Application of protein GbCET2 and coding gene thereof in regulating and controlling initial fruit branch position and initial fruit branch height of gossypium barbadense

Abstract

The invention discloses application of a protein GbCET2 and a coding gene thereof in regulation and control of an initial fruit branch position and an initial fruit branch height of gossypium barbadense. The invention belongs to the field of botany, and particularly relates to application of a protein GbCET2 and a coding gene thereof to regulation and control of the initial fruit branch position and the initial fruit branch height of island cotton. The protein GbCET2 or a substance for regulating and controlling the expression of the coding gene of the protein or a substance for regulating and controlling the activity or content of the protein can be applied to any one of the following aspects: 1) regulating and controlling the initial fruit branch knot position and the initial fruit branch height of a plant; 2) preparing a product for regulating the initial fruit branch position and the initial fruit branch height of the plant; the method comprises the following steps of 1, preparing GbCET2 in sea island cotton, 2, culturing a plant with the initial fruit branch position and the initial fruit branch height changed, 3, culturing the plant with the initial fruit branch position and the initial fruit branch height changed, 4, preparing a product for culturing the plant with the initial fruit branch position and the initial fruit branch height changed, and 5, breeding the plant. After the expression of GbCET2 is reduced in the sea island cotton by utilizing a virus-induced gene silencing technology, the initial fruit branch position and the initial fruit branch height of the sea island cotton are remarkably reduced.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of botany, and particularly relates to application of protein GbCET2 and its coding gene in regulating branch node and height of initial fruit branch of Gossypium barbadense. BACKGROUND

[0002] As a representative of high-quality long-staple cotton, the fiber quality of Gossypium barbadense is significantly better than that of other cotton varieties, and it is the core raw material of high-end textile industry. In China, due to the short frost-free period and limited climate conditions, most of the cultivated varieties have the advantage of early maturity. Generally, the branch node and height of initial fruit branch of early-maturing cotton varieties are also lower.

[0003] At the same time, the branch node and height of initial fruit branch are also the core agronomic traits of Gossypium barbadense plant architecture, which directly affect the adaptability of mechanical harvesting. The branch node and height of initial fruit branch are positively correlated, and are key variables affecting the efficiency of mechanical harvesting.

[0004] At present, the genetic regulation of the branch node and height of initial fruit branch of Gossypium barbadense is faced with the bottleneck of lack of gene resources, unknown regulation mechanism, and limitation of molecular markers. Therefore, exploring the protein and its coding gene that specifically regulate the branch node and height of initial fruit branch can not only analyze the molecular basis of plant architecture, but also provide target resources for cultivating Gossypium barbadense varieties with "suitable mechanical harvesting, high yield, and high quality". SUMMARY

[0005] The technical problem to be solved by the present application is how to regulate the branch node and height of initial fruit branch of plants.

[0006] In order to solve the problems in the prior art, the present application provides applications of a protein, a substance for regulating expression of a coding gene of the protein, or a substance for regulating activity or content of the protein in any of the following: 1) in regulating the branch node and height of initial fruit branch of plants; 2) in preparing a product for regulating the branch node and height of initial fruit branch of plants; 3) in cultivating plants with changed branch node and height of initial fruit branch; 4) in preparing a product for cultivating plants with changed branch node and height of initial fruit branch; 5) in plant breeding.

[0007] The protein is any of the following proteins: a1) a protein with an amino acid sequence of SEQ ID No: 3; a2) a protein with an amino acid sequence of SEQ ID No: 3, which has one or more substitutions, deletions, and / or additions of amino acid residues and has the same function; a3) a protein having 80% or more identity to any one of the amino acid sequences defined in a1) - (a2) and having the same function; a4) a fusion protein obtained by linking a terminal tag to any one of the proteins defined in a1) - (a3).

[0008] In the above protein, the protein tag refers to a polypeptide or protein fused and expressed with a target protein by using DNA in vitro recombination technology, so as to facilitate the expression, detection, tracing and / or purification of the target protein. The protein tag can be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag and / or a SUMO tag, etc.

