Wheat dwarf creeping allele as well as encoding protein and application thereof

Abstract

The invention discloses a wheat dwarf creeping allele and an encoding protein and application thereof, and belongs to the technical field of genetic engineering. In particular discloses a protein TaRHT-A1p (SEQ ID NO: 1), a TaRHT-A1p gene (SEQ ID NO: 2) and application of the TaRHT-A1p gene and the TaRHT-A1p gene. Experiments prove that by increasing the content and / or activity of the protein TaRHT-A1p in the target plant, the plant height of the plant can be remarkably reduced, the tillering angle of the plant can be increased, and the gravity sensing ability of the plant and / or the gravity response of the plant can be weakened. The dwarf wheat of which the plant height is remarkably reduced and the tillering angle is remarkably increased is successfully created. The dwarf wheat is not easy to lodge, and the adaptability is higher. The increased tillering angle can improve the photosynthetic utilization efficiency of wheat, enhance the field competitiveness, reduce diseases and insect pests and improve the yield. The invention provides a new way for cultivation of a new high-yield wheat variety, and provides a good gene resource for wheat breeding.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a wheat dwarf creeping allele and a coded protein and application thereof. Background Art

[0002] Wheat is an important food crop in the world. 20% of the calories and 25% of the protein consumed by the global population come from wheat. Wheat is also an important source of trace elements. Increasing wheat production is of vital importance to ensuring food security.

[0003] Plant height is one of the main components of wheat plant type and has an important influence on wheat yield and other traits. The plant height of wheat varieties closely affects the lodging resistance and yield potential of wheat, and is one of the most important agronomic traits in genetic breeding. Lodging is a major factor affecting yield and quality in wheat production. Researchers have calculated that the grain yield loss rate caused by lodging is as high as 25% to 50% (Dunca J. Mechanical properties of cereal stem[J]. Research in Agricultural Engineering - UZPI (Czech Republic), 2008.). Therefore, improving the lodging resistance of wheat varieties by regulating plant height has always been an important goal of wheat breeding.

[0004] Plant height is negatively correlated with lodging resistance within a certain range, so the lodging resistance of wheat can be improved by dwarfing the plants, thereby achieving the goal of stable yield. However, if the plant height is too low and the plant type is too short, the leaves of the plant will be too tight, which is not conducive to ventilation and light transmission, thereby reducing the yield of the variety. Therefore, the pursuit of plant dwarfing should be moderate in the breeding process. Studies have shown that the plant's own characteristics such as stem toughness will also affect the plant's lodging resistance like the stem height, so some tall-stalked varieties may also be less likely to lodging than short-stalked varieties. However, with the continuous increase in water and fertilizer use and the continuous improvement of cultivation technology in wheat production, the lodging problem of tall-stalked wheat has intensified, which not only increases the difficulty coefficient of harvesting, but also reduces the yield and processing quality. Because the stems of dwarf wheat plants are tougher than those of tall-stalked plants, they have stronger lodging resistance. Since the Green Revolution, the use of dwarfing genes has been widely used and valued in breeding work. Discovering and utilizing more new dwarfing gene loci will be a long-term research hotspot in the wheat field.

[0005] Tillering angle has a profound impact on crop yield and has always been a concern of breeders and botanists. It is one of the key traits that urgently need to be deeply studied and optimized in modern agricultural production. The formation of plant tillering angle is closely related to the gravity response. The gravity response refers to a phenomenon in which plants re-adjust their growth direction after sensing gravity stimulation to maintain the optimal angle of each organ with respect to the gravity direction, that is, plants can sense gravity and adjust their growth direction. How plants sense gravity has always been a difficult and hot topic in plant science research. By regulating the gravity perception of plants, the gravity response and tillering angle of plants can be further regulated. When the gravity perception ability is weakened, the gravity response will be weakened, resulting in a larger tillering angle. A large tillering angle enables crops to better resist diseases and compete for environmental resources, but the yield per unit area is not high; extremely compact crops have a smaller tillering angle, but the light utilization efficiency is reduced, and the ability to resist pests and diseases is also reduced, which is not conducive to crop yield increase. Elucidating the molecular mechanism of regulating crop tillering angle can breed more high-yield crop varieties suitable for planting in different environments. The prostrate trait of wheat is a wild-type trait, and different degrees of prostrate traits can be seen in the diploid ancestors of wheat, some local varieties, and the backbone parents of breeding (such as the Zhoumai series). The prostrate trait affects the plant type and density tolerance of wheat and is an important trait related to the improvement of wheat plant type. Only when the tillering angle is appropriate can high crop yields be ensured. As early as the 1990s, genetic research on rice tillering angle has been carried out in rice, and a large number of tillering angle QTLs have been mapped. Compared with rice, the genetic research on wheat tillering angle lags far behind, and the identified tillering angle genes are still very limited.

[0006] In view of this, deeply exploring genes related to plant height and tillering angle and carrying out research and utilization of functional genes using genetic transformation technology can provide valuable gene resources for the innovation of crop germplasm resources and genetic improvement, and is conducive to promoting the breeding of new high-yield wheat varieties. Summary of the Invention

[0007] The object of the present invention is to provide a new gene related to plant height and / or tillering angle, its encoded protein and applications. The technical problems to be solved are not limited to the described technical topics, and those skilled in the art can clearly understand other technical topics not mentioned herein through the following description.

[0008] To achieve the above object, the present invention first provides a protein, which can be named TaRHT-A1p, and the protein can be any one of the following: A1) A protein with an amino acid sequence as shown in SEQ ID NO:1; A2) A protein that has more than 80% identity with the protein shown in A1) and has the same function, which is obtained by substitution, deletion, and / or addition of amino acid residues to the amino acid sequence shown in SEQ ID NO:1; A3) A fusion protein with the same function obtained by connecting a tag to the N-terminus and / or C-terminus of A1) or A2).

[0009] The protein may be derived from wheat ( Triticum aestivum ).

[0010] The substitution described in A2) may be a conservative substitution.

[0011] The connection described in A3) may be directly connected through a peptide bond or connected through a linker.

[0012] In order to facilitate the separation, purification, detection, and / or localization of the protein described in A1) or A2), a tag protein may be connected to its amino terminus or carboxyl terminus. The tags include, but are not limited to: GST (glutathione S-transferase) tag protein, Trx (thioredoxin) tag protein, nitrogen utilization substance A (NusA) tag protein, His tag protein (His-tag), MBP (maltose-binding protein) tag protein, Flag tag protein, SUMO tag protein, HA (influenza hemagglutinin) tag protein, Myc tag protein, LacZ tag protein, CBD (cellulose-binding domain) tag protein, bacteriophage T7 protein kinase (T7PK) tag protein, GFP (green fluorescent protein), CFP (cyan fluorescent protein), YFP (yellow-green fluorescent protein), mCherry (monomeric red fluorescent protein), or AviTag tag protein. The use of the tag does not change the function of the target protein, and those skilled in the art know how to select a suitable tag protein according to the desired purpose.

