A plant height regulating gene GhSHMT10-A for upland cotton and its application

Abstract

The invention discloses a plant height regulating gene for upland cotton GhSHMT10‑A and its application, the gene has the nucleotide sequence shown in SEQ ID NO: 1. GhSHMT10‑ A， It is possible to regulate the plant height of upland cotton at the molecular level, thereby improving the upland cotton, dwarfing the plants, making the upland cotton more resistant to lodging, and improving the crop's group light energy utilization rate, thereby increasing crop yields.

Description

Technical Field

[0001] The present invention relates to the technical field of genetic engineering, and in particular to an upland cotton plant height regulating gene GhSHMT10-A and an application thereof. Background Art

[0002] As the most important natural textile fiber, cotton supplies about 35% of the world's fiber consumption and is also used to make feed and oil. Among them, upland cotton (Gossypium hirsutum L.) accounts for more than 90% of the global cotton production (Chen et al., 2007; Wang et al., 2015; Zhang et al., 2015; Ma et al., 2018b). Cotton production is part of the cash crop industry (Li, 2021). With the changes in the trade situation of the cotton industry and the advancement of mechanization, it is particularly important to cultivate high-yield and high-quality cotton varieties. Plant ideal plant breeding can effectively increase cotton yield and improve fiber quality, thereby affecting the economic production of cotton (Li et al., 1998, 2015; Song and Zhang, 2009; Shang et al., 2015b; Fu et al., 2019). The ideal plant architecture refers to the three-dimensional structure of the aboveground part of the plant, which is a comprehensive agronomic trait that includes factors such as plant height, number of fruit branches, angle between the main stem and fruit branches, morphological characteristics and distribution of bolls and leaves (Yang et al., 1996; Wang and Li, 2006; Wu et al., 2021). It is worth noting that the Green Revolution that emerged in the 1960s changed the plant architecture by transferring dwarf genes into crops, resulting in a significant increase in yield (Khush, 2001). For example, reducing plant height enhances crop resistance and significantly increases the yield of wheat and rice (Peng et al., 1999; Sasaki et al., 2002). In addition, the use of compact and medium-dwarf maize varieties has increased planting density and significantly improved productivity (Peiffer et al., 2014). Compared with increasing the yield potential per plant, increasing planting density has greater potential to increase yield per unit area in cotton production, which requires moderately low plant height, lodging resistance, and more compact branching organization (i.e., short fruiting branches, acute branch angles, and high photosynthetic rate per unit canopy area), which are characteristics that have obvious advantages over currently planted varieties (Su et al., 1993; Dong et al., 2018).

[0003] Plant height (PH) is one of the main factors affecting plant architecture (Jiao et al., 2010; Zhang et al., 2019). In addition, appropriate cotton plant height (80–90 cm) is also conducive to lodging resistance and mechanical harvesting (Chen, 2013; Pei et al., 2021). Therefore, studying cotton plant height will help cotton breeding for ideal plant architecture. However, plant height is a quantitative trait controlled by multiple genes and is easily affected by the environment (Cui et al., 2011; Wurschum et al., 2015; Liu et al., 2018). The molecular genetic mechanism of plant height in cotton is mainly studied through linkage analysis. Fourteen plant height QTLs were identified on 11 chromosomes in an upland cotton F2 population, of which qPH-D12-1 contributed 9.64% (Jia et al., 2021). A stable QTL qPH-Dt1-1 and a gene regulating cotton plant height, GhPIN3, were identified in an interspecific G. hirsutum × G. barbadense BIL population by SLAF-seq technology (Ma et al., 2019). Using an upland cotton RIL population, twenty-seven plant height QTLs were located on 18 chromosomes, explaining 3.81–8.54% of the phenotypic variation (Liu et al., 2018). In addition, natural dwarf cotton mutants are very important for identifying cotton plant height genes. A dwarf-related gene, EXTR-DWARF, was screened from natural dwarf mutants of cotton (Ji, 2018).

