Application of gene Bch09G013990 in controlling wax powdery traits of wax gourd peel

Abstract

The application belongs to the technical field of plant gene function identification and molecular marker assisted breeding, and discloses a gene Bch09G013990 The application is applied to controlling wax powder traits of winter melon peel; through whole genome association analysis combined with map-based cloning, a key gene for regulating wax powder formation of winter melon peel is identified Bch09G013990 ; the gene has two key loss-of-function mutation sites: one is a SNP mutation located at a splicing site, which causes abnormal splicing and produces a long transcript retaining an intron; the other is an InDel mutation causing early termination of transcription, which makes the coding protein truncated; the above two functional mutations occur in parallel selection of wax-free small fruit zucchini and wax-free large fruit winter melon; and accordingly, a dCAPS molecular marker suitable for small fruit zucchini and an InDel molecular marker suitable for large fruit winter melon are developed, which can be used for identification of winter melon peel wax powder trait related germplasm materials, molecular marker assisted selection and new variety breeding.

Description

Technical Field

[0001] This invention belongs to the field of plant gene function identification and molecular marker-assisted breeding technology, specifically relating to a gene. Bch09G013990 Application in controlling the properties of wax powder on winter melon peel. Background Technology

[0002] Winter melon ( Benincasa hispida This includes large-fruited winter melon and small-fruited wax gourd. Benincasa hispida Cogn.var. chieh-qua Winter melon (also known as mini winter melon), a variety widely cultivated in my country, is an important vegetable crop. The waxy powder on the peel is a crucial agronomical trait of winter melon, playing a vital role in fruit storage resistance and disease resistance. A visible white waxy powder forms on the peel surface, affecting its appearance. Currently, different regions have varying preferences for the type of waxy powder on winter melon peels, and both varieties with and without waxy powder are available in the market. Targeted breeding of winter melon varieties with and without waxy powder to meet different consumption needs and different planting seasons is crucial in winter melon breeding. However, the waxy powder trait only appears in the later stages of fruit development, meaning traditional breeding methods can only select for it at this stage, resulting in a long breeding cycle.

[0003] Natural population genetic analysis indicates that the waxy coating on the peel of winter melon is an important selective trait, with wax-free cultivated winter melons selected from waxy-coated groups. Segregating population genetic analysis shows that the presence or absence of waxy coating is controlled by a pair of genes, with waxy coating being dominant over wax-free. However, the genetic mechanism controlling the waxy coating remains unclear, the relevant key genes have not been cloned, and functional variation sites have not been identified. Currently developed markers for identifying the presence or absence of waxy coating are only linkage markers and have not revealed functional variation markers, resulting in incomplete and inaccurate identification, which in turn affects marker-assisted breeding. Summary of the Invention

[0004] To address the shortcomings of the existing technology, this invention aims to provide a gene... Bch09G013990 The application of controlling the wax powder trait of winter melon peel provides a precise gene target for molecular design breeding of the wax powder trait of winter melon, and accelerates the targeted breeding process of target varieties.

[0005] To solve the above problems, the present invention adopts the following technical solution: Gene Bch09G013990 In the application of controlling the wax powder trait of winter melon peel, the gene... Bch09G013990 The nucleotide sequence is shown in SEQ ID No: 1.

[0006] Furthermore, the gene Bch09G013990 The full-length nucleotide sequence containing the intron is shown in SEQ ID No: 2.

[0007] Proteins that control the properties of wax powder in winter melon peel Bch09G013990 Its amino acid sequence is shown in SEQ ID No: 3.

[0008] A vector comprising the aforementioned gene Bch09G013990 .

[0009] An engineered bacterium, comprising the aforementioned gene Bch09G013990 .

[0010] Application of SNP markers or InDel markers in winter melon variety identification and screening breeding; the SNP markers mentioned are SNP markers related to the wax powder trait of winter melon pericarp, and the SNP markers of winter melon materials with wax powder are... Bch09G013990 The splicing site of the gene is the normal splicing site of GT-AG, and the SNP marker of the waxy wax gourd material is... Bch09G013990 The splicing site of the gene is the splicing site of the GT-AA mutation; the InDel marker is an InDel marker associated with the wax powder trait of large-fruited winter melon peel, and the InDel marker of large-fruited winter melon material with wax powder is... Bch09G013990 The CDS 690-716 bp of the gene is 5'-GGTCATGGCCGTGTTTGTTGTATCGGG-3', while the InDel marker for the large-fruited winter melon material without wax powder is... Bch09G013990 A 27 bp deletion occurs in the CDS of the gene from 690 to 716 bp, while the 5'-ATGA-3' sequence is retained from 690 to 693 bp.