[0009] In the above protein, the identity refers to the identity of the amino acid sequence. The identity of the amino acid sequence can be determined by using a homology search site on the Internet, such as the BLAST webpage of the NCBI homepage. For example, the identity of a pair of amino acid sequences can be calculated by using blastp as the program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as the Matrix, setting Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default values) respectively, and performing a search in Advanced BLAST 2.1, and then the value of the identity (%) can be obtained.

[0010] In the above protein, the 80% or more identity can be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

[0011] In the above protein, SEQ ID No. 3 consists of 174 amino acid residues. It is named as GbCET2 protein, and its encoding gene is GbCET2 GbCET2.

[0012] In the above application, the protein is derived from Gossypium barbadense (Gossypium hirsutum). Gossypium barbadense L.

[0013] Herein, the substance regulating the activity and / or content of the protein can be a substance regulating the expression of a gene encoding the protein GbCET2.

[0014] In the above, the substance that regulates the expression of the gene can be a substance that performs at least one of the following six regulations: 1) regulation at the transcription level of the gene; 2) regulation after the transcription of the gene (i.e. regulation of the splicing or processing of the primary transcript of the gene); 3) regulation of the RNA transport of the gene (i.e. regulation of the transport of the mRNA of the gene from the nucleus to the cytoplasm); 4) regulation of the translation of the gene; 5) regulation of the degradation of the mRNA of the gene; and 6) post-translational regulation of the gene (i.e. regulation of the activity of the protein translated from the gene).

[0015] In the present application, the regulation can be up-regulation or enhancement or increase; the regulation can also be down-regulation or weakening or decrease.

[0016] In the present application, the enhancement, increase or up-regulation of the expression of the gene encoding the protein described above in the recipient plant, or / and the enhancement, increase or up-regulation of the activity and / or content of the gene encoding the protein described above is achieved by introducing the gene encoding the protein described above into the recipient plant.

[0017] In the present application, the regulation of the expression of the gene encoding the protein described above can be inhibition or decrease or down-regulation of the expression of the gene. The inhibition or decrease or down-regulation of the expression of the gene can be achieved by gene knockout or gene silencing.

[0018] The gene knockout refers to the phenomenon that a specific target gene is inactivated by gene editing technology. The gene knockout inactivates a specific target gene through changes in DNA sequences, including but not limited to Zinc-finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN) and CRISPR / Cas system. CRISPR (clustered regulatory interspaced short palindromic repeat) is a site containing multiple short repeat sequences in the genome. Cas9 protein can cut the target sequence recognized by crRNA-tracrRNA under the mediation of RNA.

[0019] The gene silencing refers to a phenomenon that a gene is not expressed or lowly expressed without damaging the original DNA. The gene silencing is premised on not changing the DNA sequence, and makes the gene not expressed or lowly expressed. The gene silencing can occur in two levels. One is the transcription level gene silencing caused by DNA methylation, heterochromatinization, and position effect, etc. The other is the post-transcription gene silencing, i.e. the gene is inactivated by specifically inhibiting the target RNA after the gene is transcribed, including antisense RNA, co-suppression, quelling, RNA interference (RNAi), and micro RNA (miRNA) mediated translation inhibition, etc.

