CN108384875A - A kind of and the relevant insertion/deletion site of capsicum mellow fruit color gene, molecular labeling, molecular labeling primer and application - Google Patents
A kind of and the relevant insertion/deletion site of capsicum mellow fruit color gene, molecular labeling, molecular labeling primer and application Download PDFInfo
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- CN108384875A CN108384875A CN201810243771.1A CN201810243771A CN108384875A CN 108384875 A CN108384875 A CN 108384875A CN 201810243771 A CN201810243771 A CN 201810243771A CN 108384875 A CN108384875 A CN 108384875A
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Abstract
The invention belongs to pepper breedings and molecular biology field, disclose a kind of insertion/deletion site being closely related with capsicum mellow fruit fruit color, the molecular labeling HS5 for designing that molecular labeling primer that 2 pairs are closely related with capsicum mellow fruit fruit color can be couple chain with pepper fruit color gene according to insertion/deletion site, HS6 are expanded.The present invention also provides a kind of methods of identification and screening pepper fruit color simultaneously, carry out fruit color identification using the natural population of 269 parts of materials of this method pair, coincidence rate reaches 92.5%.Result of study not only contributes to the map based cloning to controlling fruit color gene, and provides the foundation for the breeding of pepper fruit color and the parsing of molecule mechanism, has extensive promotional value.
Description
Technical field
The invention belongs to pepper breedings and molecular biology field, are related to a kind of and capsicum (Capsicum annuum L)
Application of the molecular labeling preparation method and its molecular labeling of fruit color gene linkage in pepper fruit color breeding.
Background technology
Capsicum, Latin literary fame：Capsicum annuum L., Solanaceae, Capsicum 1 year or limited herbaceos perennial.
Capsicum includes mainly 5 cultivars：C.annuum L、C.frutescens L、C.baccacum L、C.pubescens
Keep、C.chinese Jacquin.Capsicum originates in middle Latin America torrid areas, and the country of origin is Mexico.Chinese main
It is distributed in Sichuan, Guizhou, Hunan, Yunnan, Shaanxi, Henan (Xichuan County), Hebei province Jize County and Inner Mongol Tuoketuo County.According to agriculture
Industry portion bulk vegetable system statistics, China's pepper cultivation area are continuously increased, and capsicum industry achieves huge progress, in recent years
China capsicum year sown area has accounted for the 8%~10% of the total sown area of national vegetables at present in 1,500,000~2,000,000 hm2.
A part of area in China, too busy to get away capsicum in people's diet.
Capsicum is abundant as a kind of its nutritive value of important vegetables, in plantation extensively all over the world, and becomes in China
The maximum vegetables of cultivated area.Pepper fruit various colors are mainly dark green, light green, yellowish green, milky white in green fruit
Color, purple, black are divided into chromoplast with the development fruit Chloroplast and leucoplast of fruit, cause the product of carotenoid
Tired fruit color is changed into yellow, orange or red.Abundant color is conducive to that insect and birds is attracted to carry out the propagation of seed.Fruit
Solid color not still as fruit maturation whether one of important judgment criteria, and attract the important indicator of consumer, and color
Color abundant capsicum is as a kind of ornamental crops increasingly by vast welcome.Research shows that fruit color is mainly by pigment
It is determined with content, includes mainly a variety of pigments such as chlorophyll, carotenoid, flavonoids and anthocyanin in pepper fruit, and this
A little pigments can protect biologic artifact from oxidative damage, improve body immunity, and also have certain antitumaous effect.
Important character of the pepper fruit color as fruit, country variant, different regions have the fruit color of capsicum different
It is required that.In order to meet the needs in market, pepper fruit color is for a long time by the concern of breeders, traditional breeding work
Person is mainly observed by fruiting period, and the color admired and other excellent economical characters are combined, select meet it is vast
The new varieties of the consumer group.Traditional breeding technique has certain advantage, but depends on the phenotype of plant more, it is desirable that has rich
Rich experience, and time-consuming effort, it is easily protected from environmental, restrict its development.With the development of molecular biology, using point
Sub- marker assisted selection technology is modern breeding important method.Molecular mark can reach fast acquisition merit
Individual greatly improves the efficiency of selection, reduces the blindness in breeding process.
Selecting the capsicums of different fruit colors, to meet the different consumer groups be one of important breeding objective of capsicum.Profit
With the molecular labeling chain with pepper fruit color gene, by hybridize and be returned etc. technologies quickly by fruit color gene with
In other merit channel genes to plant, the new varieties for selection and breeding high quality provide genetic resources.Also it is clone's control simultaneously
Fruit color gene processed studies its molecular mechanism and lays a good foundation.
Invention content
It is insufficient it is an object of the invention to be directed to existing pepper fruit face molecular labeling available in the practices of breeding, it finds
The insertion/deletion site being closely related with capsicum mellow fruit fruit color provides a kind of chain with pepper fruit color gene
Molecular labeling HS5, HS6.
The insertion/deletion site provided by the invention being closely related with capsicum mellow fruit fruit color, it is characterised in that institute
The insertion/deletion site stated is since the gene coding regions capsicum annuum l. OC107875664 (ATG) upstream 3277bp to coding
One section of area 4513bp insertion/deletions.
By the above-mentioned insertion/deletion site design being closely related with capsicum mellow fruit fruit color can be designed that with
Capsicum mellow fruit fruit color is closely related molecular labeling amplimer.
Further, molecular labeling amplimer can be by with capsicum annuum l. OC107875664 gene delection upstream portion sequences
(including excalation sequence) and missing downstream part sequence (including excalation sequence) are designed and detect the deletion segment
Two pairs of primers.
Specifically, the molecular labeling amplimer sequence being closely related with capsicum mellow fruit fruit color is：Lack upstream
HS5 is marked：HS5-F(SEQ ID NO.1)：GCAAGTATAGCTAATACCAACATTAC, HS5-R (SEQ ID NO.2)：
AGTTCCTCAAGTGGTTAGCC.Lack downstream HS6 label HS6-F (SEQ ID NO.3)：GTGAGTCGGCCTATGTTATCG,
HS6-R(SEQ ID NO.4)：GTTTGAACAACGATGACAATGTG.
Meanwhile the present invention provides a kind of and pepper fruit color gene chain molecular labeling HS5, HS6, which is characterized in that profit
With HS5, HS6 molecular labeling amplimers, the PCR product clip size that HS5 is amplified is 500bp, nucleotide sequence such as SEQ ID
Shown in NO.5, the PCR product clip size that molecular labeling HS6 is amplified is 714bp, and nucleotide sequence is as shown in SEQ ID NO.6.
Above-mentioned capsicum mellow fruit fruit color is closely related insertion/deletion site, molecular labeling amplimer, molecule mark
Remember that HS5, HS6 can be applicable in color or the pepper fruit breeding of identification or assisting sifting pepper fruit.
Simultaneously the present invention also provides a kind of method of identification and screening pepper fruit color, it is as follows：
(1) using sample to be tested genomic DNA as template, drawn using molecular labeling HS5, the HS6 amplification described in claim 5
Object carries out PCR amplification, obtains amplified production；
(2) amplified production is detached by agarose gel electrophoresis；
(3) interpretation of result, if occurring 500bp (SEQ ID NO.5) and 714bp (SEQ ID NO.6) on Ago-Gel
Two specific bands are then determined as the material that mellow fruit fruit color is red, if only there is a 602bp (SEQID
NO.7) band is then determined as that mellow fruit fruit color is the material of yellow.
Preferably, in the method and step (1) of above-mentioned identification and screening pepper fruit color, PCR amplification program is 95.0 DEG C of pre- changes
Property 3min；95.0 DEG C of denaturation 30s, 55.0 DEG C of renaturation 30s, 72.0 DEG C of extension 45s, 32 recycle；72.0 DEG C of extension 5min, 8 DEG C of preservations.
The present invention combines BSR methods using whole-genome association (GWAS), has obtained one and fruit color chloroaniline
Because of related associated molecular labeling.It is used directly for capsicum red/yellow fruit molecular mark, it can be effective
It solves that the conventional breeding period is long, is vulnerable to the influence of environment.By early utilization, the molecular labeling can be screened quickly completely
The plant of meaning, effectively reduces planting scale, reduces the workload of later stage identification.Improve the efficiency of selection and accurate
Property.Forming Mechanism to study fruit color is of great significance.Therefore, the present invention is in pepper breeding practice and fruit pigment
Significance is all had in theoretical research.
Description of the drawings
Fig. 1 is 269 parts of material mellow fruit fruit color GWAS analysis results (A：269 parts of material kinds are indicated at Shanxi, fruit
Solid color association analysis result B：269 parts of material kinds are indicated at Sanya, fruit color association analysis result)；
Fig. 2 is two parents of BSR groups positioning：16L522,16L17
Fig. 3 is the red pond of Δ (SNP-index) analysis and the yellow pond (A of BSR methods：1-12 represents chromosome number, mellow fruit
Fruit color main effect QTL is located at No. 6 chromosome (at arrow instruction) B：Mellow fruit fruit color main effect QTL is located at No. 6 chromosomes
In the upper sections 5.3M (arrow instruction shaded side))；
Fig. 4 is to utilize HS5, analysis part results of the HS6 two to 270 parts of material fruit colors of molecular labeling pair.
Specific implementation mode
With reference to specific embodiment, the present invention will be further described, but the present invention should not be limited by the examples.
Material therefor, reagent, instrument and method in following embodiment are the routine in this field without specified otherwise
Material, reagent, instrument and method can be obtained by commercial channel.Involved capsicum germplasm is by Hunan Province in the present invention
Vegetable Research Institute provides, these germ plasm resources are collected by Inst. of Vegetables, Hunan Prov. or throughout the country from United States Department of Agriculture-state
Family Germ-plasma resources protection center (USDA-ARS-NCGRP) obtains.The genetic resources material capsicum arrived used in the present invention
(Capsicum annuum L.) saves Changsha acquisition by Ou Li armies in Hunan China.
The acquisition of molecular labeling HS5, the HS6 molecular labeling of one capsicum mellow fruit fruit color gene linkage
One whole-genome association method (GWAS) of specific embodiment
The experiment respectively executes primary respectively at Shanxi and Hainan.
1. the structure of group
Mellow fruit fruit color using 269 parts of high homogenous is that the capsicum material of red yellow builds natural population, Mei Gecai
30 plants of material kind.Planting density is 45000 plants of per hectare, and duplicate rows plantation, distance in the ranks and in the ranks is 30cM, and liquid manure, which is applied, to be shone
Often management, to ensure the healthy growth of capsicum plant.
2. the identification of phenotype
After fruit maturation, red yellow color identification is carried out to fruit color.
3. fruit color main effect QTL determines
Take plant spire in growth period, total DNA extracted using CTAB methods, using two generation sequencing technologies to material into
It goes and resurveys sequence, each material survey 10 ×.The data of acquisition extract polymorphism SNP using the perl script of independent development, using complete
The method that genome association analyzes (GWAS) obtains and the relevant SNP site of mellow fruit fruit color, determining maturation fruits face
Color main effect QTL region, as a result as shown in Figure 1A, B, the 1-12 in picture represents chromosome number, mellow fruit fruit color main effect QTL
Positioned at No. 6 chromosomes (at arrow instruction).4. the exploitation of molecular labeling
Fruit color gene candidate is carried out in the mellow fruit fruit color QTL sections determined to 3 steps, it will
LOC107875664 assignment of genes gene mapping fruit color candidate genes.By gene sequencing, candidate gene code area (ATG is found
Starting) for upstream 3277bp to code area there are the missing of large fragment (4513bp), the missing and fruit color of this large fragment are aobvious
Work property is related.According to the missing of the sheet, (include in deletion fragment upstream (including excalation) and deletion fragment downstream respectively
Excalation) HS5, HS6 two is designed to Indel labels.The primer sequence is
HS5 molecular labelings HS5-F (SEQ ID NO.1)：GCAAGTATAGCTAATACCAACATTAC
HS5-R(SEQ ID NO.2)：AGTTCCTCAAGTGGTTAGCC
HS6 molecular labelings HS6-F (SEQ ID NO.3)：GTGAGTCGGCCTATGTTATCG
HS6-R(SEQ ID NO.4)：GTTTGAACAACGATGACAATGTG
Two BSR methods of specific embodiment
1. the structure of group
Using the fruit color selected by inbreeding of more generation be red material ' 16L522 ' (shown on the right of Fig. 2) and
Fruit color is that the material ' 16L17 ' of yellow is parent (as shown in the left side Fig. 2), is to resurvey sequence hot pepper germ plasm resource, will be red
Color parent and yellow parent are hybridized to obtain F1 groups, and F2 groups are obtained after F1 selfings.
2. fruit color is identified
After fruit maturation, red yellow color identification is carried out to fruit color.
3. fruit color main effect QTL is analyzed
Take respectively fruit color be 30 plants of extremely red and yellow blades, mixed in equal amounts structure red gene pond and chloroaniline because
Pond.It is extracted using CTAB methods and mixes the total RNA of Chi.Utilize TruSeq DNA LT Sample Prep Kit (Illumina companies)
Library is built to red pond and yellow pool rna, is sequenced by Illumina HiSeq2000 platforms.The data of acquisition utilize independent development
Perl script extract polymorphism SNP, and calculate SNP indexes (SNP-index).Further calculate two pond SNP-index
Difference (i.e. Δ SNP-index) and map.As a result as shown in Figure 3A, 1-12 represents chromosome number, mellow fruit fruit color master
Effect QTL is located at No. 6 chromosomes (at arrow instruction), by further analyzing the assignment of genes gene mapping of contral ripening fruits color
On No. 6 chromosomes in the section of 5.3M (such as Fig. 3 B arrows indicate shaded side).
4. fruit color main effect QTL finely positioning
The chromosomal region of fruit color red QTL is obtained by step 3.Further analyze the DNA sequences in purpose section
Row variation designs InDel and Caps labels, analyzes the recombination single plant of target area, identified into one in conjunction with fruit color
Step reduces localization region to 700kb.
The contral ripening fruits color gene of this method candidate is consistent with whole-genome association method (GWAS).
Two capsicum mellow fruit fruit colors mark the verification of HS5, HS6
Using the molecular labeling primer of the fruit color of acquisition, 269 parts of materials to building natural population carry out fruit face
Color identifies that its step are as follows：
(1) using capsicum genomic DNA as template；Two pairs of primers that HS5, HS6 label are added in the reaction system carry out PCR
Amplification, HS5 forward primer sequences are HS5-F (SEQ ID NO.1), and reverse primer sequences are HS5-R (SEQ ID NO.2), HS6
Forward primer sequence is HS6-F (SEQ ID NO.3), and reverse primer sequences are HS6-R (SEQ ID NO.4).PCR reactants
System：Total volume is 15.8 μ l, and specific ingredient is as follows：10.4 μ l, 10 × PCR Buffer of ddH2O 1.5 μ l, 1.2 μ l dNTPs
(2mmol/L), 0.08 μ l Taq archaeal dna polymerases (5U/ μ l), two forward primers (SEQ ID NO.1 and SEQ ID NO.3)
Each 0.4 μ l (10ng/ μ l), each 0.4 μ l (10ng/ μ l) of two reverse primers (SEQ ID NO.2 and SEQ ID NO.4) and
1.0 μ l DNA profilings.
(2) PCR reactions carry out in the S1000 type PCR instruments that Bio-Rad companies of the U.S. produce.PCR amplification program：95.0
DEG C pre-degeneration 3min；95.0 DEG C of denaturation 30s, 55.0 DEG C of renaturation 30s, 72.0 DEG C of extension 45s, 32 recycle；72.0 DEG C extension
5min, 8 DEG C of preservations.
(3) as shown in figure 4, amplified production is after the separation of 2% agarose gel electrophoresis, if there are 500bp (SEQ ID
NO.5) and two specific bands of 714bp (SEQ ID NO.6) (two bands in such as Fig. 4 at middle arrow meaning), then sentence
It is set to the material that ripe fruits color is red, if only there is 602bp (SEQ ID NO.7) band (the right arrow in such as Fig. 4
A band at head meaning), then it is determined as mellow fruit fruit color as the material of yellow, left arrow meaning is in Fig. 4
Maker bands.
(table 1 is the fruit color and genotype of 269 parts of capsicum germplasm used in the present invention as shown in table 1.Gene in table
Type 1,2 is respectively represented carries out primer the amplification of PCR amplification generation using HS5, HS6 two.1：Generate 602bp (SEQ ID
NO.7) 1 band；2：Generate 500bp (SEQ ID NO.5), 714bp (SEQ ID NO.6) 2 bands；), result is：At 269 parts
In capsicum material, there are 2 specificity and carry 255 materials, it is 235 that wherein mellow fruit fruit color, which is red material,
A, coincidence rate reaches 92.1%.The material for a band occur has 14, and 14 material mellow fruit fruit colors are all yellow,
Coincidence rate reaches 100%.Result above absolutely proves that these two pair marks while having versatility and accuracy, can be applied to
Prediction, identification and the screening of capsicum mellow fruit fruit color.
Above-mentioned qualification result shows to screen by molecular markers for identification in breeding, and 500bp (SEQ ID occurs in reservation
NO.5) and the material of the special banding patterns of 714bp (SEQ ID NO.6) two, it will be able to which it is red to select mellow fruit fruit color
Material.Retain the material for 602bp (SEQ ID NO.7) Idiotype banding pattern occur, it will be able to select ripe fruits
Color is the material of yellow.The workload of later stage screening and identification can be reduced by the screening of molecular labeling, accelerate breeding process.
The corresponding fruit color of 1 269 parts of capsicum germplasm of table and genotype
Sequence table
<110>Inst. of Vegetables, Hunan Prov.
<120>It is a kind of with the relevant insertion/deletion site of capsicum mellow fruit color gene, molecular labeling, molecular labeling primer and
Using
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 26
<212> DNA
<213>Artificial sequence ()
<400> 1
gcaagtatag ctaataccaa cattac 26
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence ()
<400> 2
agttcctcaa gtggttagcc 20
<210> 3
<211> 21
<212> DNA
<213>Artificial sequence ()
<400> 3
gtgagtcggc ctatgttatc g 21
<210> 4
<211> 23
<212> DNA
<213>Artificial sequence ()
<400> 4
gtttgaacaa cgatgacaat gtg 23
<210> 5
<211> 500
<212> DNA
<213>Artificial sequence ()
<400> 5
gcaagtatag ctaataccaa cattactaat gcaagtatta ctaatacacc atattctata 60
ttaatcttat atactctacc aaacgaccct aagtgtgtat ctatatcctc cgagaatttg 120
gaatttgcaa attccaagtt ttgtatctcc ctttcccaga aattaagata attctggtgc 180
ttttagcatt agaaaagtat ttattgggta gggaaatgtc atgacttcac agcattaagc 240
atcaagggta taacttaatg aaatagtggt caatgaatta tattgagaat gacgaggtct 300
ctgttccaac tttggtagac tttggaaatg ctcgtctgga cgccgcccat tctttctagt 360
cttggtgcca ttctatttgg tctgagaatg gcatgatgcc aaattctacc ttttcacaat 420
gagcattcga cctactcttc ttttttcgac tcatttgacc tactaggcat tggccaactt 480
ggctaaccac ttgaggaact 500
<210> 6
<211> 714
<212> DNA
<213>Artificial sequence ()
<400> 6
gtgagtcggc ctatgttatc gtatatggaa gtgaaaagaa ggatggtagc aagattaaga 60
catttgggga tcaaagtgag aagtgtcctt gaggaagaga agtgtgtgat cactatggga 120
ggaccacttc cgcggattcc tcaaaatgtt atggctattg gtgggacttc agggatagtt 180
catccatcgt ctgggtacat ggtggctcgt agcatggcat tggcaccagt actggctgag 240
gccatcgtcg aaagccttgg ctcaacaaga atgataagag ggtctcaact ttaccataga 300
gtttggaatg gtttgtggcc ttcggataga agacgtgtta gagaatgtta ttgtttcgga 360
atggagactt tgttgaagct tgatttggaa ggtactagga gattgtttga tgctttcttt 420
gatgttgatc ccaagtactg gcacgggttc ctttcttcaa gattgtctgt caaagaactt 480
gctgtactca gtttgtacct ttttggacat gcctctaatt tggctaggtt ggatattgtt 540
acaaagtgca ctgtcccctt ggttaaactg ctgggcaatc tagcaataga gagcctttga 600
attaatatga tagttttgaa gcactgtttt cattttaatt tcttaggtta ttttcatctt 660
ttctcaatgc aaaagtgaaa caaaagctat acacattgtc atcgttgttc aaac 714
<210> 7
<211> 602
<212> DNA
<213>Artificial sequence ()
<400> 7
gcaagtatag ctaataccaa cattactaat gcaagtatta ctaatacacc atattctata 60
ttaatcttat atactctacc aaacgaccct aagtgtgtat ctatatcctc cgagaatttg 120
gaatttgcaa attccaagtt ttgtatctcc ctttcccaga aattaagata attctggtgc 180
tttttagagt ttggaatggt ttgtggcctt cggatagaag acgtgttaga gaatgttatt 240
gtttcggaat ggagactttg ttgaagcttg atttggaagg tactaggaga ttgtttgatg 300
ctttctttga tgttgatccc aagtactggc acgggttcct ttcttcaaga ttgtctgtca 360
aagaacttgc tgtactcagt ttgtaccttt ttggacatgc ctctaatttg gctaggttgg 420
atattgttac aaagtgcact gtccccttgg ttaaactgct gggcaatcta gcaatagaga 480
gcctttgaat taatatgata gttttgaagc actgttttca ttttaatttc ttaggttatt 540
ttcatctttt ctcaatgcaa aagtgaaaca aaagctatac acattgtcat cgttgttcaa 600
ac 602
Claims (10)
1. a kind of insertion/deletion site being closely related with capsicum mellow fruit fruit color, it is characterised in that the insertion/lack
Unsceptered point is since the gene coding regions capsicum annuum l. OC107875664 (ATG) upstream 3277bp to one section of code area 4513bp
Insertion/deletion.
2. a kind of molecular labeling amplimer being closely related with capsicum mellow fruit fruit color, it is characterised in that the molecule mark
Remember that amplimer is set for the insertion/deletion site described in claim 1 being closely related with capsicum mellow fruit fruit color
The primer of meter.
The molecular labeling amplimer 3. according to claim 2 and capsicum mellow fruit fruit color is closely related, feature
It is, it (includes part that the molecular labeling amplimer, which is with capsicum annuum l. OC107875664 gene delection upstream portions sequence,
Deletion sequence) and the detection of missing downstream part sequence (include excalation sequence) the design deletion segment two pairs of primers.
4. the molecular labeling amplimer according to claim 3 being closely related with capsicum mellow fruit fruit color, described
Molecular labeling amplimer sequence be：Lack upstream HS5 labels：HS5-F(SEQ ID NO.1)：
GCAAGTATAGCTAATACCAACATTAC, HS5-R (SEQ ID NO.2)：AGTTCCTCAAGTGGTTAGCC.Lack downstream
HS6 marks HS6-F (SEQ ID NO.3)：GTGAGTCGGCCTATGTTATCG, HS6-R (SEQ ID NO.4)：
GTTTGAACAACGATGACAATGTG。
5. a kind of and pepper fruit color gene chain molecular labeling HS5, HS6, which is characterized in that utilize claim 4 institute
The molecular labeling amplimer stated, the PCR product clip size that HS5 is amplified are 500bp, nucleotide sequence such as SEQ ID
Shown in NO.5, the PCR product clip size that molecular labeling HS6 is amplified is 714bp, nucleotide sequence such as SEQ ID NO.6 institutes
Show.
6. the insertion/deletion site that is closely related as described in claim 1 with capsicum mellow fruit fruit color is being identified or is being assisted
The color for screening pepper fruit or the application in pepper fruit breeding.
7. the molecular labeling amplimer being closely related with capsicum mellow fruit fruit color as described in claim 2-4 is any exists
The color or the application in pepper fruit breeding of identification or assisting sifting pepper fruit.
8. the molecular labeling HS5, HS6 chain with pepper fruit color gene are identifying or are assisting sieve as claimed in claim 5
The color for selecting pepper fruit or the application in pepper fruit breeding.
9. a kind of method of identification and screening pepper fruit color, is as follows：
(1) using sample to be tested genomic DNA as template, using molecular labeling HS5, the HS6 amplimer described in claim 5 into
Row PCR amplification obtains amplified production；
(2) amplified production is detached by agarose gel electrophoresis；
(3) interpretation of result, if occurring 500bp (SEQ ID NO.5) and 714bp (SEQ ID NO.6) two on Ago-Gel
Specific band is then determined as the material that mellow fruit fruit color is red, if only there is a 602bp (SEQ ID NO.7)
Band is then determined as that mellow fruit fruit color is the material of yellow.
