Melon plant high-molecular marker and application thereof in detecting height of melon plant

Abstract

The invention discloses a muskmelon plant polymer marker and application thereof in detecting the height of a muskmelon plant, and belongs to the field of biotechnology nucleotides. The technical problem to be solved by the invention is how to detect the plant height of the muskmelon or breed the muskmelon with short plant height. The muskmelon plant high-molecular marker disclosed by the invention is nucleotide at the 90th site corresponding to SEQ ID No.1 in a sequence table in a muskmelon genome, and is G or A. Experiments prove that the plant height of a homozygous muskmelon with the muskmelon plant macromolecular marker A is lower than that of a heterozygous muskmelon with the site A and G and that of a homozygous muskmelon with the site G, and the plant height of the heterozygous muskmelon is also lower than that of the homozygous muskmelon with the site A, so that the muskmelon plant macromolecular marker is related to the plant height of the muskmelon, and the muskmelon plant macromolecular marker has the advantages that the plant height is high; the method can be used for breeding dwarfed muskmelons.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology nucleotides, and particularly relates to a melon plant macromolecular marker and an application thereof in detecting melon plant macromolecular markers. Background Art

[0002] melon( Cucumis melo ) is a horticultural crop with a long cultivation history and wide distribution, and is deeply loved by consumers from all over the world.

[0003] In practical breeding and cultivation, plant shape is a crucial trait, and dwarfing is a key phenotype of plant shape. First, dwarf plants have a compact shape and small crown width, enabling efficient, dense planting and increasing yield per unit area. Furthermore, dwarf plants are resistant to lodging, facilitating mechanized harvesting and facilitating automated, intelligent agricultural production. Therefore, the identification and identification of dwarf plants has enormous economic value.

[0004] Developing molecular markers that are closely linked to dwarf plant type will help improve the efficiency of breeding dwarf melon plants and promote the development of the melon industry. Summary of the Invention

[0005] The technical problem to be solved by the present invention is how to detect the plant height of muskmelons or breed muskmelons with short plant height.

[0006] In order to solve the above technical problems, the present invention first provides an application of a substance for detecting a high molecular weight marker of a melon strain in detecting or assisting in detecting a high molecular weight melon strain; The melon strain macromolecular marker is the 90th nucleotide corresponding to SEQ ID No. 1 in the sequence list in the melon genome, which is G or A.

[0007] In the above application, the substance for detecting the high molecular marker of the melon plant can be a primer pair consisting of P1 and P2 or a set of reagents for detecting the high molecular marker of the melon plant; Said P1 and said P2 are two single-stranded DNAs shown as SEQ ID No.3 and SEQ ID No.4 in the sequence list respectively; The reagent set for detecting the high-molecular-weight markers of the melon plant includes the P1 and the P2.

[0008] In the above application, the kit may further include restriction endonuclease XhoI.

[0009] Specifically, the substance for detecting high-molecular-weight markers of melon plants can be the primer pair, or a set of reagents consisting of the primer pair and XhoI.

[0010] In the above application, the plant height of the homozygous melon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has a candidate height greater than, the plant height of the homozygous melon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A; the plant height of the homozygous melon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has a candidate height greater than, the plant height of the heterozygous melon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A; the plant height of the heterozygous melon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A is greater than, or has a candidate height greater than, the plant height of the homozygous melon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A.

[0011] The present invention also provides a method for detecting or assisting in detecting the plant height of muskmelon, the method comprising detecting the muskmelon plant height molecular marker and determining the muskmelon plant height according to the following method: The plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has the potential to be greater than, the plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A. The plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has the potential to be greater than, the plant height of a heterozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A. The plant height of a heterozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A is greater than, or has the potential to be greater than, the plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A.

[0012] In the above method, the detection of the melon plant high molecular marker is performed using the substance for detecting the melon plant high molecular marker.