[0020] In the above application, the substance for regulating the expression of the gene encoding the protein or the substance for regulating the activity or content of the protein can be a biological material related to the protein described above, and the biological material can be any one of the following: c1) a nucleic acid molecule encoding the protein described above; c2) an expression cassette containing the nucleic acid molecule of c1); c3) a recombinant vector containing the nucleic acid molecule of c1), or a recombinant vector containing the expression cassette of c2); c4) a recombinant microorganism containing the nucleic acid molecule of c1), or a recombinant microorganism containing the expression cassette of c2), or a recombinant microorganism containing the recombinant vector of c3); c5) a transgenic plant cell line containing the nucleic acid molecule of c1), or a transgenic plant cell line containing the expression cassette of c2); c6) a transgenic plant tissue containing the nucleic acid molecule of c1), or a transgenic plant tissue containing the expression cassette of c2); c7) a transgenic plant organ containing the nucleic acid molecule of c1), or a transgenic plant organ containing the expression cassette of c2); e1) a nucleic acid molecule for inhibiting or reducing or silencing the expression of the gene encoding the protein described above; e2) an expression cassette containing the nucleic acid molecule of e1); e3) a recombinant vector containing the nucleic acid molecule of e1), or a recombinant vector containing the expression cassette of e2); e4) a recombinant microorganism containing the nucleic acid molecule of e1), or a recombinant microorganism containing the expression cassette of e2), or a recombinant microorganism containing the recombinant vector of e3); e5) a transgenic plant cell line containing the nucleic acid molecule of e1), or a transgenic plant cell line containing the expression cassette of e2); e6) a transgenic plant tissue containing the nucleic acid molecule of e1), or a transgenic plant tissue containing the expression cassette of e2); e7) a transgenic plant organ comprising the nucleic acid molecule of e1), or a transgenic plant organ comprising the expression cassette of e2).

[0021] In the above uses, the nucleic acid molecule of c1) can be any one of the following DNA molecules, d1) a DNA molecule having the nucleotide sequence of SEQ ID No: 2; d2) a DNA molecule having the coding sequence of SEQ ID No: 1; d3) a DNA molecule having 90% or more identity to the nucleotide sequence defined in d1) or d2), and encoding the protein as described above; d4) a DNA molecule hybridizing to the nucleotide sequence defined in d1) or d2) under stringent conditions, and encoding the protein as described above.

[0022] In the above uses, the nucleic acid molecule of e1) can be a DNA molecule having the nucleotide sequence of SEQ ID No: 2.

[0023] The nucleic acid molecule described herein can be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule can also be RNA, such as gRNA, mRNA, siRNA, shRNA, sgRNA, miRNA or antisense RNA.

[0024] The vectors described herein are well known to those skilled in the art, and include but are not limited to: plasmids, bacteriophages (such as lambda phage or M13 filamentous phage, etc.), cosmids (i.e. cosmids), Ti plasmids or viral vectors.

[0025] The recombinant expression vector containing the gene can be constructed using existing plant expression vectors. GbCET2 The plant expression vectors include but are not limited to Agrobacterium binary vectors and vectors that can be used for plant microprojectile bombardment, etc. The plant expression vectors can also contain the 3' untranslated region of the foreign gene, i.e. containing the polyadenylation signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylation signal can guide the addition of polyadenylate to the 3' end of the mRNA precursor, such as the 3' untranslated region of the Agrobacterium crown gall induced (Ti) plasmid gene (such as the nopaline synthase Nos gene), the plant gene (such as the soybean storage protein gene) all have similar functions.

[0026] The recombinant expression vector containing the gene can be constructed using existing plant expression vectors. GbCET2In constructing a recombinant plant expression vector, any one of the enhancer promoters or constitutive promoters can be added before the transcription initiation nucleotide, including but not limited to, such as the cauliflower mosaic virus (CAMV) 35S promoter, the ubiquitin promoter of corn, which can be used alone or in combination with other plant promoters; in addition, when using the gene construction plant expression vector of the present application, enhancers, including translation enhancers or transcription enhancers, can also be used, and these enhancer regions can be ATG start codon or adjacent regions start codon, etc., but must be the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The source of the translation control signal and the start codon is wide, which can be natural or synthetic. The translation initiation region can be from the transcription initiation region or the structural gene.

[0027] The present application provides a method for changing the position of the fruiting branch and the height of the fruiting branch of a plant, which comprises the following steps M or P: The step M is to inhibit or reduce or silence the activity and / or content of the protein described above in the plant of interest, or / and, to inhibit or reduce or down-regulate the expression amount of the gene encoding the protein described above, so as to reduce the position of the fruiting branch and the height of the fruiting branch of the plant; the position of the fruiting branch and the height of the fruiting branch of the plant with reduced position of the fruiting branch and height of the fruiting branch are lower than those of the plant of interest.

[0028] The step P is to enhance, increase or up-regulate the activity and / or content of the protein described above in the plant of interest, or / and, to enhance, increase or up-regulate the expression amount of the gene encoding the protein described above, so as to increase the position of the fruiting branch and the height of the fruiting branch of the plant; the position of the fruiting branch and the height of the fruiting branch of the plant with increased position of the fruiting branch and height of the fruiting branch are higher than those of the plant of interest.

[0029] In the above method, the reduction of the expression amount and / or activity of the gene encoding the protein GbCET2 in the plant of interest can be achieved by using gene mutation, gene knockout, gene editing or gene knockdown technology to reduce or inactivate the activity of the gene encoding the protein GbCET2 in the genome of the plant of interest.

[0030] In a specific embodiment, the inhibition or reduction or silencing of the expression of the gene encoding the protein described above in the plant of interest comprises introducing an inhibitory or reducing or silencing nucleic acid molecule, expression cassette or recombinant vector described above into the plant of interest, to obtain a plant with reduced position of the fruiting branch and height of the fruiting branch.

[0031] The inhibitory or reducing or silencing nucleic acid molecule described above can be a DNA molecule as shown in SEQ ID No: 4.

[0032] As a specific embodiment, the recombinant vector is a recombinant vector pCLCrVA- GbCET2The recombinant vector pCLCrVA- GbCET2 It is the sequence of the pCLCrVA vector (starting vector). Spe I and Pac The fragments between the I recognition sites are replaced with the DNA molecule shown in SEQ ID No:4, while keeping the other nucleotides of the pCLCrVA vector (starting vector) unchanged to obtain the recombinant vector.

[0033] In this article, the breeding objectives include cultivating plants with lower initial fruiting branch node position and initial fruiting branch height; the breeding objectives also include cultivating plants with higher initial fruiting branch node position and initial fruiting branch height.

[0034] The present invention also provides a method for reducing the node position and height of the first fruiting branch of a plant, comprising inhibiting, reducing or silencing the expression level of the gene encoding the protein described above in the target plant, and / or the activity and / or content of the protein, thereby reducing the node position and height of the first fruiting branch of the plant.

[0035] In the above applications or methods, the plant is any one of the following: N1) Dicotyledons; N2) Malvales (Malva orders) N3) Malvaceae family plants; N4) Plants of the genus *Gossypium*; N5) Cotton.

[0036] The cotton mentioned above is Sea Island cotton ( Gossypium barbadense L.).

[0037] This invention identified a gene from sea island cotton. GbCET2 In genes GbCET2 The fourth exon contains a SNP site (Gbar_D07_15803388) that is highly associated with the initial fruiting branch node; this variation affects the amino acid sequence of the protein encoded by this gene. Virus-induced gene silencing (VIGS) technology was used to reduce [the gene's] in Sea Island cotton. GbCET2 After expression of this gene, the node position of the first fruiting branch was significantly reduced; simultaneously, the height of the first fruiting branch was also significantly reduced. This gene encodes a phosphatidylethanolamine-binding protein, a novel protein that regulates the node position and height of the first fruiting branch in sea island cotton, and has significant value for production applications. Attached Figure Description

[0038] Figure 1 for GbCET2 Locations of key variations in gene and protein sequences. 'a' represents two different haplotypes. GbCET2The genome sequence of the gene. Gray lines mark exons, and black lines mark introns. The C / T in the fourth exon is the key nonsynonymous SNP (Gbar_D07_15803388). The reference type (Ref) is C, corresponding to the codon CCA; the variant type (Alt) is T, corresponding to the codon TCA. b represents two different haplotypes. GbCET2 The protein sequence of the gene. The amino acid (P / S) encoded by the key nonsynonymous SNP variant (C / T) is located within the phosphatidylethanolamine-binding protein (PBP) domain. The reference translation uses proline (Pro, abbreviated as P); the variant translation uses serine (S, abbreviated as S).

[0039] Figure 2 To detect VIGS in Sea Island cotton using qRT-PCR GbCET2 A bar chart representing the levels. WT, pCLCrVA, and pCLCrVA- GbCET2 These represent wild-type control of sea island cotton, VIGS empty control, and VIGS silenced control, respectively. GbCET2 Positive single plants.

[0040] Figure 3 To utilize VIGS silence GbCET2 The initial fruiting node of the later sea island cotton. WT, pCLCrVA and pCLCrVA- GbCET2 These represent wild-type control of sea island cotton, VIGS empty control, and VIGS silenced control, respectively. GbCET2 Positive single plants.

[0041] Figure 4 To utilize VIGS silence GbCET2 The height of the first fruiting branch in post-sea island cotton. WT, pCLCrVA, and pCLCrVA- GbCET2 These represent wild-type control of sea island cotton, VIGS empty control, and VIGS silenced control, respectively. GbCET2 Positive single plants. Detailed Implementation

[0042] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0043] The experimental methods in the following examples are all routine methods, and are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained commercially, unless otherwise specified.

[0044] The quantitative tests in the following examples are all set up in triplicate, and the results are averaged, unless otherwise specified.

[0045] The pCLCrVA plasmid in the following examples has been described in Zhao N, et al. Genomic and GWAS analyses demonstrate phylogenomic relationships of Gossypium barbadense Gossypium barbadense L. in China and selection for fibre length, lint percentage and Fusarium wilt resistance. Plant Biotechnol J. 2022;20(4):691-710. The biological material is available from the applicant, and can only be used for repeating the experiments of the present application, and cannot be used for other purposes.

[0046] The Gossypium barbadense L. in the following examples has been described in Zhao N, et al. Genomic and GWAS analyses demonstrate phylogenomic relationships of Gossypium barbadense Gossypium barbadense L. in China and selection for fibre length, lint percentage and Fusarium wilt resistance. Plant Biotechnol J. 2022;20(4):691-710. The biological material is available from the applicant, and can only be used for repeating the experiments of the present application, and cannot be used for other purposes.

[0047] The data in the following examples are processed using IBM SPSS_Statistics_25 statistical software, and the experimental results are expressed as mean values, and T-Test is used for inspection, * indicates that the difference is statistically significant (P < 0.05), ** indicates that the difference is highly statistically significant (P < 0.01), and *** indicates that the difference is extremely statistically significant (P < 0.001).

[0048] Example 1: Obtaining of island cotton fruiting branch node regulation genes GbCET2 The gene is from Gossypium barbadense, GbCET2 The CDS of the gene is SEQ ID No: 1, the genome sequence is SEQ ID No: 2, the encoded protein is named as GbCET2 protein or protein GbCET2, and the amino acid sequence of the encoded protein is shown as SEQ ID No: 3.

[0049] The SNP (Gbar_D07_15803388) significantly associated with the node position of the initial fruit branch of Gossypium barbadense is located in the fourth exon of the gene GbCET2 (especially at position 337 of SEQ ID No: 1) Figure 1 , which can cause the change of the amino acid sequence of the encoded protein Figure 1 . When the position 337 of the CDS of the gene (SEQ ID No: 1) is C, the position 113 of the amino acid sequence of the encoded GbCET2 protein (SEQ ID No: 3) is proline (Pro, one-letter abbreviation P); it is found that when the position 337 of the CDS of the gene (SEQ ID No: 1) is T, the position 113 of the amino acid sequence of the encoded GbCET2 protein (SEQ ID No: 3) is serine (Ser, one-letter abbreviation S). GbCET2 GbCET2 It is speculated that the gene is a key regulatory gene significantly associated with the node position of the initial fruit branch of Gossypium barbadense. GbCET2

