CN114410826B - InDel molecular marker co-separated from cucumber peel brightness gene and application thereof - Google Patents

InDel molecular marker co-separated from cucumber peel brightness gene and application thereof Download PDF

Info

Publication number
CN114410826B
CN114410826B CN202210126440.6A CN202210126440A CN114410826B CN 114410826 B CN114410826 B CN 114410826B CN 202210126440 A CN202210126440 A CN 202210126440A CN 114410826 B CN114410826 B CN 114410826B
Authority
CN
China
Prior art keywords
cucumber
brightness
indel
molecular marker
peel
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202210126440.6A
Other languages
Chinese (zh)
Other versions
CN114410826A (en
Inventor
刘兴旺
任华中
翟许玲
董明明
尹帅
吴皓颖
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
China Agricultural University
Original Assignee
China Agricultural University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by China Agricultural University filed Critical China Agricultural University
Priority to CN202210126440.6A priority Critical patent/CN114410826B/en
Publication of CN114410826A publication Critical patent/CN114410826A/en
Application granted granted Critical
Publication of CN114410826B publication Critical patent/CN114410826B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification
    • 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
    • C12Q2600/13—Plant traits
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Genetics & Genomics (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention relates to an InDel molecular marker co-separated from a cucumber peel brightness gene and application thereof. The InDel molecular marker provided by the invention is 4895bp InDel variation existing at 20,290,734-20,295,628 position on cucumber genome chromosome 5. The InDel molecular marker provided by the invention is co-separated from the cucumber pericarp brightness gene DULL, and is named as InDel-DULL. The molecular marker InDel-DULL is obviously related to the peel brightness character of the cucumber in the commodity melon period, so that the molecular marker InDel-DULL can be used for early identification and auxiliary breeding of the cucumber peel brightness character. The InDel molecular marker can be detected through PCR amplification and further agarose gel electrophoresis, has low equipment and technical requirement thresholds, has a series of advantages of non-destructive property, simple operation, high efficiency, low cost and the like, and has higher popularization and application values.