[0013] In the above method, using the substance for detecting high-molecular-weight markers of melon plants to detect the high-molecular-weight markers of melon plants may include: performing PCR amplification on melon genomic DNA using the P1 and the P2 to obtain an amplified product; sequencing the amplified product to determine the nucleotide at position 90 in the melon genome corresponding to SEQ ID No. 1 in the sequence list, or enzymatically digesting the amplified product using the restriction endonuclease XhoI to detect the size of the digested product; a melon whose digested product has two DNA fragments is homozygous, and the nucleotide at position 90 in its genome corresponding to SEQ ID No. 1 is A; a melon whose digested product has three DNA fragments is heterozygous, and the nucleotides at position 90 in its genome corresponding to SEQ ID No. 1 are A and G; a melon whose digested product has one DNA fragment is homozygous, and the nucleotide at position 90 in its genome corresponding to SEQ ID No. 1 is G.

[0014] The substance for detecting high molecular markers of melon plants also falls within the protection scope of the present invention.

[0015] The melon plant macromolecular marker also falls within the protection scope of the present invention.

[0016] The present invention also provides any of the following applications: X1) Application of the aforementioned high-molecular-weight markers for muskmelon plants in muskmelon breeding; X2) Use of the melon strain macromolecular marker in detecting or assisting in detecting melon strain macromolecular markers; X3) Use of the substance for detecting high molecular weight markers of melon plants in melon breeding; X4) Use of the substance for detecting high-molecular-weight markers of melon plants in the preparation of melon breeding products; X5) Use of the aforementioned substance for detecting melon plant height markers in the preparation of a product for detecting or assisting in detecting melon plant height; X6) Application of the method for detecting or assisting in detecting melon plant height in melon breeding; X7) Use of the substance corresponding to nucleotide No. 90 of SEQ ID No. 1 in the sequence listing in the melon genome for the breeding of melons with shorter plant height; X8) Use of a substance corresponding to the 90th nucleotide of SEQ ID No. 1 in the sequence listing in the melon genome in preparing a product for breeding melons with short plant height.

[0017] The present invention also provides a melon breeding method, which comprises Y1) or Y2): Y1) detecting the 90th nucleotide corresponding to SEQ ID No. 1 in the sequence listing in the melon genome, and selecting a homozygous or heterozygous melon whose 90th nucleotide corresponding to SEQ ID No. 1 in the sequence listing is A as a parent for breeding; Y2) mutating the 90th nucleotide corresponding to SEQ ID No. 1 in the sequence listing of the melon genome from G to A to obtain the target melon, thereby achieving melon breeding.

[0018] In one embodiment of the present invention, the muskmelon is muskmelon ( Cucumis melo ) Progeny of the S52-8-F2 line.

[0019] Experiments have shown that the plant height of melons with the aa genotype (where the 90th nucleotide of SEQ ID No. 1 is A) is significantly lower than that of melons with the Aa genotype (where the 90th nucleotide of SEQ ID No. 1 is A or G) and the AA genotype (where the 90th nucleotide of SEQ ID No. 1 is G). The plant height of melons with the Aa genotype is also significantly lower than that of melons with the AA genotype. This indicates that the melon plant height marker of the present invention is correlated with melon plant height and can be used to breed dwarf melons. In addition, the melon plant height marker of the present invention is correlated with plant height, has little effect on stem diameter, and does not affect melon length, meaning that heterozygous melons have great application value in actual production, and can reduce production costs while maintaining production capacity.

[0020] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0021] Herein, unless otherwise defined herein, terms should be understood according to conventional usage by those of ordinary skill in the relevant art. Examples of resources describing many of the terms used herein related to molecular biology can be found in: Alberts et al., Molecular Biology of The Cell, 5th edition, Garland Science Publishing, Inc.: New York, 2007; Rieger et al., Glossary of Genetics: Classical and Molecular, 5th edition, Springer-Verlag: New York, 1991; King et al., A Dictionary of Genetics, 6th edition, Oxford University Press: New York, 2002; and Lewin, Genes IX, Oxford University Press: New York, 2007.

[0022] Description of biological material deposit Abbreviation of the depository unit: CGMCC.

[0023] Name of depository unit: General Microbiology Center of China Culture Collection Administration of Microorganisms.

