Ghuma gene for regulating fiber fineness of cotton, molecular marker and application thereof

Abstract

The application discloses a kind of regulation cotton fiber fineness GhUMA Gene, molecular marker and its application belong to the technical field of biotechnology application technology.Combining correlation analysis, near-isogenic line transcriptome sequencing and function verification, the application successfully identifies and verifies GhUMA Important function in regulating upland cotton fiber fineness;Meanwhile, a KASP marker that can be specifically used for fiber fineness screening and does not interfere with other fiber quality indicators is developed, providing valuable gene resources and practical molecular marker tools for directional improvement of cotton fiber fineness.

Description

Technical Field

[0001] This invention relates to the field of biotechnology applications, and more specifically to a method for regulating the fineness of cotton fibers. GhUMA Genes, molecular markers and their applications. Background Technology

[0002] Cotton fiber fineness (measured by micronaire value) is one of the most critical quality parameters determining its textile and economic value. It determines the number of fibers involved in cohesion in the yarn cross-section, becoming a primary factor influencing yarn strength and profoundly affecting yarn evenness, appearance smoothness, and hairiness. Simultaneously, fineness fundamentally constrains the spinning process, determining the upper limit of the spun yarn count and relating to breakage rate and yield during production. However, its importance is not a simple linear relationship; the micronaire value must be within an optimal range (typically 3.7–4.2): excessively low fineness, while increasing strength, leads to increased neps and uneven dyeing due to poor maturity; excessively high fineness weakens strength and limits spinning fineness due to insufficient cohesion. Therefore, fiber fineness is the fulcrum for striking a balance between yarn quality and process efficiency; its quality directly determines whether the final textile meets design standards and its value proposition in trade.

[0003] The core dilemma faced by traditional cotton breeding in fiber fineness selection stems from the multiple complexities at the genetic and phenotypic levels. First, the genes controlling fiber fineness (micronaire value) are tightly linked to genes determining yield on chromosomes, forming a negative correlation. This often leads breeders to unintentionally sacrifice fineness quality in pursuit of high yields, creating a bottleneck of "high quality but low yield." Second, fiber fineness is a complex quantitative trait controlled by multiple genes with minor effects, its genetic mechanism is obscure, and its final expression is highly susceptible to environmental factors such as temperature and moisture, making traditional selection methods relying on phenotypic observation both inefficient and unreliable. Furthermore, the specialized instruments and high costs required for accurate fiber fineness measurement hinder its large-scale application in early generations of breeding. Historically, the over-reliance on single indicators such as fiber length has further exacerbated the neglect of fineness characteristics.

[0004] To overcome these bottlenecks, modern cotton breeding is shifting from traditional "experience-based selection" to "precision navigation," with the core solution lying in the deep application of molecular biology techniques. Among these, marker-assisted selection (MAS) technology, by developing DNA markers closely linked to fiber fineness traits, enables breeders to efficiently and accurately identify superior genes in early generations, significantly improving selection efficiency and potentially breaking unfavorable gene linkages to achieve synergistic improvement in both quality and yield. Simultaneously, in-depth analysis of key genes and genetic networks involved in fiber fineness formation provides new genetic resources for breeding, potentially fundamentally solving the genetic challenges of fiber fineness improvement.

[0005] In recent years, through genome-wide association analysis (GWAS) and linkage map construction, researchers have successfully identified a large number of key genes and molecular markers associated with cotton fiber fineness (micronaire value). At the genetic level, in Sea Island cotton... GbSER02 The gene regulates fiber elongation by interacting with GhVOZ1 to promote gibberellin synthesis, thereby producing finer fibers (Jia et al., 2025 GbSER02 Interacts With GhVOZ1 to Promote Fiber Elongation by Modulating Gibberellin Synthesis in Cotton); our team previously discovered that in upland cotton... GhABH and GhPAPGenes can simultaneously regulate fiber length, strength, and fineness (Zhang S et al., 2025a; Zhang S et al., 2025b). An alpha / beta-hydrolase-like gene (GhABH) is identified to be responsible for fiber quality from a multi-effect quantitative trait locus (QTL) on chromosome A06 of upland cotton; a plastid lipid-associated protein-encoding gene (GhPAP) that positively regulates fiber strength was identified via genetic mapping and transcriptomic analysis of a stable QTL on chromosome D06 of upland cotton. Based on the significantly differentially expressed loci of these genes, efficient markers such as KASP have been developed, providing a direct tool for marker-assisted selection (MAS). Furthermore, recent studies, through genetic analysis of multi-level fiber quality traits (including AFIS fineness index), have discovered novel pleiotropic loci located on chromosomes A03, A05, and A07 (Journal of Integrative Agriculture, 2024 Genetic dissection and origin of pleiotropicloci underlying multi-level fiber quality traits in upland cotton). Gossypium hirsutum L.) continuously enriches the marker resources for genetic improvement of fiber fineness. However, fiber fineness (represented by micronaire value) and other quality traits such as length and strength are often synergistically regulated by pleiotropic loci or tightly linked genes. When using these genes or markers for improvement, it often leads to linked changes in other fiber quality indicators. Therefore, the focus of future research should be on deeply exploring those specific genes or markers that only affect fiber fineness and have little impact on other indicators, so as to achieve precise optimization of fiber fineness without sacrificing traits such as length and strength. This will be the key to breaking the negative correlation between quality traits and achieving synergistic improvement of fiber quality.

[0006] Therefore, how to discover specific genes or markers that only affect fiber fineness and have little impact on other indicators is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0007] In view of this, the present invention provides a method for controlling the fineness of cotton fibers. GhUMA Genes, molecular markers and their applications.

[0008] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0009] A method for regulating the fineness of cotton fibers GhUMA The gene, whose nucleotide sequence is shown in SEQ ID NO.1.

[0010] The above GhUMA Application of genes in regulating cotton fiber fineness.

[0011] Furthermore, the regulation mentioned is a positive regulation.

[0012] Knockout GhUMA Application of genes in constructing cotton models with finer fiber thickness.

[0013] One of the above GhUMA A gene-tightly linked KASP molecular marker, the nucleotide sequence of which is shown in SEQ ID NO.12.

[0014] The primer set for amplifying the above molecular markers includes the primers shown in SEQ ID NO.8 to SEQ ID NO.10.

[0015] A kit for identifying the fineness of cotton fibers, comprising the primer set described above.

[0016] The above-mentioned molecular markers, primer sets, or kits may be used in any of the following: (1) Application in identifying the fineness of cotton fibers; (2) Application in cotton breeding.

[0017] A method for determining the fineness of cotton fibers includes the following steps: Using the genomic DNA of the cotton sample to be tested as a template, PCR amplification was performed using the primer set shown in SEQ ID NO.8~SEQ ID NO.10, and the determination was based on the sequencing results of the obtained amplification products.

[0018] Furthermore, as shown in SEQ ID NO.12, this site indicates a strain with finer fibers, while the absence of this site indicates a strain with coarser fibers.

[0019] As can be seen from the above technical solution, compared with the prior art, the present invention has the following beneficial effects: This invention, combining association analysis, near-isogenic transcriptome sequencing, and functional validation, successfully identified and validated [the following]. GhUMA It plays an important role in regulating the fineness of upland cotton fibers; at the same time, it has developed a KASP marker that can be specifically used for fineness screening without interfering with other fiber quality indicators, providing valuable genetic resources and practical molecular marker tools for the targeted improvement of cotton fiber fineness. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0021] Figure 1 This study analyzes the physical regions of stable QTLs for fiber fineness (micronaire value) and the differentially expressed genes within them in Example 1 of the present invention.

[0022] Figure 2 This is a comparison of gene expression patterns of two nearly isogenetic lines within the stable equine clonal value QTL range in Example 1 of the present invention.

[0023] Figure 3 In Embodiment 2 of the present invention GhUMA Gene structure diagram.

[0024] Figure 4 In Embodiment 2 of the present invention GhUMA Spatiotemporal expression analysis of qRT-PCR at different fiber development stages in two nearly isogenic lines. Note: This means p < 0.05. This means p < 0.01.

[0025] Figure 5 VIGS-mediated in Embodiment 2 of the present invention GhUMA Silencing efficiency verification: Comparison of expression levels between silenced plants and empty CK in the 906-cell parental strain. Note: This means p < 0.01.

[0026] Figure 6 VIGS-mediated in Embodiment 2 of the present invention GhUMA Silencing efficiency verification: Comparison of expression levels between silenced plants and empty CK in the 906-crude parent line. Note: This means p < 0.05. This means p < 0.01.

[0027] Figure 7 In Example 2 of this invention, 906-cell parent material GhUMA Comparative analysis of fiber fineness (a), strength (b), and length (c) between silent plants and controls. Note: This means p < 0.01.

[0028] Figure 8 In Example 2 of this invention, 906-crude parent material GhUMA Comparative analysis of fiber fineness (a), strength (b), and length (c) between silent plants and controls. Note: This means p < 0.01.

[0029] Figure 9 The 906-fine parent and 906-coarse parent in Embodiment 2 of the present invention GhUMA Macron value after silence.

[0030] Figure 10 In Embodiment 2 of the present invention GhUMA Effect of downregulation on fibroblast cell wall thickness. Note: This means p < 0.01.

[0031] Figure 11 In Embodiment 2 of the present invention GhUMA The effect of downregulation on the length of the fiber single helix.

[0032] Figure 12 In Embodiment 3 of the present invention, and GhUMA A schematic diagram of a large fragment insertion / deletion site (D12_64657895) that is co-separated.

[0033] Figure 13 This is a KASP marker genotyping map developed based on the D12_64657895 insertion / deletion site in Embodiment 3 of the present invention, where a and b represent different samples.

[0034] Figure 14 This is the genotyping and screening effect of 278 strains based on KASP markers for fiber fineness (micronaire value) in Example 3 of the present invention. Note: represent p <0.05.

[0035] Figure 15 The results of fiber length gene typing and screening of 278 strains based on KASP markers in Example 3 of this invention are shown.

[0036] Figure 16The results of genotyping and screening of 278 strains of fiber strength based on KASP markers in Example 3 of this invention are shown.

[0037] Figure 17 This is a classification diagram of KASP markers in 137 resource materials in Embodiment 3 of the present invention, where a and b represent different samples.

[0038] Figure 18 This is an evaluation of the screening effect of the KASP marker on fiber fineness (micronaire value, a), length (b), and strength (c) in 137 resource materials in Example 3 of the present invention. Detailed Implementation

[0039] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0040] Example 1 Key genes influencing fiber fineness were identified using a stable QTL for fiber fineness. GhUMA Previously, the inventors used association analysis to locate a QTL that was stably associated with micronaire value under multiple environmental conditions near the SSR marker DPL0886 in a natural cotton population (see Zhang Sujun et al., 2021, "Association Analysis and Mining of Superior Alleles for Fiber Quality Traits in Upland Cotton"). However, due to the broad genomic region covered by this marker, the target gene affecting fiber fineness could not be precisely identified; at the same time, the complex genetic background of the natural population made it difficult to rule out the possibility that this QTL might also affect fiber length and strength.

[0041] To further identify candidate genes for fiber fineness and develop molecular markers that can specifically screen for fiber fineness without negatively affecting fiber length and strength, this invention uses the near-isogenic lines 906-fine and 906-coarse derived from Jimian 906 (Ji Shenmian 20250001), which have significant differences in fiber fineness, as materials (the three-year microclonal value test results of the two lines are shown in Table 1). A segregating population containing 278 lines was constructed (these 278 lines were used in KASP typing in Example 3).

[0042] Table 1. Comparison of clonal values ​​of near-isogenic equine strains derived from Jimian 906.

[0043] Whole-genome resequencing and transcriptome sequencing of the parents (906-fine and 906-coarse) identified 38 differentially expressed genes during fiber development near DPL0886 (Table 2). Figure 1 ).

[0044] Table 2. Annotation information of 38 differentially expressed genes in candidate regions for fiber fineness (micron value).

[0045] Among them, genes Ghi_D12G12566 It was expressed at low levels in the near-isogenic line 906-cell throughout the entire fiber development stage, and the expression difference between the two lines remained significant, especially during the secondary cell wall thickening period (20-30 DPA).

[0046] Further classification of gene expression patterns in the near-isogenic lines 906-fine and 906-coarse revealed ten expression patterns. Ghi_D12G12566 In 906-coarse fibers, this expression pattern belongs to pattern X, and its expression level first increases and then decreases with fiber development, reaching a high level at 10-15 DPA. However, this pattern is completely absent in 906-fine fibers. Figure 2 ).

[0047] Based on differences in transcriptome expression patterns, we screened Ghi_D12G12566 This gene was selected as a candidate for further functional validation. By comparing the genomes of Arabidopsis thaliana and cotton, the gene was annotated as "Usually Multiple AcidsMove In and Out Transporters 2" (UMAMIT2), and its homology in Arabidopsis thaliana is AT4G19185.1. This invention names it... GhUMA UMAMIT2 belongs to the nodulin MtN21 / EamA-like transporter family. Members of this family are mostly transmembrane proteins with transport activity, and may play a role in cotton growth and development and pathogen immunity. Currently, the relationship between the UMAMIT2 gene and fiber development is unclear.

[0048] Example 2 GhUMA Functional validation of the effect of candidate genes on cotton fiber fineness GhUMA The total length is 2976 bp, containing 7 exon regions and 6 introns. Figure 3 The mRNA is 1128 bp in length and encodes 375 amino acids.

[0049] GhUMA The full-length sequence is as follows:

[0050] GhUMA cds The sequence is as follows:

[0051] GhUMA The encoded amino acid sequence is as follows: MALVQLFNGGYHVITKVALNVGVNQLVFCVFRDLLALSLLAPVAYVREKRIRPPMTKRLLLTFFFLGLTGIFGNQLLFLIGLSYTNPTYAAAIQPAIPVFTFLLAVMMGTERVNLLKTEGQAKVGGTLICVSGAILMVLFRGPALLGQSNGDFALQNDISARGQPEPAGWLMSSFLEFGLDPWHIGV ICLIGNCICMAAFLAIQAPVLAKYPANISVTALSYFFGAILMVATAFFFTNESTDWNLTRSEIFAVVYAGVVASALNYGLLTWSNKILGPALVALYNPLQPAASAFLSRIFLGSSIYLGSVIGGFLIIAGLYTVTWASYRERCAEGMMPQNVRSSEPLIHKDESINKNPYQRARVFSEPSVLSPKSSD , SEQ ID NO.3.

[0052] This invention uses qRT-PCR technology to compare and analyze... GhUMA The spatiotemporal expression patterns of the gene during the fiber development stage (0–30 DPA) in both thinner (906-thin) and thicker (906-thick) fiber-developing lines were investigated. Experimentally, total RNA was first extracted and its purity and integrity were assessed. cDNA was then synthesized via reverse transcription. A reaction system containing the cDNA template, specific primers, SYBR Green fluorescent dye, and DNA polymerase was prepared for amplification. The fluorescence signal was monitored in real-time for each cycle. After amplification, product specificity was verified using melting curves, and relative quantification was performed based on Ct values. The histone gene His was used as an internal control to correct for systematic errors between samples and ensure the reliability of the results. The specific primer sequences used were: forward qF: ACATTCTTCTTCCTTGGGTTAA, SEQ ID NO.4; reverse qR: TCTGTTACGATGCCTTCC, SEQ ID NO.5.

[0053] The results are as follows Figure 4 As shown, during the 20-30 DPA (days after flowering) stage, GhUMA The expression level of this gene in the 906-fine line was significantly lower than that in the 906-coarse line, suggesting that the downregulation of this gene may be related to the thinning of fiber fineness.

