CN116790800B - Indel mark related to seed size of melon and application thereof - Google Patents
Indel mark related to seed size of melon and application thereof Download PDFInfo
- Publication number
- CN116790800B CN116790800B CN202310872014.1A CN202310872014A CN116790800B CN 116790800 B CN116790800 B CN 116790800B CN 202310872014 A CN202310872014 A CN 202310872014A CN 116790800 B CN116790800 B CN 116790800B
- Authority
- CN
- China
- Prior art keywords
- melon
- indel
- muskmelon
- seq
- genotype
- 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
Links
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 title claims abstract description 43
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 241000219112 Cucumis Species 0.000 title 1
- 244000241257 Cucumis melo Species 0.000 claims abstract description 58
- 235000009847 Cucumis melo var cantalupensis Nutrition 0.000 claims abstract description 14
- 238000009395 breeding Methods 0.000 claims abstract description 9
- 230000001488 breeding effect Effects 0.000 claims abstract description 8
- 239000002773 nucleotide Substances 0.000 claims abstract description 8
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 8
- 210000000349 chromosome Anatomy 0.000 claims abstract description 6
- 238000001514 detection method Methods 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 8
- 238000003780 insertion Methods 0.000 claims description 4
- 230000037431 insertion Effects 0.000 claims description 4
- 238000011144 upstream manufacturing Methods 0.000 claims description 4
- 230000003321 amplification Effects 0.000 claims description 3
- 239000003550 marker Substances 0.000 claims description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 3
- 235000009842 Cucumis melo Nutrition 0.000 claims description 2
- 239000012634 fragment Substances 0.000 claims description 2
- 238000012216 screening Methods 0.000 abstract description 7
- 241000196324 Embryophyta Species 0.000 description 11
- 239000000463 material Substances 0.000 description 10
- 238000001962 electrophoresis Methods 0.000 description 6
- 239000003292 glue Substances 0.000 description 6
- 238000011161 development Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000011521 glass Substances 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 229920000936 Agarose Polymers 0.000 description 2
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 2
- 241000353135 Psenopsis anomala Species 0.000 description 2
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000012297 crystallization seed Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000012160 loading buffer Substances 0.000 description 2
- 238000013507 mapping Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 101710134784 Agnoprotein Proteins 0.000 description 1
- 208000002109 Argyria Diseases 0.000 description 1
- 235000011273 Benincasa Nutrition 0.000 description 1
- 244000036905 Benincasa cerifera Species 0.000 description 1
- 241000219104 Cucurbitaceae Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 101710155950 Inactive receptor-like serine/threonine-protein kinase At2g40270 Proteins 0.000 description 1
- KWYHDKDOAIKMQN-UHFFFAOYSA-N N,N,N',N'-tetramethylethylenediamine Chemical compound CN(C)CCN(C)C KWYHDKDOAIKMQN-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 101710082601 Pentatricopeptide repeat-containing protein At2g30100, chloroplastic Proteins 0.000 description 1
- 241001349593 Praecitrullus Species 0.000 description 1
- 101710141687 Probable N-acetyltransferase HLS1-like Proteins 0.000 description 1
- 101710189648 Serine/threonine-protein phosphatase Proteins 0.000 description 1
- 230000009418 agronomic effect Effects 0.000 description 1
- 229910001870 ammonium persulfate Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000000546 chi-square test Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000027939 micturition Effects 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an Indel mark related to the seed size of a melon and application thereof, belonging to the technical field of molecular biology. The invention provides Indel marks related to the large-scale characters of muskmelon, a specific primer pair and a detection method, wherein the Indel marks are positioned at the 15906420 base of chromosome 10 of the muskmelon, and the nucleotide sequence shown in SEQ ID NO:1, and a pair of Indel markers (Indel 9) for screening seed sizes are developed at the same time, in the invention, the Indel markers closely linked with the seed sizes of the muskmelon seeds are finely positioned, and a primer pair for screening seed sizes is designed through the markers, so that the size of the muskmelon seeds can be accurately screened by using the primer (Indel 9), and the breeding efficiency is improved.