[0024] Address of the depository: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101.

[0025] Date of preservation: April 21, 2025.

[0026] The registration number of the CGMCC Collection Center is: CGMCC No.30706.

[0027] Classification name: Melon ( Cucumis melo ).

[0028] Plant number: S52-8-F2. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1Genetic analysis of the dwarf mutant plant 13C-S52. a. Dwarf plants were obtained by EMS mutagenesis; 13C-S52 is shown in the white box. b. The phenotypic segregation of the F2 population of 13C-S52 conforms to Mendel's 1:2:1 segregation law, indicating that the dwarf mutant phenotype is controlled by a single recessive gene. BC1F2 represents the F2 population. c. The F2 population was subjected to MutMap pooled sequencing, and the SNP indices are shown in the figure. Twenty loci with an SNP index of 1 on chromosomes 1, 3, 4, 8, and 9 of melon were selected for subsequent candidate gene analysis.

[0030] Figure 2 Candidate gene analysis. a, b. Fine mapping to four loci on melon chromosome 8. (b) The left side of panel b shows the melon strain names; 13C indicates 13C-WT, S52 indicates 13C-S52, and the rest are F2 generation plant names. The right side shows the loci. ce. Plant phenotypes of the four loci linked; c. All four loci are homozygous for dominant phenotypes; d. All four loci are heterozygous; e. All four loci are homozygous for recessive phenotypes. Scale bar = 10 cm. f, g. Phenotypes of recombinant plants; f. The chr08_34157772 locus is heterozygous, while the chr08_35184204, chr08_35240383, and chr08_35736165 loci are homozygous for dominant expression; g. The chr08_34157772 locus is homozygous for recessive expression, while the chr08_35184204, chr08_35240383, and chr08_35736165 loci are heterozygous. Scale bar = 4 cm.

[0031] Figure 3 Fine mapping of the CmTUB8 gene. BC1F3-46 represents the self-pollinated offspring of F2-46 in the F2 generation, BC1F3-118 represents the self-pollinated offspring of F2-118 in the F2 generation, A represents homozygous dominant, H represents heterozygous, and B represents homozygous recessive.

[0032] Figure 4Phenotypes and data analysis of plants with the AA, Aa, and aa genotypes. ac, scale bar = 10 cm, a. AA genotype plants show normal plant height, b. Aa genotype plants show semi-dwarfed growth, c. aa genotype plants show extreme dwarfed growth. dg, scale bar = 8 cm, from left to right, the figures show plants with the AA, Aa, and aa genotypes, respectively. d: lateral branches at the same node, e: leaves at the same node, fg: bisexual and male flowers at the day of flowering. h: internode length from the 1st to the 8th internode (cotyledonary nodes are not included in the statistics), i: internode length from the 8th to the 16th internode, j: plant height, k: stem diameter at the 8th node, l: stem diameter at the 16th node, m: bisexual flower length at the day of flowering. In hm, the horizontal axis from left to right represents AA genotype plants, Aa genotype plants, and aa genotype plants respectively.

[0033] BC1F3-A, BC1F3-H, and BC1F3-B represent plants with AA genotype, Aa genotype, and aa genotype, respectively.

[0034] In the same bar graph, the same letters indicate no significant difference, and different letters indicate significant difference, two-way ANOVA.

[0035] Figure 5 Electrophoresis results of enzyme digestion products. A represents the AA genotype, B represents the aa genotype, and H represents the Aa genotype. DETAILED DESCRIPTION

[0036] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, and instruments used in the following examples are all commercially available unless otherwise specified.