Description

InDel molecular marker co-separated from cucumber peel brightness gene and application thereof
Technical Field
The invention relates to the technical field of plant molecular genetic breeding, in particular to an InDel molecular marker coseparated with cucumber pericarp brightness genes and application thereof.
Background
Cucumber (culumis sativus l., 2n=2x=14) is an important vegetable crop of the cucurbitaceae melon genus, has high yield and good economic benefit, and is widely planted worldwide, especially in china. The brightness of the pericarp is one of important appearance quality traits of cucumber fruits, and can directly influence the commodity property of the cucumber fruits. In the fresh food consumer market, fruits with bright peel are deeply favored by consumers due to their bright, bright and shiny appearance compared with cucumber fruits with dark peel. Thus, peel brightness has become an important factor affecting the market value of fresh cucumber. Under the background, researching the heredity and molecular mark of cucumber peel brightness has important significance for breeding new cucumber varieties popular in the market.
Along with the completion of the first cucumber genome sequencing, a plurality of genes for regulating the appearance quality traits of cucumber fruits, including thorns, peel colors, fruit necks and the like, are cloned and reported sequentially. The complexity of the brightness property of the peel results in relatively slow progress in the study of the property. Since 1931, the first report of the trait has not been reported until now that there are clear regulatory genes and molecular markers for the trait that can be directly applied to molecular marker-assisted selective breeding. In this way, the cucumber breeding practitioner cannot utilize molecular markers to identify and select genotypes from the seedling stage in the process of cultivating a new variety aiming at the trait, so that great waste of manpower, material resources, financial resources and time is caused.
Disclosure of Invention
The invention provides an InDel molecular marker coseparated with the property based on the fine positioning of cucumber peel brightness genes. The marker can be used as an auxiliary selection means to be applied to the breeding process of new cucumber varieties aiming at peel brightness, so that the breeding efficiency can be greatly improved, the breeding period can be shortened, and the resources can be saved.
In a first aspect, the invention provides an InDel molecular marker co-separated from a cucumber peel brightness gene, wherein the nucleotide sequence of the InDel molecular marker is shown as SEQ ID NO. 3.
Specifically, the InDel molecular marker is positioned at 20,290,734-20,295,628 base position on chromosome 5 of the cucumber genome.
More specifically, the promoter sequence and the coding sequence of the cucumber pericarp brightness gene DULL are both located in the 4895bp indel variant sequence. If the 4895bp deletion exists in the cucumber genome, the cucumber peel brightness is achieved; if the 4895bp deletion does not exist in the cucumber genome, the cucumber peel has low brightness.
In a second aspect, the invention provides a primer pair for detecting InDel molecular markers, wherein the primer pair is shown as SEQ ID NO. 1-2.
If the cucumber genome contains a wild DULL gene sequence, the primer pair shown as SEQ ID NO.1-2 is used for amplifying an indel variation sequence shown as SEQ ID NO. 3.
When the primer pair provided by the invention is used for detection, if a single 5309bp band or double 5309bp bands are amplified, the pericarp of the cucumber material to be detected is shown to be in a dark phenotype; when only a single band of 414bp is amplified, the pericarp of the cucumber material to be tested is shown to be in a bright phenotype.
In a third aspect, the invention provides a kit containing the primer pair, wherein the kit is a rapid detection kit for identifying the brightness property of cucumber peel.
The invention also claims the use of the primer pair or kit described above for detecting the cucumber pericarp brightness gene DULL genotype, as understood by those skilled in the art.
And the application of the InDel molecular marker or the primer pair or the kit in cucumber molecular marker assisted selective breeding or germplasm resource identification.
In a fourth aspect, the invention provides a method for rapidly detecting brightness traits of cucumber peel, comprising the following steps:
step 1, extracting the whole genome DNA of a cucumber material to be detected;
step 2, performing PCR amplification by using a primer pair shown as SEQ ID NO. 1-2;
step 3, if a single 5309bp band or double 5309bp bands are amplified, indicating that the pericarp of the cucumber material to be detected presents a dark phenotype; if a single strip of 414bp is amplified, the pericarp of the cucumber material to be detected is shown to be in a bright phenotype.
Further, the extraction of the DNA of the cucumber material to be detected in the step 1 adopts an improved CTAB method;