Description
Technical Field
The invention relates to the technical field of molecular biology, in particular to an Indel marker related to the seed size of a melon and application thereof.
Background
The melon, the alias Mao Gua, mao Jie, is a variety of the white gourd species of the genus Benincasa of the family Cucurbitaceae, is one of the characteristic vegetables in our country, and has a planting history of over 300 years in our country. The melon is usually made of tender fruits, and partial varieties of old melon can be eaten. The melon is used as a common raw material for dishes, can be used for stir-frying, steaming and brewing, and is even eaten as raw food for cold dishes, and is a high-quality food material integrating delicious taste and health care due to the fact that the melon is soft and smooth in meat quality, unique in flavor, rich in various vitamins and mineral elements, low in sodium and fat content, and capable of clearing heat, relieving summer heat, promoting urination, detoxifying and the like. The melon is a main vegetable crop in the south China, is widely cultivated in Guangdong, guangxi and Hainan provinces, and is deeply favored by consumers.
Seed size and quality are important agronomic traits, and the seed size has certain influence on emergence and seedling growth, and simultaneously determines the yield of crops. Generally, larger seeds grow larger seedlings than smaller seeds. In the same growing environment, the large seeds and the small seeds can ensure that the emergence rate is not influenced or is less influenced, but the influence on the later-stage development of seedlings is larger, and especially in a unfavorable environment, the advantages of the large seeds are more obvious, compared with the small seeds, the large seeds grow more vigorously, the yield is higher, and the small seeds can be effectively dispersed and planted. Therefore, the exploration of seed traits plays an important role in improving crop germplasm, increasing yield and solving the problem of biological energy.
Although research on seed size has been conducted on many crops, there has been no report on seed size and quality of the melon. If molecular markers related to the size of the seed of the melon can be developed, the molecular markers have important significance for screening the size of the seed of the melon, and the breeding efficiency of the melon is improved.
Disclosure of Invention
The invention aims to provide Indel marks related to the seed size of a melon and application thereof, so as to solve the problems in the prior art.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides an Indel marker related to a large-scale character of a melon, which is positioned at a 15906420 base of a 10 th chromosome of the melon, and has a nucleotide sequence shown in SEQ ID NO:1, and a nucleotide sequence shown in the sequence 1.
Preferably, the genotype of the base at 15906420 of chromosome 10 of the melon comprises the nucleotide sequence as set forth in SEQ ID NO:1, and inserting a nucleotide sequence shown in the formula 1; genotype is the insertion time the melon is the individual with a large number of characteristics.
The invention also provides a specific primer pair for amplifying the Indel label, which comprises the following steps: as set forth in SEQ ID NO:2 and an upstream primer as set forth in SEQ ID NO:3, and a downstream primer shown in 3.
The invention also provides a kit for detecting the large-scale character of the muskmelon, which comprises the specific primer pair.
The invention also provides a method for detecting the Indel mark, which comprises the following steps:
(1) Extracting DNA of the melon to be detected;
(2) Using the extracted DNA as a template, and amplifying by using the specific primer pair or the kit to obtain an amplified product;
(3) And (3) identifying the genotype of the amplified product to screen out the melon individual with the genotype as the insertion, namely the melon individual with the large character.
The invention also provides application of the Indel mark, the specific primer pair or the kit in the detection of the large-scale properties of the melon or the auxiliary breeding of the melon.
The invention also provides a method for detecting the large-scale character of the melon, which is used for detecting the Indel mark of the melon to be detected so as to determine the large-scale character of the melon to be detected.
The invention also provides a melon auxiliary breeding method, which comprises the step of detecting the Indel mark.