[0013] The present invention also provides biological materials, and the biological materials may be any of the following: B1) A nucleic acid molecule encoding the protein TaRHT-A1p; B2) An expression cassette containing the nucleic acid molecule described in B1); B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2); B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing the recombinant vector described in B3); B5) A recombinant host cell containing the nucleic acid molecule described in B1), or a recombinant host cell containing the expression cassette described in B2), or a recombinant host cell containing the recombinant vector described in B3).

[0014] Furthermore, the above-mentioned biomaterials can all express B1) the nucleic acid molecule.

[0015] Among the above-mentioned biomaterials, B1) the nucleic acid molecule can be a coding sequence or a nucleotide sequence such as the DNA molecule shown in SEQ ID NO:2.

[0016] 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 mRNA or hnRNA, etc.

[0017] The nucleotide sequence shown in SEQ ID NO:2 can be TaRHT-A1p the coding sequence (CDS) of the gene, which encodes a protein TaRHT-A1p with an amino acid sequence shown in SEQ ID NO:1.

[0018] B1) the nucleic acid molecule can also include a nucleic acid molecule obtained by modifying the codon preference based on the nucleotide sequence shown in SEQ ID NO:2.

[0019] Those of ordinary skill in the art can easily use known methods, such as site-directed mutagenesis (including oligonucleotide primer-mediated site-directed mutagenesis, PCR-mediated site-directed mutagenesis and cassette mutagenesis, etc.) or directed evolution (including error-prone PCR, DNA shuffling and in vitro random priming recombination, etc.) to mutate the nucleotide sequence encoding the protein TaRHT-A1p. Those artificially modified nucleotide sequences that have 75% or more identity with the nucleotide sequence encoding the protein TaRHT-A1p (such as SEQ ID NO:2), as long as they encode the protein TaRHT-A1p and have the same function as the protein TaRHT-A1p, are nucleotide sequences derived from and equivalent to the sequences of the present invention.

[0020] Among the above-mentioned biomaterials, the recombinant vector can be a cloning vector or an expression vector.

[0021] Furthermore, the recombinant vector can be a recombinant expression vector obtained by cloning the coding gene of the protein TaRHT-A1p into an expression vector (such as a prokaryotic expression vector, a eukaryotic expression vector or a viral expression vector). Although the expression vector used in the examples provided by the present invention is the pWMB111 vector, the present invention is not limited to this specific vector. Those skilled in the art can use other suitable vectors as long as the vector can express the nucleic acid molecule encoding the protein TaRHT-A1p.

[0022] Existing plant expression vectors can be used to construct those containing TaRHT-A1pRecombinant expression vector of the gene. The plant expression vector includes but is not limited to binary expression vectors (such as pBI series vectors (such as pBI121), pBIN series vectors (such as pBin19), pCAMBIA series vectors (such as pCAMBIA1300 vector), pPZP series vectors, pGreen series vectors, pBIBAC series vectors, pSKI015 vector, pSKI074 vector, pRI101-AN vector, etc.) and co-integrating vectors (which can be constructed by inserting a segment homologous to the Ti plasmid or its segment into the intermediate vector by homologous recombination or cloning). The plant expression vector contains elements required for the expression of the foreign gene, such as a promoter, a multiple cloning site, a terminator, a ribosome binding site, etc. The plant expression vector may also contain the 3'-untranslated region of the foreign gene, that is, it contains a polyadenylation signal and any other DNA fragment involved in mRNA processing or gene expression. The polyadenylation signal can direct the addition of polyadenylic acid to the 3' end of the mRNA precursor. For example, the non-translated regions transcribed from the genes of Agrobacterium tumefaciens (Ti) plasmid (such as the nopaline synthase Nos gene) and plant genes (such as soybean storage protein gene) at the 3' end have similar functions. When the encoding gene of the protein TaRHT-A1p of the present invention is introduced by the Agrobacterium method, it is preferably to use an expression vector suitable for the Agrobacterium method, such as a binary vector or its modified vector. Examples of these plant expression vectors include pBI121, pBIN19, pSMAB704, pCAMBIA series vectors, and pGreen series vectors, etc.

[0023] When using TaRHT-A1p a gene to construct a recombinant plant expression vector, any one of enhanced promoters or constitutive promoters can be added before the transcription start nucleotide, including but not limited to cauliflower mosaic virus (CaMV) 35S promoter, maize ubiquitin promoter (Ubi), rice actin promoter (Actin), Emu promoter, maize Adhl gene promoter, rice rbcS gene promoter, tomato rbcS gene promoter, potato pinⅡ gene promoter, which can be used alone or in combination with other plant promoters; in addition, when using the gene of the present invention to construct a plant expression vector, an enhancer can also be used, including a translation enhancer or a transcription enhancer. These enhancer regions can be the ATG start codon or the adjacent region start codon, etc., but need to be in the same reading frame as the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signal and the start codon are extensive and can be natural or synthetic. The translation initiation region can be from the transcription initiation region or the structural gene.

[0024] For the convenience of identifying and screening transgenic plant cells or plants, the plant expression vectors used can be processed, such as adding genes that can be expressed in plants and encode enzymes or luminescent compounds that can produce color changes (GUS gene, luciferase gene, GFP gene, etc.), antibiotic resistance genes (kanamycin resistance gene kanr, neomycin resistance gene neo, hygromycin resistance gene hyg, chloramphenicol resistance gene cat, streptomycin resistance gene str, bleomycin resistance gene ble, etc.) or herbicide resistance genes (bar gene, glyphosate resistance marker gene epsps, chlorsulfuron resistance marker gene als, etc.). Considering the safety of transgenic plants, no selective marker gene can also be added, and the transformed plants can be directly screened.

[0025] In one or more embodiments of the present invention, the recombinant vector is pWMB111- TaRHT-A1p The recombinant vector pWMB111- TaRHT-A1p is a recombinant expression vector obtained by cloning the DNA fragment with the nucleotide sequence of SEQ ID NO:2 into the BamHI recognition site of the pWMB111 vector, while keeping other nucleotide sequences of the pWMB111 vector unchanged.

[0026] The present invention also provides the application of the protein TaRHT-A1p or the biological material in any of the following: C1) Application in regulating plant plant height; C2) Application in regulating plant tiller angle; C3) Application in regulating plant gravity perception ability or gravity response; C4) Application in cultivating plants with altered plant height, tiller angle, gravity perception ability and / or gravity response; C5) Application in molecular breeding or germplasm resource improvement related to plant plant height, tiller angle, gravity perception ability and / or gravity response.

[0027] The above application can be achieved by up-regulating or down-regulating the content and / or activity of the protein TaRHT-A1p.

