CN114107549A - KASP molecular marker related to rose fragrance of grape fruits and application - Google Patents
KASP molecular marker related to rose fragrance of grape fruits and application Download PDFInfo
- Publication number
- CN114107549A CN114107549A CN202111525072.4A CN202111525072A CN114107549A CN 114107549 A CN114107549 A CN 114107549A CN 202111525072 A CN202111525072 A CN 202111525072A CN 114107549 A CN114107549 A CN 114107549A
- Authority
- CN
- China
- Prior art keywords
- grape
- seq
- vvdxs
- genotype
- nucleotide
- 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.)
- Pending
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
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Botany (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses application of a KASP molecular marker related to rose fragrance of grape fruits. The invention provides a technical scheme for applying a composition for detecting the polymorphism or genotype (namely allele) of VVDXS-422G/T in a grape genome to preparation of products for identifying or assisting in identifying fruit rose fragrance. The composition for detecting the VVDXS-422G/T polymorphism and genotype can be combined with other substances (such as substances for detecting single nucleotide polymorphism or genotype of other molecular markers related to the rose fragrance of the grape fruits) to prepare a product for identifying the grape fruit variety with the rose fragrance in high throughput.
Description
Technical Field
The invention relates to application of KASP molecular markers, in particular to KASP molecular markers related to the rose fragrance of grape fruits and application thereof.
Background
Grapes are one of the main fruits, and for grapes, the aroma and flavor of the fruits are important fruit shapes for consumers.
At present, in grape breeding work, a traditional crossbreeding means is mainly adopted by breeding personnel, crossbreeding seeds are cultivated into seedlings, after years of childhood, the growth result habit and the fruit character of each hybrid plant are evaluated, and the hybrid plants are selected and eliminated.
Disclosure of Invention
The invention aims to solve the technical problem of how to identify the rose fragrance of the grape fruits in a high-throughput manner or in an auxiliary manner.
In order to solve the technical problem, the invention provides any one of the following applications A1-A3:
a1, application of a composition for detecting the polymorphism or genotype (i.e. allele) of VVDXS-422G/T in grape genome in identification or auxiliary identification of grape fruit rose fragrance;
a2, application of a composition for detecting the polymorphism or genotype (i.e. allele) of VVDXS-422G/T in grape genome in preparing products for identifying or assisting in identifying the rose fragrance of grape fruits;
a3, the application of the composition for detecting the polymorphism or genotype (i.e. allele) of VVDXS-422G/T in grape genome in grape breeding or the application in preparing grape breeding products; the breeding aim comprises breeding grapes with rose fragrance;
the VVDXS-422G/T is an SNP site in a grape genome, the nucleotide type is G or T, and the nucleotide is 422 th nucleotide of SEQ ID No. 1; the composition comprises the PCR primer, and the PCR primer is a primer composition consisting of single-stranded DNA with a nucleotide sequence of SEQ ID No.2, single-stranded DNA with a nucleotide sequence of SEQ ID No.3 and single-stranded DNA with a nucleotide sequence of SEQ ID No. 4.
The invention also provides a method for identifying or assisting in identifying whether the grape fruits have rose fragrance, which comprises the steps of detecting the genotype of the grapes to be detected, and identifying or assisting in identifying whether the grape fruits have rose fragrance according to the genotype of the grapes to be detected; the genotype is the genotype of VVDXS-422G/T in grape genome; the VVDXS-422G/T is an SNP site in a grape genome, the nucleotide type is G or T, and the nucleotide is 422 th nucleotide of SEQ ID No. 1; the composition comprises the PCR primer, and the PCR primer is a primer composition consisting of single-stranded DNA with a nucleotide sequence of SEQ ID No.2, single-stranded DNA with a nucleotide sequence of SEQ ID No.3 and single-stranded DNA with a nucleotide sequence of SEQ ID No. 4.
The method comprises the following steps of identifying or assisting in identifying whether the grape fruit has rose fragrance according to the genotype of the grape to be detected, wherein the probability that the grape fruit to be detected with the genotype of GT or TT has rose fragrance is higher or the candidate is higher than the probability that the grape fruit to be detected with the genotype of GG has rose fragrance.
The invention also provides application of the method for identifying or assisting in identifying the rose fragrance of the grape fruits in grape breeding.
Another technical problem to be solved by the present invention is how to perform grape breeding, including: detecting the genotype of the VVDXS-422G/T in a grape genome, and selecting grapes with the genotype GT or TT in the grape genome as parents to breed; the GT is heterozygote of the VVDXS-422G/T in grape genome for G and T, and the TT is homozygote of the VVDXS-422G/T in grape genome for T.
The purpose of the breeding includes breeding grapes with rose fragrance.
Any of the following products 1) -3) comprising a composition for detecting the polymorphism or genotype (i.e., allele) of VVDXS-422G/T in the grape genome are also within the scope of the present invention:
1) products for detecting single nucleotide polymorphisms or genotypes associated with rose aroma in grape fruits;
2) identifying or assisting in identifying whether a grape has a fruit rose-scented product;
3) a product for grape breeding.
In the application, the method and the product, the VVDXS-422G/T is an SNP site in the grape genome, the nucleotide type is G or T, and the SNP site is 422 th nucleotide of SEQ ID No. 1; the polymorphism or genotype (i.e. allele) of VVDXS-422G/T in the gene group for detecting grape can be specifically the nucleotide species for detecting VDXS-422G/T. The genotype of VVDXS-422G/T in the grape genome can be GG, TT or GT. The GG is homozygous for the VVDXS-422G/T being G in grape genome, the TT is homozygous for the VVDXS-422G/T being T in grape genome, and the GT is homozygous for the VDXS-422G/T being G and T in grape genome.
In the application, the method and the product, the grape breeding is to breed and breed the grape with the rose fragrance.
In the above applications, methods and products, the composition for detecting the polymorphism or genotype (i.e., allele) of VVDXS-422G/T in the grape genome can be a reagent and/or an apparatus for determining the polymorphism or genotype of VDXS-422G/T by at least one of the following methods: DNA sequencing, restriction enzyme fragment length polymorphism, single-strand conformation polymorphism, denaturing high performance liquid chromatography and SNP chip. The SNP chip comprises a chip based on nucleic acid hybridization reaction, a chip based on single base extension reaction, a chip based on allele-specific primer extension reaction, a chip based on one-step reaction, a chip based on primer connection reaction, a chip based on restriction endonuclease reaction, a chip based on protein DNA binding reaction and a chip based on fluorescent molecule DNA binding reaction.
In the above applications, methods and products, the composition for detecting the polymorphism or genotype (i.e. allele) of VVDXS-422G/T in grape genome is 1), 2) or 3):
D1) the composition for detecting the polymorphism or genotype of VVDXS-422G/T in grape genome comprises PCR primers for amplifying grape genome DNA fragments including the VVDXS-422G/T;
D2) the composition for detecting the polymorphism or genotype of VVDXS-422G/T in the grape genome is a PCR reagent containing the PCR primer;
D3) a kit containing the PCR primer described in D1) or the PCR reagent described in D2).
In the above applications, methods and products, the PCR primers may be labeled with a label. The label refers to any atom or molecule that can be used to provide a detectable effect and that can be attached to a nucleic acid. Labels include, but are not limited to, dyes; radiolabels, e.g.32P; binding moieties such as biotin (biotin); haptens such as Digoxin (DIG); a luminescent, phosphorescent, or fluorescent moiety; and a fluorescent dye alone or in combination with a portion of the emission spectrum that can be suppressed or shifted by Fluorescence Resonance Energy Transfer (FRET). Labels can provide signals detectable by fluorescence, radioactivity, colorimetry, gravimetry, X-ray diffraction or absorption, magnetism, enzymatic activity, and the like. Labels can be charged moieties (positive or negative) or alternatively, can be charge neutral. The label may comprise or be combined with a nucleic acid or protein sequence, provided that the sequence comprising the label is detectable. In some embodiments, the nucleic acid is detected directly (e.g., direct sequence read) without a label.
The PCR primer is a primer composition consisting of a single-stranded DNA shown by SEQ ID No.2 in a sequence table, a single-stranded DNA shown by SEQ ID No.3 in the sequence table and a single-stranded DNA shown by SEQ ID No.4 in the sequence table. Wherein, SEQ ID No.2 in the sequence table consists of 23 nucleotides, the 1 st to 21 st nucleotides are FAM linker sequences (as markers), and the 22 nd to 44 th nucleotides are specific sequences; SEQ ID No.3 in the sequence table is composed of 25 nucleotides, the 1 st to 21 st nucleotides are HEX linker sequences (as markers), and the 22 nd to 46 th nucleotides are specific sequences.
In the above applications, methods and products, the product may be a reagent or a kit or a system, and the system may comprise a combination of reagents or kits, instruments and analytical software, such as a product consisting of PCR primers, PARMS master mix reagent, microplate reader and on-line software SNP decoder (http:// www.snpway.com/snpdecoder01/), a combination consisting of PCR primers, PARMS master mix reagent, on-line software SNP decoder and a fluorescence quantitative PCR instrument. The product can comprise the composition for detecting the polymorphism or genotype of VVDXS-422G/T in grape genome.
The invention discloses a novel KASP marker for detecting the rose fragrance of grape fruits. The specific primer group provided by the invention consists of single-stranded DNA shown in SEQ ID No.2, single-stranded DNA shown in SEQ ID No.3 and single-stranded DNA shown in SEQ ID No.4, wherein the single-stranded DNA shown in SEQ ID No.2 and the single-stranded DNA shown in SEQ ID No.3 are provided with fluorescent labeling joints. In one embodiment of the invention, the primer set with the fluorescence labeling joint is used for amplifying two groups of grape genomic DNA of a plurality of samples including VVDXS-422G/T, fluorescence signal processing is carried out by utilizing a high-throughput genotyping detector PHERAStar SNP, the nucleotide type of VDXS-422G/T is determined, and the germination index of each sample to be detected is determined. Experiments prove that in a group consisting of 210 grape varieties, 41 varieties are GT or TT genotypes, wherein 35 varieties have rose fragrance, 159 varieties are GG genotypes, and all varieties have no rose fragrance, so that the probability that fruits of the grape varieties with the genotypes of GT (plumbum virgatum) or TT have the rose fragrance is remarkably higher than that of grape varieties with the genotypes of GG at the SNP loci of DXVVVS-422G/T. The experimental results of 194 hybrid progeny plants also verify the results, which indicates that VVDXS-422G/T is an SNP molecular marker related to the rose fragrance of grape fruits, and the specific primer group provided by the invention can be used for identifying or assisting in identifying the rose fragrance of the grape fruits, screening grape fruit rose fragrance varieties, assisting in breeding grape molecular markers, and breeding and cultivating grapes with the rose fragrance of fruits. The invention has important theoretical significance and economic value for utilizing molecular markers to assist in selecting grape varieties with rose fragrance.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples are conventional unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The grape variety used in the following examples is a known variety and is described in non-patent documents such as "Zhangxiaoli, Liu Chong Huai, Liu Qiang, Fan Xiucai, Zhang Ying, Sun Lei, Niuhuang, Jianfu with ginger.
Example 1 primers for amplifying SNP molecular markers developed based on KASP technique and verification thereof
The invention provides a KASP primer and a reaction system based on a grape fruit rose fragrance related SNP locus, wherein the SNP locus is positioned on a grape VvDXS gene, specifically positioned at 422 th nucleotide of SEQ ID No.1, the type of the nucleotide is G or T, and the nucleotide is represented by a letter N. The SNP site was named VVDXS-422G/T.
One allele of VVDXS-422G/T is GG (i.e. homozygous for G at nucleotide 422 of SEQ ID No. 1), and the genotype corresponds to grape fruits without rose scent. The other allele of VVDXS-422G/T is TT (i.e. homozygous for T at 422 of SEQ ID No. 1), the third allele of VVDXS-422G/T is GT (i.e. heterozygous for G and T at 422 of SEQ ID No. 1), and grape fruits corresponding to the GT genotype or TT genotype have rose fragrance.
The method comprises the following steps of utilizing KASP (Kasp mark) to detect 210 grape varieties in grape resource gardens of fruit tree scientific research institute of forestry in Beijing city in batches, and specifically comprising the following steps:
design of specific primers useful for KASP technology:
forward primer F1: 5' -GAAGGTGACCAAGTTCATGCTGAGAATTACGAGAGGTTGCCAAG-3 '' (shown in SEQ ID No.2, wherein the FAM linker sequence is underlined);
forward primer F2:
(as shown in SEQ ID No.3, wherein double underlining indicates a HEX linker sequence);
reverse primer R: 5'-CCAATTCATGCATCGGTCCGCC-3' ' (shown in SEQ ID No. 4).
Amplifying a segment of which the 422 th nucleotide of the SEQ ID No.1 is G by the single-stranded DNA molecules shown in the SEQ ID No.2 and the SEQ ID No.4, and reading a fluorescent signal of a fluorescent group combined with the FAM sequence by using an enzyme-labeling instrument or a fluorescent quantitative PCR instrument;
the single-stranded DNA molecules shown in SEQ ID No.3 and SEQ ID No.4 amplify a segment of which the 422 th nucleotide of SEQ ID No.1 is T, and a fluorescence signal of a fluorescent group combined with the HEX sequence can be read by an enzyme-labeling instrument or a fluorescent quantitative PCR instrument.
The KASP reaction was performed in 384 microwell plates (cat # 04729749001, Roche) using the PCR reaction system: total 3. mu.L, DNA template 1. mu.L, concentration 4 ng/. mu.l, KASP Primer mix 1. mu.L, 2 XKASP Mastermix 1. mu.L; wherein the concentration of the forward primers F1, F2 and the reverse Primer R in the KASP Primer mix is 10. mu.M, KASP Mastermix (available from LGC, www.lgcgroup.com, cat. KBS-1016-002).
The PCR reaction conditions are shown in Table 1:
TABLE 1
Reading fluorescence data of a PARMS reaction product by using a microplate reader or a fluorescence quantitative PCR (polymerase chain reaction) instrument, performing fluorescence signal processing by using an online software SNP decoder (http:// www.snpway.com/snpdecoder01/), and if the fluorescence signal of only a fluorophore combined with an FAM sequence is displayed, determining that the genotype of VVDXS-422G/T of the grape to be detected is GG (namely the homozygous type of the VDXS-422G/T in the grape genome is G, which is hereinafter referred to as genotype GG); if the fluorescent signal of the fluorophore combined with the HEX sequence is displayed, the genotype of the VVDXS-422G/T of the grape to be detected is TT (namely the genotype of the VVDXS-422G/T in the grape genome is homozygote of T, which is hereinafter referred to as genotype TT); if the fluorescence signal of the fluorophore combined with the FAM sequence and the fluorescence signal of the fluorophore combined with the HEX sequence are displayed, the genotype of the VVDXS-422G/T of the grape to be detected is GT (namely the VVDXS-422G/T in the grape genome is a hybrid type of G and T, and is hereinafter referred to as genotype GT).
The KASP typing results and phenotypes for 210 grape varieties in the grape resource garden of the forestry fruit tree science research institute of Beijing City are shown in Table 2, and the results show that 41 varieties are GT or TT genotypes, 35 of the varieties have rose fragrance, 159 of the varieties have GG genotypes, and all the varieties have no rose fragrance. The probability that the fruit with the genotype of GT or TT grape variety as the VDXS-422G/T gene is rose fragrance is obviously higher than that of GG grape variety as the VDXS-422G/T gene at the SNP locus.
TABLE 2
Example 2 primers for amplifying SNP molecular markers developed based on KASP technique and use
In order to further prove that the SNP site VVDXS-422G/T can be used for selection and cultivation of grape pericarp aroma characters, 194 hybrid progeny (the female parent is Tamiana, the male parent is Rudunyi and the hybrid progeny is F1 generation) plants stored in a hybrid garden of a research institute of forestry and fruit trees of the agroforestrial academy of sciences in Beijing are verified, and the details are as follows:
1. DNA extraction
Selecting young leaves of the variety to be detected, wherein the young leaves are about 0.1g, using a novel plant genome DNA extraction kit (aidlab, product number DN15), detecting the quality of DNA, wherein A260/280 is between 1.7 and 1.9, and diluting the DNA into 10ng/L for later use.
2. PCR reaction based on KASP technique
The entire experimental procedure was performed according to the standard experimental protocol provided by LGC corporation, genetic assay, manual part # 15004070. The KASP reaction was performed in 384 microwell plates with a reaction system of 3 μ Ι _: DNA template 1. mu.L, concentration 4 ng/. mu.l, KASP Primer mix 1. mu.L, 2 XKASP Mastermix 1. mu.L; wherein the concentration of the forward primers F1, F2 and the reverse Primer R in the KASP Primer mix is 10. mu.M, KASP Mastermix (available from LGC company, www.lgcgroup.com, cat. KBS-1016-002).
The PCR reaction conditions are shown in Table 3:
TABLE 3
The KASP typing results and phenotypes are shown in table 4, wherein the 117 progeny were genotyped as GG, and none of the 117 progeny had a rose scent. The 77 progeny genotypes were GT, 64 of which had rose scent and 13 of which had no rose scent. The probability that the fruit with the genotype of GT or TT grape variety is rose-scented is obviously higher than that of GG grape variety with the genotype of VDXS-422G/T at the SNP locus.
TABLE 4
The above results show that the efficiency of selection of rose-scented lines in a population can be effectively improved by using the marker.
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
Sequence listing
Scientific research institute for forestry fruit trees in Beijing City
<120> KASP molecular marker related to rose fragrance of grape fruits and application thereof
<130> GNCSY213380
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1050
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
cttacccaca caaaattcta actgggagaa gagatcaaat gcataccatg aggcaaacag 60
atgggttagc gggattcacc aagcgctcgg agagtgaata tgactgcttt ggaaccggcc 120
acagttctac taccatctca gcaggcttgg gaatggcggt cggccgggat ctaaaaggaa 180
aaaacaacaa cgttattgct gtcataggtg atggagccat gactgcaggg caagcttatg 240
aagcaatgaa caatgctggg tacctggatt ctgacatgat tgttatcctt aatgacaaca 300
agcaggtttc tttacccact gctactctag atgggcccat accacctgta ggagctttga 360
gcagtgctct tagtaggtta caatcaaaca gacctcttag agaattacga gaggttgcca 420
anggcgttac caaacagatt ggcggaccga tgcatgaatt ggctgcaaaa gttgatgaat 480
atgctcgtgg gatgatcagt ggttctggat caacactttt tgaagagctt ggactctatt 540
atataggtcc tgttgatggc cacaacatag atgaccttgt tgccattctc aaggaggtta 600
agagtaccaa gacaacaggt ccagttctga tccatgttgt cacagagaaa ggccgcggat 660
atccatatgc tgagaaagct gcagataagt accatggagt gaccaagttc gatcctgcta 720
ctggaaaaca attcaaatcc agtgctccta ctcagtccta cacaacatat tttgcagagg 780
ctttgattgc agaagcagag gtggacaagg atattgttgc aattcatgca gcaatggggg 840
gtggaacggg cttgaatctc ttccatcgcc ggttccccac acgatgcttt gatgttggga 900
tagcagaaca gcatgctgtt acctttgctg ctggtctagc ctgtgaaggc attaaacctt 960
tttgtgcaat ctactcatct ttcatgcaga gagcttatga ccaggtggtg catgatgtag 1020
atttgcagaa gctgccagtg aaatttgcaa 1050
<210> 2
<211> 44
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
gaaggtgacc aagttcatgc tgagaattac gagaggttgc caag 44
<210> 3
<211> 46
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
gaaggtcgga gtcaacggat ttagagaatt acgagaggtt gccaat 46
<210> 4
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
ccaattcatg catcggtccg cc 22
Claims (10)
1. The application of the composition for detecting the polymorphism or genotype of VVDXS-422G/T in grape genome in identifying or assisting in identifying whether grape fruits have rose fragrance; the VVDXS-422G/T is an SNP site in a grape genome, the nucleotide type is G or T, and the nucleotide is 422 th nucleotide of SEQ ID No. 1; the composition comprises the PCR primer, and the PCR primer is a primer composition consisting of single-stranded DNA with a nucleotide sequence of SEQ ID No.2, single-stranded DNA with a nucleotide sequence of SEQ ID No.3 and single-stranded DNA with a nucleotide sequence of SEQ ID No. 4.
2. The application of the composition for detecting the polymorphism or genotype of VVDXS-422G/T in grape genome in preparing products for identifying or assisting in identifying the rose fragrance of grape fruits; the VVDXS-422G/T is an SNP site in a grape genome, the nucleotide type is G or T, and the nucleotide is 422 th nucleotide of SEQ ID No. 1; the composition comprises the PCR primer, and the PCR primer is a primer composition consisting of single-stranded DNA with a nucleotide sequence of SEQ ID No.2, single-stranded DNA with a nucleotide sequence of SEQ ID No.3 and single-stranded DNA with a nucleotide sequence of SEQ ID No. 4.
3. The application of the composition for detecting the polymorphism or genotype of VVDXS-422G/T in grape genome in grape breeding or the application in preparing grape breeding products; the VVDXS-422G/T is an SNP site in a grape genome, the nucleotide type is G or T, and the nucleotide is 422 th nucleotide of SEQ ID No. 1; the composition comprises the PCR primer, and the PCR primer is a primer composition consisting of single-stranded DNA with a nucleotide sequence of SEQ ID No.2, single-stranded DNA with a nucleotide sequence of SEQ ID No.3 and single-stranded DNA with a nucleotide sequence of SEQ ID No. 4.
4. The method for identifying or assisting in identifying whether the grape fruits have the rose fragrance comprises the steps of detecting the genotype of the grapes to be detected, and identifying or assisting in identifying whether the grape fruits have the rose fragrance according to the genotype of the grapes to be detected; the genotype is the genotype of VVDXS-422G/T in grape genome; the VVDXS-422G/T is an SNP site in a grape genome, the nucleotide type is G or T, and the nucleotide is 422 th nucleotide of SEQ ID No. 1; the composition comprises the PCR primer, and the PCR primer is a primer composition consisting of single-stranded DNA with a nucleotide sequence of SEQ ID No.2, single-stranded DNA with a nucleotide sequence of SEQ ID No.3 and single-stranded DNA with a nucleotide sequence of SEQ ID No. 4.
5. Use of the method of claim 4 in grape breeding.
6. A method of grape breeding comprising: detecting the genotype of the VVDXS-422G/T of claim 1 in grape genome, and selecting grapes with the genotype of GT or TT as parents of the VVDXS-422G/T in the grape genome for breeding; the GT is heterozygote of the VVDXS-422G/T in grape genome for G and T, and the TT is homozygote of the VVDXS-422G/T in grape genome for T.
7. The product containing the composition for detecting polymorphism or genotype of VVDXS-422G/T in grape genome as claimed in claim 1, is any one of products C1) -C3):
C1) products for detecting single nucleotide polymorphisms or genotypes associated with rose aroma in grape fruits;
C2) identifying or assisting in identifying whether a grape fruit has a rose-scented product;
C3) a product for grape breeding.
8. The use according to any one of claims 1-3 and 5, the method of claim 4 or 6, or the product of claim 7, wherein: the grape breeding is to breed grapes with rose fragrance in fruits or breed grapes with rose fragrance in fruits.
9. The use according to any one of claims 1-3, 5 and 8, the method of claim 4, 6 or 8 or the product of claim 7 or 8, wherein: the composition for detecting the polymorphism or genotype of VVDXS-422G/T in the grape genome is D1), D2) or D3):
D1) the composition for detecting the polymorphism or genotype of VVDXS-422G/T in the grape genome comprises PCR primers for amplifying grape genome DNA fragments including the VVDXS-422G/T;
D2) the composition for detecting the polymorphism or genotype of VVDXS-422G/T in the grape genome is a PCR reagent containing the PCR primer;
D3) a kit containing the PCR primer described in D1) or the PCR reagent described in D2).
10. The use, method or product according to claim 9, wherein: the PCR primer is a primer composition consisting of a single-stranded DNA shown by SEQ ID No.2 in a sequence table, a single-stranded DNA shown by SEQ ID No.3 in the sequence table and a single-stranded DNA shown by SEQ ID No.4 in the sequence table.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111525072.4A CN114107549A (en) | 2021-12-14 | 2021-12-14 | KASP molecular marker related to rose fragrance of grape fruits and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111525072.4A CN114107549A (en) | 2021-12-14 | 2021-12-14 | KASP molecular marker related to rose fragrance of grape fruits and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114107549A true CN114107549A (en) | 2022-03-01 |
Family
ID=80364480
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111525072.4A Pending CN114107549A (en) | 2021-12-14 | 2021-12-14 | KASP molecular marker related to rose fragrance of grape fruits and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114107549A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103429612A (en) * | 2010-10-20 | 2013-12-04 | 格诺普朗特-巴洛尔公司 | 1-deoxy-d-xylulose 5-phosphate synthase alleles responsible for enhanced terpene biosynthesis |
| CN112301148A (en) * | 2020-11-23 | 2021-02-02 | 中国农业科学院果树研究所 | SNP molecular marker related to content of grape ester aroma substances and application thereof |
-
2021
- 2021-12-14 CN CN202111525072.4A patent/CN114107549A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103429612A (en) * | 2010-10-20 | 2013-12-04 | 格诺普朗特-巴洛尔公司 | 1-deoxy-d-xylulose 5-phosphate synthase alleles responsible for enhanced terpene biosynthesis |
| CN112301148A (en) * | 2020-11-23 | 2021-02-02 | 中国农业科学院果树研究所 | SNP molecular marker related to content of grape ester aroma substances and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| FRANCESCO EMANUELLI等: "A candidate gene association study on muscat flavor in grapevine (Vitis viniferaL.)", BMC PLANT BIOLOGY, vol. 10, no. 241, pages 3 - 5 * |
| 孙磊等: "‘亚历山大\'葡萄果实单萜生物合成相关基因转录及萜类物质积累规律", 中国农业科学, vol. 47, no. 7, pages 1379 - 1386 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113584216B (en) | Development and application of KASP marker of wheat grain weight gene TaCYP78A16 | |
| CN114657277B (en) | KASP molecular marker related to wheat grain length and application thereof | |
| CN113584215B (en) | Development and application of KASP marker of wheat powdery mildew resistance gene pmCH7015 | |
| CN109609687B (en) | KASP marker primer combination for detecting watermelon fusarium wilt resistance and application thereof | |
| CN112322772B (en) | Haplotype molecular marker of corn grain cadmium content related gene ZmCD9 and application thereof | |
| CN111793712A (en) | Application of single nucleotide polymorphism substance of wheat SNP IWA586 site | |
| CN115927718B (en) | KASP molecular marker for identifying grain weight and grain width of wheat and application | |
| CN116121445A (en) | KASP molecular marker for detecting grain weight and grain width of wheat in multiple environments and application | |
| CN114606335A (en) | Development and application of KASP molecular marker of sugarcane mosaic virus disease resistance gene of corn | |
| CN114107549A (en) | KASP molecular marker related to rose fragrance of grape fruits and application | |
| CN111607664A (en) | Application of SNP molecular marker on 1DS chromosome related to wheat stripe rust | |
| CN116121433B (en) | KASP molecular marker for detecting wheat powdery mildew resistance and application thereof | |
| CN115873975B (en) | SNP molecular marker for identifying wheat powdery mildew resistance and application thereof | |
| CN113637790B (en) | KASP molecular marker of stripe rust resistance gene YrAS2388R, primer, kit and application | |
| KR102668083B1 (en) | Molecular markers for selecting TMV resistance or susceptible pepper cultivar and use thereof | |
| CN113699273B (en) | SNP locus combination for detecting resistance of tomato root-knot nematode and application thereof | |
| CN114561488B (en) | Functional molecular marker of gene OsNPF3.1 with rice nitrogen fertilizer utilization efficiency and application | |
| CN115820914A (en) | SNP molecular marker of sorghum SbDW3 gene and application thereof | |
| CN115896335A (en) | SNP molecular marker of sorghum SbSOS1 gene and application | |
| CN118147357A (en) | Application of molecular marker combination in detecting purity of bottle gourd type watermelon stock variety 'Su stock 1' seed | |
| CN117187432A (en) | Haplotype molecular marker of gene SbPH14 related to sorghum plant height and application thereof | |
| CN106755493B (en) | SNP marker primer combination for identifying leymus chinensis varieties and identification method | |
| CN115807116A (en) | SNP molecular marker of gene SbDW1 related to overground part dry weight of sorghum and application | |
| CN114032322A (en) | Development and application of SNP molecular marker of maize flowering phase gene | |
| CN116814826A (en) | Application of molecular marker combination in detecting purity of watermelon variety Su Mi 518 seed |
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 |