The invention discloses the following technical effects:
the invention provides Indel marks, specific primer pairs and detection methods related to the large-scale characters of the muskmelon, wherein the Indel marks are positioned at the 15906420 th base of the chromosome 10 of the muskmelon, TTCGTACTATTCTA bases are inserted into the Indel marks, a pair of Indel marks (Indel 9) for screening the size of seeds are developed at the Indel marks, the Indel marks closely linked with the size of the seeds of the muskmelon are finely positioned, and the primer pairs for screening the size of the seeds are designed through the marks, so that the size of the seeds of the muskmelon can be accurately screened by using the primers, and the breeding efficiency is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the comparison of seed sizes of melon seeds, each including a large female parent (P 1 ) Male parent of miniascape (P) 2 ) Large species F 1 A population;
FIG. 2 shows the fine positioning of the seed size of the melon, wherein a is the BSA-seq result, b is the preliminary positioning result, and c is the fine positioning result;
FIG. 3 is Indel marking portion F 2 A population glue pattern;
FIG. 4 is Indel mark 70F 2 A single plant parting result;
FIG. 5 is Indel mark 70F 2 A single plant parting result;
FIG. 6 shows Indel-labeled 70 different germplasm typing results;
FIG. 7 shows the results of Indel labeling of 70 different germplasm typing.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
The technical scheme of the invention is conventional in the field, and the reagents or raw materials are purchased from commercial sources or are disclosed.
Example 1
1. Experimental materials: the large seed melon inbred line J16 is used as a female parent (P 1 ) The inbred line 18FJ5 of the small seed melon is taken as the male parent (P 2 ) The materials are all multi-generation stable inbred lines bred by Guangzhou city agricultural science institute. J16 (P) 1 ) The seed length, width and mass of 30 seeds were 11.44 (+ -0.27) mm, 5.97 (+ -0.14) mm and 1.34 (+ -0.1) g, respectively. 18FJ5 (P) 2 ) The seed length, width and mass of 30 seeds are 6.5 (+ -0.24) mm, 3.43 (+ -0.15) mm and 0.62 (+ -0.07) g respectively.
2. Seed size gene mapping comprising the steps of:
2022 autumn, strain 236F 2 Performing phenotype identification on the population, taking 10 full seeds from each plant, measuring the length and the width of each seed by using a vernier caliper, and weighing the total mass of 30 full seeds by using a ten-thousandth balance for genetic analysis of the size and the mass of the seeds to obtain F 1 The seed size is close to that of large seed parentThe present invention, F 2 Separation occurs, with 172 parts of the major species, 64 parts of the minor species, meeting 3 by chi-square test: 1 the desired separation ratio. Each 20 plants were then selected to construct an extreme pool of blends, finding a region above the threshold on Chr10, with a total length of 1.31Mb (as shown in fig. 2 a).
A plurality of pairs of Indel markers (Table 1) were designed in this region based on the sequencing results for strain 363F 2 DNA analysis was performed to initially position the seed size gene at about 141kb in the range of Indel2-Indel4 (15,857,520-15,998,480) (shown in FIG. 2 b).
TABLE 1Indel primer pairs
| Primers | Forward | Reverse |
| Indel1 | GAGAGTAAAAAGTTCTAAATATCCG | CTATCTCCACTGTACTAGAAAAATC |
| Indel2 | GTGTCAAAAATATCCGTAGACTTTC | AAAGGTATAGGTACCAAAGAAGAGG |
| Indel3 | CACTACATTACTCATTTGGGAGTAG | CAGAGGCTACTATTGCTCACTCTAT |
| Indel4 | GTCTATGTAATCCACCTCTCACTTG | AAGGAGTACACGATGTACAACAAGA |
| Indel5 | GGTGTAACTTACGTAATTTCTAACC | AGTCTCAATTTTACGCATATTACTG |
| Indel6 | AAAAAAAAGAGGGGCCTATATACAA | TAATCTGCTGTTCTTCAAAACCAAT |
Subsequently, on the basis of initial positioning, development of a plurality of pairs of Indel markers (Table 2) for 1483 strain F was continued 2 Individuals were genotyped and strain 1493F was identified by markers in the table 2 Individuals were genotyped and the interval was finally narrowed to a 109kb region (as shown in figure 2 c) based on phenotypic data, containing 4 genes (table 3).
TABLE 2Indel primer pairs
TABLE 3 mapping Interval candidate genes and functional annotations
| Gene name | Gene function annotation |
| Bch10G006400 | probable N-acetyltransferase HLS1-like |
| Bch10G006410 | serine/threonine-protein phosphatase 7long form-like protein |
| Bch10G006420 | Pentatricopeptide repeat-containing protein At2g30100,chloroplastic |
| Bch10G006430 | PREDICTED:inactive receptor-like serine/threonine-protein kinase At2g40270 |
According to the parental resequencing results, at base 15906420, the base occurs at SEQ ID NO:1, and using Primer Premier 5 software, indel markers were designed at this point for screening seed sizes of cucumis melo.
SEQ ID NO：1：TTCGTACTATTCTA。
The specific primer sequences are as follows:
upstream primer (SEQ ID NO: 2): 5'-CTTTTTGTACGTGGATATATCTGGA-3';
downstream primer (SEQ ID NO: 3): 5'-TAGACACGGCGAATAACAACTAATA-3'.
Indel mark validation:
using 70 parts F 2 The markers were verified by individual plants and by different germplasm melon materials (35 parts of each of the large and small species, all from the Guangzhou agricultural sciences).
3.1 extraction of DNA by cetyltrimethylammonium bromide method (CTAB method);
3.2 PCR amplification by using the primer;
3.3PCR reaction System (10. Mu.L): 5 mu LMix (2X), 1.0 mu L forward primer, 1.0 mu L reverse primer R1,1.0 mu L DNA,2.0 mu L ddH 2 O；
3.4PCR amplification procedure is shown in Table 4;
TABLE 4PCR amplification procedure
3.5 this experiment was run on 6% polyacrylamide gel and 0.5 XTBE buffer. The specific steps are as follows:
(1) And (3) glue preparation: the bottom of the glass plate was sealed with 1% agarose. 100mL of acrylamide solution was added with 800. Mu.L of 12% ammonium persulfate and 35. Mu.L of TEMED and mixed well. And slowly pouring the uniformly mixed solution into a glass plate sealed by agarose along one side of a concave groove, wherein the flow speed is uniform so as to prevent bubbles. After filling the glue, observing whether bubbles exist in the glue, if so, gently picking out the bubbles by using a plastic strip, and inserting a comb with the diameter of 1.0 mm. And (5) waiting for solidification. (gel time is temperature dependent, about 25-40 min in summer and about 40-60 min in winter).
(2) Spotting: after the glue is completely solidified, the comb is gently pulled out, and the sample application hole is kept complete (ddH can be reached) 2 And O, preventing bubbles from forming in the sample application holes). Fixing the gel on a vertical electrophoresis tank, and adding a proper amount of 0.5 XTBE buffer; 3-5 mu L of PCR amplified product (4 mu L of 10×loading buffer is added to each PCR product and mixed) is taken by a microsyringe and injected into a sample application hole.
(3) Electrophoresis: and (3) electrophoresis is carried out for 30-60 min under the condition of constant voltage of 300V, so that the 10×loading buffer blue indicator reaches the middle lower part of the gel. When the fragments of part of PCR products are larger, electrophoresis is needed for 2-3 hours.
(4) And (3) glue removal: after electrophoresis, the electrophoresis apparatus is closed, the glass plate is disassembled, carefully pried open, one corner of the gel is gently lifted, the glass plate with the gel is immersed into the prepared deionized water downwards, and the gel can fall down. Each gel was noted as marked.
(5) Color development: (1) rinsing: gel in ddH 2 Rinsing in O for 1-2 times each timeRinsing for no more than 20s; (2) silver staining: the gel was carefully transferred to 0.1% agno 3 Dyeing in the solution, and shaking for 10min on a decolorizing shaker at 100 rpm; (3) rinsing: transfer of gel to ddH 2 Rinsing in O for 1-2 times, wherein each rinsing time is not longer than 20s; (4) color development: transferring the gel into a color development liquid, and shaking for 5-10 min on a decolorizing shaking table at 100 rpm; until the strip is clearly identifiable; (5) flushing and recording: with ddH 2 O washes the developed gel, then records the banding results and photographs for storage.
And (3) result statistics: (1) 70F 2 The KASP typing results on individual plants are shown in FIGS. 4 and 5, 35 parts of ministrain F 2 The genotypes of the single plants are the B genotypes and are consistent with the genotypes of the male parent; 35 strains of large variety F 2 The genotype of the single plant is 27 plants which are consistent with the genotype A and the female parent, and the remaining 8 plants are consistent with the large variety F 1 Genotype (H). Thus, 70 parts of F 2 The genotype of the single plant accords with the phenotype, and the accuracy is 100%. (2) The results of 70 parts of melon materials with different germplasm show that the phenotype of 35 parts of small materials and 3 parts of large materials are not matched with the genotype (figure 6), and the accuracy rate is 95.7 percent.
The results show that the Indel marks and primers developed by the research can be used for screening the seed size according to the genotype of the melon seeds more quickly, so that the breeding efficiency is greatly improved, and the method has important application value.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (7)
1. The application of Indel marks related to the melon macrophyte is characterized in that the Indel marks are positioned at the 15906420 base of the chromosome 10 of the melon and are shown in SEQ ID NO:2 and an upstream primer as set forth in SEQ ID NO:3, where the amplified fragment of the downstream primer shown in SEQ ID NO:1, and inserting a nucleotide sequence shown in the formula 1;
the Indel marked reference gene is Bch10G006400.
2. The use according to claim 1, wherein the genotype of base 15906420 of chromosome 10 of cucumis melo comprises the nucleotide sequence set forth in SEQ ID NO:1, and inserting a nucleotide sequence shown in the formula 1; genotype is the insertion time the melon is the individual with a large number of characteristics.
3. An application of a specific primer pair in the detection of the large-scale character of the muskmelon or the auxiliary breeding of the muskmelon, which is characterized in that the specific primer pair comprises: as set forth in SEQ ID NO:2 and an upstream primer as set forth in SEQ ID NO:3, and a downstream primer shown in 3.
4. The application of a kit for detecting the large-scale properties of the muskmelon in the detection of the large-scale properties of the muskmelon or the auxiliary breeding of the muskmelon, which is characterized by comprising the specific primer pair as claimed in claim 3.
5. A method of detecting the Indel label of any one of claims 1 to 2, comprising the steps of:
(1) Extracting DNA of the melon to be detected;
(2) Amplifying the extracted DNA serving as a template by using the specific primer pair of claim 3 or the kit of claim 4 to obtain an amplification product;
(3) And (3) identifying the genotype of the amplified product to screen out the melon individual with the genotype as the insertion, namely the melon individual with the large character.
6. A method for detecting the large-scale character of a melon, which is characterized in that the Indel mark of claim 1 is detected on the melon to be detected so as to determine the large-scale character of the melon to be detected.
7. A method for breeding a melon in an assisted manner, comprising the step of detecting the Indel marker according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310872014.1A CN116790800B (en) | 2023-07-14 | 2023-07-14 | Indel mark related to seed size of melon and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310872014.1A CN116790800B (en) | 2023-07-14 | 2023-07-14 | Indel mark related to seed size of melon and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116790800A CN116790800A (en) | 2023-09-22 |
| CN116790800B true CN116790800B (en) | 2024-03-08 |
Family
ID=88040259
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310872014.1A Active CN116790800B (en) | 2023-07-14 | 2023-07-14 | Indel mark related to seed size of melon and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116790800B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111334606A (en) * | 2020-03-04 | 2020-06-26 | 青岛农业大学 | Molecular marking method for peanut seed kernel soluble sugar content related sites and application thereof |
| CN112251529A (en) * | 2020-10-22 | 2021-01-22 | 广东省农业科学院蔬菜研究所 | Indel molecular marker closely linked with female shape of towel gourd and application thereof |