[0028] Furthermore, the above application may include reducing the plant height, increasing the tiller angle of the plant, weakening the plant gravity perception ability and / or weakening the plant gravity response by up-regulating the content and / or activity of the protein TaRHT-A1p (for example, overexpressing TaRHT-A1p gene). The above application may also include increasing the plant height, reducing the tiller angle of the plant, enhancing the plant gravity perception ability and / or enhancing the plant gravity response by down-regulating the content and / or activity of the protein TaRHT-A1p (for example, knocking out or silencing TaRHT-A1p gene).

[0029] The present invention also provides a method for cultivating transgenic plants, the method comprising increasing the content and / or activity of the protein TaRHT-A1p in a target plant to obtain a transgenic plant, wherein the plant height of the transgenic plant is lower than that of the target plant, the tillering angle is greater than that of the target plant, and / or the gravity perception ability or gravity response is weaker than that of the target plant.

[0030] In the above method, the increase in the content and / or activity of the protein TaRHT-A1p in the target plant can be achieved by increasing the expression level of the coding gene of the protein TaRHT-A1p in the target plant.

[0031] The increase in the expression level of the coding gene of the protein TaRHT-A1p in the target plant can be achieved by at least one of the following methods: (1) increasing the copy number of the coding gene of the protein TaRHT-A1p; (2) expressing the coding gene of the protein TaRHT-A1p under the drive of a strong promoter; (3) increasing the regulatory elements of the coding gene of the protein TaRHT-A1p to overexpress it, and the regulatory elements include enhancer elements, elements for improving mRNA stability, elements for enhancing translation efficiency, and / or elements for enhancing protein secretion; (4) increasing the ribosome binding site of the coding gene of the protein TaRHT-A1p; (5) optimizing the codons of the coding gene of the protein TaRHT-A1p; (6) upregulating the expression of the gene (the coding gene of the protein TaRHT-A1p) by changing epigenetic modifications such as DNA methylation or histone acetylation.

[0032] Further, the method (2) can be achieved by replacing the natural promoter of the coding gene of the protein TaRHT-A1p with a strong promoter, or by operably linking a second promoter to the coding gene of the protein TaRHT-A1p.

[0033] The strong promoter includes but is not limited to T7 promoter, CaMV promoter, SV40 promoter, SFFV promoter, ubq promoter, ubi promoter, RBCS promoter, Actin promoter, Emu promoter, CYP450 promoter, Adhl promoter, and pinⅡ promoter.

[0034] The enhancer in the method (3) includes but is not limited to CMV enhancer, SV40 enhancer, and RSV enhancer.

[0035] In the above method, increasing the expression level of the coding gene of the protein TaRHT-A1p in the target plant can be achieved by introducing the coding gene of the protein TaRHT-A1p into the target plant.

[0036] In the above method, the nucleotide sequence of the coding gene of the protein TaRHT-A1p can be as shown in SEQ ID NO:2.

[0037] The present invention also provides a method for reducing the plant height of a target plant, increasing the tillering angle of the target plant, weakening the gravity perception ability of the target plant or weakening the gravity response of the target plant, the method comprising increasing the content and / or activity of the protein TaRHT-A1p in the target plant.

[0038] The method for cultivating transgenic plants described herein may include the following steps: (1) Constructing a recombinant vector containing a nucleic acid molecule encoding the protein TaRHT-A1p; (2) Introducing the recombinant vector constructed in step (1) into the target plant; (3) Obtaining transgenic plants through screening and identification.

[0039] Furthermore, the nucleic acid molecule encoding the protein TaRHT-A1p may be a coding sequence or a DNA molecule with a nucleotide sequence as shown in SEQ ID NO:2.

[0040] Furthermore, in the above method, after step (3), step (4) may further be included: crossing the transgenic plant with a plant to be improved to obtain progeny transgenic plants, and the progeny transgenic plants are substantially identical in phenotype to the transgenic plant (i.e., the transgenic plant as the parent). The substantially identical phenotype may mean that compared with the plant to be improved, the plant height is reduced, the tillering angle is increased, the gravity perception ability is weakened and / or the gravity response is weakened.

[0041] Furthermore, the introduction method includes but is not limited to: Agrobacterium-mediated method, plant virus vector-mediated transformation method, gene gun method (also known as particle bombardment method or biolistic method), chemical stimulation method, electrotransformation method, liposome-mediated method, microinjection method, laser microbeam method, pollen tube pathway method, ultrasonic method, pneumatic gun method, and vortex method, etc.

[0042] Furthermore, the introduction method may be the Agrobacterium-mediated method.

[0043] Furthermore, the Agrobacterium-mediated method may include the following steps: introducing the recombinant vector constructed in step (1) into Agrobacterium (such as by Ca ion-induced transformation method, polyethylene glycol-mediated transformation method, metal cation-mediated transformation method, electroporation transformation method, phage transduction method, etc.) to obtain recombinant Agrobacterium, and infecting the callus or explant of the target plant with the recombinant Agrobacterium; inducing and culturing the obtained positive callus or explant through identification to obtain a regenerated plant.

[0044] The explants include, but are not limited to, seeds, roots, leaves, petioles, cotyledons, cotyledon petioles, hypocotyls, stem segments, shoot apical meristems, epidermal parenchyma cells, tubers, stolon segments, embryogenic suspension cells, and protoplasts, etc.

[0045] The methods of screening and identification are known to those skilled in the art. For example, the transformed transgenic plants (including transgenic progeny materials) can be identified by techniques such as PCR detection, Southern hybridization, immunoblotting, Northern hybridization, enzyme-linked immunosorbent assay (ELISA), functional identification (testing for the presence of selectable marker genes and target genes), and / or in situ hybridization, etc.

[0046] In this article, the plant can be any one of the following: D1) Monocotyledonous plants or dicotyledonous plants; D2) Gramineous plants; D3) Triticum plants; D4) Wheat.

[0047] The regulation described in this article can be up-regulation (such as increase or enhancement) or down-regulation (such as decrease, reduction or attenuation).

[0048] In this article, the transgenic plants are understood to include not only the first-generation transgenic plants obtained by introducing the coding gene of the protein TaRHT-A1p into the target plant, but also their progeny. The transgenic plants include seeds, callus, whole plants and cells.

[0049] Through extensive and in-depth research, the inventors of the present invention discovered the gene shown in SEQ ID NO:2, which is TaRHT-A1 a new allele of the TaRHT-A1p gene, named TaRHT-A1p Compared with the wild-type gene ( TaRHT-A1 gene), there is a difference at the 131st base in the CDS region (SEQ ID NO:2), changing from G in the TaRHT-A1 gene to TaRHT-A1pA in the gene, the base change causes glycine (G) at the 44th position in the corresponding amino acid sequence to mutate into glutamic acid (E). The mutation is located between the DELLA domain and the TVHYNP domain, adjacent to the DELLA domain, and this mutation can regulate the plant height and tillering angle of wheat.

[0050] The present invention discovers and confirms a new gene that regulates the plant height and tillering angle of wheat (wheat dwarf creeping allele TaRHT-A1p ), and for the first time reveals TaRHT-A1p the application of the gene and its encoded protein TaRHT-A1p in regulating plant height, tillering angle, gravity perception ability, and / or gravity response. The results of plant genetic transformation experiments and phenotypic analyses show that by increasing the content and / or activity of the protein TaRHT-A1p in the target plant (such as overexpressing TaRHT-A1p the gene), the plant height of the plant can be significantly reduced, the tillering angle of the plant can be increased, the gravity perception ability of the plant can be weakened, and / or the gravity response of the plant can be weakened; correspondingly, by reducing the content and / or activity of the protein TaRHT-A1p in the target plant (such as knocking out or silencing TaRHT-A1p the gene), the plant height of the plant can be significantly increased, the tillering angle of the plant can be decreased, the gravity perception ability of the plant can be enhanced, and / or the gravity response of the plant can be enhanced.

[0051] The present invention successfully creates dwarf wheat with significantly reduced plant height and significantly increased tillering angle. The dwarf wheat is not easily lodged, has stronger adaptability, can better resist the invasion of strong winds, reduce losses caused by natural disasters such as typhoons and tornadoes, and ensure the harvest rate and quality. The increased tillering angle can improve the photosynthetic utilization efficiency of wheat, enhance the competitiveness in the field, reduce pests and diseases, and increase the yield. The TaRHT-A1p gene of the present invention can be applied to the improvement of wheat plant type, thereby providing a new way for the cultivation of new high-yield wheat varieties and laying a foundation for improving wheat yield and traits.

[0052] The present invention provides material accumulation and an efficient and safe technical method for the improvement of wheat germplasm resources and breeding, provides a new and good gene resource for wheat breeding, and is conducive to promoting the commercial breeding process of wheat.

[0053] Term Definition In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. At the same time, for a better understanding of the present invention, the definitions and explanations of relevant terms are provided below.

[0054] The term "expression cassette" generally refers to a nucleic acid construct containing nucleic acid elements sufficient to express a gene of interest. A typical expression cassette contains a promoter, an MCS (multiple cloning site), and a terminator. The expression cassette may also include a gene of interest, marker genes (such as the TK gene, DHFR gene, CAT gene, and NEO gene), ribosome recognition and binding sites (SD), transcription factor binding sites (TFBS), enhancers, silencers, repressors, introns, poly(A) signal sequences, and / or mRNA splicing signal sequences, etc. The elements in the expression cassette can be directly linked or indirectly linked through a linker.

[0055] The term "vector" generally refers to a vector that can carry foreign DNA or a gene of interest into a host cell for amplification and / or expression. The vector can be a cloning vector or an expression vector. The vector can be introduced into the host cell by transformation, transduction, or transfection, so that the genetic material elements it carries are amplified and / or expressed in the host cell. Those skilled in the art can select a suitable vector according to the purpose of genetic engineering and the nature of the recipient cell. The vectors include but are not limited to: plasmids, phages (such as λ phage or M13 phage), cosmids (i.e., cosmid plasmids), phagemids, shuttle vectors (such as yeast expression vectors), Ti plasmids, artificial chromosomes (such as yeast artificial chromosomes (YAC), bacterial artificial chromosomes (BAC), P1 artificial chromosomes (PAC), or Ti plasmid artificial chromosomes (TAC)), viral vectors (such as baculovirus vectors, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, poxviruses, papillomaviruses, polyomaviruses (such as SV40), herpesviruses (such as herpes simplex virus)). A vector can contain multiple elements controlling expression, including but not limited to promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. In addition, the vector may also contain an origin of replication.

[0056] The term "microorganism" generally includes bacteria, viruses, fungi, actinomycetes, rickettsiae, mycoplasmas, chlamydiae, spirochetes, algae, etc. For example, the bacteria can be from the genus Escherichia ( Escherichia sp. ), such as Escherichia coli, the genus Erwinia ( Erwinia sp. ), the genus Agrobacterium ( Agrobacterium sp. ), such as Agrobacterium tumefaciens, the genus Flavobacterium ( Flavobacterium sp. ), the genus Alcaligenes ( Alcaligenes sp. ), the genus Pseudomonas ( Pseudomonas sp. ), and the genus Bacillus ( Bacillus sp.), such as Bacillus spp. The viruses may include rotavirus, baculovirus, retrovirus (such as lentivirus), adenovirus, adeno-associated virus, poxvirus, papillomavirus, influenza virus, papovavirus (such as SV40), and herpesvirus (such as herpes simplex virus), etc. The fungi may be from the genus Saccharomyces ( Saccharomyces sp. ), such as Saccharomyces cerevisiae, Pichia methanolica, Pichia pastoris), Fusarium ( Fusarium sp. ), Rhizoctonia ( Rhizoctonia sp. ), Verticillium ( Verticillium sp. ), Penicillium ( Penicillium sp. ), Aspergillus ( Aspergillus sp. ), and Cephalosporium ( Cephalosporium sp. ), etc. The actinomycetes may be from the genus Streptomyces ( Streptomyces sp. ), such as Streptomyces). The algae may be from the phylum Cyanophyta (such as cyanobacteria), Fucus ( Fucus sp. ), Achnanthes ( Achnanthes sp. ), Amphiprora ( Amphiprora sp. ), Amphora ( Amphora sp. ), Ankistrodesmus ( Ankistrodesmus sp. ), Asterococcus ( Asteromonas sp. ), and Chrysosphaera ( Boekelovia sp. ), etc.

[0057] The term "host cell" is also referred to as recipient cell, and generally refers to any type of cell that can be used to introduce a vector, such as plant cells and animal cells. The host cell can be understood to refer not only to a specific recipient cell, but also to the progeny of such a cell, and due to natural, accidental or intentional mutations and / or alterations, the progeny may not have to be exactly the same as the original parental cell, but is still included in the scope of the host cell. Suitable host cells are known in the art, among which: the plant cells may be Arabidopsis thaliana ( Arabidopsis thaliana ), tobacco ( Nicotiana tabacum ), maize ( Zea mays ), rice ( Oryza sativa ), wheat ( Triticum aestivum ), etc., but not limited to these; the animal cells may be mammalian cells (such as Chinese hamster ovary cells (CHO cells), Chinese hamster ovary cell sub-line (CHO-K1 cells), African green monkey kidney cells (Vero cells), SV40-transformed African green monkey kidney cells (COS cells), baby hamster kidney cells (BHK cells), mouse breast cancer cells (C127 cells), human embryonic kidney cells (HEK293 cells), human HeLa cells, fibroblasts, bone marrow cell lines, T cells or NK cells, etc.), avian cells (such as chicken or duck cells), amphibian cells (such as Xenopus laevis ( Xenopus laevis )) cells or Andrias davidianus ( Andrias davidianus), cells), fish cells (such as grass carp, common carp, rainbow trout or catfish cells), insect cells (such as Sf21 cells, Sf-9 cells or Hi-5), etc., but not limited to this.

[0058] The term "recombinant vector" generally refers to a recombinant DNA molecule constructed by ligating an exogenous target gene and a vector in vitro, which can be constructed in any suitable manner as long as the constructed recombinant vector can carry the exogenous target gene into the recipient cell and provide the ability for the exogenous target gene to replicate, integrate, amplify and / or express in the recipient cell.

[0059] The term "recombinant microorganism" generally refers to a microorganism whose genes are manipulated and modified to obtain a recombinant microorganism with changed functions. For example, an exogenous target gene or a recombinant vector is introduced into the target microorganism, or the endogenous genes of the target microorganism are directly gene-edited.

[0060] The term "recombinant host cell" generally refers to a host cell whose genes are manipulated and modified to obtain a recombinant host cell with changed functions. For example, an exogenous target gene or a recombinant vector is introduced into the host cell, or the endogenous genes of the host cell are directly gene-edited.

[0061] The term "ligation" generally refers to the association of two or more molecules. The ligation can be covalent or non-covalent. The ligation described herein can be directly ligated through a peptide bond or ligated through a linker (adapter).

[0062] The term "identity" generally refers to the degree to which two (nucleotide or amino acid) sequences have the same residues at the same positions in an alignment, and is usually expressed as a percentage. The identity described herein can refer to the identity of an amino acid sequence or a nucleotide sequence. Two copies with exactly the same sequence have 100% identity. Those skilled in the art know that the identity of an amino acid sequence or a nucleotide sequence can be determined using identity search sites on the Internet, such as the BLAST page of the NCBI homepage website. For example, in Advanced BLAST 2.1, by using blastp as the program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as the Matrix, setting the Gap existence cost, Perresidue gap cost, and Lambda ratio to 11, 1, and 0.85 (default values) respectively and performing a search, the identity value (%) of the amino acid sequence can be calculated, and then the identity value can be obtained. In addition, it can be determined using sequence analysis software (such as CLC Main Workbench and MegAlignTM), for example, using the computer program BLAST with default parameters, especially BLASTP or TBLASTN. The 75% or more identity described herein can be 75%, 80%, 85%, 90%, or 95% or more identity. In this article, the identity of more than 80% can be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more identity.

[0063] The term "conservative substitution" generally refers to the replacement of an amino acid residue with another amino acid residue having a side chain with similar physicochemical properties. For example, conservative substitutions can be made between hydrophobic side chain amino acid residues (such as Met, Ala, Val, Leu, and Ile), between neutral hydrophilic side chain residues (such as Cys, Ser, Thr, Asn, and Gln), between acidic side chain residues (such as Asp, Glu), between basic side chain amino acids (such as His, Lys, and Arg), or between aromatic side chain residues (such as Trp, Tyr, and Phe). It is known in the art that conservative substitutions generally do not cause significant changes in the conformational structure of the protein and basically do not change the biological activity of the protein. Conservative substitutions that are expected to have only a minimal or no effect on the protein structure or function in a protein sequence can be easily designed by those of ordinary skill in the art.

[0064] The term "overexpression" generally refers to increasing or upregulating the level and / or activity of a target protein or gene. Overexpression can be achieved by regulation at the gene level (such as gene replication, transcription, translation, post-transcriptional modification, and / or post-translational modification), or by promoting or increasing the content, activity, and / or function of the target protein at the protein level. The means of overexpression are not particularly limited, and numerous ways to achieve overexpression are well-known to those skilled in the art. For example, a nucleic acid molecule to be overexpressed or a nucleic acid molecule encoding a protein to be overexpressed can be placed under the control of a strong promoter; the copy number of one or more genes encoding the protein of the present invention can also be increased; or the strength of the ribosome binding site or Kozak sequence can be increased, the stability of mRNA can be enhanced, codon usage can be altered, inhibitory elements can be knocked out, etc.

[0065] The term "promoter" generally refers to the site specifically recognized and bound by RNA polymerase, which is located upstream of the transcription start point of the structural gene, has strict directionality, and initiates transcription. Since the strength of the promoter determines the efficiency of transcription, different types of promoters can be used in genetic engineering to regulate the expression of key genes. It is known to those skilled in the art that overexpression of a target gene can be achieved by using a constitutive strong promoter. To further enhance the expression of the target gene, multiple promoters can also be used in tandem.

[0066] The term "enhancer" generally refers to a DNA sequence that can enhance the transcriptional activity of a gene, which is located upstream or downstream of the structural gene far away, or can also be located in an intron.

[0067] The term "regulatory element" generally refers to a DNA molecule with gene regulatory activity. Regulatory elements that function in plants include promoters, leader sequences, enhancers, introns, and 3' UTRs, etc.

[0068] The term "operably linked" generally refers to the physical and / or functional connection of a DNA segment to another DNA segment, and the connection mode allows the segments to function in their intended manner. The DNA encoding a gene product is operably linked to a regulatory element, which can directly or indirectly regulate the transcription of the DNA. For example, when an enhancer is operably linked to the DNA encoding a gene product, the DNA transcription can be enhanced, and the enhancer can be located upstream, downstream, or embedded in the coding region of the DNA.

[0069] The term "codon optimization" generally refers to a technique for improving the protein expression level in an organism by increasing the translation efficiency of a target gene. Codon optimization usually redesigns the gene by avoiding rare codons, using preferred codons, simplifying the secondary structure of mRNA, optimizing repetitive sequences, eliminating restriction enzyme cleavage sites, adjusting the GC content, etc., to improve the translation efficiency and thus increase the protein expression level.

[0070] The term "introduction" generally refers to the transfer of foreign genes into recipient cells such as eukaryotic recipient cells or prokaryotic recipient cells. There is no particular limitation on the introduction method, and any known transformation method can be used as long as it can transfer the target gene (such as the DNA molecule of the present invention) into the recipient cell. The introduced DNA molecule can be single-copy or multi-copy. The introduction can be the integration of foreign genes into the host chromosome or the expression of plasmids outside the chromosome. The introduction methods may include any of the following: (1) introducing the target gene or the recombinant vector containing the target gene into the host bacterium by chemical transformation methods (such as Ca ion-induced transformation method, polyethylene glycol-mediated transformation method, or metal cation-mediated transformation method, etc.) or physical transformation methods (such as electroporation transformation method). (2) Transducing the target gene into the host bacterium by phage transduction method. (3) Transferring the target gene into plant recipient cells by physical or chemical methods, such as gene gun method (also known as particle bombardment method or biolistic method), chemical stimulation method, electroshock method, liposome-mediated method, microinjection method, laser microbeam method, pollen tube pathway method, ultrasonic method, pneumatic gun method, and eddy current method, etc. (4) Transferring the target gene into plant recipient cells with a vector as a medium, such as Agrobacterium Ti plasmid vector (including Ti plasmid-derived vectors such as co-integrated vector system and binary vector system) mediated method (Agrobacterium-mediated method), plant virus vector-mediated transformation method, etc.

[0071] The term "explant" generally refers to a part of a plant body used as an in vitro culture material in plant tissue culture. After appropriate treatment and under suitable conditions, it can regenerate into a whole plant. In actual operation, those skilled in the art select appropriate explants for transformation according to different plants.

[0072] The term "callus" generally refers to the new tissue formed on the wound surface after local trauma stimulation of the original plant body. It is composed of living parenchyma cells and can originate from living cells of various tissues in any organ of the plant body. In plant tissue culture, it can refer to a mass of parenchyma cells with strong division ability that grow disorderly formed from explants. Culturing callus on an appropriate medium can induce it to form a whole plant. Brief Description of the Drawings

[0073] Figure 1 For TaRHT-A1p Screening of positive seedlings of gene overexpression plants.

[0074] Figure 2 For TaRHT-A1p Statistical analysis of phenotypes, tiller angles, and plant heights of gene overexpression plants during the filling stage.

[0075] Figure 3 For TaRHT-A1p Statistical analysis of phenotypes and tiller angles of gene overexpression plants at the jointing stage.

[0076] Figure 4 Under normal light culture conditions TaRHT-A1p Results of the gravitropism experiment of gene overexpression plants at the seedling stage.

[0077] Figure 5 Under dark conditions TaRHT-A1p Results of the gravitropism experiment of gene overexpression plants at the seedling stage.

[0078] Figure 6 Schematic diagram of the pWMB111 vector. Specific implementation manners

[0079] The present invention will be further described in detail below in conjunction with specific implementation manners. The provided embodiments are only for clarifying the present invention, rather than limiting the scope of the present invention. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements, and do not limit the present invention in any way.

[0080] The experimental methods in the following embodiments are all conventional methods unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in this field or according to the product specifications. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0081] The following embodiments use GraphPad Prism statistical software to process data. The experimental results are expressed as mean ± standard deviation. The Student's t test is used for the test. P < 0.05 (*) indicates a statistically significant difference, P < 0.01 (**) indicates a significant statistical difference, and P < 0.001 (***) indicates an extremely significant statistical difference. For the quantitative experiments in the following embodiments, unless otherwise specified, three biological replicate experiments are set, and the results are averaged.

[0082] Source of the dwarf and prostrate mutant 2246 of wheat Liangxing 99 in the following embodiments: Liangxing 99 is a wheat variety selected by Shandong Liangxing Seed Industry Co., Ltd., and the parental source is ((Ji 91102 × Lumai 14) × PH85-16). 2246 is a dwarf mutant obtained by EMS mutagenesis of Liangxing 99, and was kindly provided by Teacher Zhang Mingyi of the Shanxi Academy of Agricultural Sciences. The public can obtain it from the applicant. This biological material is only used for repeating the relevant experiments of the present invention and cannot be used for other purposes.

[0083] The pWMB111 vector in the following examples was gifted by the research group of Teacher Ye Xingguo from the Institute of Crop Sciences, Chinese Academy of Agricultural Sciences and is described in the following literature: Riaz B, Chen HQ et al. Overexpression of Maize ZmC1 and ZmR Transcription Factors in Wheat Regulates Anthocyanin Biosynthesis in a Tissue-Specific Manner[J]. International Journal of Molecular Sciences, 2019, 20, 5806. The vector map is shown in Figure 6 .

[0084] Example 1, TaRHT-A1p Discovery and Cloning of Genes After extensive and in-depth research, the inventors of this application first used the dwarf and prostrate mutant 2246 of wheat Liangxing 99 as the research material and conducted phenotypic identification on it. Then, it was crossed with its wild type Liangxing 99 and Jing 411 to construct a segregating population of the F2 generation and its derived offspring for the initial mapping and fine mapping of the dwarf and prostrate gene, and TaRHT-A1p was listed as a candidate gene. Through screening and identification, genes regulating wheat plant height and tiller angle were obtained. TaRHT-A1p The cloning and sequence of the gene are as follows: The sequence of the wheat TaRHT-A1 gene was found on the Ensembl Plants website. According to the TaRHT-A1 base sequence of the gene on the website, primers for its specific fragment were designed. The stems of mutant 2246 wheat at the heading stage were sampled, and after sampling, the wheat stems were frozen in liquid nitrogen and ground. RNA was extracted by the Trizol method, and cDNA was obtained by reverse transcription using RNA as a template. Using the stem cDNA of mutant 2246 wheat as a template, PCR amplification was carried out, and the obtained amplification product was sequenced.

[0085] The sequencing results showed that TaRHT-A1p the full length of the CDS of the gene is 1863 bp, encoding 620 amino acid residues. The CDS nucleotide sequence is shown in SEQ ID NO:2, and its amino acid sequence is shown in SEQ ID NO:1. Compared with the wild type, TaRHT-A1p the 131st base in the CDS region of the gene mutated, changing from TaRHT-A1 G in TaRHT-A1p to

[0086] Amino acid sequence of protein TaRHT-A1p: MKREYQDAGGSGGGGGMGSSEDKMMVSAAAGEGEEVDELLAALEYKVRASDMADVAQKLEQLEMAMGMGGVGAGAAPDDSFATHLATDTVHYNPTDLSSWVESMLSELNAPPPPLPPAPQQLNASTSSTVTGGGYFDLPPSVDSSCSTYALRPIPSPAGAVGPADLSADSVRDPKRMRTGGSSTSSSSSSSSSLGGGARSSVVEAAPPVAAGANAPALPVVVVDTQEAGIRLVHALLACAEAVQQENFSAAEALVKQIPLLAASQGGAMRKVAAYFGEALARRVFRFRPQPDSSLLDAAFADLLHAHFYESCPYLKFAHFTANQAILEAFAGCRRVHVVDFGIKQGMQWPALLQALALRPGGPPSFRLTGVGPPQPDETDALQQVGWKLAQFAHTIRVDFQYRGLVAATLADLEPFMLQPEGEEDPNEEPEVIAVNSVFEMHRLLAQPGALEKVLGTVRAVRPRIVTVVEQEANHNSGTFLDRFTESLHYYSTMFDSLEGGSSGGPSEVSSGAAAAPAAAGTDQVMSEVYLGRQICNVVACEGAERTERHETLGQWRNRLGNAGFETVHLGSNAYKQASTLLALFAGGDGYKVEEKEGCLTLGWHTRPLIATSAWRLAAP (SEQ ID NO:1).