[0004] Serine hydroxymethyltransferase (SHMT) is a conserved enzyme involved in the one-carbon metabolic pathway that is widely present in animals and plants. It not only converts serine to glycine by transferring the β-carbon of serine to tetrahydrofolate (THF), but also produces serine by transferring the one-carbon unit of methylene THF to glycine. In addition, SHMT catalyzes the cleavage of various 3-hydroxyamino acids and the decarboxylation of amidinomalonic acid. At the same time, it plays a key role in the coenzyme metabolic pathway, promoting the conversion of 5,10-CH±H4PteGlu to 5-CHO-H4PteGlu, which is of great significance for folate metabolism (Schirch and Szebenyi, 2005). Functionally, SHMT is involved in one-carbon metabolism, methionine synthesis, and maintaining redox homeostasis during photorespiration (Hanson and Roje, 2001). Structurally, SHMT contains pyridoxal phosphate (PLP) tightly bound to a lysine residue, forming an aldehyde-amine bond.

[0005] Currently, there are seven SHMTs in Arabidopsis thaliana (Nogues et al., 2022; Zhang et al., 2010), seven in Medicago truncatula (Ruszkowski et al., 2018), five in rice (Oryza sativa) (Pan et al., 2024), 64 in wheat (Triticum aestivum) (Hu et al., 2022), and at least twelve in soybean (Glycine max) (Lakhssassi et al., 2019). Mitochondrial-localized SHMTs (mSHMTs) play a conserved role in catalyzing the photorespiratory conversion of glycine to serine and work in concert with the glycine decarboxylase complex (Douce and Neuburger, 1999). In addition, mSHMT is also thought to play a role in regulating cellular damage caused by abiotic stress factors such as high light exposure, salt stress, and pathogen-induced cell death (Fang et al., 2020; Liu et al., 2019; Mishra et al., 2019; Moreno et al., 2005; Zhou et al., 2012). Cytoplasmic localized SHMT (cSHMT) plays a key function in soybean resistance to cyst nematodes by regulating global methylation levels (Lakhssassi et al., 2019; Liu et al., 2012; Wu et al., 2016). In plastids, SHMT is thought to catalyze the transfer of hydroxymethyl groups from serine to the H4PteGlun-bound one-carbon unit metabolic pathway. Genes encoding potential plastid SHMTs have been detected in the genomes of various plant species, mainly located in chloroplasts (Zhang et al., 2010), but the biological functions of plastid-localized SHMTs remain largely unclear. In the nucleus, OsSHMT4 regulates the methylation levels of genes involved in ion transport, storage material synthesis, and other pathways. After mutation of the non-conservative domain, it enhanced rice tolerance to cadmium and increased the selenium content in seeds (Chen et al., 2020b). Our previous study found that after mutation of the conserved domain, it severely affected the normal development of rice endosperm (Yan et al., 2022a, 2024). Therefore, SHMT isozymes in plants play a key role in seed development and in providing resistance to biotic and abiotic stresses.

[0006] In upland cotton, only the mitochondrial-localized GhSHMT11-A has been reported to alleviate salt stress by reducing ROS accumulation in Arabidopsis thaliana. However, as a key component of cellular one-carbon metabolism and epigenetic modification, the regulation of cytoplasmic-localized SHMT in cotton has not been reported. Summary of the invention

[0007] The purpose of the present invention is to provide an upland cotton plant height regulating gene GhSHMT10-A and its application. By regulating GhSHMT10-A, the upland cotton plant height can be regulated at the molecular level, thereby improving the upland cotton, dwarfing the plants, making the upland cotton have a stronger lodging resistance, improving the crop group light energy utilization rate, and thus increasing the crop yield.

[0008] The technical solution adopted by the present invention to solve its technical problem is:

[0009] A plant height regulating gene GhSHMT10-A of upland cotton has a nucleotide sequence shown in SEQ ID NO:1.

[0010] The present invention first discovered that the GhSHMT10-A gene in upland cotton has the function of regulating plant height. The present invention screened out the gene GhSHMT10-A that may regulate plant height from the GhSHMT family. Experiments have shown that GhSHMT10-A is mainly expressed at a high level in cotton stems. Subcellular localization analysis of GhSHMT10-A in tobacco showed that GhSHMT10-A is located in the cytoplasm. Through heterologous overexpression of Arabidopsis and virus-induced gene silencing (VIGS) experiments, it was found that GhSHMT10-A positively regulates plant height, laying a foundation for cultivating ideal plant types, high-yield varieties, and in-depth research on the relevant mechanisms of GhSHMT10-A regulating plant height.