further, the DNA polymerase used for PCR amplification in the above step 2 is Takara Primer STAR Max, the amplification rate is 5-10 seconds/1 kb; the procedure used for PCR amplification included:
pre-denaturation: 95 ℃ for 5 minutes;
and (3) circulation: denaturation at 98℃for 15 sec; annealing at 60 ℃ for 25 seconds; extending at 72 ℃ for 1 minute and 20 seconds; a total of 35 cycles;
extension: extending at 72 ℃ for 5 minutes;
and (3) preserving: preserving at 4 ℃.
The invention has the beneficial effects that:
(1) The molecular markers commonly used at present are SSR, SNP, caps, dcaps, inDel and the like. The detection of the SSR molecular markers requires polyacrylamide gel electrophoresis, the operation is complex, and the used reagents are harmful to human bodies and the environment; although the SNP molecular marker has high density, the instrument and equipment are expensive, and the application threshold is high; the detection steps of caps and Dcaps molecular markers are more, various restriction enzymes are needed, and the cost is high. In contrast, detection of the InDel molecular marker can be realized only through PCP amplification and agarose gel electrophoresis, and the method is simple and convenient to operate, low in cost, high-efficiency, nontoxic and low in application threshold.
(2) 150F pairs using InDel molecular markers of the invention 2 Experimental results of peel brightness prediction or identification of the segregating population and 50 natural populations show that the molecular marker is 100% linked with the cucumber peel brightness trait, and the molecular marker is specifically expressed as follows: when a single 5309bp strip or a double strip of 414bp and 5309bp is amplified by using the molecular marker, the brightness of the pericarp of the tested cucumber material is extremely low; when the molecular marker is used for amplifying a single 414bp band, the brightness of the pericarp of the tested cucumber material is extremely high. Therefore, the InDel molecular marker reported by the invention can be used as an auxiliary selection means to be applied to the breeding process of new cucumber varieties aiming at the brightness of the pericarps, the breeding efficiency can be greatly improved, the breeding period is shortened, the resources are saved, and in addition, the molecular marker can also be used for identifying the DULL genotype or the brightness of the pericarps in large-scale cucumber germplasm resources.
Drawings
FIG. 1 is a chart showing the fruit phenotype of near isogenic line materials with only a significant difference in peel brightness in example 1 of the present invention.
FIG. 2 is a gel electrophoresis pattern of the identification of genotypes of the pericarp brightness gene DULL in cucumber materials 2073-1, 2073-2 and F1 using InDel-DULL molecular markers in example 2 of the present invention.
Detailed Description
The following examples are illustrative of the invention and are not intended to limit the scope of the invention.
Example 1 acquisition of InDel molecular markers coseparated with cucumber pericarp Brightness Gene DULL
The embodiment provides a method for obtaining an InDel molecular marker coseparated with a cucumber peel brightness gene DULL, which comprises the following specific steps:
1) Materials and reagents
2073-1 and 2073-2 are a pair of near isogenic material with a significant difference in peel-only brightness formed by saturated backcrossing (fig. 1). The fruit of 2073-1 is dark and the fruit of 2073-2 is bright. Both of these materials are disclosed in journal articles (Liu B, guan D, zhai X, et al selection footprints reflect genomic changes associated with breeding efforts in 56 cucumber inbred lines[J ]. Horticulture research 2019,6 (127)). All primers used in this example were biosynthesized by Shanghai workers, were PAGE grade purified and were checked by HPLC. The rest reagents are all commonly used molecular biological reagents.
2) Genetic rule statistics of cucumber peel brightness traits
In the invention, in order to ascertain the genetic rule of cucumber peel brightness character, near isogenic line 2073-1 (P 1 ) And 2073-2 (P) 2 ) Construction of F 1 Hybridization population, and based thereon, further creates backcross population and F 2 Isolating the population. F (F) 1 The fruit exhibits a dark-grey phenotype like 2073-1. At F 2 In the population, the ratio of dark to bright plants was 3.05:1 (1119:367), proved by chi-square test to be 3:1 (χ) 2 ＝0.06<χ 2 0.05 ＝3.841)。BC 1 P 2 The ratio of dark to bright plants in the population was 1.17:1 (123:105), proved by chi-square test to be 1:1 (χ) 2 ＝1.27<χ 2 0.05 = 3.841), all plants BC 1 P 1 The backcross population exhibited a dull phenotype. The genetic analysis indicated that the cucumber peel brightness trait was controlled by a single recessive gene (table 1).
TABLE 1 genetic rule statistics of cucumber pericarp brightness Properties
3) Fine localization of cucumber peel brightness gene