[0087] TaRHT-A1p Coding sequence (CDS) of the gene:

[0088] Example 2 TaRHT-A1p Application of Gene in Regulating Wheat Plant Height and Tiller Angle 1. Construction of Recombinant Expression Vector According to the CDS sequence of wheat TaRHT-A1p gene, primers were designed using DNAman software. Amplify TaRHT-A1p the CDS sequence of, recover the PCR product by gel extraction, ligate the obtained gel extraction product to the pWMB111 vector digested with BamHI by seamless cloning enzyme, transform Escherichia coli competent T1, pick single colonies for shaking culture and send for sequencing, select the bacteria with correct sequencing for shaking culture to extract plasmids, and obtain the wheat TaRHT-A1p overexpression vector, named pWMB111- TaRHT-A1p . The recombinant vector pWMB111- TaRHT-A1p is obtained by cloning the DNA fragment with the nucleotide sequence of SEQ ID NO:2 into the BamHI recognition site of the pWMB111 vector, keeping the other nucleotide sequences of the pWMB111 vector unchanged, and obtaining the recombinant expression vector.

[0089] The designed primer sequences are as follows: 111-A1p-F: 5’-AGGTCGACTCTAGAGGATCCATGAAGCGCGAGTACCAGGA-3’ (SEQ ID NO:3); 111-A1p-R: 5’-AGCTCGGTACCCGGGGATCCTCACGGCGCGGCCAGGCGCC-3’ (SEQ ID NO:4).

[0090] 2. Obtaining of Transgenic Wheat Take immature ears of wheat Fielder, wash them 3 times with 5% sodium hypochlorite for 20 minutes each time in a laminar flow hood, and wash them thoroughly 4 - 5 times with sterile ddH2O. Then, take the intact embryos of the seeds in the laminar flow hood and put them into the corresponding liquid medium; transform the constructed wheat TaRHT-A1p overexpression vector (pWMB111- TaRHT-A1p ) into Agrobacterium competent (EHA105), use the young embryos of wild Fielder wheat as experimental materials, and carry out wheat genetic transformation by Agrobacterium-mediated method. The specific steps are as follows: The embryo and Agrobacterium were co-cultured in WLS1 liquid medium (containing 1 / 10 Linsmaier and Skoog (LS) salts, 1 / 10 Murashige and Skoog (MS) vitamins, 10 g / L glucose, 0.5 g / L 2-(N-morpholino)ethanesulfonic acid (MES), and 100 μM acetosyringone (AS), pH 5.8) at room temperature for 5 minutes, and then transferred to a solid co-culture medium supplemented with vitamins (WLS liquid medium supplemented with 0.85 mg / L silver nitrate, 1.25 mg / L copper sulfate pentahydrate, and 8 g / L agarose). The scutellum was facing up, and the culture was carried out at 25°C in the dark for 2 days. After the co-culture, the embryonic axis was removed with a scalpel, and the remaining scales were transferred to a callus induction medium (containing LS salts, MS vitamins, 0.5 mg / L 2,4-D, 2.2 mg / L picloram, 0.85 mg / L silver nitrate, 100 mg / L ascorbic acid, 250 mg / L carbenicillin, 100 mg / L cefotaxime, 1.95 g / L MES, and 5 g / L agarose), and the culture was continued under the same conditions for 5 days. Subsequently, the tissue was transferred to a selection medium (callus induction medium supplemented with 5 mg / L glufosinate-ammonium (PPT, Sigma, no. 45520), without cefotaxime) for callus induction. After 2 weeks, the callus was transferred to a selection medium containing 10 mg / L PPT, and the culture was carried out in the dark for 3 weeks to induce the formation of embryonic callus. The embryogenic callus was placed on a 1 / 2MS (without zeatin) medium containing 5 mg / L PPT, and the differentiation culture was carried out at 25°C under a light intensity of 100 μmol m-2 s-1. The regenerated shoots were transferred to a rooting medium containing 5 mg / L PPT for elongation and rooting culture. After the roots were fully developed, the plants were transplanted into flower pots and cultured in a growth chamber to finally obtain T0 generation transgenic seedlings.

[0091] 3. Molecular Detection of Transgenic Wheat For the obtained transgenic lines, DNA was extracted respectively (CTAB method). The TaRHT-A1p-OE transgenic wheat plants were qualitatively identified by PCR reaction. The electrophoresis detection results showed that the target gene band could be amplified in TaRHT-A1p-OE positive plants, while there was no target band in the wild Fielder control group (see Figure 1 , and a total of about 800 bp of a sequence on the vector and a sequence on the target fragment were selected, so as to indicate that the amplified band was the target gene on the introduced overexpression vector rather than the gene of wheat itself). A total of 25 TaRHT-A1p-OE positive plants ( TaRHT-A1p gene overexpression plants) were identified. The identification primers are as follows: UBI-F (vector sequence): 5’-TAGCCCTGCCTTCATACGCT-3’ (SEQ ID NO:5); A1p-R (CDS sequence): 5’-TCCGCGGCAGAGAAGTTCTC-3’ (SEQ ID NO:6).

[0092] The CTAB method was used for extracting wheat genomic DNA. Fresh leaves of wheat seedlings were used as materials, and the main steps were as follows: First, take fresh wheat tissue samples and put them into a 2.0 mL centrifuge tube containing 6 mm diameter steel beads, and quickly freeze them in liquid nitrogen. Subsequently, place the samples in a grinding machine and grind them into powder. Then, add 800 μL of preheated CTAB extraction buffer to the ground samples, mix well, and incubate them in a 65 °C water bath or oven for 30 minutes, gently shaking and mixing them every 15 minutes during this period. After incubation, cool the samples to room temperature, add 800 μL of chloroform / isopentanol (24:1) mixture, gently shake and mix for 5 minutes, let it stand at room temperature for 5 minutes, and then centrifuge at 12,000 rpm for 15 minutes at room temperature. After centrifugation, pipette 600 μL of the supernatant and transfer it to a new 1.5 mL centrifuge tube, add 0.8 - 1 volume of isopropanol, gently invert and mix, and place it at -20 °C for at least 30 minutes (can be overnight). Subsequently, centrifuge at 10,000 rpm for 10 minutes at 4 °C, and discard the supernatant. Add 1 mL of 75% ethanol to the precipitate, gently shake to wash the DNA precipitate, and centrifuge at 10,000 rpm for 2 minutes at 4 °C. Repeat the washing step once. After discarding the supernatant, air-dry the DNA precipitate at room temperature. Finally, add an appropriate amount of ddH2O to dissolve the DNA, store it in a -20 °C refrigerator for subsequent identification and analysis.

[0093] 4. Phenotypic identification of transgenic wheat The tiller angle is the included angle formed by the outermost tillers, and the plant height is the height from the base of the stem to the top of the highest spike (excluding the awn length).