[0011] Furthermore, the specific dCAPS primer pairs for the SNP marker include forward and reverse primers such as SEQ ID No: 4-SEQ ID No: 5; the specific primer pairs for the InDel marker include forward and reverse primers such as SEQ ID No: 6-SEQ ID No: 7.

[0012] A method for identifying whether the peel of wax gourd contains wax powder includes the following steps: Genomic DNA was extracted from the wax gourd to be tested; Using the genomic DNA template of the wax gourd to be tested, PCR amplification was performed using the specific dCAPS primer pair with the SNP markers described above; The obtained PCR product was digested with the restriction enzyme DdeI; Detection of enzyme digestion products using polyacrylamide gel electrophoresis; If the detection result is a band of 114 bp, then the gene of the tested wax gourd is... Bch09G013990 It is a wild-type, waxy wax gourd variety; if the test result shows an 84bp band, then the gene... Bch09G013990It is a mutant variety, belonging to the wax gourd variety without wax powder.

[0013] A method for identifying whether the peel of a large-fruited winter melon contains wax powder includes the following steps: Genomic DNA was extracted from the large-fruited winter melon to be tested; Using the genomic DNA template of the large-fruited winter melon to be tested, PCR amplification was performed using the InDel-labeled specific primer pair described above; PCR amplification products were detected using polyacrylamide gel electrophoresis; if the detection result showed a band of 178 bp, then the large-fruited winter melon being tested... Bch09G013990 The gene is wild-type, belonging to the large-fruited winter melon variety with waxy powder; if the test result shows a band of 155bp, then... Bch09G013990 It is a mutant gene and belongs to the large-fruited winter melon variety without wax powder.

[0014] The beneficial effects of this invention are as follows: This invention is the first to clone and identify the key gene controlling the wax powder trait of winter melon peel. Bch09G013990 This invention reveals the molecular genetic mechanism of the waxy powder trait in winter melon. Two loss-of-function mutation sites in this gene were identified, and these two types of mutations were observed in parallel selection in small-fruited wax gourds and large-fruited winter melons without waxy powder, respectively. This provides precise gene targets for molecular design breeding of the waxy powder trait in winter melon. Molecular markers developed based on functional mutation sites can directly detect gene functional variations, with significantly higher identification accuracy than traditional linkage markers. Using the molecular markers of this invention, genotypes of materials can be rapidly identified at the seedling stage, eliminating the need to wait for phenotypic identification in the later stages of fruit development. This significantly shortens the breeding cycle, greatly improves the efficiency of marker-assisted selection breeding for the waxy powder trait in winter melon and wax gourd, and accelerates the targeted breeding process of target varieties. Attached Figure Description

[0015] Figure 1 The key gene controlling the wax powder trait of winter melon peel in Example 1 of this invention. Bch09G013990 The localization results are as follows: Figure a shows the population structure analysis of 163 winter melon materials, Figure b shows the GWAS analysis of the wax powder characteristics of winter melon pericarp, Figure c shows the initial BSA localization of the wax powder characteristics of winter melon pericarp, and Figure d shows the fine localization of the wax powder characteristics of winter melon pericarp.

[0016] Figure 2 In Embodiment 2 of the present invention Bch09G013990 The identification results of SNPs and InDel functional variant sites in the gene; where, Figure a shows the sequence characteristics of SNP variants, and Figure b shows the effects of SNP variants at alternative splicing sites. Bch09G013990 Gene transcript differences, Figure c shows the sequence characteristics of the InDel variant, and Figure d shows... Bch09G013990 The predicted protein structures of wild-type and indel mutants are shown in Figure e. Bch09G013990The relationship between different types of gene variation and wax powder phenotype. Figure f shows the distribution of different types of variation in the winter melon subgroup.

[0017] Figure 3 This is a schematic diagram showing some of the results of the verification of SNP markers in the germplasm resources of Cucurbita esculenta in Embodiment 3 of the present invention.

