CN117821474A

CN117821474A - Gene for regulating and controlling early ripening property of upland cotton and application thereof

Info

Publication number: CN117821474A
Application number: CN202410184373.2A
Authority: CN
Inventors: 宿俊吉; 王彩香; 李颖; 袁文敏; 杨琦文; 魏楚
Original assignee: Gansu Agricultural University
Current assignee: Gansu Agricultural University
Priority date: 2024-02-19
Filing date: 2024-02-19
Publication date: 2024-04-05

Abstract

The invention discloses a gene for regulating and controlling early ripening traits of upland cotton and application thereof, and belongs to the technical field of genetic engineering. The genes comprise at least one of GhGAA 9 gene and GhGAA 14 gene; the nucleotide sequence of the GhGAA 9 gene is shown in SEQ ID NO. 1; the nucleotide sequence of the GhGAA 14 gene is shown in SEQ ID NO. 3. According to the invention, through constructing a VIGS silencing vector of a target gene and researching the biological function of regulating and controlling early ripening, the forward regulation of GhGAA 9 and GhGAA 14 genes on upland cotton early ripening response gibberellin is verified, the phenotype growth and development characters of cotton blooming, bud emergence, plant height and the like of upland cotton are improved, and important gene resources are provided for upland cotton early ripening breeding.

Description

Gene for regulating and controlling early ripening property of upland cotton and application thereof

Technical Field

The invention relates to the technical field of genetic engineering, in particular to a gene for regulating and controlling early ripening traits of upland cotton and application thereof.

Background

Cotton (Gossypium spp) is an economic crop that provides natural fibers, oil and vegetable proteins and plays a very important role in national economic development. The northwest inland is the region where the current Chinese cotton planting area is most dominant, and the air temperature in spring rises slowly and the final frost ends late, the air temperature in autumn falls quickly and the initial frost begins early, so that the accumulated temperature is insufficient, and the method is more suitable for planting early-maturing cotton varieties. The early maturing trait is one of the most important breeding target traits of cotton, and it not only determines whether cotton can mature normally during the harvest period, but also determines whether the variety is suitable for mechanical harvesting. In the early-maturing related traits, flowering time is one of the important features for evaluating cotton early maturing.

Gibberellin (GA) ₃ ) Is a tetracyclic diterpenoid plant growth factor which influences the induction of flowers and the development of flower organs. Whereas GASA (GA-Stimulated inArabidopsis) is a family of gibberellin regulation located downstream of DELLA, widely found in monocot and dicot species, has been extensively studied in some plant species, and therefore it has been found that GASAs are of great significance in response to gibberellin regulating flowering.

Disclosure of Invention

The invention aims to provide a gene for regulating and controlling early ripening traits of upland cotton and application thereof, so as to solve the problems in the prior art. The invention proves that the GhGAA 9 and GhGAA 14 genes positively regulate and control the early ripening response gibberellin of upland cotton, improve the phenotype growth and development characters of cotton bloom, bud, plant height and the like, and provide important gene resources for the early ripening breeding of upland cotton.

In order to achieve the above object, the present invention provides the following solutions:

the invention provides a gene for regulating and controlling the early ripening property of upland cotton, wherein the gene comprises at least one of GhGASA9 gene and GhGASA14 gene; the nucleotide sequence of the GhGAA 9 gene is shown in SEQ ID NO. 1; the nucleotide sequence of the GhGAA 14 gene is shown in SEQ ID NO. 3.

The invention also provides a biological material for silencing the gene.

Further, the biological material comprises a recombinant expression vector for silencing the gene and a recombinant bacterium comprising the recombinant expression vector.

The invention also provides an application of the gene or the biological material in regulating and controlling the early ripening of upland cotton, and the early ripening of upland cotton is delayed by silencing the gene.

Further, the delaying of upland cotton early ripening includes delaying bud time and flowering time of upland cotton.

The invention also provides an application of the gene or the biological material in reducing upland cotton plant height.

The invention also provides a method for delaying the bud time of upland cotton, which comprises the step of transforming the biological material into upland cotton and constructing a transgenic upland cotton plant for silencing the gene.