Segregating population mix analysis is an effective way to rapidly locate genes. At 1486F 2 In plants, 50 single plants with dark fruits and 50 single plants with glossy fruits are selected, and two extreme DNA pools are respectively constructed: dull cell and Glossy cell. The invention also uses P 1 (2073-1) and P 2 (2073-2) two parental pools were constructed. Subsequently, the present invention performed high throughput sequencing on the 4 pools described above. Based on the BSA results, 1 region associated with cucumber peel brightness trait, chr5, was obtained: 17,358,895 ～ 20,945,460, total length 3.59Mb, contains 435 genes in total.
Subsequently, a series of InDel markers and SNP markers were developed in this interval for fine localization and finally the gene DULL controlling the brightness of cucumber pericarp was located in a 24.5kb interval on chromosome 5.
4) Development of Indel molecular marker co-separated with cucumber peel brightness gene
The genomic DNA sequences from the above-described localization regions of 2073-1 and 2073-2 were cloned and sequenced in the present invention. By sequence alignment, a 4895bp deletion fragment was found to exist in this region of the 2073-2 genome. Subsequently, an InDel molecular marker, i.e., inDel-DULL, was designed based on the deletion fragment. The nucleotide sequence of the detected molecular marker is shown as SEQ ID NO. 1-2.
Example 2 application of InDel molecular marker in identification of cucumber pericarp Brightness Gene DULL genotype
The implementation provides a method for identifying the cucumber pericarp brightness gene DULL genotype by using InDel molecular markers, which comprises the following steps: firstly, extracting whole genome DNA of cucumber materials to be detected; secondly, performing PCR amplification by using the InDel molecular marker; finally, the length of the amplified band is detected by agarose gel electrophoresis and the genotype of the material to be detected is judged, wherein the DULL genotype consistent with the sequence of the reference variety 2073-1 can amplify a single band of 5309bp, the DULL allele genotype with the 4895bp deletion variation can amplify a single band of 414bp, and the heterozygote can amplify double bands of 414bp and 5309bp (figure 2).
Example 3 application of InDel molecular marker in predicting or identifying cucumber pericarp brightness
This example uses InDel-DULL markers for 200F 2 The brightness of the peel was identified for the segregating population (50 bright peel, 150 dark peel) and 60 natural population (20 bright peel, 40 dark peel).
Indel-DULL molecular marker at F 2 The banding pattern and phenotype statistics in the segregating populations are shown in Table 2. Band-type and phenotype statistics for Indel-DULL molecular markers in natural populations are shown in table 3. Experimental results show that the Indel-DULL molecular marker is 100% linked with the brightness property of cucumber peel, and the specific expression is as follows: when a single 5309bp band or a double band of 414bp and 5309bp is amplified by using Indel-DULL molecular markers, the pericarp of the tested cucumber material presents a dark phenotype; when a single 414bp band is amplified by using the molecular marker, the pericarp of the tested cucumber material presents brightness.
Therefore, the Indel molecular marker reported by the invention is separated from the cucumber peel brightness character, and early, damage-free, simple and efficient prediction or identification of the cucumber peel brightness can be carried out through simple PCR amplification and agarose gel electrophoresis detection. Therefore, the Indel molecular marker reported by the invention can be used as an auxiliary selection means to provide convenience for genetic improvement of cucumber peel brightness, greatly improve breeding efficiency, shorten breeding period and save resources, and can be used for rapid identification of DULL genotype or peel brightness in large-scale cucumber germplasm resources.
TABLE 2 Indel-DULL molecular markers at F 2 Banding and phenotype statistics in isolated populations
TABLE 3 band and phenotype statistics of Indel-DULL molecular markers in Natural populations
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Sequence listing
<110> Chinese university of agriculture
<120> InDel molecular marker co-separated from cucumber peel brightness gene and application thereof
<130> KHP221110813.9
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 1
catgaattta attagtcaag gg 22
<210> 2
<211> 22
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
caatatgtgc aaattaacgg ga 22
<210> 3
<211> 4895
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 3
ttgtttcgat gtttctttac ttttctttct tgttttttag agaatacgca ccattattta 60
tgcaatttaa gctattaatt gacacaatta atgattcatt ttaatttgaa atgatataat 120
aatggaatgt tgactaatta aattgttaaa gaatacttga tatatattaa catataatct 180
tcatagtata tgttttaatt cttcttttga gggatcttat ttgattttgt ttgatagaaa 240
tatattatat aaaactccca attgtggttc atattgacta tttgaatgct ttaattatta 300
ttatctaaaa atcttcctaa atatattttt cactattgtt taatttactt ggatcactaa 360
ttcaaatgtt taatggaaaa tatcattaac ttgacttcct cacattaaat atctttctcc 420
aaaattatgt tcattagttg tcttcaacaa atttatatta taattctcac ttttattgaa 480
gtttacccaa aacacatcct actaattttg ctccaaagaa aaaagaacat ctgtattgat 540
