Gene for regulating and controlling differentiation of plant spikelet primordium to seta and application

Abstract

The invention provides a gene for regulating and controlling differentiation of a plant spikelet primordium to seta and application, and belongs to the technical field of molecular biology. The invention provides a protein for regulating and controlling differentiation of a plant spikelet primordium to seta. The protein comprises an amino acid sequence as shown in SEQ ID No. 1. The invention also provides a gene sequence for coding the protein, and a biological material for regulating and controlling the differentiation of the plant spikelet primordium to the seta is prepared on the basis of the gene sequence or the protein, and the transformation of the plant seta to the grain can be regulated and controlled by utilizing the biological material. After the coding gene of the protein is knocked out, differentiation of a part of spikelet primordia to seta can be successfully inhibited, and the part of spikelet primordia which are not differentiated into seta is differentiated into spikelet with stems. The protein and the gene are excavated, so that the molecular mechanism of seta development of the millet can be filled, and meanwhile, the high-yield breeding process of the millet can be promoted.

Description

Technical Field

[0001] This invention belongs to the field of molecular biology technology, specifically relating to genes that regulate the differentiation of plant spikelet primordia into setae and their applications. Background Technology

[0002] Millet (Setaria italica L. Beauv.), also known as ji or su in ancient times, is characterized by its drought resistance, high light efficiency, tolerance to poor soil, and wide adaptability. Millet is a diploid C4 crop with a relatively small genome of approximately 490 Mb and a short growth cycle. It is rich in protein, fatty acids, and vitamins, making it highly nutritious. With the continuous improvement of people's living standards and the diversification and nutritional focus of food, millet has become a popular choice for health maintenance.

[0003] The first T2T reference genome for millet has been published, and the most complete multi-omics database for millet has been established. This database contains much-needed multi-omics data for millet research, including T2T genomes, pangenomes, large-population variant genomes, transcriptomes, metabolomes, multi-environment phenotypic genomes of various populations, and germplasm information. Related online tools have also been developed, providing the most comprehensive sharing platform for millet functional gene theory and breeding research. Millet has become a highly promising C4 cereal model plant. In 1998, Diao Xianmin et al. conducted electron microscopy observations on millet and explored the origin of millet setae, concluding that millet setae originate from spikelet primordia and speculating that the setae are undeveloped spikelets.

[0004] The differentiation process of millet spikes can be divided into five stages: the pre-elongation stage of the growth cone (vegetative growth stage), the elongation stage of the growth cone, the branch differentiation stage, the spikelet and bristle differentiation stage, and the floret differentiation stage. Millet is a panicle-like inflorescence. A mature millet inflorescence consists of a main axis, primary, secondary, and tertiary branches, spikelets, and bristles. The spikelets and bristles are attached to the tertiary branches. The appearance of bristles occurs during the spikelet differentiation stage of millet growth. After the differentiation of tertiary branches, many protrusions will appear. These protrusions are spikelet primordia. The differentiation sequence of spikelet primordia is from top to bottom. Initially, the differentiation morphology of these spikelet primordia is not different. However, during further development, some of these spikelet primordia begin to swell and enlarge, and glumes differentiate, becoming spikelets with stalks. Others do not swell but increase in length and become bristles, i.e., undeveloped spikelets. If millet encounters drought during the spikelet differentiation stage, it will affect the swelling of the spikelet primordia, leading to most spikelet primordia developing into bristles, thus reducing yield. Therefore, research on how to inhibit the differentiation of millet spikelet primordia into setae and instead transform them into spikelets is expected to make a significant contribution to increasing millet yield. Summary of the Invention

[0005] This invention provides a gene that regulates the differentiation of plant spikelet primordia into setae and its application, wherein the gene can regulate the transformation of plant setae into grains.

[0006] This invention provides a protein that regulates the differentiation of plant spikelet primordia into setae, wherein the protein is any one of the following:

[0007] (1) A protein with an amino acid sequence as shown in SEQ ID No. 1;

[0008] (2) A protein obtained by substituting, deleting and / or adding one or more amino acid residues of the protein described in (1) that has more than 80% identity with the protein shown in (1) and has the function of regulating the differentiation of plant spikelet primordia into setae.