[0018] Figure 4 This is a schematic diagram showing some of the results of the InDel marker verification in winter melon germplasm resources in Example 4 of the present invention. Detailed Implementation

[0019] The present invention will be further described in detail below with reference to specific embodiments.

[0020] It should be noted that these embodiments are only used to illustrate the present invention and are not intended to limit the present invention. Simple improvements to the method under the premise of the present invention are all within the scope of protection claimed by the present invention.

[0021] Example 1: Key genes controlling the wax powder trait in winter melon peel Bch09G013990 Positioning This invention utilizes SNP and InDel variant datasets from 163 natural populations of winter melon (including 20 wax gourd varieties, also known as small winter melon) and wax powder phenotypic data for GWAS analysis. A significant signal was obtained at position 48.09 Mb on chromosome 9, with the p-value of the highest-signal SNP being 2.11 × 10⁻⁶. -12 ( Figure 1 (a, b) Simultaneously, genetic populations were constructed using GF-711 (with wax powder) and GX-19 (without wax powder). The F1 generation exhibited the wax powder phenotype, and in the F2 population, the ratio of individuals with to those without the wax powder phenotype was 3:1, demonstrating that the wax powder trait is a dominant trait controlled by a single gene. Fifty individuals with and 50 individuals without the wax powder phenotype were selected from each F2 population for pooled sequencing. Preliminary BSA analysis revealed overlap with GWAS signals (…). Figure 1 (c) Further expanding the F2 population to 7,352 plants, the mapping interval was successfully narrowed to a 53.8 kb interval between the two markers K4806 and K48.1 ( Figure 1 (d) in this interval. This interval contains only d). Bch09G013990 An acyltransferase gene, a key gene controlling the waxy powder trait of winter melon peel. Bch09G013990 The nucleotide sequence of the gene is shown in SEQ ID No: 1. Bch09G013990 The full-length nucleotide sequence containing the intron is shown in SEQ ID No: 2, protein Bch09G013990 The amino acid sequence is shown in SEQ ID No: 3.

[0022] SEQ ID No: 1: ATGGGTGGCGAAATCATAAGATTCATCAAGATGTTGCCCATATTCATTATGTCCTCTTTGTATTGCTATTTCATAGCTTCAAAGTTTTCCAAAGGCAAACTTAGGCTCCTCTCTCTGCTCCCCGTCTTCTCTCTCTTCACCGTCCTCCCTCTCTACTTCTCCTCCATCCTCTTCACGAGCAACATCGCCTTATTCGTCACCTGGCTTACGACCTTCAAGCTCCTTCTTTTCTCATTCGATTTGGGTCCTCTAGCTTCGGACCCACCATTGTCATTCCCTCTTTTTGCTTCTATTGCTTTCTTTCCAACTAGAGTCAAGCATAACAAAACAACCCCACCTGACCAAGAACCACCTCCAAAGTTGCCTCTAAACTTGCCAAGCAAAGTGGTGATACTTGCCATATTGATCTTAGGAAACAGCTATATTGTCCTTGTTTGTCCCAATAATGTAAAGTTCTGGATCAATTGTGTTACAATATATTTCTACTTAGACGTAATTATGAGTCTATCCAATGTTTTTGTTCGATCAACTTTCGGTGTCGAGGTCAGACAAGCATTTAACGAGCCATATCTATCGACATCGCTTCAAAATTTATGGGGTCGGAGGTGGAATCTATTAGTATCTGAAACTTTGCACAACACAATCTACAAACCAGTTCGTTACAAAGTAGGGATGCCACGGTGGGTGGCGGTCATGGCCGTGTTTGTTGTATCGGGGCTTATGCATGAGCTTTTGTATTACTACATTGTCAGAGCGAGTCCGACATGGGAAGCCACGTGGTTCTTTGTGATGCATGGAGTGTGTTTGGCCGTGGAGATTGAATTGAAGAGGGCGTATGGGAAAAGGTGGCCGTTGCATTGGGCAGTTTCTGGACCATTAACGACGGTGTTTATGGTAACCACCACTTATTGGTTGCTTTTTCCTCTAATGTTGAGGTCTTGA。

[0023] SEQ ID No:2:

[0024] SEQ ID No: 3: MGGEIIRFIKMLPIFIMSSLYCYFIASKFSKGKLRLLSLLPVFSLFTVLPLYFSSILFTSNIALFVTWLTTFKLLLFSFDLGPLASDPPLSFPLFASIAFFPTRVKHNKTTPPDQEPPPKLPLNLPSKVVILAILILGNSYIVLVCPNNVKFWINCV TIYFYLDVIMSLSNVFVRSTFGVEVRQAFNEPYLSTSLQNLWGRRWNLLVSETLHNTIYKPVRYKVGMPRWVAVMAVFVVSGLMHELLYYYIVRASPTWEATWFFVMHGVCLAVEIELKRAYGKRWPLHWAVSGPLTTVFMVTTTYWLLFPLMLRS*.

