orange products;
respectively carrying out PCR amplification on the extracted genome DNA by using a primer pair of a molecular marker Cschr8-M28 to obtain an amplification product;
respectively carrying out electrophoresis on the amplified products to obtain an electrophoresis gel diagram;
and screening out the target navel orange according to the pattern of the electrophoresis gel.
7. Use of a molecular marker according to any one of claims 1-3, or a method according to any one of claims 5-6, or a kit according to claim 4, for breeding easy-to-peel navel orange germplasm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202310918381.0A CN117051149A (en) | 2023-07-25 | 2023-07-25 | Molecular marker for identifying navel orange M28 easy to peel and application thereof |
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| CN202310918381.0A CN117051149A (en) | 2023-07-25 | 2023-07-25 | Molecular marker for identifying navel orange M28 easy to peel and application thereof |
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- 2023-07-25 CN CN202310918381.0A patent/CN117051149A/en active Pending
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