[0009] (3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of the protein described in (1) or (2).

[0010] The present invention also provides a nucleic acid molecule encoding the above-mentioned protein.

[0011] In a preferred embodiment of the present invention, the nucleic acid molecule is any one of the following:

[0012] 1) Nucleic acid molecules with nucleotide sequences as shown in SEQ ID No. 2;

[0013] 2) A nucleic acid molecule obtained by substituting, deleting and / or adding one or more nucleotides to the nucleic acid molecule described in 1), having more than 80% identity with the nucleic acid molecule shown in 1) and encoding the protein of claim 1;

[0014] 3) A fusion gene obtained by linking the 3' end and / or 5' end of the nucleic acid molecule described in 1) or the nucleic acid molecule described in (2) to the coding gene of a protein tag.

[0015] The present invention also provides the use of the above-mentioned protein, substances that regulate the activity or content of the above-mentioned protein, or substances that regulate the expression of the above-mentioned nucleic acid molecules in any of the following:

[0016] (a) Application in regulating plant setae differentiation;

[0017] (b) Application in the preparation of products that regulate plant setae differentiation.

[0018] In a preferred embodiment of the present invention, the type of plant includes monocotyledonous plants.

[0019] In a preferred embodiment of the present invention, the regulation includes reducing the content or activity of the protein to inhibit the differentiation of plant spikelet primordia into setae.

[0020] The regulation also includes inhibiting the expression of the above-mentioned nucleic acid molecules or knocking out the above-mentioned nucleic acid molecules to inhibit the differentiation of plant spikelet primordia into setae.

[0021] The present invention also provides a method for regulating the differentiation of plant spikelet primordia into setae, including regulating the activity and / or content of the above-mentioned proteins, or regulating the expression of the above-mentioned nucleic acid molecules.

[0022] The present invention also provides a biomaterial for inhibiting the differentiation of plant spikelet primordia into setae, comprising any one of the following: a biomaterial that reduces the content or activity of the aforementioned protein, a biomaterial that inhibits the expression of the aforementioned nucleic acid molecules, or a biomaterial that knocks out the aforementioned nucleic acid molecules.

[0023] The present invention also provides a method for breeding monocotyledonous plants, including reducing the content or activity of the above-mentioned proteins in the target monocotyledonous plant to obtain monocotyledonous plants with few or no bristles.

[0024] Alternatively, the expression of the aforementioned nucleic acid molecules in the target monocotyledonous plant can be inhibited or the aforementioned nucleic acid molecules can be knocked out to obtain monocotyledonous plants with few or no bristles.

[0025] This invention also provides the application of the above-mentioned protein, nucleic acid molecule, method, biological material or breeding method in increasing the yield of monocotyledonous plants.

[0026] Beneficial Effects: This invention provides a protein that regulates the differentiation of plant spikelet primordia into setae, the protein comprising the amino acid sequence shown in SEQ ID No. 1. This invention also provides the gene sequence encoding the protein, and based on the gene sequence or protein, prepares biological materials that regulate the differentiation of plant spikelet primordia into setae. These biological materials can be used to regulate the transformation of plant setae into grains. Specifically, in an embodiment of this invention, by knocking out the gene encoding the protein, the differentiation of a portion of spikelet primordia into setae was successfully suppressed, causing these undifferentiated spikelet primordia to differentiate into spikelets with pedicels. The discovery of the protein and gene described in this invention not only fills the gap in the molecular mechanism of millet setae development but also promotes the breeding process for high-yield millet. Attached Figure Description

[0027] Figure 1 This diagram illustrates the BSL gene editing sites. A: The structure of the SiBSL gene and the editing types of the two edited plants, SiBSL-KO-1 and SiBSL-KO-2; B: The sequence alignment of the protein at editing site 1 in the homozygous mutant SiBSL-KO-1 and the Sanger sequencing peak diagram; C: The sequence alignment of the protein at editing site 1 in the homozygous mutant SiBSL-KO-2 and the Sanger sequencing peak diagram; D: Comparison of protein sequences between the homozygous mutants SiBSL-KO-1 and SiBSL-KO-2.