[0011] A plasmid comprising the upland cotton plant height regulating gene GhSHMT10-A.

[0012] A plant expression vector comprising the upland cotton plant height regulating gene GhSHMT10-A.

[0013] A host cell, comprising the upland cotton plant height regulatory gene GhSHMT10-A.

[0014] A protein encoded by an upland cotton plant height regulating gene GhSHMT10-A, wherein the protein has an amino acid sequence shown in SEQ ID NO:2.

[0015] Application of an upland cotton plant height regulating gene GhSHMT10-A in regulating the height of upland cotton plants.

[0016] GhSHMT10-A positively regulates plant height of upland cotton.

[0017] A method for dwarfing upland cotton, which reduces the plant height of upland cotton and achieves dwarfing by silencing the plant height regulating gene GhSHMT10-A of upland cotton.

[0018] The beneficial effects of the present invention are as follows: by regulating GhSHMT10-A, the plant height of upland cotton can be regulated at the molecular level, thereby improving the upland cotton, dwarfing the plants, making the upland cotton have a stronger lodging resistance, improving the group light energy utilization rate of the crop, and thus increasing the crop yield; GhSHMT10-A has breeding utilization potential, and lays a foundation for cultivating ideal plant types, high-yield varieties and in-depth research on the relevant mechanisms of GhSHMT10-A regulating plant height. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Expression pattern analysis diagram of GhSHMT family genes; DPA stands for days after anthesis.

[0020] Figure 2 is the spatiotemporal expression pattern of GhSHMT10-A and GhSHMT10-D analyzed by qRT-PCR; DPA stands for days after anthesis;

[0021] Figure 3 This is a subcellular localization analysis diagram of GhSHMT10-A; GFP is the protein expressed after the vector connected to the GFP tag was injected into tobacco, empty represents no vector, GhSHMT10-A-GFP represents that the gene is connected to the GFP tag, bright field is used to indicate the cell background when taking pictures with a laser confocal microscope, merged is a picture of the fluorescence of the fusion tag protein and the cell background, and OsAlaAT1-mCherry is a marker;

[0022] Figure 4 is the pBI121 vector map;

[0023] Figure 5 is the pTRV2 vector map;

[0024] Figure 6 is the pTRV1 vector map;

[0025] Figure 7This is an analysis of the effect of overexpression of GhSHMT10-A on the height of Arabidopsis plants; A: Analysis of the expression level of GhSHMT10-A in transgenic Arabidopsis. OE1-OE3 represent independent GhSHMT10-A overexpression lines. B: Phenotype and statistical analysis of rosette leaves of 5-week-old Arabidopsis, Bar = 5 cm. C: Plant height and statistical analysis of mature Arabidopsis, Bar = 10 cm. D: Length and statistical analysis of mature Arabidopsis siliques, Bar = 1 cm. E: Phenotype and statistical analysis of Arabidopsis seeds, Bar = 0.5 mm. Error bars represent ±SD, * p <0.05, ** p <0.01, *** represents significant difference P < 0.001;

[0026] Figure 8 This is an analysis of the effect of silencing GhSHMT10-A on cotton plant height; A: Plant phenotype after VIGS injection, Bar = 5cm. TRV:00 represents the transformed empty-load cotton line as the experimental control; 1~3 represent the transformed independent TRV: GhSHMT10-A lines, using the GhPDS gene as a positive control, and after VIGS in cotton, an albino phenotype appears. B: Analysis of the expression level of GhSHMT10-A in cotton VIGS lines. CD: Plant height (C) and different internode lengths (D) at different stages after injection. dpi represents the number of days after the annotation of the infection solution; error bars represent ±SD, * represents a significant difference P < 0.05, ** represents a significant difference P < 0.01, and *** represents a significant difference P < 0.001;