10. the method for identification as claimed in claim 9 and screening pepper fruit color, it is characterised in that the identification and screening
In the method and step (1) of pepper fruit color, PCR amplification program is 95.0 DEG C of pre-degeneration 3min；95.0 DEG C denaturation 30s, 55.0
DEG C renaturation 30s, 72.0 DEG C of extension 45s, 32 cycles；72.0 DEG C of extension 5min, 8 DEG C of preservations.
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| CN201810243771.1A CN108384875A (en) | 2018-03-23 | 2018-03-23 | A kind of and the relevant insertion/deletion site of capsicum mellow fruit color gene, molecular labeling, molecular labeling primer and application |
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| CN201810243771.1A CN108384875A (en) | 2018-03-23 | 2018-03-23 | A kind of and the relevant insertion/deletion site of capsicum mellow fruit color gene, molecular labeling, molecular labeling primer and application |
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| CN201810243771.1A Pending CN108384875A (en) | 2018-03-23 | 2018-03-23 | A kind of and the relevant insertion/deletion site of capsicum mellow fruit color gene, molecular labeling, molecular labeling primer and application |
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| CN109207622A (en) * | 2018-10-24 | 2019-01-15 | 湖南省蔬菜研究所 | A kind of molecular labeling and application with the stagnant green gene linkage of capsicum |
| CN110373487A (en) * | 2019-04-24 | 2019-10-25 | 华南农业大学 | One kind InDel label relevant to capsicum pungent character and its application |
| CN111088383A (en) * | 2019-11-29 | 2020-05-01 | 绿亨科技集团股份有限公司 | Molecular marker for identifying purple genes of capsicum olivum and development method and application thereof |
| CN111394508A (en) * | 2020-05-25 | 2020-07-10 | 湖南省蔬菜研究所 | Molecular marker linked with capsicum frutescens gene and application thereof |
| CN112725516A (en) * | 2021-02-25 | 2021-04-30 | 中国农业科学院蔬菜花卉研究所 | Specific primer of molecular marker closely related to orientation of pepper fruits and application |
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| CN109207622A (en) * | 2018-10-24 | 2019-01-15 | 湖南省蔬菜研究所 | A kind of molecular labeling and application with the stagnant green gene linkage of capsicum |
| CN109207622B (en) * | 2018-10-24 | 2021-07-23 | 湖南省蔬菜研究所 | Molecular marker linked with capsicum green-staying gene and application thereof |
| CN110373487A (en) * | 2019-04-24 | 2019-10-25 | 华南农业大学 | One kind InDel label relevant to capsicum pungent character and its application |
| CN111088383A (en) * | 2019-11-29 | 2020-05-01 | 绿亨科技集团股份有限公司 | Molecular marker for identifying purple genes of capsicum olivum and development method and application thereof |
| CN111088383B (en) * | 2019-11-29 | 2022-12-06 | 绿亨科技集团股份有限公司 | Molecular marker for identifying purple genes of capsicum olivum and development method and application thereof |
| CN111394508A (en) * | 2020-05-25 | 2020-07-10 | 湖南省蔬菜研究所 | Molecular marker linked with capsicum frutescens gene and application thereof |
| CN111394508B (en) * | 2020-05-25 | 2023-03-28 | 湖南省蔬菜研究所 | Molecular marker linked with capsicum frutescens gene and application thereof |
| CN112725516A (en) * | 2021-02-25 | 2021-04-30 | 中国农业科学院蔬菜花卉研究所 | Specific primer of molecular marker closely related to orientation of pepper fruits and application |
| CN112725516B (en) * | 2021-02-25 | 2022-04-08 | 中国农业科学院蔬菜花卉研究所 | Specific primer of molecular marker closely related to orientation of pepper fruits and application |
| CN112831589A (en) * | 2021-03-02 | 2021-05-25 | 中国农业科学院蔬菜花卉研究所 | Specific primer of molecular marker closely related to fruit shape index and application |
| CN112831589B (en) * | 2021-03-02 | 2022-05-31 | 中国农业科学院蔬菜花卉研究所 | Specific primer of molecular marker closely related to fruit shape index and application |
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