[0037] Example 1: The melon plant height marker is a molecular marker associated with melon plant height. 1. Genetic analysis of the dwarf mutant plant 13C-S52 The melon 13C inbred line (13C-WT) is a wild-type thin-skinned melon variety. 13C-S52 was obtained from melon 13C-WT by EMS mutagenesis and is an M1 generation ( Figure 1The M1 generation was phenotypically distinct from the wild-type 13C-WT, characterized by dwarfing, reduced stem length, fewer nodes, smaller leaves (both width and length), and smaller floral organs. The M1 generation was self-pollinated to produce the M2 generation. To remove background noise, the M2 generation was hybridized with the 13C-WT to produce the F1 generation, which was then self-pollinated to produce the F2 generation. Segregation of traits was observed in the F2 generation population. Of the 160 F2 generation plants, 55 had normal height, 75 had intermediate height, and 30 had dwarf mutants, conforming to the Mendelian law of 1:2:1 segregation (χ 2 =8.4<χ 2 0.01 =9.21), indicating that melon dwarfism is controlled by a recessive single gene ( Figure 1 (b)

[0038] Among them, one of the strains in the F1 generation is named S52-BC1F1-8, and its self-pollinated progeny is recorded as S52-8-F2 strain (i.e. BC1F2). The S52-8-F2 strain was deposited in the General Microbiology Center of the China Culture Collection Administration on April 21, 2025, with the deposit number CGMCC No.30706.

[0039] 2. Candidate gene analysis and fine mapping of the CmTUB8 gene The inventors selected 13 dwarf mutants and 25 normal plant height plants with extreme phenotypic differences in the BC1F2 generation, extracted DNA from young leaves at the stem tip, and used the MutMap method and the MGI-DNBSeq platform to perform mixed pool sequencing on the samples. The sequencing read length was PE150, and the sequencing data volume was 10 G. The raw reads in the original sequencing sequence were filtered, and the filtered clean reads were further quality controlled and interpreted using the advanced quality control FastQC software to obtain the variant SNP information of the W-pool (normal plant height pool) and the M-pool (dwarf mutant pool). For the SNPs in the two mixed pools, a total of 1195 SNPs were extracted that mutated from guanine to adenine or cytosine to thymine. SNPs with SNP-index ≤ 0.5 in the W-pool and SNP-index ≥ 0.3 in the M-pool were retained. Then, five consecutive SNPs were used as a sliding window, and one SNP was moved at a time to obtain the regression line of the SNP index graph ( Figure 1 (c)

[0040] The inventors selected 25 sites with SNP-index = 1, which are located on chromosomes 1, 3, 4, 8, and 9 of melon. They used Primer Premier 5 to design primers for sequencing and identification, excluding sites that were inconsistent with the dwarf mutation phenotype. Finally, the target was located on four candidate sites, all of which are located on chromosome 8 of melon ( Figure 2 Then, CAPs and dCAPS molecular markers designed using the dCAPSFinder 2.0 (http: / / helix.wustl.edu / dcaps / dcaps.html) tool were used to identify four candidate loci: chr08_34157772, chr08_35184204, chr08_35240383, and chr08_35736165. These four loci are often inherited together as dominant, heterozygous, or recessive genotypes, but have a recombination rate of 4.38%, indicating an incomplete linkage phenomenon. Figure 2 The performance of the recombinant plants is as follows: (1) When the chr08_34157772 locus is dominant homozygous and the chr08_35184204, chr08_35240383 and chr08_35736165 loci are heterozygous, the plant height is normal; (2) When the chr08_34157772 locus is heterozygous and the chr08_35184204, chr08_35240383 and chr08_35736165 loci are recessive homozygous, the plant height is semi-dwarf; (3) When the chr08_34157772 locus is heterozygous and the chr08_35184204, chr08_35240383 and chr08_35736165 loci are dominant homozygous, the plant height is semi-dwarf ( Figure 2 f); (4) When the chr08_34157772 locus is recessive homozygous and the chr08_35184204, chr08_35240383 and chr08_35736165 loci are heterozygous, the plant becomes dwarf ( Figure 2 Here, the inventors found that only the chr08_34157772 site was completely co-segregated with the dwarf phenotype, and chr08_34157772 was predicted to be a synonymous mutation, the chr08_35184204 site was in the intergenic region, chr08_35240383 was a synonymous mutation, and chr08_35736165 was a mutation located downstream of the gene. Therefore, the inventors speculated that chr08_34157772 was the target site ( Figure 3 ). Dominant homozygosity means that the site is the same homozygous type as the wild type, and recessive homozygosity means that the site is a homozygous type different from the wild type (i.e., the homozygous type after mutation).