The invention also provides a method for delaying the flowering time of upland cotton, which comprises the step of transforming the biological material into upland cotton and constructing a transgenic upland cotton plant for silencing the gene.

The invention also provides a method for reducing the height character of upland cotton plants, which comprises the step of transforming the biological material into upland cotton to construct transgenic upland cotton plants for silencing the genes.

The invention discloses the following technical effects:

in order to further define the molecular mechanism of flowering regulation of GhGAA 9 and GhGAA 14 genes, the invention clones the target genes, constructs a VIGS silencing vector of the target genes, researches the biological functions of regulating precocity, and discovers that the GhGAA 9 and GhGAA 14 gene silencing leads to bud emergence and flowering time delay of upland cotton and influences the growth and development of upland cotton. After the gibberellin is sprayed from an external source, the plant height is obviously changed, the concentration of bud is improved, and flowering is promoted, so that the positive regulation of the GhGASA9 and GhGAA 14 genes to the early-maturing response gibberellin of the upland cotton is proved, the phenotype growth and development characters of the cotton, such as flowering, bud emergence, plant height and the like, are improved, and important gene resources are provided for the early-maturing breeding of the upland cotton.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a map of pEASY-T5 Zero vector and TRV vector;

FIG. 2 shows the result of amplification of a target gene;

FIG. 3 shows the result of PCR detection of the objective genetic bacteria solution;

FIG. 4 shows the results of double cleavage assay;

FIG. 5 shows the sequencing results of the cloning products of the genes GhGAA 9 and GhGAA 14;

FIG. 6 is a graph showing the detection of silencing efficiency of a gene of interest; wherein A is TRV, ghCLA1 positive control phenotype; b is the detection of the silencing efficiency of the target gene;

FIG. 7 is a phenotypic assay of GhGAA 9 and GhGAA 14 silenced plants; wherein A is seedling height; b is plant height; c is gibberellin treatment Group (GA) ₃ ) And plant height in control group (MOCK);

FIG. 8 comparison of flowering phenotypes for gene silencing; wherein A is flowering statistics result; b is bud to flowering period statistics; c is gibberellin treatment Group (GA) ₃ ) And flowering phenotype control under control group (MOCK).

Detailed Description

Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.

Examples

1. Materials and reagents

1.1 test materials

The material used in this example was cotton-in-cotton 113 (ZM 113), the seed was provided by cotton institute, national academy of agricultural sciences.

1.2 test reagents

The reagents used in this example are shown in Table 1:

table 1 reagents used in the experiments

1.3 Strain and vector

GV3101 Agrobacterium competent cells and pEASY-T5 Zero cloning vector (CT 501-01, DH 5. Alpha. Containing E.coli competent cells) were purchased from Shanghai Weidi Biotechnology Inc. and full gold Biotechnology Inc., respectively, and the VIGS vector systems for gene silencing (TRV: 00 (PYL 156), PYL192 and TRV: ghCLA 1) were given away by the cotton institute transgenic subject group of the national academy of agricultural sciences, and pEASY-T5 Zero vector maps are shown in FIG. 1.

1.4 preparation of solutions

a. The formulation of the medium is shown in Table 2:

table 2 culture medium formulation

Note that: the culture medium is sterilized at 121deg.C for 20min

b.100mL 50 xTAE buffer: 24.2g Tris+3.72g Na ₂ EDTA·2H ₂ O+5.71mL glacial acetic acid;

50mg/mL rifampicin (Rif) and 20mg/mL Acetosyringone (AS): respectively dissolving 5g of Rif powder and 2g of acetosyringone powder in 100mLDMSO solution, respectively filtering and sterilizing with 0.22 filter membrane, packaging, and preserving at-20deg.C;

d.0.5M MES: 10.65g MES powder was dissolved in 50mL ddH ₂ In O, regulating the pH value to 5.6 by NaOH, then fixing the volume to 100mL, filtering and sterilizing by using a filter membrane of 0.22, packaging, and preserving at 4 ℃;

e.1M MgCl ₂ : with 100mL ddH ₂ O-dissolution 9.521g MgCl ₂ Powder (amplified heat), 0.22 filter membrane for sterilization, and storing at 4deg.C;

f.500ml heavy suspension: 10mL 0.5M MES+1mL 20mg/mLAS+5mL of 1M MgCl ₂ Sterile ddH ₂ O was made up to 500mL.