[0028] Figure 2 Figure A shows the statistical results of the phenotypic and agronomical traits of SiBSL mutant plants. Figure A: Phenotypic diagram of SiBSL-edited plants and WT at the jointing stage; Figure B: Phenotypic diagram of SiBSL-edited plants and WT at the heading stage; Figure C: Statistical diagram of plant height of SiBSL-edited plants and WT at the jointing stage; Figure D: Statistical diagram of tiller number of SiBSL-edited plants and WT at the heading stage. *: p<0.05, ****: p<0.0001, ns: no significance.

[0029] Figure 3 This is a phenotypic difference diagram of the SiBSL mutant ear. In the diagram, A: whole ear phenotype of mutant and wild type; B: middle ear phenotype of mutant and wild type; C: ear code phenotype of mutant and wild type; D: two types of grain phenotype observed in the mutant species; EF: ear length (E) and ear width (F) data statistics of mutant and wild type; G: bristle length data statistics of type 2; H: proportion of type 1 and type 2 in the mutant; *: p<0.05, **: p<0.01, ****: p<0.0001, ns: no significance;

[0030] Figure 4 The figure shows the phenotypic diagram of the mutant grains. A: Comparison of grain width between mutant and WT grains; B: Comparison of grain length between mutant and WT grains; C: Grain width data statistics; D: Grain length data statistics; E: Thousand-grain weight statistics. Detailed Implementation

[0031] This invention provides a protein that regulates the differentiation of plant spikelet primordia into setae, wherein the protein is any one of the following:

[0032] (1) A protein with an amino acid sequence as shown in SEQ ID No. 1;

[0033] (2) A protein obtained by substituting, deleting and / or adding one or more amino acid residues of the protein described in (1), which has more than 80% identity with the protein shown in (1) and has the function of regulating the differentiation of plant spikelet primordia into setae.

[0034] (3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of the protein described in (1) or (2).

[0035] The protein tag described in this invention refers to a polypeptide or protein fused with a target protein using in vitro DNA recombination technology for expression, to facilitate the expression, detection, tracing, and / or purification of the target protein. The protein tag may be a Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag, and / or SUMO tag, etc.

[0036] The identity referred to in this invention refers to the identity of amino acid sequences. The identity of amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST page on the NCBI homepage. For example, in Advanced BLAST 2.1, using blastp as the program, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as the matrix, setting Gapexistencecost, Perresiduegapcost, and Lambdaratio to 11, 1, and 0.85 (default values) respectively, and performing an identity search on a pair of amino acid sequences to calculate the identity value (%).

[0037] The 80% or more identity mentioned in this invention can be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99%, or 100% identity.

[0038] In one embodiment of the present invention, the amino acid sequence of the protein is shown in SEQ ID No. 1:

[0039] MMAGELALAALAILLSSLLALVLSHFLPLLLNPKAPRGSFGWPLIGETLRFLTPHASNTLGGFLEDHCARYGRVFKSHLFCTPTVVSCDQDLNHFILQNEERLFQCSYPRPIHGILGKSSM LVVLGEDHKRLRNLALALVTSTKLKPSYLGDIEKIALHVVGSWRQAAGGKECGGGGCVKVITFCEEARKFAFSVIVKQVLGLSPEEPVTARILEDFLAFMKGLISFPLYIPGTPYAKAVQAR ERISSTVKGIIEERRSAGSCKKGDFLDVLLSSNELSDEEKVSFVLDSLLGGYETTSLLISMVVYFLGQSAEDLDLVKREHDSIRSNKGKEECLTSEDYKKMEYTQHVINEALRCGNIVKFVHRKALKDVRYKEYLIPSGWKVLPVFSAVHLNPSLHGNAQHFQPCRWEGSSQGASKRFTPFGGGPRLCPGSELAKVEAAFFLHHLVLNYRWRIDGDDVPMAYPYVEFQRGLPIEIEPICPES. The protein described in this invention is derived from millet and is named protein SiBSL.

[0040] The present invention also provides a nucleic acid molecule encoding the above-mentioned protein.