aattgtaagt ttaaaaattg gctaattaag tacaatataa aagatttcag atgacagact 600
aaagttgttt ctcaaatata tagaccaaaa tacttttctt ccattagtta ggttttaaaa 660
caaagattag tgttaatctc gttttacttt tatttatcac attttacaaa tattattcac 720
taatgataat tttgatagag taatttagtc aatataaaca taattcaact aatcaactta 780
tgaatacaaa atttgataaa cttaaattga aaaataactg ccatcataat aaaacttttt 840
cacgagcaca aggcaagata taaattaaca ttaacaataa ataaaaatta ggaacttcaa 900
tctaataaca attttcagta gaataattaa aatcaatcca ttttattagg ttttatataa 960
atgaaagagg catatgttga ttgaccttat tttaatattt agcaagcttg acatgaagca 1020
catgaataca cttcactgat tgattaaact ttttattaac aataaaaaat ctgtttttta 1080
ttcatctgaa atctaaaatt attaggacaa gtagtagtca tgaatcctca agacccaaat 1140
caaaaagtcc ttcacttgaa gtatttaaaa gaattgttga ttgaataatt aattcgtata 1200
accaataatt agtgtcaaaa ttcattgaat gtaacattta aatcaacata tattaatatt 1260
tgtataataa taatttcagc ttgtatatgc ctagaaaaac aatttcagta tgtacgtgtt 1320
ggtaatatat tcttcaaatt aaactatata tctagttgtg aatgattttg gattaaagga 1380
tataattgat atttaaaaat gacaaaaata agaaataaca ttatatagat tgacaatctt 1440
tgccaaatgt gcaacaaggc atgtgtttat gcatgcctcc accaaaagta gcactttaag 1500
cctaaggaaa agaaaagccc aaccaatgtc gacaagaaat cgaatctaat tatcttaaga 1560
cgactcataa actaggtcaa ctttaatttt ttttctataa aatatattat tctaaaatca 1620
tataaaataa caacttaagt cttactcaca aactctttta agctttcgca atctttcgtt 1680
gtttgacaga cgaaatgtct gtgatcgata aattttttta taaaacgtca cttattcttt 1740
tctttaaaca aatttattgt ttgagtcacc acaccaacat atatgtttac tatgtaaaaa 1800
aacttaacac atacatgtta aaaaaagaaa cttaggtgca agtagctaga tctatgcatt 1860
tttttttttc acgttagaaa aagtatataa gaaattgatt ttttgagaaa attttttttg 1920
tgaggtgaca atttatgatt aaatagtgga tggactagta ttttttttag aaaattataa 1980
actctttatt ctctaaatat gtcccaataa tagtaaagct ttaaactttg aatattgtcc 2040
ctatcacatc catcacacac attcttcaat cacggtagaa acatctatat atatatatat 2100
aaataaaaga tattcaattt atgaaaattt gtaattcaga agaaattaaa atatcttcca 2160
cttaattttc tccccatatc tacttcctct ctaactctct tgcattctcc ctactctcct 2220
cttttgactt ctcctactat gattgacata tcattacaag acttatatta ataatatttc 2280
gagttgcatc tatcttttag ttttagtttt gagttagcaa catatcaata taagaaattt 2340
ataatagatc taataaaata aggatcgatt tcttaattga tatcgacaat ttaaatttgg 2400
atcgattatg acacatccta atactaaatt acttaaccta ttctttagtt catttcaaac 2460
tattgtttaa aaaatatatt tttgttaaga aagtgaaacc cacttttatt attaaaaaaa 2520
aaaagaagaa ctagatttaa aaacaagaaa taaaatcata aaaatgaaaa aagaaaggat 2580
atatatgaag agatggatag tattaaagaa gtgcaatcct ttgtgtccca aaaatgtttg 2640
ttgagttgtg ttcccatatt acttgacttt cttctcattt tatttggttt gttgttccct 2700
cctctatctc tttccccaca ttcttaattg aaaaaaaaac aaattgagtg cactaatatg 2760
catgtgactt ttataaatat ttatttttat ttttctatat tatatatttt ataacaaaaa 2820
tgattcattc aaacaaatgt tatagtctat ttatgataga tagcaacaaa atacaataaa 2880
ctaccatata tatctataag tttatcgcaa gtataatggt aatattttta ttatatttaa 2940
aaatattttg aatggtttta ataatttttg ggctctctag aaaatatgag tacgtccatt 3000
tgtatgaatt cttgttagat tttttttggg tggaaatgta gaaagataga tgctaacaca 3060
aaatattttt tttatttttg caagaaaact ccatttattt tattttattt gttaagctat 3120
atatacctct actcatgcat taacctcccc ttgcatttcc taccttcctt cctttctctc 3180
tctcactctt aaccctttta ttttatttta ttttttctct ctctctctta tcatcttcca 3240
tttttccatg gcagctctag aaaaccatta ccaaaccaaa caaaacaaca ataatccggc 3300
caccactcgc ttaaagcttt ttgggtttga cgttcaagaa gatcttgatc aagacgattc 3360
cacccccacc tcctccgact caggcgccgc cgttccatcc tccggcgacc gcaaatacga 3420
gtgtcagtac tgctatagag agtttgccaa ttcccaagct ctcggcggcc accaaaatgc 3480
ccacaagaaa gagcgtcaac agctcaaacg cgcccagtta caagcttccc gaaacgccgc 3540
cgtttttaga aaccccatcg ttgctgcctt tgctcctccg cctcacctcc ttccctccgc 3600
cgccgttacg gcttcctctt cttggccggt ttatatccca cgggctccac ctcaatttca 3660
agtgtctcat ggttgtgtgt ttactccgtc ctcgtcttac ggcggcggcg gcgacgctcc 3720
accaccaccg tcgccgccgg atttctttac gatggggtct cggtcaagtc atgggatggt 3780
ggatgctccg atgtcgctga gtcggttttc taaagtggac ggtggaaccg cgttcgacga 3840
tggacccggt ttggatttgc atcttagcct cgctccggcc gccccatgaa aattaaagtc 3900
tccaactttt tttaaaacaa tttatttttt ttcttagaaa ttaatggtta gtgttttgct 3960
ctttaagcca ctggtttttt ttttcccttt ttgtatttga gaaatagatg aaggaattta 4020
agttagagat tgaaaccaga tgggctaaga aaaacaaaga tgaaaaaaaa gaaaaagaaa 4080
atcttgaaca tgttcatgat gattcttcca tcatcatcca tccatcaacc ttattgtata 4140
tttttcaaat ttatttattt atttttaatt tttgtcgact attcaaaagg tttctttggt 4200
aatcaataat gctcgcaatt gatgattgat atgggtattg ggtaggttca aagatttaat 4260
tgttttctct tcttttgtta ttggttttga aaaattaagt atacaatact atttatttgt 4320
ctgttttgtt gtttattttt ttacacatgt agtaaaaagt tagctctagt tttgaaaatt 4380
ataatcatgc tatagggaat tgaagtggta aagaaaacca acaccctctt taaaatttaa 4440
agaaaaataa ttaaatgctt atcaatcaca catgttagtt tcttttctat attttgagtt 4500
ttctaaattt gtacttattt tctcacagtt tctctatatc ttcttttttt gagtagtcaa 4560
agtttaattt ataagtaaat tcaaagctag aagtatatgt taaaaaatca tatattaatc 4620
atgtgaaagc ataataattt tgagtagaca ttaaagtggg tcaatgaatt tcaattagta 4680
tgacaatatg gatgagttgg ctttaggagt agtgctattg agtatatata taggcaccaa 4740
atataatctg aaatatttca atctacatca attctatact tttttttctc tcttcttttc 4800
attttgtatt acaaaattaa cgacctttta gatatctcta cgataatatg atattgttca 4860
cgtaaaaaat aattctcatc actttgtttt tcgtt 4895