[0050] Example 2, VIGS silencing GbCET2 of the initial fruit branch of Gossypium barbadense 1. Construction of VIGS recombinant vector and obtaining of recombinant strain GbCET2 According to the CDS of the gene , primers were designed for VIGS vector construction, an GbCET2 I restriction site (ACTAGT) was added at the 5' end of the forward primer F, and the nucleotide sequence of the forward primer F was: 5'-GTTGGGGAGAGTGATTGGGG-3'; an Spe I restriction site (TTAATTAA) was added at the 5' end of the reverse primer R, and the nucleotide sequence of the reverse primer was: 5'-GCGTCTTCTAGCAGCTGTTTC-3'. Pac

[0051] The structure of the recombinant plasmid pCLCrVA- GbCET2 is described as follows: to insert the pCLCrVA vector sequence Spe I and Pac ​​​I The fragment between the recognition sites was replaced with the DNA molecule shown in SEQ ID No: 4, and the other nucleotides of the pCLCrVA vector were kept unchanged to obtain a recombinant vector.

[0052] Recombinant vector pCLCrVA- GbCET2 After construction, it was transformed into Agrobacterium EHA105 strain (Shengong Bioengineering Co., Ltd., product number B528432) by heat shock to obtain Agrobacterium EHA105 containing pCLCrVA- GbCET2 plasmid, named Agrobacterium tumefaciens EHA105 / pCLCrVA- GbCET2 .

[0053] The empty vector pCLCrVA was heat-shocked into Agrobacterium EHA105 strain to obtain EHA105 / pCLCrVA as an empty control.

[0054] The vector pCLCrVB was heat-shocked into Agrobacterium EHA105 strain to obtain EHA105 / pCLCrVB. Before transformation, they were mixed with Agrobacterium containing EHA105 / pCLCrVA- GbCET2 , EHA105 / pCLCrVA in equal volume, respectively.

[0055] 2、 GbCET2 Obtaining of silenced Gossypium barbadense plants and identification of primordial fruit branch node and primordial fruit branch height Gossypium barbadense was planted by soil culture method as a transgenic receptor. Two weeks later, the cotyledons were completely flat, and genetic transformation was performed.

[0056] The strain EHA105 / pCLCrVA- GbCET2 , EHA105 / pCLCrVA and EHA105 / pCLCrVB obtained in step 1 were cultured at 28°C to the logarithmic growth phase; centrifuged at 8000 rpm for 5 min, the bacterial cells were collected, and the bacterial cells were resuspended with VIGS infection solution (10 mM MES, 200 µM AS, 10 mM MgCl2), and the bacterial solution concentration was adjusted to OD 600 =1.0 or so; EHA105 / pCLCrVA- GbCET2 , EHA105 / pCLCrVA were mixed with EHA105 / pCLCrVB bacterial solution at a volume ratio of 1:1, and after standing at room temperature for 3 h, they were used for transformation of Gossypium barbadense leaves. The mixed bacterial solution of EHA105 / pCLCrVA and EHA105 / pCLCrVB was used as an empty control.

[0057] Using 1 mL sterile syringe, the bacterial solution was taken up, a tiny wound was first made on the back of cotyledon with the needle without penetrating the leaf, and then the inoculation was performed with the syringe without needle. The G. hirsutum plants that had finished the injection were placed in the 28℃ greenhouse with 16 h / 8 h light-dark cycle. The experiment was repeated three times, and 20 plants were planted for each strain each time.