[0025] Example 2 Bch09G013990 Identification of SNPs and InDel functional variant sites in genes The winter melon materials involved in this invention include two cultivation types: small-fruited wax gourd and large-fruited winter melon. Figure 1 a) Both are winter melons, but belong to different cultivation types recognized in this field. Those skilled in the art can clearly distinguish between them based on characteristics such as fruit morphology, fruit size, cultivation methods, and breeding classification. Small-fruited winter melons typically have smaller fruits, which are short cylindrical or cylindrical in shape, and are mainly harvested for consumption as young fruits. Representative varieties include, but are not limited to, widely cultivated local varieties of winter melon and their bred varieties. Large-fruited winter melons typically have larger fruits and are mainly harvested and stored as mature fruits. Representative varieties include, but are not limited to, black-skinned winter melons, pink-skinned winter melons, green-skinned winter melons, and their improved varieties. Those skilled in the art can clearly determine which of the above cultivation types the tested material belongs to based on existing breeding and variety classification standards.

[0026] This invention discovers Bch09G013990 The gene consists of two exons and a large intron of 3447 bp in length, with its coding region located only in the second exon, encoding a protein containing 313 amino acids. Comparison was made between two parental materials: GF-711 (with wax powder) and GX-19 (without wax powder). Bch09G013990 A differentially expressed site of the gene was found to contain an SNP variant at the splice site. Figure 2 In the text a), SNP is a G-to-A mutation located at an alternative splicing site. Materials with wax powder have a normal GT-AG splicing site, producing short transcripts without introns; materials without wax powder have a GT-AA mutation splicing site, producing long transcripts that retain introns. Figure 2 (b) In this context, the SNP mutation is located at the conserved motif of the 3' splice acceptor in the Bch09G013990 gene intron, corresponding to the G→A base at position 3623 (the 3' end of the intron) in SEQ ID NO: 2 of the genome sequence. The wild-type intron has a conserved AG dinucleotide at the 3' end, which is the core signal for spliceosome recognition; after mutation, it becomes AA, and the spliceosome cannot properly recognize the 3' splice site, resulting in the intron not being properly excised, producing an abnormally long transcript that retains the full-length intron, ultimately leading to the loss of function of the encoded protein and the formation of the non-wax powder phenotype.

[0027] This invention also discovered a method in winter melon population materials that enables... Bch09G013990 The InDel mutation, which prematurely terminates gene transcription, causes the protein sequence of this gene to be shortened from 313 amino acids to 230 amino acids. Figure 2 In the case of c), the protein structure changes from 11 helices to 8 helices, leading to loss of gene function. Figure 2 (d in the text). The characteristic of this InDel marker is that the CDS 690-716 bp of the Bch09G013990 gene in waxy materials has the sequence 5'-GGTCATGGCCGTGTTTGTTGTATCGGG-3' (defined as the Insertion type), while the InDel marker in large-fruited winter melon materials without wax is... Bch09G013990 A 27 bp deletion occurs in the CDS of the gene from 690 to 716 bp, while the 5'-ATGA-3' sequence (defined as a deletion type) is retained from 690 to 693 bp. This InDel marker is a special InDel marker of sequence substitution type.

[0028] More importantly, this invention also discovered that two key variants, SNP and InDel, can be selected in parallel in small-fruited wax gourds and large-fruited winter melons without wax powder. Figure 2 (e, f in the text). In the small-fruited wax gourd population, all materials with the normal splice site type of GT-AG had wax powder, while all materials with the GT-AA mutant splice site type did not have wax powder. In the large-fruited winter melon population, all materials with the Deletion type did not have wax powder, all materials with the Insertion type except for one had wax powder, and all materials with the Deletion / Insertion heterozygous type had wax powder.