Claims (2)

1. The application of an InDel molecular marker coseparated with a cucumber peel brightness gene in detecting cucumber peel brightness characters is provided, and the nucleotide sequence of the InDel molecular marker is shown as SEQ ID NO. 3.
2. A method for detecting brightness characteristics of cucumber peel is characterized by comprising the following steps:
extracting the whole genome DNA of the cucumber material to be detected; PCR amplification is carried out by using a primer pair shown as SEQ ID NO. 1-2; the primer pair is used for detecting InDel molecular markers coseparated with cucumber pericarp brightness genes, and the nucleotide sequence of the InDel molecular markers is shown as SEQ ID NO. 3;
agarose gel electrophoresis analysis is carried out on the amplified bands, and if a single 5309bp band or double 5309bp bands are amplified, the pericarp of the cucumber material to be detected is shown to be in a dark phenotype; if a single strip of 414bp is amplified, the pericarp of the cucumber material to be detected is shown to be in a bright phenotype.
CN202210126440.6A 2022-02-10 2022-02-10 InDel molecular marker co-separated from cucumber peel brightness gene and application thereof Active CN114410826B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202210126440.6A CN114410826B (en) 2022-02-10 2022-02-10 InDel molecular marker co-separated from cucumber peel brightness gene and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210126440.6A CN114410826B (en) 2022-02-10 2022-02-10 InDel molecular marker co-separated from cucumber peel brightness gene and application thereof