| CN112760396A (en) * | 2020-12-01 | 2021-05-07 | 广东省农业科学院蔬菜研究所 | Indel molecular marker gyIndel3 closely linked with full female shape of jigua and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108048595B (en) * | 2017-12-26 | 2018-11-02 | 广东省农业科学院蔬菜研究所 | Indel molecular labelings with pumpkin photoperiod insensitive character close linkage and its application |
-
2023
- 2023-07-14 CN CN202310872014.1A patent/CN116790800B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111334606A (en) * | 2020-03-04 | 2020-06-26 | 青岛农业大学 | Molecular marking method for peanut seed kernel soluble sugar content related sites and application thereof |
| CN112251529A (en) * | 2020-10-22 | 2021-01-22 | 广东省农业科学院蔬菜研究所 | Indel molecular marker closely linked with female shape of towel gourd and application thereof |
| CN112760396A (en) * | 2020-12-01 | 2021-05-07 | 广东省农业科学院蔬菜研究所 | Indel molecular marker gyIndel3 closely linked with full female shape of jigua and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Development and application of indel markers linked to fruit-shape and peel-colour genes in wax gourd;Huang, X.等;《 Genes》;第13卷(第9期);1-13 * |
| 节瓜种子大小的主基因+多基因遗传分析;高银等;《广东农业科学》;第50卷(第4期);51-59 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116790800A (en) | 2023-09-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| BRPI0708486A2 (en) | HIGH PERFORMANCE NON-DESTRUCTIVE METHODS FOR ANALYZING INDIVIDUAL SEEDS IN A SEED POPULATION, HIGH PERFORMANCE FOR ANALYSIS OF A HAPLID SEED POPULATION, AND HIGH PERFORMANCE TO GROUP A DOUBLE HAPLID SEED POPULATION | |
| CN114182045B (en) | Soybean high-protein content related molecular marker located on chromosome 14 and method for identifying soybean high-protein content | |
| CN108315439A (en) | It is a kind of to grow relevant SNP marker and its application with Pelteobagrus vachelli | |
| Szilagyi et al. | Evaluation of genetic diversity in common bean (Phaseolus vulgaris L.) using RAPD markers and morpho-agronomic traits | |
| CN113637794A (en) | SSR molecular marker of new variety of mulberry, namely Guangdong mulberry 201, and core primer group, kit and application thereof | |
| CN109182534A (en) | Jian carp breeding of new variety method | |
| CN109722486B (en) | Watermelon seed navel spot character major gene, molecular marker for detecting major gene and application | |
| CN105838785A (en) | SSR molecular marker tightly linked to sesame black seed coat gene and application | |
| CN112592996B (en) | Molecular marker ZMM1776 closely linked with sesame seed sesamin content major gene locus and application thereof | |
| CN111057771B (en) | SNP molecular marker for distinguishing 'Zhongyang No. 1' from common fugu obscurus and application thereof | |
| CN116790800B (en) | Indel mark related to seed size of melon and application thereof | |
| CN109652587B (en) | Method for identifying holly germplasm by using SSR molecular marker of transcriptome sequencing | |
| CN101845487A (en) | Construction method of Apostichopus japonicus standard microsatellite marker | |
| CN103966316B (en) | A kind of diploid loach micro-satellite Parentage determination method and application thereof | |
| CN114736972B (en) | Reagent for evaluating eleutheronema tetradactylum growth-related characters | |
| CN111793698B (en) | SNP locus related to rapid growth of large-sized fries of fugu rubripes and application thereof | |
| CN114350847A (en) | SNP (Single nucleotide polymorphism) site for identifying early tea trees and application thereof | |
| WO2016171344A1 (en) | Saengjang, which is novel aloe variety, and molecular marker for classifying same | |
| CN106435006A (en) | SNP (Single Nucleotide Polymorphism) marker relevant to growth rate of schizothorax prenanti and application thereof | |
| Sepahian et al. | Authentication, genetic fingerprinting and assessing relatedness of rice (Oryza Sativa) genotypes by SSR molecular markers | |
| CN100376675C (en) | Method for structuring standard mini-satellite marks for ctenidium scallop and its application | |
| CN101942512B (en) | Development and application of molecule marker for corn with considerable number of kernels and excellent allele function in low-nitrogen adverse environment | |
| CN1793358A (en) | Process for structure of standard microstatellite mark of bay scallop and application thereof | |
| CN114292954B (en) | Molecular marker closely linked with green petal gene Clgf of watermelon and application thereof | |
| CN109161599B (en) | Kit for screening Yili Ma Bi milk characteristics by PITX2 gene 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 |