[0058] Two weeks later, the leaf samples of G. hirsutum wild type control (WT), VIGS empty control (pCLCrVA) and VIGS silenced GbCET2 pCLCrVA- GbCET2 were taken, and the RNA was extracted and reverse transcribed into cDNA, and then the Real-time PCR was used to detect the silencing efficiency of the gene GbCET2 (Forward primer 5'-ATATCCCCGGCACAACAGATGC-3'; Reverse primer 5'-CACTGTTTGCCTGCCTTTTTGC-3'). The results showed that the transcriptional expression level of GbCET2 in the pCLCrVA- GbCET2 plants was extremely significantly reduced after the VIGS treatment. Figure 2 .

[0059] The primordial fruit branch node of G. hirsutum plant was defined as the main stem node where the first fruit branch was born from the bottom to the top after the cotton flower bud appeared.

[0060] The primordial fruit branch height of G. hirsutum plant was defined as the height from the cotyledon node to the main stem where the first fruit branch was born.

[0061] GbCET2 The primordial fruit branch node of the silenced G. hirsutum plant (average 3 nodes) was extremely significantly shortened compared with the primordial fruit branch node of the control plant, and was reduced by one node compared with the wild type control (average 4 nodes) and the VIGS empty control (average 4 nodes). Figure 3 Therefore, it was confirmed that GbCET2 positively regulated the primordial fruit branch node of G. hirsutum.

[0062] In addition, GbCET2 the primordial fruit branch height of the silenced G. hirsutum plant (average 13.6 cm) was significantly reduced by 6.8 cm and 7.8 cm compared with the primordial fruit branch height of the wild type control (average 20.4 cm) and the VIGS empty control (average 21.4 cm) respectively. Figure 4 Therefore, it can be seen that GbCET2 positively regulated the primordial fruit branch height of G. hirsutum.

[0063] The application has been described in detail. For those skilled in the art, the application can be implemented in a wider range under the same parameters, concentrations and conditions without departing from the spirit and scope of the application and without unnecessary experiments. Although the application gives a special example, it should be understood that the application can be further improved. In summary, according to the principle of the application, the application intends to include any change, use or improvement of the application, including the change made by the conventional technology known in the art, which is out of the range disclosed in the application.

Claims

1. The use of a protein or a substance that regulates the expression of the gene encoding the protein or a substance that regulates the activity or content of the protein in any of the following: 1) Application in regulating the node position and height of the first fruiting branch in plants; 2) Application in the preparation of products that regulate the node position and height of the first fruiting branch in plants; 3) Application in cultivating plants with altered node positions and heights of the initial fruiting branches; 4) Application in the preparation of products from plants that have altered the node position and height of the initial fruiting branch; 5) Applications in plant breeding; The protein is any of the following proteins: a1) A protein with the amino acid sequence SEQ ID No:3; a2) A protein having the same function as the amino acid sequence shown in SEQ ID No:3, but with one or more amino acid residues substituted and / or deleted and / or added. a3) Proteins that share more than 80% identity with the amino acid sequence defined by a1) or a2) and have the same function; a4) A fusion protein obtained by attaching a tag to the end of any of the proteins defined in a1)-a3).

2. The application according to claim 1, characterized in that, The protein is derived from cotton.

3. The application according to claim 1 or 2, characterized in that, The substance that regulates gene expression or the substance that regulates the activity or content of the protein is a biological material related to the protein in the application of claim 1 or 2, and the biological material is any one of the following: c1) The nucleic acid molecule encoding the protein; c2) An expression cassette containing the nucleic acid molecule described in c1); c3) A recombinant vector containing the nucleic acid molecule described in c1), or a recombinant vector containing the expression cassette described in c2); c4) Recombinant microorganisms containing the nucleic acid molecules described in c1), or recombinant microorganisms containing the expression cassette described in c2), or recombinant microorganisms containing the recombinant vector described in c3); c5) A transgenic plant cell line containing the nucleic acid molecule described in c1), or a transgenic plant cell line containing the expression cassette described in c2); c6) Transgenic plant tissue containing the nucleic acid molecules described in c1), or transgenic plant tissue containing the expression cassette described in c2); c7) A transgenic plant organ containing the nucleic acid molecule described in c1), or a transgenic plant organ containing the expression cassette described in c2); e1) Nucleic acid molecules that inhibit, reduce, or silence the expression of the protein-encoding gene; e2) An expression cassette containing the nucleic acid molecule described in e1); e3) A recombinant vector containing the nucleic acid molecule described in e1), or a recombinant vector containing the expression cassette described in e2); e4) Recombinant microorganisms containing the nucleic acid molecules described in e1), or recombinant microorganisms containing the expression cassette described in e2), or recombinant microorganisms containing the recombinant vector described in e3); e5) A transgenic plant cell line containing the nucleic acid molecule described in e1), or a transgenic plant cell line containing the expression cassette described in e2); e6) Transgenic plant tissue containing the nucleic acid molecules described in e1), or transgenic plant tissue containing the expression cassette described in e2); e7) A transgenic plant organ containing the nucleic acid molecule described in e1) or a transgenic plant organ containing the expression cassette described in e2).