[0041] In a preferred embodiment of the present invention, the nucleic acid molecule is any one of the following:

[0042] 1) Nucleic acid molecules with nucleotide sequences as shown in SEQ ID No. 2;

[0043] 2) A nucleic acid molecule obtained by substituting, deleting and / or adding one or more nucleotides to the nucleic acid molecule described in 1), having more than 80% identity with the nucleic acid molecule shown in 1) and encoding the protein of claim 1;

[0044] 3) A fusion gene obtained by linking the 3' end and / or 5' end of the nucleic acid molecule described in 1) or the nucleic acid molecule described in (2) to the coding gene of a protein tag.

[0045] In the nucleic acid molecules described in this invention, those skilled in the art can easily mutate the nucleotide sequence encoding the SiBSL protein of this invention using known methods, such as directed evolution or point mutation. Those artificially modified nucleotides that have 80% or more identity with the nucleotide sequence of the SiBSL protein isolated in this invention, as long as they encode and function the SiBSL protein, are all derived from and equivalent to the nucleotide sequence of this invention.

[0046] The 80% or higher identity of the nucleic acid molecules described in this invention can be 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity.

[0047] In this invention, the nucleic acid molecule may refer to a gene. In one embodiment of this invention, the nucleotide sequence of the nucleic acid molecule is shown in SEQ ID No. 2:

[0048]

[0049] The present invention also provides the use of the above-mentioned protein, substances that regulate the activity or content of the above-mentioned protein, or substances that regulate the expression of the above-mentioned nucleic acid molecules in any of the following:

[0050] (a) Application in regulating plant setae differentiation;

[0051] (b) Application in the preparation of products that regulate plant setae differentiation.

[0052] In a preferred embodiment of the present invention, the type of plant includes monocotyledonous plants, such as grasses, specifically plants of the genus Setaria, and more specifically, millet. In one embodiment, millet is used as an example, wherein the variety of millet is Ci846.

[0053] In a preferred embodiment of the present invention, the regulation includes reducing the content or activity of the protein to inhibit the differentiation of plant spikelet primordia into setae; the regulation also includes inhibiting the expression of the above-mentioned nucleic acid molecules or knocking out the above-mentioned nucleic acid molecules to inhibit the differentiation of plant spikelet primordia into setae.

[0054] The present invention also provides a method for regulating the differentiation of plant spikelet primordia into setae, including regulating the activity and / or content of the above-mentioned proteins, or regulating the expression of the above-mentioned nucleic acid molecules.

[0055] In a preferred embodiment of the present invention, the regulation includes reducing the content or activity of the protein to inhibit the differentiation of plant spikelet primordia into setae; the regulation also includes inhibiting the expression of the above-mentioned nucleic acid molecules or knocking out the above-mentioned nucleic acid molecules to inhibit the differentiation of plant spikelet primordia into setae.

[0056] The present invention also provides a biomaterial for inhibiting the differentiation of plant spikelet primordia into setae, comprising any one of the following: a biomaterial that reduces the content or activity of the aforementioned protein, a biomaterial that inhibits the expression of the aforementioned nucleic acid molecules, or a biomaterial that knocks out the aforementioned nucleic acid molecules.

[0057] The regulation described in this invention, at the gene level, includes at least one of the following types: regulation at the gene transcription level; post-transcriptional regulation, such as regulation of splicing or processing of the primary transcript of the gene; regulation of RNA transport of the gene, such as regulation of mRNA transport of the gene from the nucleus to the cytoplasm; regulation of gene translation; regulation of mRNA degradation of the gene; and post-translational regulation of the gene, such as regulation of the activity of the protein translated from the gene.

[0058] This invention does not specifically limit the type of biological material used to regulate the expression of the nucleic acid molecules, and it can be any of the following:

[0059] C1) Nucleic acid molecules that inhibit, reduce, downregulate, or knock out the expression of the gene encoding the protein;

[0060] C2) Expresses the gene encoding the nucleic acid molecule described in C1);

[0061] C3), an expression cassette containing the gene described in C2);

[0062] C4) a recombinant vector containing the gene described in C2), or a recombinant vector containing the expression cassette described in C3);

[0063] C5) recombinant microorganisms containing the gene described in C2), or recombinant microorganisms containing the expression cassette described in C3), or recombinant microorganisms containing the recombinant vector described in C4);

[0064] C6) A transgenic plant cell line containing the gene described in C2), or a transgenic plant cell line containing the expression cassette described in C3), or a transgenic plant cell line containing the recombinant vector described in C4);

[0065] C7) Transgenic plant tissue containing the gene described in C2), or transgenic plant tissue containing the expression cassette described in C3), or transgenic plant tissue containing the recombinant vector described in C4);

[0066] C8) a transgenic plant organ containing the gene described in C2), or a transgenic plant organ containing the expression cassette described in C3), or a transgenic plant organ containing the recombinant vector described in C4).

[0067] In one embodiment of the present invention, CRISPR / Cas9 single-gene knockout technology was used to design two editing target sites at two base positions (221bp-241bp and 345bp-364bp) of the first CDS sequence in the target genome. A gene knockout vector was constructed and transformed into the millet variety Ci846 using Agrobacterium infection technology, thereby achieving the knockout of the SiBSL gene. In one embodiment of the present invention, two primer pairs were designed for the two editing target sites:

[0068] 1F (SEQ ID No. 3): 5'-CGCTCCTAGCCCTGGTGCT-3';

[0069] 1R (SEQ ID No. 4): 5'-AGCACCAGGGCTAGGAGCG-3';

[0070] 2F (SEQ ID No. 5): 5'-ACACGCTGGGTGGCTTCCTG-3';

[0071] 2R (SEQ ID No. 6): 5'-CAGGAAGCCACCCAGCGTGT-3'.

[0072] The present invention also provides a method for breeding monocotyledonous plants, including reducing the content or activity of the above-mentioned proteins in the target monocotyledonous plant to obtain monocotyledonous plants with few or no bristles.

[0073] Alternatively, the expression of the aforementioned nucleic acid molecules in the target monocotyledonous plant can be inhibited or the aforementioned nucleic acid molecules can be knocked out to obtain monocotyledonous plants with few or no bristles.

[0074] In one embodiment of the present invention, after annealing the 1F / R and 2F / R primers respectively, they were ligated into a functional vector and transformed into competent DH5α cells in the large intestine. The plasmid was then extracted and sent to Suzhou Genewiz Biotechnology Co., Ltd. for Sanger sequencing. The plasmid with correct sequencing was named BSL-CRISPR / Cas9.

[0075] This invention also provides the application of the above-mentioned protein, nucleic acid molecule, method, biological material or breeding method in increasing the yield of monocotyledonous plants.

[0076] Using the method described in this invention, mutants SiBSL-KO-1 and SiBSL-KO-2, which knock out the SiBSL gene, can be constructed from wild-type millet Ci846 with relatively long bristles. The ears of these two mutants are without bristles. Furthermore, the kernels of SiBSL-KO-1 and SiBSL-KO-2 change from single kernels in the wild type to double kernels, and the ear length and tiller number are significantly increased compared to the wild type.

[0077] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, of the genes and applications provided by the present invention for regulating the differentiation of plant spikelet primordia into setae, should not be construed as limiting the scope of protection of the present invention.

[0078] Example 1: Obtaining SiBSL gene knockout plants

[0079] 1. Construction of SiBSL gene knockout vector

[0080] Two editing target sites were designed at two base positions in the first CDS sequence of the target genome, namely 221bp-241bp and 345bp-364bp. Seitla.7G132100 of millet Ci846 (long bristles) was edited using CRISPR-Cas9 technology. The vector construction and method are as follows:

[0081] 1.1 The primers shown below were synthesized by Sangon Biotech (Shanghai) Co., Ltd.:

[0082] 1F (SEQ ID No. 3): 5'-CGCTCCTAGCCCTGGTGCT-3';

[0083] 1R (SEQ ID No. 4): 5'-AGCACCAGGGCTAGGAGCG-3';

[0084] 2F (SEQ ID No. 5): 5'-ACACGCTGGGTGGCTTCCTG-3';

[0085] 2R(SEQ ID No.6):5'-CAGGAAGCCACCCAGCGTGT-3';

[0086] BSL-F (SEQ ID No. 7): GCGAGTGGTGGTCGATGATG;

[0087] BSL-R (SEQ ID No. 8): GCACGGATGGTGGTGGTAA.

[0088] 1.3 Obtaining SiBSL gene knockout plants

[0089] Using the Ci846 millet cultivar as plant material, the material was preserved in the Breeding Research Laboratory of the Millet Research Institute, Hebei Academy of Agricultural and Forestry Sciences. Mature seeds of the Ci846 cultivar were dehulled, aseptically treated, and induced to develop embryogenic callus. Then, embryogenic callus transformation was mediated by Agrobacterium containing the gene-editing vector BSL-CRISPR / Cas9. Hygromycin was used for screening to obtain BSL gene-edited millet lines.

[0090] 1.4 Identification of Edited Plants

[0091] DNA was extracted from the transformed seedlings and PCR amplification was performed using primers BSL-F and BSL-R. The system consisted of 1 μL DNA template, 1 μL primer BSL-F (10 μmol / L), 1 μL primer BSL-R (10 μmol / L), 10 μL 2×Taq MasterMix, and ddH2O to make up to 20 μL.

[0092] The program was as follows: 95℃ for 10 min; 35 cycles: 95℃ for 30 s, 58℃ for 30 s, 72℃ for 40 s; 72℃ for 10 min, and storage at 4℃. The PCR products with bands were sent to Suzhou Genewiz Biotechnology Co., Ltd. for Sanger sequencing, which yielded two homozygous edited plants of two editing types, named SiBSL-KO-1 and SiBSL-KO-2, respectively.

[0093] The results are as follows Figure 1As shown, the first type of editing was named SiBSL-KO-1. Compared with the wild type, a guanine (G) nucleotide was added after the 3 bases of editing site 1, resulting in a frameshift mutation in the SiBSL gene. The second type of editing was named SiBSL-KO-2. A cytosine (C) nucleotide was added after the 6th base of editing site 1, and a thymine (T) nucleotide was added after the 3rd base of editing site 2. Both of these base changes caused frameshift mutations in the SiBSL gene.

[0094] 2. Conduct phenotypic observation

[0095] Wild-type millet Ci846, gene-edited materials SiBSL-KO-1 and SiBSL-KO-2 were sown normally, and the millet ears were observed after the grain filling was completed.

[0096] The results are as follows Figures 2-4 As shown, there was no difference in plant height between the two edited types and the wild type at the jointing stage. However, at the heading stage, the plant height of SiBSL-KO-1 and SiBSL-KO-2 was significantly lower than that of WT (P<0.0001), with the decrease in plant height being more pronounced in SiBSL-KO-1, showing a significant difference compared to SiBSL-KO-2. These results indicate that SiBSL-KO has a significant impact on plant height, and there are also phenotypic differences between the different edited types.

[0097] Observation of the ear characteristics of the mutants revealed significant changes in ear morphology in both edited plant types. The ear length of both SiBSL-KO-1 and SiBSL-KO-2 was significantly longer than that of WT, showing a marked difference; the ear width was also significantly narrower than that of WT. Therefore, the ears of both mutants exhibited a slender, club-like shape. There were no significant differences in ear length and width between the two mutants. Observation of the upper part of the ear revealed that the ear cotyledons of SiBSL-KO-1 and SiBSL-KO-2 were tightly distributed on the rachis, while those of WT were relatively loosely distributed. This tight arrangement seemed to exist not only between ear cotyledons but also between kernels. However, the ear cotyledon arrangement was extremely sparse in the lower part of the ear for both mutants. Under a stereomicroscope, the kernels on individual ear cotyledons in the mutants were also tightly clustered together, presenting a spherical shape, while the kernels on individual ear cotyledons in WT were slightly dispersed, presenting a fan shape. Further observation revealed two morphologies in the ears of SiBSL-KO-1 and SiBSL-KO-2: type 1, compound kernels, and type 2, single kernels. Normally, the ear of wild-type Ci846 should exhibit a structure with two long bristles sandwiching a single kernel on a single branch. This phenomenon seemed to confirm that the degenerated bristles had transformed into kernels. However, statistical analysis of bristle traits showed that approximately 94% of the kernels changed from single to compound kernels, while about 6% of the kernels still had bristles, indicating they were not completely degenerated, but their length was significantly shorter than that of WT kernels. This morphology was only present in the lower part of the ears of the mutants. Further analysis of agronomic traits such as kernel length, kernel width, and thousand-kernel weight in the mutants and WT kernels showed that the kernel width of the edited plants was significantly narrower (P<0.01) than that of WT kernels, and the kernel length was significantly shorter (P<0.0001) than that of WT kernels, resulting in the overall kernel volume of SiBSL-KO-1 and SiBSL-KO-2 kernels being smaller than that of WT kernels. Ultimately, this also affected the thousand-grain weight of the mutant seeds, resulting in the thousand-grain weight of both SiBSL-KO-1 and SiBSL-KO-2 being significantly lower than the WT.

[0098] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A protein that regulates the differentiation of plant spikelet primordia into setae, characterized in that, The protein is any one of the following: (1) A protein with an amino acid sequence as shown in SEQ ID No. 1; (2) A protein obtained by substituting, deleting and / or adding one or more amino acid residues of the protein described in (1), which has more than 80% identity with the protein shown in (1) and has the function of regulating the differentiation of plant spikelet primordia into setae. (3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of the protein described in (1) or (2).

2. A nucleic acid molecule encoding the protein of claim 1.

3. The nucleic acid molecule according to claim 2, characterized in that, The nucleic acid molecule is any one of the following: 1) Nucleic acid molecules with nucleotide sequences as shown in SEQ ID No. 2; 2) A nucleic acid molecule obtained by substituting, deleting and / or adding one or more nucleotides to the nucleic acid molecule described in 1), having more than 80% identity with the nucleic acid molecule shown in 1) and encoding the protein of claim 1; 3) A fusion gene obtained by linking the 3' end and / or 5' end of the nucleic acid molecule described in 1) or the nucleic acid molecule described in (2) to the coding gene of a protein tag.

4. The use of the protein of claim 1, the substance that regulates the activity or content of the protein of claim 1, or the substance that regulates the expression of the nucleic acid molecule of claim 2 or 3, in any of the following: (a) Application in regulating plant setae differentiation; (b) Application in the preparation of products that regulate plant setae differentiation.

5. The application according to claim 4, characterized in that, The types of plants mentioned include monocotyledonous plants.

6. The application according to claim 4 or 5, characterized in that, The regulation includes reducing the content or activity of the protein to inhibit the differentiation of plant spikelet primordia into setae. The regulation also includes inhibiting the expression of the nucleic acid molecule described in claim 2 or 3, or inhibiting the differentiation of plant spikelet primordia into setae after knocking out the nucleic acid molecule described in claim 2 or 3.

7. A method for regulating the differentiation of plant spikelet primordia into setae, characterized in that, This includes regulating the activity and / or content of the protein described in claim 1, or regulating the expression of the nucleic acid molecule described in claim 1 or 2.

8. A biomaterial for inhibiting the differentiation of plant spikelet primordia into setae, characterized in that, Includes any of the following: biological materials that reduce the content or activity of the protein of claim 1, biological materials that inhibit the expression of the nucleic acid molecule of claim 2 or 3, and biological materials that knock out the nucleic acid molecule of claim 2 or 3.

9. A method for breeding monocotyledonous plants, characterized in that, This includes reducing the content or activity of the protein described in claim 1 in the target monocotyledonous plant to obtain monocotyledonous plants with few or no bristles. Alternatively, the expression of the nucleic acid molecule described in claim 2 or 3 may be inhibited or the nucleic acid molecule described in claim 2 or 3 may be knocked out in the target monocotyledonous plant to obtain a monocotyledonous plant with few or no bristles.

10. The application of the protein of claim 1, the nucleic acid molecule of claim 2 or 3, the method of claim 7, the biological material of claim 8, or the breeding method of claim 9 in increasing the yield of monocotyledonous plants.

Images