[0027] Fig. 9 The cell size statistics of the GhSHMT10-A silenced strain; AD: Staining observation of the second (A) and third (D) internode stem cells of plants after VIGS injection, Bar = 50 μm. BC: Length (B) and width (C) of the second internode stem cells of plants after VIGS injection. EF: Length (E) and width (F) of the third internode stem cells of plants after VIGS injection. Error bars represent ±SD, * represents significant difference P < 0.05, ** represents significant difference P < 0.01, *** represents significant difference P < 0.001. DETAILED DESCRIPTION

[0028] The technical solution of the present invention is further described in detail below through specific embodiments.

[0029] In the present invention, unless otherwise specified, the raw materials and equipment used can be purchased from the market or are commonly used in the art. The methods in the following embodiments, unless otherwise specified, are all conventional methods in the art.

[0030] Example 1: Cloning and expression pattern analysis of the plant height regulating gene GhSHMT10-A in upland cotton

[0031] The transcription levels of the 20 identified GhSHMT family members in different tissues and developmental stages were compared, and it was found that compared with other family members, the expression levels of two homologous genes, GhSHMT10-A and GhSHMT10-D, were higher in the stem, and also higher in different developmental stages of ovules and fibers ( Figure 1 ). By designing specific quantitative primers (Table 1, SEQ ID No.5-SEQ ID No.8), the expression of GhSHMT10-A and GhSHMT10-D in different tissues at different stages of cotton was analyzed. It was found that both genes were constitutively expressed and detected in all tissues at different stages, but the expression level of GhSHMT10-A was significantly higher than that of GhSHMT10-D ( Figure 2 ), indicating that GhSHMT10-A plays a key role in various tissues of cotton, so we cloned and analyzed the function of GhSHMT10-A. First, the RNA of the standard line of upland cotton "TM-1" was used as a template to synthesize the cDNA template by reverse transcription, and PCR amplification was performed using the primers GhSHMT10-AF and GhSHMT10-AR (Table 1, SEQ ID No.3- SEQ ID No.4), and the amplified product was sequenced. The sequencing results showed that it consisted of 1416bp bases (SEQ ID No.1), and the protein it encoded was shown in SEQ ID No.2, consisting of 472 amino acid residues.

[0032] The SHMT family mainly functions in the nucleus, mitochondria, cytoplasm and plastids. GhSHMT10-A is predicted to be located in the cytoplasm. In order to further clarify the localization of GhSHMT10-A in cells, the 35S::GhSHMT10-A-GFP vector was constructed by designing specific primers and transformed into Agrobacterium (primer sequences are shown in Table 1, SEQ ID No.9-SEQ IDNo.10), and it was connected with the cytoplasmic localization marker (OsAlaAT1 gene was connected to the mCherry tag vector to form a marker, OsAlaAT1 gene see GhCDPK60 positively regulates drought stress tolerance inboth transgenic Arabidopsis and cotton by regulating proline content and ROSlevel, 01 December 2022, 10.3389 / fpls.2022.1072584,) were simultaneously injected into tobacco for transient expression. The results showed that GhSHMT10-A co-localized with the cytoplasmic marker in the cytoplasm, indicating that it functions in the cytoplasm ( Figure 3 ).

[0033] Example 2: Application of the Upland Cotton Plant Height Control Gene GhSHMT10-A

[0034] 1. Construction of vector

[0035] The pBI121 vector (see vector map for details) Figure 4 ) was replaced with the double-stranded cDNA molecule shown in SEQ ID No.1 to obtain the pBI121-GhSHMT10-A overexpression vector (primer sequences are shown in Table 1, SEQ ID No.13-SEQ ID No.14).

[0036] The CDS sequence of GhSHMT10-A (SEQ ID No. 1) was used to perform BLAST in the entire cotton genome to determine the specific target sequence of 150-400 bp, and primers were designed to amplify the target fragment (primer sequences are shown in Table 1, SEQ ID No. 11-SEQID No. 12), and the target fragment was ligated to the pTRV2 vector (vector map is shown in Figure 5 ) to obtain pTRV2: GhSHMT10-A recombinant vector, of which pTRV1 (vector map see Figure 6 ) is the auxiliary vector and pTRV2:PDS is the positive control.