[0041] chr08_34157772 is located in the coding region of the MELO3C003218 gene, which encodes β-tubulin in melon. This gene is homologous to TUB8 in Arabidopsis thaliana and is designated CmTUB8. At chr08_34157772, the CmTUB8 gene has a mutation from cytosine to thymine at position 1366 (i.e., position 422 of the CDS), resulting in a mutation from proline to leucine at position 141 of the amino acid chain.

[0042] Next, for further verification, the inventors retained chr08_34157772 as heterozygous, and chr08_35184204, chr08_35240383 and chr08_35736165 as dominant homozygous BC1F2 recombinant plants BC1F2-46 and BC1F2-118, and self-pollinated them to obtain BC1F3 generations, separating out three phenotypes: dwarf, intermediate, and normal plant height, thus proving that the chr08_34157772 locus is indeed co-segregated with the dwarf phenotype ( Figure 4 ac), and the site was recorded as a high-molecular-weight marker for melon plants.

[0043] 3. Analysis of molecular markers of plant height in BC1F3 generation and melon plant height 3.1 Primer design In the melon 13C-WT genome, the upstream and downstream sequences of the melon strain high molecular weight marker are shown as SEQ ID No. 1, and in the 13C-S52 genome, the upstream and downstream sequences of the melon strain high molecular weight marker are shown as SEQ ID No. 2. SEQ ID No. 1 and SEQ ID No. 2 differ at position 90. The complementary nucleotide corresponding to position 90 of SEQ ID No. 1 in the melon 13C-WT genome is C, and the complementary nucleotide corresponding to position 90 of SEQ ID No. 2 in the 13C-S52 genome is T. The upstream and downstream sequences of the melon strain high molecular weight marker were designed to design primers for amplifying the melon strain high molecular weight marker. The primers are as follows: P1: 5'-ACATAGCAGAGGCGGTTAAGTAGCG-3' (SEQ ID No. 3); P2: 5'-TGACTTGCTGTTTGAGATTCCCGG-3' (SEQ ID No. 4).

[0044] 3.2 Detection of high-molecular-weight markers in BC1F3 generation melon plants Genomic DNA was extracted from the self-pollinated progeny of the S52-8-F2 line (i.e., the BC1F3 generation, a total of 128 plants). PCR amplification was performed using primers P1 and P2. The PCR reaction system consisted of 7 μL of 2 × Rapid Taq Master Mix (Vazyme), 0.5 μL of primer P1, 0.5 μL of primer P2, 0.5 μL of genomic DNA, and 5.5 μL of ddH2O. The concentrations of primers P1 and P2 in the system were both 10 μM.

[0045] The obtained PCR reaction system was subjected to PCR amplification under the following conditions: pre-denaturation at 95°C for 3 min; denaturation at 95°C for 15 s, annealing at 56°C for 15 s, extension at 72°C for 30 s, 30 cycles, and extension at 72°C for 5 min.

[0046] The resulting PCR product was digested with XhoI. The digestion reaction mixture for 14 μL was as follows: 7 μL of PCR product, 1.4 μL of rcutsmart buffer, 0.2 μL of XhoI, and 5.4 μL of ddH2O. Both rcutsmart buffer and XhoI are products from New England Biolabs.

[0047] The enzyme digestion reaction system was digested at 37° C. for 6 hours to obtain the digestion product.

[0048] The digested products were then electrophoresed on a 4% agarose gel at 120 V for 50 min, stained with M5 Gelred Plus nucleic acid, and observed and recorded under a gel imaging system.

[0049] The enzyme digestion product has two DNA fragments in total. The melon with two bands shown by electrophoresis is homozygous. The nucleotide corresponding to the 90th position of SEQ ID No.1 in its genome is A. It is recorded as aa genotype melon. The size of each band is 86 and 157 bp. The enzyme digestion product has three DNA fragments in total. The melon with three bands shown by electrophoresis is heterozygous. The nucleotide corresponding to the 90th position of SEQ ID No.1 in its genome is A and G. It is recorded as Aa genotype melon. The size of each band is 86, 157 and 243 bp. The enzyme digestion product has one DNA fragment in total. The electrophoresis result shows one band. The melon with homozygous. The nucleotide corresponding to the 90th position of SEQ ID No.1 in its genome is G. It is recorded as AA genotype melon. The size of the band is 243 bp. The electrophoresis results of some plants are as follows. Figure 5 shown.

[0050] The results showed that among the 128 plants tested, there were 21 aa genotype melons, 65 Aa genotype melons, and 42 AA genotype melons. The melons with the aa genotype all showed extreme dwarfing, the melons with the Aa genotype all showed semi-dwarfing, and the melons with the AA genotype were of normal height. The height of the entire plant was measured after the 128 plants were planted and allowed to climb naturally for 2.5 months. The results were ( Figure 4 Figure j) shows that the average plant height of AA genotype melons is 212.5 cm, the average plant height of Aa genotype melons is 139.0 cm, and the average plant height of aa genotype melons is 31.7 cm. The plant height of aa genotype melons is significantly lower than that of Aa and AA genotype melons, and the plant height of Aa genotype melons is also significantly lower than that of AA genotype melons, indicating that the melon plant height molecular marker of the present invention is correlated with melon plant height and can be used for breeding dwarf melons.

[0051] 4. Morphological and cytological analysis of CmTUB8 single nucleotide mutation offspring Field observations showed that the CmTUB8 single nucleotide mutation caused the plant to be extremely dwarfed, with reduced lateral branch length, fewer stem nodes, smaller leaves including leaf width and leaf length, and smaller floral organs ( Figure 4 Then, the inventors selected 13 AA genotype melons, 14 Aa genotype melons, and 14 aa genotype melons from 128 plants of the BC1F3 generation that had been planted for two months, and measured the internode lengths of nodes 1-8 and 8-16 from the bottom to the top, and the stem diameter lengths of nodes 8 and 16 from the bottom to the top of the plants that had been planted for 2.5 months ( Figure 4 h, i, k, l). Two-way ANOVA followed by Bonferroni multiple comparison test was performed using GraphPad Prism version 9.5.1 for Windows (GraphPad Software, Boston, Massachusetts, www.graphpad.com). The results showed that the average internode lengths of the AA, Aa, and aa genotypes from nodes 1 to 8 were 29.4, 26.1, and 12.7 cm, respectively, and the average internode lengths of nodes 8 to 16 were 49.8, 38.5, and 14.9 cm, respectively. The average diameters of the stem at node 8 were 5.0, 4.9, and 3.5 mm, respectively, and the average diameters of the stem at node 16 were 5.4, 4.9, and 2.9 mm, respectively. The internode length of the aa genotype plants was significantly reduced, while the internode length of the Aa genotype plants was between the aa and AA genotypes, but there were significant differences between the internode lengths of the AA and aa genotypes ( Figure 4h, i). The stem diameter of the aa genotype plants was significantly smaller than that of the Aa and AA genotype plants. Similarly, the stem diameter of the Aa genotype was between the aa and AA genotypes. However, there was no significant difference in stem diameter between the Aa and AA genotypes in Section 8 ( Figure 4 (k,l).

[0052] In addition, the inventors selected 16 bisexual flowers from AA genotype plants, 13 bisexual flowers from Aa genotype plants, and 13 bisexual flowers from aa genotype plants on the day of flowering, and measured the length of the bisexual flowers (i.e., the length from the base of the pedicel to the base of the petals). It was found that the average lengths of the bisexual flowers of the AA genotype, Aa genotype, and aa genotype were 27.0, 25.1, and 18.5 mm, respectively. The bisexual flowers of the aa genotype plants were significantly smaller, while there was no significant difference in length between the bisexual flowers of the Aa genotype and the AA genotype plants ( Figure 4 Chinese m).

[0053] The results showed that the heterozygous mutation from C to T at the chr08_34157772 site of the CmTUB8 gene only affected plant height, had little effect on stem thickness, and did not affect melon length, which means that the heterozygote has great application value in actual production, reducing production costs while maintaining production capacity.

[0054] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced over a wide range of parameters, concentrations, and conditions without departing from the spirit and scope of the present invention and without unnecessary experimentation. Although specific embodiments have been given herein, it should be understood that further modifications may be made to the present invention. In summary, this application is intended to encompass any variations, uses, or improvements to the present invention, including those made by conventional techniques known in the art that depart from the scope of the present invention. Applications of the essential features may be made within the scope of the following claims.

Claims

1. Application of high molecular weight markers for detecting melon strains in detecting or assisting in the detection of melon strains; The melon strain macromolecular marker is the 90th nucleotide corresponding to SEQ ID No. 1 in the sequence list in the melon genome, which is G or A.

2. The use according to claim 1, characterized in that: The substance for detecting the high-molecular-weight marker of the melon strain is a primer pair consisting of P1 and P2 or a set of reagents for detecting the high-molecular-weight marker of the melon strain; Said P1 and said P2 are two single-stranded DNAs shown as SEQ ID No.3 and SEQ ID No.4 in the sequence list respectively; The reagent set for detecting the high-molecular-weight markers of the melon plant includes the P1 and the P2.

3. The use according to claim 2, characterized in that: The kit also includes the restriction endonuclease XhoI.

4. The use according to any one of claims 1 to 3, characterized in that: The plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has the potential to be greater than, the plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A. The plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has the potential to be greater than, the plant height of a heterozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A. The plant height of a heterozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A is greater than, or has the potential to be greater than, the plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A.

5. A method for detecting or assisting in detecting the height of a melon plant, characterized in that: The method comprises detecting the melon plant height molecular marker of claim 1 and determining the melon plant height according to the following method: The plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has the potential to be greater than, the plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A. The plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G is greater than, or has the potential to be greater than, the plant height of a heterozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A. The plant height of a heterozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is G and A is greater than, or has the potential to be greater than, the plant height of a homozygous muskmelon in which the nucleotide at position 90 corresponding to SEQ ID No. 1 in the sequence listing is A.

6. The method according to claim 5, characterized in that: The detection of the melon plant high molecular weight marker described in claim 1 is carried out using the substance for detecting the melon plant high molecular weight marker described in any one of claims 1-3.

7. The substance for detecting high molecular markers of melon plants according to any one of claims 1 to 3.

8. The high molecular weight marker of the melon plant according to claim 1.

9. Any of the following applications: X1) Use of the high-molecular-weight marker for melon plants as claimed in claim 1 in melon breeding; X2) Use of the melon strain high molecular marker of claim 1 in detecting or assisting in detecting high-molecular-weight melon strains; X3) Use of the substance for detecting high-molecular-weight markers of melon plants according to any one of claims 1 to 3 in melon breeding; X4) Use of the substance for detecting high-molecular-weight markers of melon plants according to any one of claims 1 to 3 in the preparation of melon breeding products; X5) Use of the substance for detecting melon plant height marker according to any one of claims 1 to 3 in the preparation of a product for detecting or assisting in detecting melon plant height; X6) Use of the method according to claim 5 or 6 in melon breeding; X7) Use of the substance corresponding to nucleotide No. 90 of SEQ ID No. 1 in the sequence listing in the melon genome for the breeding of melons with shorter plant height; X8) Use of a substance corresponding to the 90th nucleotide of SEQ ID No. 1 in the sequence listing in the melon genome in preparing a product for breeding melons with short plant height.

10. Melon breeding methods, including Y1) or Y2): Y1) detecting the 90th nucleotide corresponding to SEQ ID No. 1 in the sequence listing in the melon genome, and selecting a homozygous or heterozygous melon whose 90th nucleotide corresponding to SEQ ID No. 1 in the sequence listing is A as a parent for breeding; Y2) mutating the 90th nucleotide corresponding to SEQ ID No. 1 in the sequence listing of the melon genome from G to A to obtain the target melon, thereby achieving melon breeding.

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