1.5 use of instruments

The main instrument is as follows: visible spectrophotometers, high-speed refrigerated centrifuges, water baths, electrophoresis apparatuses, artificial climate boxes, gel cutting apparatuses, ultra-low temperature refrigerators, fluorescence quantitative apparatuses, gradient PCR apparatuses, sterilization pans, electronic balances, microwave ovens, table-type constant temperature shaking tables, ice-making machines, ultra-clean benches, biochemical incubators, ultra-micro spectrophotometers, ultra-pure water apparatuses and mini vortex mixing apparatuses.

2 test method

2.1 primer design

Cloning primers, qRT-PCR primers, and VIGS silencing primers (the size of the VIGS silencing primer product fragment is 300-500 bp) of GhGAA 9 and GhGAA 14 genes were designed by using NCBIPrimer-BLAST (https:// www.ncbi.nlm.nih.gov/tools/primer-BLAST /), and the primer sequences are shown in Table 3:

TABLE 3 list of primers used in the present invention

2.2 extraction of RNA

2.2.1RNA extraction and reverse transcription

RNA is extracted according to the instruction of the polysaccharide polyphenol plant total RNA extraction kit.

Reverse transcription (synthesis of the first strand of cDNA):

a. the RNA was removed from the-80℃refrigerator and thawed on ice as required, and 5 XFastKing-RT Supermix reagent and RNase-Free ddH were removed in the-20℃refrigerator ₂ O is melted on ice and is gently mixed;

b. the reaction system is shown in Table 4:

TABLE 4 reverse transcription reaction system

c. The reaction procedure is shown in Table 5:

table 5 reaction procedure

d. After the reaction, the purity and concentration of cDNA were measured, and the cDNA was packaged and stored at-20 ℃.

2.3 amplification of fragments of interest and ligation and transformation of cloning vectors

2.3.1 amplification of fragments of interest

The cDNA of Zhongcotton 113 (ZM 113) is used as a template, and Taq 2× PCR Mix with Dye V2 premix (containing dye) kit is used for amplifying the target gene, and the amplification system is shown in Table 6:

TABLE 6 amplification System

After the reaction solution is added according to the system, the mixture is gently mixed, centrifuged for a short time, and the reaction is carried out according to the reaction procedure of Table 7:

TABLE 7 amplification reaction procedure

After completion of the reaction, the size of the target gene was determined to be appropriate by agarose gel electrophoresis at 1.8%.

Cloning to obtain gene segment of upland cotton GhGAA 9, its nucleotide sequence is shown in SEQ ID NO. 1.

The cloned and obtained silent fragment in GhGAA 9 gene sequence has the nucleotide sequence shown in SEQ ID NO. 2.

SEQ ID NO.1：

ATGGCCATCTTTAAGGCTTTCATTGCTTCACTTCTCATCTCTCTTCTCCTTCTCCAACTTGTTGCTGC TGATCAACTGGTGACAAGTGCCAGCAAGAACAAGGGAACTGCTCCCCCTAAGAAAATTGACTGCGGCGGGGCTTGT GCGGCGAGGTGCCGATTATCATCGAGGCCACACCTATGCAAGAGGGCATGCGGGACATGTTGTGCACGCTGCAACT GTGTTCCACCGGGGACAGCCGGTAACCAAGAAATGTGTCCTTGCTATGCTAGCTTGACCACCCACGGTGGCCGACG CAAGTGCCCTTGA wherein the underlined part is a silent fragment.