[0037] Table 1 Primer sequences

[0038] Primer name Primer sequence (5'-3') GhSHMT10-AF ATGGATCCAGTAAGCAGTTGG (SEQ ID No.3) GhSHMT10-AR CTAATCCTTGTACTTCATTTC (SEQ ID No.4) qPCR-SHMT10-AF TGGTGCTCTGTTGTTGTGTG (SEQ ID No.5) qPCR-SHMT10-AR GCTTTGGTCCCTTGCGATAG (SEQ ID No. 6) qPCR-SHMT10-DF CGTTACTACGGGTGGAAACGAATTC (SEQ ID No. 7) qPCR-SHMT10-DR CCATTTGGTAGGGTCGAGGTGGA (SEQ ID No. 8) 1305-GhSHMT10-A-GFP-F CGGAGCTAGCTCTAGAATGGATCCAGTAAGCAGTTGG (SEQ ID No. 9) 1305-GhSHMT10-A-GFP-R TGCTCACCATGGATCCATCCTTGTACTTCATTTCGGAC (SEQ ID No. 10) pTRV2-GhSHMT10-AF TAAGGTTACCGAATTCATGGATCCAGTAAGCAGTTGG (SEQ ID No. 11) pTRV2-GhSHMT10-AR GAGCTCGGTACCGGATCCGTAAGCAGCGAAGTTGGC (SEQ ID No. 12) PBI121-GhSHMT10-AF ACACGGGGGACTCTAGAATGGATCCAGTAAGCAGTTGG (SEQ ID No. 13) PBI121-GhSHMT10-AR TCGGGGAAATTCGAGCTCCTAATCCTTGTACTTCATTTCGGAC (SEQ ID No. 14)

[0039] 2. Obtaining transgenic Arabidopsis overexpression and cotton gene silencing lines

[0040] (1) Steps for obtaining overexpression transgenic Arabidopsis

[0041] 1. Transform the recombinant expression vector pBI121-GhSHMT10-A into Agrobacterium GV3101 competent cells and obtain recombinant Agrobacterium by freeze-thaw method;

[0042] 2. Prepare 1 to 2 days before infection, cut off the opened flowers and fruit pods of Arabidopsis thaliana (Columbia-0 variety), and water the plants before infection depending on the plant's condition to keep the plants in a good physiological state;

[0043] 3. Add the positive monoclonal Agrobacterium liquid with the correct size band to a 50 mL conical flask containing 0.1% kanamycin and 0.1% rifampicin antibiotics LB liquid culture medium, and culture it in a constant temperature shaker at 28°C and 200 rpm. Use a spectrophotometer to measure the OD value. It is best to culture it until the OD value is between 1.2 and 1.8;

[0044] 4. Pour the bacterial solution into a 50 mL centrifuge tube, centrifuge at 5000 rpm for 10 min, discard the supernatant, retain the bacteria, repeat the above steps to enrich the bacteria;

[0045] 5. Prepare 1 / 2MS suspension (Table 2). Use 1 / 2MS suspension to suspend the bacteria. Shake the suspension up and down until the OD value of the permeate is between 0.8 and 1.0.

[0046] 6. Immerse the Arabidopsis buds in the infiltration solution for 45-60 seconds, return them to the tray, and keep them in the dark for 24 hours;

[0047] 7. Open the tray, continue to cultivate it, harvest the T1 generation seeds after they are fully mature, and dry them for storage;

[0048] 8. The harvested T1 generation seeds were cultured on 1 / 2MS solid culture dishes containing corresponding resistance (kanamycin resistance, concentration 25 mg / L) for the first step of screening. After one week of growth, the seeds that still maintained normal growth were preliminarily identified as positive seedlings. The Arabidopsis seedlings were screened and identified, and then DNA was extracted for PCR. Agarose gel electrophoresis was used to observe whether there were bands of the correct size. Homozygous transgenic Arabidopsis strains were obtained after the T3 generation, and RNA was finally extracted to measure their expression levels.

[0049] Table 2 Suspension formulation

[0050] drug Working fluid configuration / 1L Remark sucrose 50 g MES 0.5 g pH=5.6 MS salt 2.2 g 6-BA (1 mg / mL) 9.9 μL Silwet77 200 μL KOH Adjust pH to 5.7

[0051] (2) Steps for obtaining cotton gene-silenced lines

[0052] 1. Transform the recombinant vector pTRV2:GhSHMT10-A into competent Agrobacterium GV3101, place the plate in a 28°C incubator for 48 h, and select positive single clones for subsequent experiments by colony PCR detection;

[0053] 2. Plant the "TM-1" seeds in the soaked seedling blocks, place the seedling blocks evenly in the tray, and culture them in the greenhouse. The culture conditions are set at 25 / 23℃ and 16 / 8 h of light. The Agrobacterium solution can be injected only when the two cotyledons of the seedlings are flattened;

[0054] 3. Two days in advance, pick a single clone from the prepared bacteria pTRV2: GhSHMT10-A, pTRV1, pTRV2, and pTRV2:GhPDS, shake it in a 28°C shaker for 24 h, shake it slowly and then shake it vigorously. Cultivate it in 50 mL LB liquid medium containing the corresponding antibiotics (0.1% kanamycin + 0.1% rifampicin) at a ratio of 1:50, place it in a shaker, set the conditions to 200 rpm, 28°C, until the OD600 value of the bacterial solution is between 1.0 and 1.2, at which time the activity of the bacteria is the highest;

[0055] 4. Transfer the shaken bacterial solution to a sterilized 50 mL centrifuge tube, centrifuge at 4000 rpm for 10 min, and quickly discard the supernatant;

[0056] 5. Add an equal volume of the following resuspension solution (Table 3) to the centrifuge tube, vortex until the bacteria are completely dissolved in the resuspension solution, then adjust the OD600 to between 1.2-1.5 with the resuspension solution, and let it stand at room temperature in the dark for 2-3 hours;

[0057] Table 3 Resuspension formula (1L)

[0058] drug Working concentration Working fluid configuration / 1L Remark MgCl2.6H2O 10mM 2.033g MES 10mM 2.132g pH=5.6 AS (Acetosyringone) 200uM 39.24mg Dissolve in 2 mL DMSO

[0059] 6. Mix the resuspensions containing pTRV2:GhSHMT10-A, pTRV2, and pTRV2:GhPDS with the resuspension containing pTRV1 in a 1:1 ratio and shake gently;

[0060] 7. Select seedlings with consistent growth conditions and intact cotyledons for injection. Gently cut a hole on the back of the cotyledons and use a sterile syringe to slowly infiltrate the prepared solution into the cotyledons until it occupies more than 90% of the leaf area.

[0061] 8. After the injection is completed, the seedlings are placed in dark conditions for 24 hours of dark treatment at a temperature of 23°C. After the dark treatment, the seedlings are uncovered and placed in a culture room with a temperature of 25 / 23°C and a light intensity of 16 / 8 h for continued cultivation.

[0062] 9. About two weeks after infection (three-leaf stage), albino phenotype was observed in the leaves of pTRV2:GhPDS, indicating that the silencing system was successful; qRT-PCR was used to detect the expression level of GhSHMT10-A in pTRV2:GhSHMT10-A and pTRV2 lines.

[0063] 3. Phenotypic identification

[0064] Three homozygous lines with stable overexpression of GhSHMT10-A were obtained through genetic transformation of Arabidopsis thaliana, and the expression level of the overexpression lines was significantly increased ( Figure 7 The whole growth and development process was observed, and the results showed that the rosette leaf area and number of the three overexpressing Arabidopsis lines were greater than those of the wild type ( Figure 7 Statistical analysis showed that the number of rosette leaves, leaf length, leaf width and leaf area of ​​the overexpression line were significantly larger than those of the wild type ( Figure 7 In addition, the plant height of the overexpression line was significantly higher than that of the wild type at maturity, but the stem width was not significantly different from that of the wild type ( Figure 7 C in the figure); further statistics on fresh and dry weights revealed that the fresh and dry weights of the overexpression strains were greater than those of the wild type ( Figure 7 C in Figure ), indicating that overexpression of GhSHMT10-A may have accumulated more biomass. In addition, no significant difference was found in the silique length between the wild type and the three overexpression lines ( Figure 7 D in the figure). Further statistical analysis of Arabidopsis yield-related traits revealed that the seed length, seed width, and 1000-grain weight of the three overexpression lines were significantly higher than those of the wild type ( Figure 7 Therefore, these results suggest that overexpression GhSHMT10-A After injection, the plant height of Arabidopsis thaliana was increased and the biomass accumulation of Arabidopsis thaliana was increased. In the cotton gene-silenced strain, TRV was observed 14 days after injection: GDJ The strain showed albinism ( Figure 8 A in GhSHMT10-A Transcription levels were significantly reduced in the silenced strain ( Figure 8 B), indicating that the gene silencing system is successful. GhSHMT10-A The height difference between silent plants and empty plants was found GhSHMT10-A The height of the silent strain is shorter than that of the empty strain, and the dynamic statistical results also show that GhSHMT10-A The height of the silenced strain was significantly shorter than that of the empty vector ( Figure 8A and C in Figure 1). Dynamic statistics of each internode of different strains revealed that TRV: GhSHMT10-A The internodes of the plants except the first internode were significantly lower than those of the control plants 21 days after injection ( Figure 8 We further performed staining observation and statistical analysis on the stem cells of the second and third internodes of the silenced strains and empty-loaded plants 21 days after injection ( Fig. 9 The results show that the length and width of the stem cells in the second internode of the GhSHMT10-A silenced strain were significantly smaller than those in the control ( Fig. 9 B and C in the figure); while the width of the stem cells in the third internode did not show significant difference, the length was significantly smaller than that in the control ( Fig. 9 The above results show that silence GhSHMT10-A Genes can reduce cotton plant height by regulating culm cell size.

[0065] SEQ ID No.1:

[0066]

[0067] SEQ ID No.2:

[0068] MDPVSSWGNSSLDSVDPEIHDLIEKEKRRQCRGIELIASENFTSFAVIEALGSALTNKYSEGMPGNRYYGGNFIDEIENLCRSRAIQAFHLDPTKWGVNVQPYSGSPANFAAYTAVL QPHDRIMGLDLPSGGHLTHGYYTSGGKKISATSIYFESLPYKVNSTTGYIDYDKLEEKALDFRPKLIICGGSAYPRDWDYARFRAVADKCGALLLCDMAHISGLVAAQEANNPFEFCD IVTTTTHKSLRGPRAGMIFYRKGPKPPKKGQPEDAVYDFEDKINFAVFPSLQGGPHNHQIGALAVALKQSMTPGFKAYAKQVKANAVALGKYLMGKGYQLVTGGTENHLVLWDLRPLG LTGNKVEKLCDLCNITVNKNAVFGDSSALAPGGVRIGTPAMTSRGLVEKDFEQIGEFLHRAVTITLNIQKQYGKLLKDFNKGLDNNKEIQELKVDVEKFASSFDMPGFKMSEMKYKD*.

[0069] The above-described embodiment is only a preferred solution of the present invention and does not limit the present invention in any form. There are other variations and modifications without exceeding the technical solution described in the claims.

Claims

1. A plant height regulating gene in upland cotton GhSHMT10-A The application in regulating the growth of upland cotton is characterized by: Plant height regulatory genes in upland cotton GhSHMT10-A The encoded protein sequence is the amino acid sequence shown in SEQ ID NO:

2.

2. The use according to claim 1, characterized in that: GhSHMT10-A Positively regulate the plant height of upland cotton.

3. A method for dwarfing upland cotton, characterized in that: Plant height regulatory genes in upland cotton GhSHMT10-A Gene silencing is performed to reduce the plant height of upland cotton and achieve dwarfing; Upland cotton plant height regulation gene GhSHMT10-A The encoded protein sequence is the amino acid sequence shown in SEQ ID NO:2.

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