4. The application according to claim 3, characterized in that: c1) The nucleic acid molecule is any of the following DNA molecules. d1) The nucleotide sequence is the DNA molecule shown in SEQ ID No:2; d2) The coding sequence is the DNA molecule shown in SEQ ID No:1; d3) has 90% or more identity with the nucleotide sequence defined by d1) or d2) and is a DNA molecule encoding the protein of claim 1; d4) Hybridizes under stringent conditions to a nucleotide sequence defined by d1) or d2) and encodes a DNA molecule that encodes the protein of claim 1.

5. A method for altering the node position and height of the initial fruiting branch of a plant, characterized in that: The method includes step M or P, wherein step M is to inhibit or reduce or silence the activity and / or content of the protein described in claim 1 or 2 in the target plant, or / and, inhibit or reduce or downregulate the expression level of the gene encoding the protein described in claim 1 or 2, so as to reduce the node position and height of the first fruiting branch of the plant. The method includes step P, which is to enhance, increase or upregulate the activity and / or content of the protein described in claim 1 or 2 in the target plant, or / and enhance, increase or upregulate the expression level of the gene encoding the protein described in claim 1 or 2, so as to increase the node position and height of the first fruiting branch of the plant.

6. A method for cultivating plants with reduced initial fruiting branch node position and initial fruiting branch height, characterized in that, This includes inhibiting, reducing, or silencing the expression level of the gene encoding the protein described in claim 1 or 2 in the target plant, and / or, the activity and / or content of the protein, to obtain a plant with a reduced initial fruiting branch node position and initial fruiting branch height, wherein the initial fruiting branch node position and initial fruiting branch height of the plant with the reduced initial fruiting branch node position and initial fruiting branch height are lower than those of the target plant.

7. The method according to claim 6, characterized in that, The inhibition, reduction, or silencing of the expression of the protein encoding gene of claim 1 or 2 in the plant comprises introducing the nucleic acid molecule of claim 4 (e1), the expression cassette of claim 4 (e2), or the recombinant vector of claim 4 (e3) into the target plant to obtain a plant with reduced node position and height of the initial fruiting branch.

8. A method for cultivating plants with increased node position and height of the initial fruiting branch, characterized in that, This includes enhancing, increasing, or upregulating the expression level of the gene encoding the protein described in claim 1 or 2 in the target plant, and / or, the activity and / or content of the protein to obtain a plant with an increased initial fruiting branch node position and initial fruiting branch height, wherein the initial fruiting branch node position and initial fruiting branch height of the plant with the increased initial fruiting branch node position and initial fruiting branch height are higher than those of the target plant.

9. The protein as described in claim 1 or 2 and / or the biomaterial as described in claim 3 or 4.

10. The method according to any one of claims 5-8, characterized in that, The plant is any one of the following: N1) Dicotyledons; N2) Malvales (Malva orders) N3) Malvaceae family plants; N4) Plants of the genus *Gossypium*; N5) Cotton.

Images