SEQ ID NO.2：

GCCATCTTTAAGGCTTTCATTGCTTCACTTCTCATCTCTCTTCTCCTTCTCCAACTTGTTGCTGCTGATCAACTGGTGACAAGTGCCAGCAAGAACAAGGGAACTGCTCCCCCTAAGAAAATTGACTGCGGCGGGGCTTGTGCGGCGAGGTGCCGATTATCATCGAGGCCACACCTATGCAAGAGGGCATGCGGGACATGTTGTGCACGCTGCAACTGTGTTCCACCGGGGACAGCCGGTAACCAAGAAATGTGTCCTTGCTATGCTAGCTTGACCACCCACGGTGGCCGACGCAAGTGCCCTT。

Cloning fragment of GhGAA 14 gene of upland cotton, and its nucleotide sequence is shown in SEQ ID NO. 3.

The cloned and obtained silent fragment in GhGAA 14 gene sequence has the nucleotide sequence shown in SEQ ID NO. 4.

SEQ ID NO.3：

ATGGCTAAACTTGTTGCTACTTTCTTCTTGGCTCTCATTGCCATCTCCATGGTTCAAACTTTGGTTAT GGCATCACATGGACATGGTGGTCATCATTATGATAACAAACGTTATGGACCTGGAAGTCTCAAGCCTTACCAATGT CCATAACTCAGTGCACAAGGAGGTGTAGCCAGACACAGTACCATAAACCATGCATGTTCTTCTGTCAGAAATGTTG CAGGACATGCCTTTGTGTTCCTCCAGGGTATTATGGGAACAAACAAGTTTGCCCTTGCTACAACAACTGGAAAACC AAGGAAGGAGGCCCCAAATGCCCTTAA wherein the underlined section is a silent fragment.

SEQ ID NO.4：

TGCTACTTTCTTCTTGGCTCTCATTGCCATCTCCATGGTTCAAACTTTGGTTATGGCATCACATGGACATGGTGGTCATCATTATGATAACAAACGTTATGGACCTGGAAGTCTCAAGCCTTACCAATGTCCATAACTCAGTGCACAAGGAGGTGTAGCCAGACACAGTACCATAAACCATGCATGTTCTTCTGTCAGAAATGTTGCAGGACATGCCTTTGTGTTCCTCCAGGGTATTATGGGAACAAACAAGTTTGCCCTTGCTACAACAACTGGAAAACCAAGGAAGGAGGCCCCAAATG。

2.3.3 ligation of the Gene of interest to the cloning vector pEASY-T5 Zero

a. The pEASY-T5 Zero vector was removed from the-80℃refrigerator and thawed on ice.

b. The volume of the added target fragment (molar ratio of carrier to target fragment=1:5) was calculated and the reaction system shown in table 8 (whole procedure completed on ice) was added to a sterile 1.5mL centrifuge tube:

table 8 connection system

c. The mixture was gently shaken and mixed, and after a short centrifugation, it was connected at 25℃for 5min.

2.3.4 transformation of DH 5. Alpha. E.coli competent cells

The vector connected with the target gene is transformed into DH5 alpha escherichia coli competent cells by adopting a heat shock method, and bacterial liquid PCR verification and sequencing (completed by Shanghai biological engineering Co., ltd.) are carried out by using a target gene sequence primer.

2.4 construction of silencing vectors

Amplification of the silencing fragments was performed by adding restriction enzyme EcoRI and KpnI cleavage sites and silencing primers protecting bases to positive plasmid with 2.3.4 sequencing success as template, and the silencing fragments of GhGAA 9 and GhGAA 14 were inserted into silencing vectors by double cleavage method to construct TRV: ghGAA 9 and TRV: ghGAA 14 silencing vectors, the specific cleavage system being as shown in Table 9:

table 9 enzyme digestion System

And (3) after reacting for 15min at 37 ℃ according to the system, recovering a target gene fragment PCR product, recovering a large fragment by enzyme digestion of a carrier, connecting the target fragment with a silencing carrier, converting the connecting product into competent cells of escherichia coli, carrying out bacterial liquid PCR and double enzyme digestion identification, sequencing positive plasmids (Shanghai) after completion, and transferring the positive plasmids into the competence of agrobacterium GV 3101.

The procedure for transformation of Agrobacterium GV3101 was as follows:

a. GV3101 competent cells were removed from the-80℃ultra-low temperature refrigerator, thawed on ice, split into two tubes, 2. Mu.L of the successfully sequenced plasmid was aspirated, and added to centrifuge tubes for transformation.

b. After the above steps are completed, add350. Mu.L of LB liquid medium (without antibiotics), shaking culture for 2h (28 ℃,200 rpm), and uniformly coating the bacterial liquid on a solid medium (Kan addition) ⁺ And Rif antibiotics), in the dark, at 28℃for 2d.

c. After the completion of the culture, single colonies were picked into 5mL of LB liquid medium (Kan was added ⁺ And an if antibiotic), culturing for 16 hours according to the condition of shaking culture in the step b;

d. after the cultivation is completed, the bacterial liquid is preserved by using 50% glycerol (bacterial liquid: glycerol=1:1, v: v), and is preserved at 80 ℃ for standby; and (5) performing bacterial liquid PCR to confirm that the vector is positive.

2.5 upland cotton VIGS silencing target Gene

VIGS silencing of cotton 113 seedlings was performed as follows:

a. cotton 113 seeds are planted, when the seeds grow to the seventh day and cotyledons are fully unfolded, the seeds are soaked in water until nutrient soil in the flowerpot absorbs water to the surface, and then the soaking is stopped, and the seeds are placed for standby.

b. Adding Kan to LB liquid Medium ⁺ And Rif standby, wherein Kan ⁺ And the final concentration of Rif was 50 μg/mL and 25 μg/mL, respectively. The VIGS vector system and the objective gene bacterial liquid taken out from-80 ℃ were thawed on ice, at 28 ℃, activated at 200rpm for 16h (bacterial liquid: LB liquid medium=1:10, v:v). And after the activation is finished, the propagation is carried out according to the same proportion.

c. After the bacterial liquid is propagated, centrifuging for 10min at 5000rpm, pouring out supernatant, retaining bacterial cells, suspending bacterial cells by using a spectrophotometer with heavy suspension, and adjusting OD ₆₀₀ 1.8.

d. After resuspension was completed, the cells were left in the dark for 3 hours, and after resuscitating, PYL192 was mixed with 1:1 of the cell mass suspension containing TRV:00 (as a blank control group), TRV: ghCUA 1 (as a positive control group), TRV: ghGAA 9, and TRV: ghGAA 14 (as an experimental group), and thoroughly mixed.

e. On day 7 of the growth of upland cotton seedlings, they were soaked in water according to the method of step a. VIGS injections were performed on the eighth day of upland cotton seedling growth, with specific manipulations: and d, cutting the back of the cotyledon by using a 1mL syringe needle (the wound is not too large and the needle point is just large), injecting the mixed bacterial liquid in the step d into the upland cotton cotyledon, and filling the whole cotyledon with the bacterial liquid as much as possible.

f. After injection, in order to achieve better infection effect, the seeds are wrapped by a plastic bag, and are cultivated under normal growth conditions after being placed in darkness at 25 ℃ for 24 hours.

2.6 identification of silenced plants

After the positive control upland cotton seedlings whiten, adopting experimental groups and blank group upland cotton young leaves to carry out fluorescent quantitative experiments, detecting the silencing efficiency of the upland cotton young leaves, and carrying out phenotype observation at proper time.

3 results and analysis

3.1 construction of silencing vectors and Positive identification of silencing lines

3.1.1 construction of the Gene fragment of interest and silencing vector

The primer was designed using NCBI blast-primer, and the target fragment was amplified using Taq 2XPCRMix with Dye V2 enzyme of Eboltaaceae using cDNA of Zhongcotton 113 as a template, and recovered using a universal DNA purification recovery kit (Tiangen). The recovered target fragment was ligated with pEASY-T5 Zero vector to select for plasmid extraction. The target fragment (FIG. 2) was ligated with TRV:00 vector by double digestion and cultured, and positive plasmids were obtained by bacterial liquid PCR (FIG. 3), double digestion and sequencing (FIGS. 4 and 5). The target fragment ligated to the vector was obtained by sequencing without base deletion and substitution, and thus sequencing was successful. The positive plasmid was transformed into Agrobacterium by freeze thawing for cultivation.

3.1.2 detection of silencing efficiency of the Gene of interest and phenotypic analysis

Screening out candidate genes for the growth and development of upland cotton based on gene family data analysis. Therefore, the effect of the target gene in the flowering regulation of upland cotton is researched by utilizing the VIGS technology. The invention selects GhGAA 9 gene and GhGAA 14 gene for VIGS test. Silencing the target gene in cotton 113 in upland cotton by the method of epidermis injection under cotyledons, and finding that the positive control (TRV: ghCLA 1) plant starts to whiten on the 7 th day after infection, and the positive control whitens obviously on the 9 th day after silencing, which indicates that the upland cotton is successfully infected by agrobacterium. The silencing efficiency of the target gene is detected, and compared with a control group, the expression of GhGASA9 genes and GhGAA 14 genes in TRV: ghGASA9 and TRV: ghGAA 14 plants is obviously inhibited. Analysis of the expression levels of VIGS-silenced plants and control plants indicated that the gene of interest had been silenced (fig. 6).

3.1.3 phenotypic Effect of silenced plants

4-week-old plants of TRV: ghCUA 1, TRV:00, TRV: ghGASA9 and TRV: ghGASA14 were selected and cultured at 25 ℃. After that, the plant height, the bud and the flowering time were observed, and it was found that there was no significant difference in the plant height after silencing the GhGAA 9 gene or the GhGAA 14 gene, but the TRV:00 plant height was significantly increased after the gibberellin was applied externally (FIG. 7). After silencing the GhGAA 9 gene or GhGAA 14 gene, the bud and flowering times of upland cotton were delayed, affecting upland cotton growth (FIG. 8). After silencing the GhGAA 9 gene and the GhGAA 14 gene, the average bud time of upland cotton TRV: ghGAA 9 and TRV: ghGAA 14 plants was delayed for 4.8 days and 6.0 days, respectively, and flowering time was delayed for 4.23 days and 5.0 days, respectively, to reach significant levels. The bud time after gibberellin spraying is concentrated for 10-12 days, and the bud time has no obvious difference from TRV 00 flowering time, and the results show that the GhGAA 9 gene and the GhGAA 14 gene can regulate and control the flowering time of upland cotton and respond to gibberellin.

The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims

1. A gene for regulating early ripening of upland cotton, characterized in that the gene comprises at least one of the gene ghasa 9 and the gene ghasa 14; the nucleotide sequence of the GhGAA 9 gene is shown in SEQ ID NO. 1; the nucleotide sequence of the GhGAA 14 gene is shown in SEQ ID NO. 3.

2. A biological material that silences the gene of claim 1.

3. The biomaterial of claim 2, comprising a recombinant expression vector that silences a gene of claim 1 and a recombinant bacterium comprising the recombinant expression vector.

4. Use of a gene according to claim 1 or a biomaterial according to claim 2 or 3 for regulating the early ripening behaviour of upland cotton, characterized in that early ripening of upland cotton is delayed by silencing the gene.

5. The use of claim 4, wherein delaying early ripening of upland cotton comprises delaying bud time and flowering time of upland cotton.

6. Use of the gene of claim 1 or the biomaterial of claim 2 or 3 for reducing upland cotton height traits.

7. A method of delaying the emergence of upland cotton, comprising the step of transforming the biological material of claim 2 or 3 into upland cotton to construct a transgenic upland cotton plant that silences said gene.

8. A method for delaying flowering time of upland cotton, comprising the step of transforming the biological material of claim 2 or 3 into upland cotton to construct a transgenic upland cotton plant that silences said gene.

9. A method for reducing upland cotton plant height traits comprising the step of transforming the biological material of claim 2 or 3 into upland cotton to construct a transgenic upland cotton plant that silences said gene.