[0029] Example 3: Obtaining SNP markers for controlling the wax powder trait in the pericarp of small-fruited wax gourd and validating these markers in wax gourd germplasm resources. To further verify Bch09G013990 SNP variations in genes are the cause of the waxy powder-free trait in small-fruited wax gourds. dCPAS markers were developed targeting SNP variation sites and validated in core wax gourd germplasm resources. Figure 3 ).

[0030] 3.1 SNP-labeled dCAPS-specific primer pairs Specific primer pairs labeled with dCAPS were designed for the above-mentioned SNP sites. Their sequence characteristics are: forward primer F5'-CAATGTTTGATTACATGTCGCAAT-3' (SEQ ID No: 4) and reverse primer R5'-CTCATTCTTTTATTATTATTTTCTCAC T A-3' (SEQ ID No: 5).

[0031] 3.2 PCR reaction system and reaction conditions Using the genomic DNA of the wax gourd to be tested as a template, PCR amplification was performed using the above primer pairs: PCR amplification system: The system is 20 μL, containing 10 μL of 2× Taq PCR Mix, 0.5 μL each of forward and reverse primers, 1 μL of DNA template from wax gourd, and the remainder is ddH2O.

[0032] PCR reaction conditions: 94℃ pre-denaturation for 5 minutes; 94℃ denaturation for 30 seconds, 58℃ annealing for 10 seconds, 72℃ extension for 30 seconds, for a total of 35 cycles; 72℃ for 5 minutes.

[0033] 3.3 Enzyme digestion of PCR amplified fragments The obtained PCR product was digested with the restriction enzyme DdeI.

[0034] The reaction system (20 μL) consisted of 10 U / μL of 0.5 μL of DdeI restriction enzyme, 10 μL of PCR product, 2 μL of 10× rCutSmart buffer, and the remainder being ddH2O.

[0035] The reaction conditions were: incubation at 37 degrees Celsius for 4 hours.

[0036] 3.4 Genotype determination and association analysis of wax powder traits Polyacrylamide gel electrophoresis was used to detect the enzyme digestion products. If the detection result showed a band of 114 bp, then the winter melon to be tested... Bch09G013990 The gene is wild-type, belonging to the waxy wax gourd variety. If the test result shows an 84bp band, then... Bch09G013990 The gene is a mutant type, belonging to the wax gourd variety without wax powder. Figure 3 The results of testing 10 waxy wax gourd germplasm samples with and 10 without waxy wax gourd germplasm samples are presented. The waxy materials all showed bands of 114 bp, while the non-waxy materials all showed bands of 84 bp. This further demonstrates... Bch09G013990Genes are key genes controlling the waxy powder trait of wax gourd peel, and SNP variations are the cause of the absence of waxy powder trait.

[0037] Example 4: Obtaining the InDel marker for controlling the wax powder trait of large-fruited winter melon pericarp and validating the marker in winter melon germplasm resources. To further verify Bch09G013990 The InDel mutation in the gene is the cause of the waxy powder-free trait in large-fruited winter melon. We developed an InDel marker targeting the InDel mutation site and validated it in core germplasm resources of winter melon. Figure 4 ).

[0038] 4.1 InDel-labeled specific primer pairs Specific primer pairs for the InDel marker were designed for the InDel site mentioned above. Their sequence characteristics are forward primer F 5'-TCTGAAACTTTGCACAACACAATC-3' (SEQ ID No: 6) and reverse primer R 5'-CTCCATGCATCACAAAGAACCAC-3' (SEQ ID No: 7).

[0039] 4.2 PCR reaction system and reaction conditions Using the genomic DNA template of the winter melon to be tested, PCR amplification was performed using the primer pairs described above.

[0040] PCR amplification system: The system is 20 μL, containing 10 μL of 2× Taq PCR Mix, 0.5 μL each of forward and reverse primers, 1 μL of DNA template from various germplasms of winter melon, and the remainder is ddH2O.

[0041] PCR reaction conditions: 94℃ pre-denaturation for 5 minutes; 94℃ denaturation for 30 seconds, 58℃ annealing for 10 seconds, 72℃ extension for 30 seconds, for a total of 35 cycles; 72℃ for 5 minutes.