Publications (2)

Publication Number Publication Date
CN114410826A CN114410826A (en) 2022-04-29
CN114410826B true CN114410826B (en) 2023-08-11

Family

ID=81279835

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210126440.6A Active CN114410826B (en) 2022-02-10 2022-02-10 InDel molecular marker co-separated from cucumber peel brightness gene and application thereof

Country Status (1)

Country Link
CN (1) CN114410826B (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103184219A (en) * 2013-01-05 2013-07-03 上海交通大学 Molecular markers closely linked to gloss gene d of cucumber fruit
CN112458200A (en) * 2021-01-13 2021-03-09 河南农业大学 InDel molecular marker closely linked with cucumber fruit spine base size gene and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103184219A (en) * 2013-01-05 2013-07-03 上海交通大学 Molecular markers closely linked to gloss gene d of cucumber fruit
CN112458200A (en) * 2021-01-13 2021-03-09 河南农业大学 InDel molecular marker closely linked with cucumber fruit spine base size gene and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
黄瓜果实形态建成的遗传及分子基础研究进展;刘兴旺等;《园艺学报》;20200925(第09期);第1793-1809页 *

Also Published As

Publication number Publication date
CN114410826A (en) 2022-04-29

Similar Documents

Publication Publication Date Title
CN106191101B (en) Corn event 5307
US9745596B2 (en) Identification and use of KRP mutants in wheat
CN106148353B (en) Rice brown planthopper resistant gene Bph6 and closely linked molecular marker thereof
CN108239647A (en) A kind of gene, molecular labeling and application for controlling rape plant type
CN109762921B (en) SNP (Single nucleotide polymorphism) marker for detecting color of cucumber pulp and application thereof
CN101962640A (en) Specific molecular markers of related genes of brassica napus grain weight and application thereof
CN108300801B (en) Molecular marker closely related to rape grain weight and silique length and application
CN111926097A (en) Insect-resistant herbicide-resistant corn transformation event and creation method and detection method thereof
CN112080515A (en) UP gene and application thereof in plant improvement
CN117070658A (en) InDel molecular marker primer for identifying white gourd peel color and application thereof
Papan et al. Validation of molecular markers linked to cercospora leaf spot disease resistance in mungbean (Vigna radiata [L.] wilczek).
US10036029B2 (en) Molecular markers for low palmitic acid content in sunflower (Helianthus annus), and methods of using the same
KR101822995B1 (en) DNA marker for selecting Jubilee-type stripe pattern of watermelon
CN114350655B (en) Exogenous radish fragment specific marker and preparation method and application thereof
RU2717017C2 (en) Molecular markers for blackleg resistance gene rlm2 in brassica napus and methods of use thereof
CN110468229B (en) Coseparation molecular marker Hxjy-1 of rice broad-spectrum high-resistance bacterial leaf blight gene Xa45(t)
CN108239675B (en) Molecular marker TJcM02 for identifying melon unisexual flower and application thereof
CN114410826B (en) InDel molecular marker co-separated from cucumber peel brightness gene and application thereof
Olalekan et al. Analysis of recurrent parent genome recovery in marker-assisted backcross breeding programme in Watermelon
CN111172317B (en) Molecular marker HSRC3911 closely linked with major QTL site in flowering phase of sesame and application thereof
CN110358862B (en) Molecular marker Hxjy-14 closely linked with rice broad-spectrum high-resistance bacterial blight gene Xa45(t)
RU2718584C2 (en) Molecular markers of rlm4 gene of brassica napus black stem resistance and methods of using them
CN108315463B (en) Primer group for detecting corn 1768 gene and application thereof
CN112029895A (en) InDel molecular marker and method for identifying porphyra haitanensis thin leaf strain HR-5
CN106834527B (en) Molecular marker closely linked with wheat seedling sheath blight resistance QTL and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant