Application of GhFRC1 gene in regulating plant fertility
By heterologously expressing the GhFRC1 gene in Arabidopsis thaliana, tomato, and rice to suppress pollen activity, the problem of the narrow applicability of fertility genes was solved, enabling precise regulation of fertility in multiple crops and efficient breeding.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHAANXI INST OF BIOLOGICAL AGRI
- Filing Date
- 2026-05-19
- Publication Date
- 2026-06-30
AI Technical Summary
Existing fertility genes have a narrow scope of application and are difficult to broadly regulate the fertility of multiple crops. Traditional hybridization breeding is time-consuming, labor-intensive, and results in low purity of hybrids.
By heterologously expressing the GhFRC1 gene in Arabidopsis thaliana, tomato, and rice, pollen activity was inhibited, leading to abnormal fruit set. This was achieved through recombinant vectors to negatively regulate plant fertility.
It enables broad-spectrum regulation of fertility in multiple crops using a single gene, improves the efficiency of hybridization breeding, reduces operational difficulty and cost, and expands the scope of breeding applications.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of plant genetic engineering technology, and more particularly to... GhFRC1 The application of genes in regulating plant fertility. Background Technology
[0002] Crop fertility regulation is one of the core technologies of hybridization breeding, and the creation of male-sterile lines is key to utilizing hybrid vigor and improving breeding efficiency. In traditional hybridization breeding, hybrids are obtained through manual or mechanical emasculation, which is not only time-consuming, labor-intensive, and costly, but also prone to affecting the purity of hybrids due to incomplete emasculation, thus restricting the improvement of breeding efficiency.
[0003] With the development of plant genetic engineering technology, the creation of transgenic male-sterile lines using fertility-related genes has become a research hotspot in the breeding field, offering advantages such as precision, efficiency, and controllability compared to traditional methods. Currently, most publicly available plant fertility-related genes focus on the fertility regulation of single crops, such as soybeans. GmMS1 Genes are mainly used for fertility regulation in soybeans, while those in cotton are publicly known. GhRALF4L and GhRALF30L The gene clusters focus on the creation of male sterility under high temperature stress and lack gene resources that can broadly regulate the fertility of multiple crops.
[0004] As an important economic crop, cotton's fertility-related gene functional studies have largely focused on its own fertility regulation. There are no reports showing that cotton fertility genes can be heterologously expressed and play a fertility-regulating role in distantly related crops (such as Arabidopsis, tomato, and rice). Due to the significant differences in the genetic backgrounds of different crops, it is generally believed that cotton genes are unlikely to function properly in distantly related crops. Therefore, screening cotton fertility genes that can broadly regulate the fertility of multiple crops and establishing efficient fertility regulation techniques are of great significance for promoting the development of multi-crop hybrid breeding technology.
[0005] This invention clones a substance that affects fertility from cotton. GhFRC1 The gene was experimentally verified to inhibit pollen activity and cause abnormal fruit set when expressed heterologously in Arabidopsis, tomato, and rice. In Arabidopsis, a male sterility phenotype with completely undeveloped pollen sacs can be observed. This sterility phenotype can be restored to fertility through normal pollination. This solves the problem of the narrow applicability and limited breeding application of existing fertility genes, and provides new gene resources and technical solutions for fertility regulation and hybridization breeding in multiple crops. Summary of the Invention
[0006] The purpose of this invention is to overcome the shortcomings of existing technologies, such as the narrow applicability of fertility genes and the difficulty in broadly regulating the fertility of multiple crops, and to provide... GhFRC1The application of genes and their encoded proteins, as well as recombinant vectors, in the regulation of fertility in multiple crops, the creation of male-sterile lines, and hybridization breeding enables precise regulation of the fertility of multiple crops by a single gene, thereby improving the efficiency of hybridization breeding.
[0007] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides GhFRC1 The application of genes in regulating plant fertility (1) The above GhFRC1 The nucleotide sequence of the gene is shown in SEQ ID NO.1; (2) Or a nucleotide sequence that has more than 90% homology with the nucleotide sequence shown in SEQ ID NO.1 and encodes the same fertility function protein; (3) Or, based on the nucleotide sequence shown in SEQ ID NO.1, a nucleotide sequence obtained by base substitution, deletion or insertion, which still has the function of regulating plant fertility.
[0008] Furthermore, by the aforementioned GhFRC1 The amino acid sequence encoded by the gene is shown in SEQ ID NO.2; Or an amino acid sequence that has more than 90% homology with the amino acid sequence shown in SEQ ID NO.2 and has the same fertility function.
[0009] Furthermore, the aforementioned GhFRC1 The method by which genes regulate plant fertility is as follows: [The text abruptly ends here, likely due to an incomplete sentence or missing information.] GhFRC1 Heterologous expression of genes in recipient plants inhibits pollen activity in the recipient plants, leading to abnormal fruit set and achieving negative regulation of plant fertility.
[0010] Furthermore, the recipient plants include Arabidopsis thaliana, tomato, and rice.
[0011] A type containing the above GhFRC1 A gene recombinant vector, wherein the recombinant vector uses a plant expression vector as its backbone.
[0012] The present invention also provides the above. GhFRC1 Application of genes in the creation of transgenic male-sterile plant lines or crop hybridization breeding.
[0013] Furthermore, utilizing the aforementioned GhFRC1 The method for creating transgenic male-sterile plant lines is as follows: constructing a line containing the aforementioned... GhFRC1 A gene recombinant vector; the recombinant vector is introduced into recipient plant cells; the transformed plant cells are screened and cultivated to obtain positive transgenic lines and their fertility is identified. Male-sterile lines with inactive pollen and abnormal fruit setting are screened out, thus obtaining the target transgenic male-sterile plant lines.
[0014] Compared with the prior art, the present invention has the following significant advantages: 1. This invention is the first to clone cotton. GhFRC1 The gene was identified and verified to be heterologously expressed in three different genera and families of crops: Arabidopsis thaliana, tomato, and rice, and to play a role in fertility regulation. This breakthrough overcomes the limitation of existing fertility genes that can only regulate the fertility of a single crop, and realizes "broad-spectrum regulation of fertility of multiple crops by a single gene", filling the gap in the application of distant heterologous expression of cotton fertility genes.
[0015] 2. The present invention GhFRC1 The effect of gene regulation on fertility is clear. Experimental verification shows that the transgenic plant lines of this invention all exhibit obvious abnormal fertility phenotypes: among 30 positive Arabidopsis lines, 5 lines showed no fruit set and no pollen activity; paraffin section observation showed that the pollen sacs of these sterile lines were completely undeveloped; among 33 positive tomato lines, 9 lines showed no fruit set and no pollen activity; among 29 positive rice lines, 3 lines showed extremely low fruit set rate and no pollen activity; and the Arabidopsis GhFRC1 sterile line, after being pollinated with Col-0 wild-type pollen, could regain fertility and produce normal pods. Therefore, GhFRC1 Heterologous gene expression can stably lead to inactive pollen and abnormal fruit set in recipient crops. In Arabidopsis thaliana, the sterility trait can be restored to fertility through normal pollination, which facilitates the creation of male-sterile lines, as well as the propagation of sterile lines and the production of hybrid varieties.
[0016] 3. Utilizing the present invention GhFRC1 The creation of male-sterile lines through gene technology is simple and efficient, requiring no complex gene editing operations and can be achieved solely through conventional genetic transformation techniques. It is low-difficulty and low-cost, and can rapidly produce transgenic male-sterile lines. This addresses the pain points of traditional hybridization breeding, such as the cumbersome, costly, and low-purity artificial emasculation, significantly improving hybridization breeding efficiency and meeting the current industrial needs of the bio-breeding field. Furthermore, this gene can be extended to the fertility improvement of other economic crops, showing broad prospects for industrial application. Attached Figure Description
[0017] Figure 1 : GhFRC1 Electrophoresis diagram of PCR amplification of the gene; the white arrow points to... GhFRC1 The target band of the gene.
[0018] Figure 2 Electrophoresis diagram of PCR identification of positive Arabidopsis thaliana strains.
[0019] Figure 3 Arabidopsis wild-type (Col-0) and GhFRC1 Anatomical comparison of petals of transgenic sterile lines.
[0020] Figure 4 Arabidopsis wild-type (Col-0) and GhFRC1 Comparison of pollen viability of transgenic sterile lines; left side: wild-type Arabidopsis pollen (red, viable); right side: pollen from transgenic sterile lines. GhFRC1 Pollen from transgenic sterile lines (colorless, non-viable).
[0021] Figure 5 Arabidopsis wild-type (Col-0) and GhFRC1 Comparative images of paraffin sections of pollen sacs from transgenic sterile lines; the left side shows pollen sacs from wild-type Arabidopsis thaliana (normally developed, containing mature pollen), while the right side shows pollen sacs from the GhFRC1 transgenic sterile line (completely undeveloped, without pollen).
[0022] Figure 6 : GhFRC1 Results of pollination recovery experiment of transgenic Arabidopsis sterile lines; the left side shows the unpollinated sterile lines (shriveled siliques), and the right side shows the overall fertility recovery of the sterile lines pollinated with wild-type pollen (normal pod formation).
[0023] Figure 7 Electrophoresis diagram of PCR identification of transgenic tomato lines.
[0024] Figure 8 Example 4: Wild-type tomato (MT) and GhFRC1 Comparison of fruit set in transgenic sterile lines: A: Floral organs and inflorescences of wild-type / MT tomatoes (hairy, normally developed); B: GhFRC1 C: Floral organs and inflorescences of sterile tomato lines (abnormal trichome morphology); D: Phenotype of mature wild / MT tomato plants (normal flowering and fruit set); GhFRC1 The phenotype of adult male-sterile tomatoes (stunted plants, inhibited reproductive development).
[0025] Figure 9 Electrophoresis diagram of PCR identification of positive transgenic rice lines.
[0026] Figure 10 Example 4: Wild-type rice (Nip) and GhFRC1 Comparison of seed setting in transgenic sterile lines; A is the overall view, and B is a comparison of individual spikelets. Detailed Implementation
[0027] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0028] Example 1: GhFRC1 Gene cloning and sequence analysis
[0029] 1.1 Experimental Materials
[0030] Cotton variety: Texas Marker-1 (TM-1), the genetic standard line of upland cotton, was grown in a greenhouse. Flower buds during the full bloom period were taken, flash-frozen in liquid nitrogen, and stored at -80℃ for total RNA extraction.
[0031] 1.2 Total RNA extraction and cDNA synthesis
[0032] Total RNA was extracted from cotton buds using Trizol reagent (Invitrogen) following the manufacturer's instructions. DNase I was used to remove genomic DNA contamination from the RNA. First-strand cDNA was synthesized using the total RNA as a template and stored at -20°C for later use.
[0033] 1.3 GhFRC1 PCR amplification of genes
[0034] Based on the predicted CLE gene sequence from the cotton genome database, a design was developed. GhFRC1 Gene-specific primers were synthesized by Nanjing Qingke Biotechnology Co., Ltd.
[0035] Using cotton petal cDNA as a template, PCR amplification was performed: PCR reaction system (25 μL): 1 μL cDNA template, 1 μL upstream primer, 1 μL downstream primer, 12.5 μL 2×Taq PCR MasterMix, 9.5 μL ddH2O; PCR reaction program: 94℃ pre-denaturation for 5 min; 94℃ denaturation for 30 s, 58℃ annealing for 30 s, 72℃ extension for 1 min, for a total of 35 cycles; 72℃ final extension for 10 min, and storage at 4℃.
[0036] 1.4 Gene Cloning and Sequence Analysis
[0037] The PCR amplification products were subjected to 1% agarose gel electrophoresis to recover the target fragment (using an agarose gel recovery kit, TaKaRa). The recovered target fragment was ligated into the pMD19-T vector (TaKaRa), transformed into E. coli DH5α competent cells, plated on LB medium containing ampicillin (100 μg / mL), and incubated overnight at 37°C. White single colonies were selected, plasmids were extracted, and enzyme digestion and sequencing were performed for identification and verification (Nanjing Qingke Biotechnology Co., Ltd.).
[0038] Sequencing results showed that the cloned GhFRC1The complete coding region nucleotide sequence of the gene is shown in SEQ ID NO.1, which is 255 bp in length and encodes 84 amino acids. The amino acid sequence is shown in SEQ ID NO.2. Sequence alignment analysis shows that this gene has certain homology with other plant CLE family genes, but both the nucleotide and amino acid sequences are unique and have not been disclosed in the prior art.
[0039] SEQ ID NO.1: GhFRC1 nucleotide sequence of a gene
[0040] ATGGTTTGCAGTGCTCAAAGAGTGATCATATTGCTCATCTTTTTGGGGTTATTAGCACTTCAACCAGTTGAAGTGTCTGGTTTGAGAAATATAGACTTTGTCTTCAGGCATAGTCAACGTGTTCTTA AGGCTGTTGATACGAAGGGAATGAGTACTGAGAAGAAAACAGCTTTCGTAAACAACAAATTCGATCCGAATCGATCAAGCAAACGAAGAGTTCGAAGAGGATCAGATCCTATCCACAACAGGTCTTGA
[0041] SEQ ID NO.2: GhFRC1 The amino acid sequence encoded by the gene
[0042] MVCSAQRVIILLIFLGLLALQPVEVSGLRNIDFVFRHSQRVLKAVDTKGMSTEKKTAFVNNKFDPNRSSKRRVRRGSDPIHNRS
[0043] SEQ ID NO.3: GhFRC1 Gene-coding region specific primers, upstream primers
[0044] 5'-ATGGTTTGCAGTGCTCAAAGAGT-3'; SEQ ID NO.4: GhFRC1 Gene-coding region-specific primers, downstream primers 5'-AGACCTGTTGTGGATAGGATCTGAT-3'.
[0045] Example 2: Contains GhFRC1 Construction of gene recombination vectors
[0046] 2.1 Experimental Materials
[0047] Vector: pBINPLUS.GFP4 vector (containing kanamycin resistance marker); Target gene: the gene cloned in Example 1 GhFRC1 Gene; Enzymes: Sal I. Bam H I restriction endonuclease (NEB); Recombinant Cloning Kit: ClonExpress® II One Step Cloning Kit (C112) (Vazyme).
[0048] 2.2 Construction of Recombinant Vectors
[0049] (1) Vector linearization and target gene amplification: The plant expression vector is linearized using... Sal I and Bam H I was used for double digestion, followed by purification as needed to obtain linearized vectors; simultaneously, specific primers containing homologous sequences and corresponding restriction sites were designed, and high-fidelity PCR amplification was performed to obtain... GhFRC1 Insertion of complete coding region of gene ( Figure 1 After amplification, the fragment is quantified to ensure that its molar ratio with the linearized vector meets the recommended requirement of 1:2.
[0050] (2) Recombination reaction: Prepare a 20 μL recombination reaction system on ice according to the ratio, and add sterile ddH2O, 5×CEII Buffer, linearized vector, insert fragment, and Novizan Exnase II recombinase in sequence. After gently mixing, briefly centrifuge to allow the liquid to settle to the bottom, and incubate at 37°C for 30 min to complete the recombination reaction.
[0051] (3) Transformation of recombinant products: The recombinant reaction products were transformed into Escherichia coli DH5α competent cells. After being subjected to ice bath for 30 min, heat shock at 42℃ for 45 s, ice bath cooling for 2-3 min, and recovery at 37℃ for 1 h, the bacterial solution was spread on LB plates containing the corresponding resistance and incubated upside down at 37℃ for 12-16 h.
[0052] (4) Recombinant vector identification: Single clones on the plate were picked for preliminary identification by colony PCR. After incubating the PCR-positive clones overnight, plasmids were extracted and further verified by first-generation sequencing to confirm the recombinant vector. GhFRC1 The gene is correctly inserted into the vector and the sequence is free of mutations, which ultimately ensures that the recombinant vector is constructed correctly.
[0053] Example 3: GhFRC1 Heterologous expression and fertility analysis of genes in Arabidopsis thaliana
[0054] 3.1 Genetic transformation
[0055] (1) Agrobacterium transformation: The pBINPLUS.GFP4-GhFRC1 recombinant vector constructed in Example 2 was transformed into Agrobacterium GV3101 competent cells, plated on LB medium containing 50 μg / mL kanamycin and 50 μg / mL rifampin, cultured at 28℃ for 2 days, single colonies were selected, and positive Agrobacterium strains were identified by PCR. (2) Arabidopsis transformation: Arabidopsis (Col-0 ecotype) was transformed using the flower-dipping method. Positive Agrobacterium strains were inoculated into LB liquid medium (containing corresponding resistance) and cultured at 28°C with shaking at 200 rpm until OD. 600 =0.8-1.0; Centrifuge the bacterial culture, discard the supernatant, resuspend the bacterial culture in a buffer solution containing 5% sucrose and 0.02% Silwet L-77, and adjust the OD value to 0.8-1.0. 600 =0.6; Soak Arabidopsis inflorescences in bacterial solution for 10 seconds, cover with plastic wrap after soaking, and incubate in the dark for 1 day, then incubate normally. After the plants bloom again, repeat the flower dipping once to improve the conversion efficiency. (3) Seed harvesting and screening: After Arabidopsis thaliana matures, the seeds (T0 generation) are harvested and sown on MS medium containing 50 μg / mL kanamycin. After vernalization at 4℃ for 3 days, the seeds are placed in a greenhouse for cultivation (16h light / 8h dark, 22℃). T0 generation positive plants with kanamycin resistance are screened and transplanted to a greenhouse for cultivation.
[0056] 3.2 Identification of positive strains
[0057] DNA was extracted from the leaves of T0 generation resistant plants and used... GhFRC1 Gene-specific primers were used for PCR identification. The universal primers for the pBINPLUS.GFP4 empty vector were: pbfp4F (SEQ ID NO.5): CAAGCAATCAAGCATTCTAC; pbfp4R (SEQ ID NO.6): CGGACACGCTGAACTTGTGG. Plants that amplified the target gene fragment were considered successful. GhFRC1 Positive Arabidopsis thaliana lines expressing genes heterologously. For example... Figure 2 As shown, bands 1-36 represent 36 strains, from which 30 T0 generation positive strains were obtained, representing the wild-type Arabidopsis thaliana and... GhFRC1 Anatomical comparison of petals of transgenic sterile lines, such as... Figure 3 As shown, GhFRC1 The stamens of the transgenic sterile line are not fully developed.
[0058] 3.3 Fertility Analysis
[0059] (1) Pollen viability detection: After the positive lines flower, mature anthers are taken, placed on a glass slide, and 0.5% TTC staining solution is added. The slide is incubated at 37℃ for 15 min, and the pollen staining is observed under a microscope.Figure 4 As shown, red pollen is active pollen, and colorless pollen is inactive pollen; the results showed that among the 30 positive lines, the pollen of 5 lines was almost entirely colorless (inactive), while the pollen of the remaining lines had normal activity. (2) Statistics on fruit set: The fruit set of all positive lines was observed and counted. The results showed that the five lines with no pollen activity were all infertile, with short, shriveled siliques and no mature seeds, while the lines with normal pollen activity had normal fruit set. (3) Observation of pollen sac development: flower buds of pollen-inactive strains and wild-type Arabidopsis thaliana were taken and paraffin sections were prepared: flower buds were fixed in FAA fixative, dehydrated in a gradient, embedded in paraffin, sectioned (8 μm thick), stained with toluidine blue staining solution, dehydrated and cleared, mounted with neutral resin, and the development of pollen sacs was observed under a microscope. Figure 5 The results showed that the pollen sacs of wild-type Arabidopsis thaliana were normally developed and contained a large amount of mature pollen, while GhFRC1 The pollen sacs of the sterile lines with heterologous gene expression did not develop at all, and no pollen was formed. (4) Pollination recovery experiment: Mature pollen from wild-type Arabidopsis thaliana (Col-0) was used for pollination until... GhFRC1 After normal culture, observe the fruit set on the stigma of the sterile line; Figure 6 The results showed that the siliques of the unpollinated sterile lines were shriveled, while the sterile lines pollinated with wild-type pollen could form pods normally and produce mature seeds, indicating that the sterility trait can be restored to fertility through normal pollination.
[0060] Example 4: GhFRC1 Heterologous expression and fertility analysis of genes in tomatoes and rice
[0061] 4.1 Heterologous expression and fertility analysis in tomatoes
[0062] The Agrobacterium-mediated leaf disc method was used to transform tomatoes (variety "Micro-Tom") with the pBINPLUS.GFP4-GhFRC1 recombinant vector and the empty vector: Sterile tomato seedling leaves were taken, cut into 0.5cm × 0.5cm leaf discs, and immersed in Agrobacterium-positive bacterial solution (OD200). 600 =0.6) for 10 min, after aspirating the bacterial solution, inoculate into co-medium (MS + 2 mg / L 6-BA + 0.2 mg / L IAA), and incubate in the dark at 25℃ for 2 days; transfer the leaf disc to selection medium (MS + 2 mg / L 6-BA + 0.2 mg / L IAA + 50 mg / L kanamycin + 250 mg / L cephalosporin), and culture until resistant shoots appear; cut off the resistant shoots, inoculate into rooting medium (MS + 0.1 mg / L IAA + 50 mg / L kanamycin), and culture until roots appear to obtain resistant plants; extract DNA from the resistant plants, and identify positive lines by PCR, such as Figure (As shown, bands 1-34 represent 34 strains, and a total of 33 positive strains were obtained.
[0063] Fertility analysis showed that among the 33 positive lines, 9 lines exhibited abnormal fruit setting, producing small, seedless fruits; the remaining lines showed normal fertility, consistent with the expression results in Arabidopsis thaliana. Tomato wild-type (MT) and GhFRC1 Comparison of seed setting in transgenic sterile lines (see figure) Figure 8 As shown, the wild-type / MT tomato has hairy floral organs and inflorescences, develops normally, flowers normally, and sets fruit normally, while... GhFRC1 The sterile tomato line exhibits abnormal morphology of its floral organs and inflorescence trichomes, resulting in stunted plants and impaired reproductive development.
[0064] 4.2 Heterologous expression and fertility analysis in rice
[0065] Agrobacterium-mediated callus transformation method was used to transform rice (variety "Nipponbare") with the pBINPLUS.GFP4-GhFRC1 recombinant vector and an empty vector: Mature rice seeds were dehulled, sterilized, and inoculated onto MS medium to induce callus formation; positive Agrobacterium-positive bacterial suspension was co-cultured with rice callus for 3 days, then transferred to selection medium (containing 50 μg / mL hygromycin) to screen for resistant callus; resistant callus was transferred to differentiation medium to induce differentiation into shoots and roots, obtaining resistant plants; DNA was extracted from resistant plants, and positive lines were identified by PCR, such as... Figure 9 As shown, bands 1-31 represent 31 strains, and a total of 29 positive strains were obtained.
[0066] Fertility analysis showed that among the 29 positive lines, 3 lines exhibited extremely low seed setting rate (<5%) and shriveled grains, while the remaining lines showed normal fertility, indicating that... GhFRC1 Genes also play a role in fertility regulation in rice. Rice wild-type (Nip) and GhFRC1 For example, the seed setting rate of transgenic sterile lines Figure 10 As shown, GhFRC1 It should be noted that there seems to be an error in "Figure ( " in the original text. It might be a typo. The above translation is based on the given text as accurately as possible. The transgenic sterile lines have extremely low seed setting rate and shriveled grains.
[0067] Example 5: Stable genetic verification of transgenic male-sterile lines
[0068] The male-sterile lines of Arabidopsis, tomato, and rice selected in Examples 3 and 4 were backcrossed (using wild-type plants as male parents) to obtain T1 generation seeds. The T1 generation seeds were sown, and positive lines were obtained through resistance screening and PCR identification. The fertility of the T1 generation positive lines was identified, and the results showed that the T1 generation positive lines still exhibited the male-sterile phenotype of inactive pollen and abnormal fruit setting. After repeated propagation to the T3 generation, the abnormal fertility phenotype was stably inherited, indicating that a stably inherited transgenic male-sterile line had been obtained.
[0069] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. GhFRC1 The application of genes in regulating plant fertility is characterized by, (1) The above GhFRC1 The nucleotide sequence of the gene is shown in SEQ ID NO.1; (2) Or a nucleotide sequence that has more than 90% homology with the nucleotide sequence shown in SEQ ID NO.1 and encodes the same fertility function protein; (3) Or, based on the nucleotide sequence shown in SEQ ID NO.1, a nucleotide sequence obtained by base substitution, deletion or insertion, which still has the function of regulating plant fertility.
2. The application according to claim 1, characterized in that, From the above GhFRC1 The amino acid sequence encoded by the gene is shown in SEQ ID NO.2; Or an amino acid sequence that has more than 90% homology with the amino acid sequence shown in SEQ ID NO.2 and has the same fertility function.
3. The application according to claim 1, characterized in that, The GhFRC1 The method by which genes regulate plant fertility is as follows: [The text abruptly ends here, likely due to an incomplete sentence or missing information.] GhFRC1 Heterologous expression of genes in recipient plants inhibits pollen activity in the recipient plants, leading to abnormal fruit set and achieving negative regulation of plant fertility.
4. The application according to claim 1, characterized in that, The recipient plants include Arabidopsis thaliana, tomato, and rice.
5. A device comprising the contents of claim 1 GhFRC1 A gene recombination vector, characterized in that, The recombinant vector uses a plant expression vector as its backbone.
6. The claim 1 GhFRC1 The application of the gene or the recombinant vector of claim 5 in the creation of transgenic male-sterile plant lines or in crop hybridization breeding.
7. A method of utilizing GhFRC1 A method for creating transgenic male-sterile plant lines, characterized in that, Includes the following steps: Construct containing the GhFRC1 A gene recombinant vector; the recombinant vector is introduced into recipient plant cells; The transformed plant cells were screened and cultured to obtain positive transgenic lines and their fertility was identified. Male-sterile lines with inactive pollen and abnormal fruit set were screened out, thus obtaining the target transgenic male-sterile plant lines.