[0042] 4.3 Genotype determination and association analysis of wax powder traits The PCR amplification products were detected by polyacrylamide gel electrophoresis, and the result was a band of 178 bp. Bch09G013990 The CDs for the gene, 690-716 bp, are 5'-GGTCATGGCCGTGTTTGTTGTATCGGG-3', indicating a winter melon variety with a waxy coating. The detection result shows a 155 bp band, i.e. Bch09G013990 The 27-base mutation in the CDs 690-716 bp gene is 5'-ATGA-3', which belongs to the winter melon variety without wax powder. Figure 4The results of testing 10 winter melon germplasm samples with and 10 samples without wax powder are presented. The wax-powdered materials all showed bands of 178 bp, while the non-wax-powdered materials all showed bands of 155 bp. This further strongly proves... Bch09G013990 Genes are key genes controlling the wax powder trait in winter melon peel, and the InDel mutation is the reason for the absence of wax powder.

[0043] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described with reference to preferred embodiments, those skilled in the art should understand that various changes in form and detail can be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Genes Bch09G013990 In the application of controlling the properties of wax powder from winter melon peel, the characteristic is... The gene Bch09G013990 The nucleotide sequence is shown in SEQ ID No:

1.

2. The application according to claim 1, characterized in that, The gene Bch09G013990 The full-length nucleotide sequence containing the intron is shown in SEQ ID No:

2.

3. Proteins that control the waxy powder properties of winter melon peel Bch09G013990 Its amino acid sequence is shown in SEQ ID No:

3.

4. A carrier, characterized in that, Including the gene described in claim 1 Bch09G013990 .

5. An engineered bacterium, characterized in that, Including the gene described in claim 1 Bch09G013990 .

6. Application of SNP markers or InDel markers in winter melon variety identification and screening breeding; the SNP markers are those related to the wax powder trait of winter melon pericarp, and the SNP markers of winter melon materials with wax powder are... Bch09G013990 The splicing site of the gene is the normal splicing site of GT-AG, and the SNP marker of the waxy wax gourd material is... Bch09G013990 The splicing site of the gene is the splicing site of the GT-AA mutation; the InDel marker is an InDel marker associated with the wax powder trait of large-fruited winter melon peel, and the InDel marker of large-fruited winter melon material with wax powder is... Bch09G013990 The CDS 690-716 bp of the gene is 5'-GGTCATGGCCGTGTTTGTTGTATCGGG-3', while the InDel marker for the large-fruited winter melon material without wax powder is... Bch09G013990 A 27 bp deletion occurs in the CDS of the gene from 690 to 716 bp, while the 5'-ATGA-3' sequence is retained from 690 to 693 bp.

7. The application according to claim 6, characterized in that, The specific dCAPS primer pairs for the SNP markers include forward and reverse primers such as SEQ ID No: 4-SEQ ID No: 5; the specific primer pairs for the InDel markers include forward and reverse primers such as SEQ ID No: 6-SEQ ID No:

7.

8. A method for identifying the presence or absence of wax powder on the peel of wax gourd, characterized in that, Includes the following steps: Genomic DNA was extracted from the wax gourd to be tested; Using the genomic DNA template of the cucumber to be tested, PCR amplification was performed using the specific dCAPS primer pair with SNP markers as described in claim 7; The obtained PCR product was digested with the restriction enzyme DdeI; Detection of enzyme digestion products using polyacrylamide gel electrophoresis; If the detection result is a band of 114 bp, then the gene of the tested wax gourd is... Bch09G013990 It is a wild-type, waxy wax gourd variety; if the test result shows an 84bp band, then the gene... Bch09G013990 It is a mutant variety, belonging to the wax gourd variety without wax powder.

9. A method for identifying the presence or absence of wax powder on the peel of large-fruited winter melon, characterized in that, Includes the following steps: Genomic DNA was extracted from the large-fruited winter melon to be tested; Using the genomic DNA template of the large-fruited winter melon to be tested, PCR amplification was performed using the InDel-labeled specific primer pair described in claim 7; PCR amplification products were detected using polyacrylamide gel electrophoresis; if the detection result showed a band of 178 bp, then the large-fruited winter melon being tested... Bch09G013990 The gene is wild-type, belonging to the large-fruited winter melon variety with waxy powder; if the test result shows a band of 155bp, then... Bch09G013990 It is a mutant gene and belongs to the large-fruited winter melon variety without wax powder.

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