[0094] 4-1. Tiller angle and plant height at the filling stage Plant lines 1# and 2# of the constructed TaRHT-A1p-OE transgenic wheat were planted in the greenhouse, with the wild type Fielder as the control, and the tiller angle and plant height phenotypes at the filling stage were statistically analyzed. The results showed that at the filling stage, the tiller angles of TaRHT-A1p-OE plants were significantly larger than those of the wild type (see Figure 2 ). At the filling stage, the average tiller angle of wild type plants was 22°, and the average tiller angle of TaRHT-A1p-OE plants reached 45°, among which the tiller angle of plant line 1# had the most obvious difference (see Figure 2); At the filling stage, the average plant height of the wild type was 72 cm, while that of the TaRHT-A1p-OE plants at the filling stage was 48 cm. The plant height of the TaRHT-A1p-OE plants in both lines was significantly lower than that of the wild type (see Figure 2 ). In summary, TaRHT-A1p Upregulation of gene expression levels led to an increase in the tiller angle and a decrease in plant height in wheat.

[0095] 4-2, Tiller angle at the jointing stage Since the phenotypic characteristics of line 1 of the TaRHT-A1p-OE transgenic wheat constructed were more obvious, we statistically analyzed the tiller angle phenotype at the jointing stage. The results showed that compared with the mature stage, at the jointing stage, the tiller angle of the TaRHT-A1p-OE plants increased more significantly compared with the wild type and showed a prostrate shape (see Figure 3 ). At the jointing stage, the average tiller angle of the wild type plants was 20°, and the average tiller angle of the TaRHT-A1p-OE plants reached 70°.

[0096] 4-3, Gravity perception ability or gravity response The degree of gravity perception of the above-ground part is an important way to regulate the tiller angle of wheat. An increase in the tiller angle in plants is often caused by defects in the gravity response. To verify the changes in the gravity perception of the above-ground part of the TaRHT-A1p-OE plant seedlings, we conducted a gravity sensing experiment on the stems of the seedlings. The specific method was as follows: Take a petri dish, lay germination paper in the dish, moisten it with deionized water, and evenly distribute the Fielder seeds and the seeds of lines 1 and 2 of the constructed TaRHT-A1p-OE transgenic wheat on the germination paper. Incubate at room temperature for 1-2 days, select seeds with consistent growth and plant them at the mouth of a 15 ml centrifuge tube containing 0.4% agar vertically in an artificial climate chamber. The culture conditions were 16 h of light, 8 h of darkness, and the temperature was set at 25°C. Incubate for 3 days, then place the test tube horizontally for gravity stimulation and incubate under conditions of 16 h of light, 8 h of darkness, and complete darkness, respectively. Take pictures every 12 h for recording. The method for measuring the bending angle was as follows: Take pictures of the wheat seedlings at each gravity sensing treatment time point, and then use the software ImageJ to measure the angle.

[0097] The results showed that whether under light culture conditions or complete darkness conditions, there were significant differences in the bending angle of the stems of the TaRHT-A1p-OE plants compared with the wild type, and they were significantly smaller than those of the wild type plants (see Figure 4 , Figure 5 ). Therefore, TaRHT-A1p An increase in gene expression levels can lead to a decrease in the gravity perception of the stem, resulting in a smaller bending angle of the stem and a prostrate phenotype.

[0098] The present invention has been described in detail above. For those skilled in the art, without departing from the spirit and scope of the present invention and without unnecessary experiments, the present invention can be implemented within a relatively wide range under equivalent parameters, concentrations and conditions. Although specific embodiments of the present invention are given, it should be understood that the present invention can be further improved. In short, according to the principle of the present invention, this application intends to cover any modifications, uses or improvements of the present invention, including those that depart from the scope disclosed in this application and are made by conventional techniques known in the art.

Claims

1. A protein, characterized in that, The protein is any one of the following: A1) a protein having an amino acid sequence as shown in SEQ ID NO:1; A2) a protein having more than 80% identity with the protein shown in A1) and having the same function, which is obtained by substitution, deletion and / or addition of amino acid residues to the amino acid sequence shown in SEQ ID NO:1; A3) a fusion protein having the same function, which is obtained by connecting a tag to the N-terminus and / or C-terminus of A1) or A2).

2. A biological material, characterized in that, The biological material is any one of the following: B1) a nucleic acid molecule encoding the protein recited in claim 1; B2) an expression cassette containing the nucleic acid molecule recited in B1); B3) a recombinant vector containing the nucleic acid molecule recited in B1), or a recombinant vector containing the expression cassette recited in B2); B4) a recombinant microorganism containing the nucleic acid molecule recited in B1), or a recombinant microorganism containing the expression cassette recited in B2), or a recombinant microorganism containing the recombinant vector recited in B3); B5) a recombinant host cell containing the nucleic acid molecule recited in B1), or a recombinant host cell containing the expression cassette recited in B2), or a recombinant host cell containing the recombinant vector recited in B3).

3. The biomaterial according to claim 2, characterized in that, The nucleic acid molecule recited in B1) is a DNA molecule having a coding sequence or nucleotide sequence as shown in SEQ ID NO:

2.

4. Use of the protein recited in claim 1, or the biological material recited in claim 2 or 3, in any one of the following: C1) use in regulating plant plant height; C2) use in regulating plant tillering angle; C3) use in regulating plant gravity perception ability or gravity response; C4) use in cultivating plants with altered plant height, tillering angle, gravity perception ability and / or gravity response; C5) use in molecular breeding or germplasm resource improvement related to plant plant height, tillering angle, gravity perception ability and / or gravity response.

5. A method for cultivating transgenic plants, characterized in that, The method includes increasing the content and / or activity of the protein recited in claim 1 in a target plant to obtain a transgenic plant, wherein the plant height of the transgenic plant is lower than that of the target plant, the tillering angle is greater than that of the target plant and / or the gravity perception ability or gravity response is weaker than that of the target plant.

6. The method according to claim 5, characterized in that The increase in the content and / or activity of the protein recited in claim 1 in the target plant is achieved by increasing the expression level of the coding gene of the protein in the target plant.

7. The method according to claim 6, characterized in that, The increase in the expression level of the coding gene of the protein in the target plant is achieved by introducing the coding gene of the protein recited in claim 1 into the target plant.

8. The method according to claim 7, characterized in that The nucleotide sequence of the coding gene of the protein is as shown in SEQ ID NO:

2.

9. A method for reducing the plant height of a target plant, increasing the tillering angle of the target plant, weakening the gravity perception ability of the target plant, or weakening the gravity response of the target plant, characterized in that, The method includes increasing the content and / or activity of the protein recited in claim 1 in a target plant.

10. The application according to claim 4, or the method according to any one of claims 5-9, characterized in that The plant is any one of the following: D1) a monocotyledonous plant or a dicotyledonous plant; D2) a gramineous plant; D3) a plant of the genus Triticum; D4) wheat.

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