[0054] To further verify this hypothesis, we used VIGS (virus-induced gene silencing) technology to inhibit gene silencing in two different strains. GhUMA The expression will GhUMA VIGS carrier—pCLCrVA: GhUMA pCLCrVA: 00 (Negative control) and pCLCrVA: GhCLA1 (Albinism indicator trait) (The vector construction method is described in Zhang J, Jia X, Guo X, Wei H, Zhang M, Wu A et al (2021) QTL and candidate gene identification of the node of the first fruiting branch (NFFB) by QTL-seq in upland cotton (Gossypium hirsutum L.). BMC Genomics 22(1):1-14. https: / / doi.org / 10.1186 / s12864-021-08164-2) — transformed into Agrobacterium strain GV3101. Among them, pCLCrVA: GhUMA Carrying a 300 bp amplified from 906-cDNA GhUMA Product(GCCTGCTGGATGGCTTATGTCTAGTTTTCTCGAGTTTGGACTTGACCCTTGGCATATTGGAGTTATATGCTTGATAGGGAACTGTATATGCATGGCTGCTTTTCTGGCCATTCAGGCTCCAGTTTTAGCCAAGTATCCTGCCAATATTTC TGTCACCGCGCTTTCCTATTTTTTTGGCCGCTATATTGATGGTAGCAACAGCATTTTTTTTCACCAATGAGTCAACAGATTGGAATCTGACACGGTCTGAGATTTTTGCTGTTGTGTATGCTGGAGTTGTAGCATCAGCTCTTAACTACGG, SEQ. ID NO.6), while pCLCrVA:00 and pCLCrVA: GhCLA1These served as negative and positive controls, respectively. Agrobacterium strain GV3101 carrying the aforementioned vector was immersed in the two fully expanded cotyledons of 10-day-old seedlings according to the method described by Zhang et al. (2021) (Zhang J, JiaX, Guo X, Wei H, Zhang M, Wu A et al (2021) QTL and candidate gene identification of the node of the first fruiting branch (NFFB) by QTL-seq inupland cotton (Gossypium hirsutum L.). BMC Genomics 22(1):1-14. https: / / doi.org / 10.1186 / s12864-021-08164-2). The plants were then planted in 7-gallon pots and cultured in a greenhouse with suitable growing conditions (light / dark cycle: 16 hours light (28°C) / 8 hours darkness (22°C)) until the cotton bolls opened. To maintain the desired silencing effect, the shoot tips were injected with Agrobacterium every two weeks when the plants entered the flowering stage. When pCLCrVA: GhCLA1 When the new leaves of the plant turn white, take 2-3 VIGS-treated cotton bolls from each plant (5-15 days after flowering) for qRT-PCR verification. Collect cotton bolls from the lower part of 4-5 plants that flower within 7 days, mix them as a biological replicate; set up 3 biological replicates for each silenced gene and the negative control (pCLCrVA:00).

[0055] Silence effect Figure 5 , Figure 6 As shown, in the two strains GhUMA The expression of all of them decreased significantly.

[0056] After that GhUMA Fiber fineness, strength, and length were compared between silent plants and controls. Fiber length was measured using an AFIS PRO 2 instrument (Urst Technologies, Switzerland). In each biological replicate, 3000 fibers were tested individually, with three independent biological replicates. Fiber strength and fineness-related traits—including breaking strength (fiber strength) and fineness (linear density, Lin. Den)—were assessed using a FAVIMAT instrument (Textechno, Germany). In each biological replicate, 100–120 fibers were tested individually, with three independent biological replicates.

[0057] In both strains GhUMAAfter silencing, fiber fineness (measured by linear density Lin. Den., with lower values ​​indicating finer fibers) was significantly reduced. Figure 7 a, 8a), while fiber strength (indicated by Tenac) did not change significantly ( Figure 7 (b, 8b). Notably, in the coarse 906 line, fiber length was not significantly affected; while in the fine 906 line, it was silenced. GhUMA The fiber length actually increased later. Figure 7 c, 8c).

[0058] Using HVI1000 for testing, it was found that... GhUMA The Macron value decreases after silencing ( Figure 9 ).

[0059] Meanwhile, we observed and statistically analyzed the cell wall thickness and surface spiral number of fibers in silent plants of the 906-fine strain. We compared the differences in cell wall thickness of mature fibers between the treatment and control groups using paraffin sections and optical microscopy, and observed the fiber spiral morphology using scanning electron microscopy (SEM). Data visualization was performed using ImageJ, and SigmaPlot 14.0 was used for professional chart creation and basic statistical analysis.

[0060] The results showed that GhUMA After the expression level decreased, the thickness of the fibroblast cell wall decreased. Figure 10 ), while the number of individual spirals remains basically unchanged ( Figure 11 ).

[0061] Based on the above results, the reduction GhUMA The expression level can significantly improve fiber fineness while maintaining fiber length and strength essentially unchanged, indicating that this gene is a potential specific target for fiber quality improvement, providing an important basis for subsequent targeted improvement of fiber fineness using gene editing technology.

[0062] Example 3 and GhUMA Development and validation of gene-linked KASP markers Based on the whole-genome resequencing results of the 906-fine and 906-coarse lines, we... GhUMA A large insertion / deletion (InDel) was identified near the gene, and the variant is located in... GhUMA Downstream at approximately 2.8 Mb ( Figure 12 ).

[0063] To further verify the authenticity of the insertion / deletion, we used homologous cloning to amplify cDNA from leaves of 906-fine and 906-coarse lines. The results confirmed the presence of the insertion / deletion in both lines. Furthermore, in a segregating population of 278 lines constructed using 906-fine and 906-coarse lines as parents, the insertion / deletion site exhibited significant segregation and recombination. Based on this, we developed a KASP marker for this linkage site and designed specific primers using Primer3 software based on the flanking sequences. Primer sequence information and genotyping information are shown in Table 3.

[0064] Table 3 KASP marker genotyping information and primers used

[0065] Using an HVI1000 analyzer, we measured the micronaire value, fiber length, and fiber strength of a segregating population comprising 278 lines under three different growing conditions, and used BLUP analysis to eliminate environmental influences. Simultaneously, genomic DNA was extracted from the 278 lines for KASP genotyping.

[0066] The results show that this marker can accurately identify TCCCGACGACACGAAACATGACGTCCCAACGAGGGATATACGACGTTGCGATGTGAGAACATGTTTCC:TCCCGACGACACGAAACATGACGTCCCAACGAGGGATATACGACGTTGCGATGTGAGAACATGTTTCC, SEQ ID NO.12 / -type ( Figure 13 Furthermore, the above typing is significantly correlated with micronaire value. The locus is TCCCGACGACACGAAACATGACGTCCCAACGAGGGATATACGACGTTGCGATGTGAGAACATGTTTCC:TCCCGACGACACGAAACATGACGTCCCAACGAGGGATATACGACGTTGCGATGTGAGAACATGTTTCC, SEQ ID NO. 12, which indicates a finer fiber line. If this locus sequence is deleted, it indicates a coarser fiber line. Figure 14 ), and has no significant screening effect on fiber length and strength ( Figures 15-16 ).

[0067] To verify the applicability of this locus in cotton, we used the KASP marker to perform genotyping on 137 cotton resource materials (the names of the above materials are published in the Cotton Journal, but this locus was used for the first time). [The following is a separate section: Verification of SSR-associated loci for major breeding traits in upland cotton and identification of superior materials, Cotton Journal, 2022, Vol. 34, Issue (2): 120-136. DOI: 10.11963 / cs20210066] Figure 17 The micronaire value, fiber length, and fiber strength of the above resources were measured under two years of planting conditions, and BLUP analysis was used to eliminate environmental impact.

[0068] The results showed that the marker insertion type (TCCCGACGACACGAAACATGACGTCCCAACGAGGGATATACGACGTTGCGATGTGAGAACATGTTTCC:TCCCGACGACACGAAACATGACGTCCCAACGAGGGATATACGACGTTGCGATGTGAGAACATGTTTCC, SEQ ID NO.12) indicates a finer fiber material, while the deletion of this site indicates a coarser fiber line. Figure 18 a), and regarding fiber length ( Figure 18 b) and intensity ( Figure 18 c) No obvious screening effect.

[0069] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0070] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for regulating the fineness of cotton fibers GhUMA Genes, characterized by, Its nucleotide sequence is shown in SEQ ID NO.

1.

2. The claim 1 GhUMA Application of genes in regulating cotton fiber fineness.

3. The application as described in claim 2, characterized in that, The regulation mentioned is a positive regulation.

4. Knockout GhUMA Application of genes in constructing cotton models with finer fiber thickness.

5. A device as described in claim 1 GhUMA KASP molecular markers that are tightly linked to genes are characterized by, The nucleotide sequence of the KASP molecular marker is shown in SEQ ID NO.

12.

6. The primer set for amplifying the molecular marker of claim 5, characterized in that, The primer set includes the primers shown in SEQ ID NO.8 to SEQ ID NO.

10.

7. A reagent kit for identifying the fineness of cotton fibers, characterized in that, Includes the primer set as described in claim 6.

8. The use of the molecular marker of claim 5, the primer set of claim 6, or the kit of claim 7 in any of the following: (1) Application in identifying the fineness of cotton fibers; (2) Application in cotton breeding.

9. A method for determining the fineness of cotton fibers, characterized in that, Includes the following steps: Using the genomic DNA of the cotton sample to be tested as a template, PCR amplification was performed using the primer set shown in SEQ ID NO.8~SEQ ID NO.10, and the determination was based on the sequencing results of the obtained amplification products.

10. The method as described in claim 9, characterized in that, If this site is shown in SEQ ID NO.12, it indicates a strain with finer fibers; if this site is missing, it indicates a strain with coarser fibers.

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